Comparative molecular analyses of Eimeria Schneider (Apicomplexa: Eimeriidae) species from rock ptarmigan in Iceland, Svalbard-Norway, and Japan.

The rock ptarmigan (Lagopus muta) has a Holarctic breeding distribution and is found in arctic and sub-arctic regions. Isolated populations and glacial relicts occur in alpine areas south of the main range, like the Pyrenees in Europe, the Pamir mountains in Central Asia, and the Japanese Alps. In recent decades considerable effort has been made to clarify parasite infections in the rock ptarmigan. Seven Eimeria spp. have been reported parasitizing rock ptarmigan. Two of those species, E. uekii and E. raichoi parasitizing rock ptarmigan (L. m. japonica) in Japan, have been identified genetically. Here we compare partial sequences of nuclear (18S rRNA) and mitochondrial (COI) genes and we detail the morphology of sporulated oocysts of E. uekii and E. raichoi from Japan, E. muta and E. rjupa, from the rock ptarmigan (L. m. islandorum) in Iceland, and two undescribed eimerian morphotypes, Eimeria sp. A, and Eimeria sp. B, from rock ptarmigan (L. m. hyperborea) in Norway (Svalbard in the Norwegian Archipelago). Two morphotypes, ellipsoidal and spheroidal, are recognized for each of the three host subspecies. Our phylogenetic analysis suggests that the ellipsoidal oocyst types, E. uekii, E. muta, and Eimeria sp. A (Svalbard-Norway) are identical and infects rock ptarmigan in Japan, Iceland, and Svalbard-Norway, respectively. Eimeria uekii was first described in Japan in 1981 so that E. muta, described in Iceland in 2007, and Eimeria sp. A in Svalbard-Norway are junior synonyms of E. uekii. Also, phylogenetic analysis shows that the spheroidal oocyst types, E. rjupa and Eimeria sp. B (Svalbard-Norway), are identical, indicating that rock ptarmigan in Iceland and Svalbard-Norway are infected by the same Eimeria species and differ from E. raichoi in Japan.

Evaluation of the host specificity of Eimeria uekii and Eimeria raichoi for Japanese rock ptarmigans by oocyst transfer to taxonomically related birds.

Eimeria spp. are protozoan parasites that are commonly found in a broad range of vertebrate hosts. These parasites generally exhibit strict host specificity, but some Eimeria spp. can infect groups of closely related species such as species within a genus or family. Compared with Eimeria spp. that infect livestock, limited information is available about such infections in wild animals including data on host specificity, virulence, and prevalence. The Japanese rock ptarmigan, Lagopus muta japonica, is an endangered bird belonging to the family Phasianidae, order Galliformes, and inhabits only alpine areas of Japan. In conservation efforts for these birds, two Eimeria spp., E. uekii and E. raichoi, were frequently detected. Here, we examined cross-transmission of the parasites to other bird species to characterize their infectivity as well as the development of experimental bird models to contribute to conservation programs by the oocyst transfer. Consequently, among the examined eight bird species (chickens, Japanese pheasants, turkeys, chukar partridges, quails, helmeted guineafowls and ducks), only turkeys (family Phasianidae, order Galliformes) could be infected with E. raichoi. However, the number of oocysts per feces was relatively low, and few parasites in the intestinal mucosa could be found by histopathological analyses. These results might indicate that E. uekii and E. raichoi are highly adapted to Japanese rock ptarmigans that inhabit the alpine zone although further studies are anticipated.

First detection of coccidia species in blue peafowl (Pavo cristatus) in China via both microscopic and molecular methods.

Coccidia are protozoan parasites in the class Conoidasida. To determine the prevalence and molecular characteristics of coccidia species in blue peafowl (Pavo cristatus) in Henan, China, 240 fecal specimens were collected and screened for the presence of Eimeria spp. and Isospora spp. The overall prevalence was 65.0% (156/240), and seven different coccidia species were identified: E. pavonis (51.3%, 123/240), E. arabic (40.0%, 96/240), E. riyadhae (37.1%, 89/240), E. mandali (22.9%, 55/240), E. mayurai (14.2%, 34/240), I. mayuir (10.9%, 26/240), and I. lacazei (8.5%, 21/240). E. arabic and E. riyadhae were detected for the first time in China. Additionally, we provide molecular data of the seven different coccidia species at the 18S and 28S ribosomal RNA and the COI loci. Sequence homology percentages among the five species of Eimeria at the 18S rRNA, 28S rRNA, and COI loci were 96.0%-98.6%, 90.7%-98.2%, and 85.0%-94.9%, respectively, whereas for two species of Isospora the sequence homology percentages were 98.8%, 99.1%, and 95.4% at three corresponding loci. This is the first report of the molecular data of the seven coccidia species in blue peafowl in China.

PREVALENCE AND MOLECULAR CHARACTERISTICS OF BLASTOCYSTIS SP. FROM PEAFOWL (PAVO CRISTATUS) IN CHINA.

This study is the first description of Blastocystis infection in peafowls in China. In total, 143 fecal specimens collected from a peafowl breeding farm in Henan Province were tested for Blastocystis infection by PCR assay targeting the small subunit ribosomal RNA (SSU rRNA) gene, and a total of 50 specimens (35.0%) were positive. Based on sequences and phylogenetic analysis, 2 genetically distinct subtypes (STs) were determined: ST9 and ST7. ST9 was the predominant subtype, accounting for 82% (41/50). The rare zoonotic subtype ST7 was also identified in peafowls, with the infection rate of 18% (9/50). Altogether, the present study is the first report of the prevalence and molecular characteristics of Blastocystis in peafowls in central China. The presence of zoonotic subtypes in peafowls suggests the potential risk of zoonotic transmission of Blastocystis to workers at peafowl farms.

DESCRIPTION OF A NEW EIMERIA SPECIES (APICOMPLEXA: EIMERIIDAE) RESPONSIBLE FOR CLINICAL COCCIDIOSIS IN COMMERCIAL CHUKAR PARTRIDGE (ALECTORIS CHUKAR).

Recurrent coccidiosis affecting a commercial chukar partridge (Alectoris chukar) farm in Ontario, Canada was investigated. The responsible pathogenic Eimeria species was isolated for biological characterization. The uniformity of oocyst morphometrics supported that only a single Eimeria sp. was present. Experimental infections with coccidia-free chukars were used to describe exogenous and endogenous developmental stages of the parasite. The prepatent period of the causative Eimeria species was 5 days and patency lasted 11 days; fecundity was 1,573 to 30,057, with the highest fecundity recorded with the lowest challenge dose. Endogenous development was elucidated histologically from samples collected at 8 locations along the intestinal tract at 26 time points throughout prepatency. The parasite had 5 asexual generations before oocyst formation that were located from the mid-jejunum to the mid-rectum and in the ceca. Sporulation of oocysts suspended in potassium dichromate at room temperature (22 C) occurred within 24 hr. Oocysts (n = 50) averaged 21.8 by 18.6 µm and featured a polar granule; sporocysts (n = 50) averaged 10.9 by 7.1 µm and possessed a Stieda body, sub-Stieda body, sporozoite refractile bodies, and sporocyst residuum. Comparisons with described Eimeria spp. infecting partridges suggest that the biological features of this pathogenic species are unique; similarly, sequences from both mitochondrial and nuclear loci support the naming of this new Eimeria species.

Gastrointestinal nematodes in two galliform birds from South Africa: patterns associated with host sex and age.

Parasite ecology has recently focused on elucidating patterns and processes that shape helminth communities in avian hosts. However, helminths parasitizing gamebirds are still poorly understood. Here we describe the gastrointestinal nematode fauna of Swainson's spurfowl, Pternistis swainsonii (Phasianidae) and helmeted guineafowl, Numida meleagris (Numididae), collected at three and four localities, respectively, in South Africa and analyze the prevalence, mean abundance and diversity of their helminth communities. Eleven nematode species were collected from spurfowl, which had a mean number of nematode species per host of 3.01 ± 0.18, whereas guineafowl harboured 15 nematode species, with a mean number of nematode species per host of 3.93 ± 0.12. Focusing on the most prevalent species, we also asked if host sex and/or age were associated with infracommunity structure with regard to nematode counts and species richness, as well as the species and taxonomic composition of infracommunities. While pooling data of nematode species masked the influence of host characteristics on helminth communities, analysis of individual nematode species revealed a number of patterns. In particular, adult female bias was seen in Tetrameres swainsonii in spurfowl and in Allodapa dentigera and Gongylonema congolense in guineafowl; Acuaria gruveli reached higher numbers in adult spurfowl than in juveniles, and helminth infracommunities in juvenile male guineafowl were more species rich than those in adult males. Combined, our results suggest that helminth communities of spurfowl and guineafowl are associated with a complex interplay of numerous factors, including host characteristics, parasite traits and environmental conditions.

GAPEWORM (SYNGAMUS SPP.) PREVALENCE IN WISCONSIN GREATER PRAIRIE CHICKENS (TYMPANUCHUS CUPIDO PINNATUS).

Under Wisconsin state law, the greater prairie chicken (GRPC; Tympanuchus cupido pinnatus) has been listed as a threatened species since 1976. In 2014-15, we conducted a pilot study to determine the prevalence and intensity of gapeworms (Syngamus spp.) in female Wisconsin GRPCs collected from 2 monitored populations. We captured 62 female GRPCs using walk-in-style traps for females and night lighting for juveniles ≥45 days of age. From these individuals, we collected 15 carcasses of radio-marked birds, most of whom died due to predation events. Through dissection, we identified gapeworm in 20% of examined carcasses and report an intensity ranging between 4 and 74 worms.

Morphological and molecular identification of Eimeria tetartooimia oocysts from a Japanese green pheasant (Galliformes; Phasianidae; Phasianus versicolor) at a zoo in Japan.

We detected Eimeria oocysts from Japanese green pheasants (Phasianus versicolor) at a zoo in Osaka, Japan. The oocyst isolates were subspherical or ovoidal shaped and measured 17.2 (range 14.7-20.0) µm in length and 14.8 (13.3-16.7) µm in width with a length/width (L/W) ratio of 1.2 (1.0-1.4) and each had one polar granule. The oocysts lacked a residuum and micropyle. Sporocysts measured 9.8 (6.7-13.3) µm in length and 5.9 (4.7-7.3) µm in width, with a L/W ratio of 1.2 (1.1-1.4). Compared to previously published values, this strain shows morphological similarities with an isolate of E. teetartooimia from ring-necked pheasants from other countries. Phylogenetic analysis of the 18S rRNA and mitochondrial cytochrome c oxidase subunit I genes places the isolate in a clade related to chicken Eimeria spp., such as E. acervulina or E. brunetti. Although further analysis is needed, this information can be helpful for the diagnosis and determination of virulence of Eimeria spp. in pheasants.

New species of ischnoceran chewing lice (Phthiraptera: Philopteridae) from Chinese birds.

Five new species of chewing lice (Phthiraptera: Ischnocera) are described and illustrated from Chinese birds, as follows: Cuculicola calyptocamptus new species ex Hierococcyx sparverioides (Vigors, 1832) (Cuculiformes), Goniocotes kristinae new species ex Lophura swinhoii (Gould, 1863) (Galliformes), Goniocotes rolandi new species ex Crossoptilon harmani Elwes, 1881 (Galliformes), Rallicola (Rallicola) tibetana new species ex Zapornia bicolor Walden, 1872 (Gruiformes), and Resartor elugeus new species ex Alcippe fratercula yunnanensis Harington, 1913 (Passeriformes).

Gastrointestinal parasites of black grouse Lyrurus tetrix: A long-term study (1986-2019) in the French Alps.

This study investigates the gastroenteric parasites community of 336 black grouse Lyrurus tetrix, covering a wide part of the totality of the species range in the French Alps and characterized by a long-term sampling (1986-2019). Parasite community was described using common epidemiological descriptors. Additionally, for the most prevalent parasite, a species distribution model (SDM) was built to understand the main factors influencing parasite occurrence. Ascaridia compar, Capillaria caudinflata, cestodes (Raillietina urogalli, R.cesticillus, and Hymenolepis microps), Eimeria sp. (E. lyruri and E.nadsoni), Corrigia corrigia and Acanthocephalus sp. The most prevalent species was C.caudinflata (P: 33%,CI95%: 28-38), whereas the lowest prevalence was recorded for Acantocephalus sp. (P:0.3%,CI95%:-0.3-0.9).The selected SDM for C.caudinflata includes only 3 variables: BIO11 (Mean Temperature of Coldest Quarter), the latitude of sampled animals and the distance from ski tour way. The distribution of C.caudinflata in the French Alps presents sharply defined areas where there are optimal environmental conditions for maintaining the life cycle of this parasite. These are the colder areas, characterized by higher latitude values and close to the ski tour ways. This study provides a valuable contribution to the scarce literature on wild Galliformes, describing the composition of the gastrointestinal parasite community of the black grouse.

Helminthological studies in francolins (Galliformes: Phasianidae) of Sindh, Pakistan with two new species and epidemiological parameters.

Francolins are among the very gregarious variety of game birds severely infected by various parasites that may cause an important source for infection transmission in humans by eating them very fondly in the country. During this study, two commonly found francolins species of the region; Francolinus francolinus (Black Francolin) and F. pondicerianus (Grey Francolin), were examined for helminthes fauna. The study deals with the primary survey of helminthic infection occur in the francolin birds (Phasianidae: Perdicinae) with reference to their forage in gut content. It is the first helminthological and epidemiological study in Sindh, Pakistan. At present, a total of 20 birds were examined, out of which 17 were found infected with cestode larvae of two species of genus Cotugnia and genus Raillietina, one species of trematode, Prosthogonimus potentially new species; one new species of acanthocephalan, Mediorhynchus francolinae sp. nov.; one species of nematode, Subulura brumpti (López-Neyra, 1922) were recovered, with new host records. The forage content and comparative incidence of helminth species were also observed and discussed in the current study, which revealed the high prevalence of infection in grey francolins than that of black francolin. The mean intensity of infection relative to the forage in guts of hosts was found significant (P < 0.05), observed 9.14 ± 1.65 in black francolins than in grey francolins (5.8 ± 0.51). The parasitic abundance was compared using Fisher's Exact Test, which showed no significant difference between cestodes and nematodes in the two hosts, however trematode and acanthocephalan was found only in grey francolin. The parasitic frequencies along with the confidence intervals were recorded higher in black francolins by cestodes and nematodes than the black francolins during the present study.

Some gastrointestinal nematodes and ixodid ticks shared by several wildlife species in the Kruger National Park, South Africa.

Parasite surveys were conducted for 1­2 years in the Kruger National Park (KNP), South Africa on blue wildebeest, impalas, greater kudus, common warthogs and scrub hares. The host associations of some of the gastrointestinal nematode species infecting ≥60% of at least one of the five host species, were determined. These were Agriostomum gorgonis, Cooperia acutispiculum, Cooperia connochaeti, Cooperia hungi, Cooperia neitzi, Cooperioides hamiltoni, Gaigeria pachyscelis, Haemonchus bedfordi, Haemonchus krugeri, Haemonchus vegliai, Impalaia tuberculata, Longistrongylus sabie, Strongyloides papillosus, Trichostrongylus deflexus and Trichostrongylus thomasi. Although the prevalence of Trichostrongylus falculatus did not exceed 50% in any host species, it was present in all five hosts. Nematodes in the KNP range from those exhibiting strict host associations to generalists. Nematode-host associations may be determined by host feeding patterns and habitat use. Eight ixodid tick species were commonly collected from the same animals and in 2­3 year long surveys from plains zebras and helmeted guinea fowls: Amblyomma hebraeum, Amblyomma marmoreum, Hyalomma truncatum, Rhipicephalus appendiculatus, Rhipicephalus decoloratus, Rhipicephalus evertsi evertsi, Rhipicephalus simus and Rhipicephalus zambeziensis. Host specificity was less pronounced in ixodid tick species than in nematodes and the immature stages of five tick species infested all host species examined.

Syngamus asphyxiation in a captive ring-necked pheasant.

A deceased ring-necked pheasant (Phasianus colchicus) presented for necropsy following a history of chronic wasting. Necropsy revealed nematodes consistent with the genus Syngamus partially obstructing the trachea. Phylogentic analysis failed to reveal conclusive results regarding the species. Syngamus spp. can cause obstruction of the trachea in several different hosts. Additional genetic data from this taxon would aid in the more precise identification of diagnostic specimens.

Pathology of Trombiculosis caused by Neoschoengastia simonovichi in Wild Red-Legged Partridges (Alectoris rufa).

Lesions caused by trombiculid mite larvae ('chiggers') in birds have been inadequately described. A juvenile red-legged partridge (Alectoris rufa) presented with multifocal, crater-like lesions of necrotizing dermatitis, which often contained multiple orange mites. Microscopically, there was a nodular necrotizing and pyogranulomatous dermatitis with occasional intralesional arthropods. Histologically, the mites were readily identified by the presence of stylostomes within the necrotic areas. Morphological and morphometrical studies on isolated mites identified them as Neoschoengastia (Hypogastia) simonovichi. This is the first report of N. simonovichi as an aetiological agent of severe trombiculosis in wild birds in Western Europe. Infestation with this parasite requires further study to understand its role in animal and human trombiculosis and its potential role as a vector of infectious, including zoonotic, agents.

In vitro and in vivo activity of cinnamaldehyde against Eimeria kofoidi in chukar partridge (Alectoris chukar).

One hundred and twenty one-day-old chukar partridges were randomly divided into eight groups which received diets with different supplementations. There were four unchallenged groups. One group received salinomycin (50 ppm), two groups received cinnamaldehyde (CINN) (100 and 200 mg/kg of diet), and another one received only the basal diet from the 1st to the 31st day. There were also four corresponding groups orally challenged by 3 × 105Eimeria kofoidi sporulated oocysts at the 21st day. Three samplings were done at the 24th, 26th, and 31st days of rearing for pathological and biochemical assessments. Fecal samples were daily taken to check the pattern of oocyst shedding from the 26th to 31st day. The body weight of birds was measured at 21st and 31st days. Along with the in vivo experiment, an in vitro sporulation inhibition test was carried out. The in vitro results showed that CINN decreased sporulation rate at 1 and 0.5 mg/ml. In vivo, it was found that CINN did not prevent the oocyst shedding. Furthermore, the histopathological findings revealed that CINN and salinomycin had no effect on infection establishment. However, our findings showed that CINN (200 mg/kg of diet) could enhance the body weight and improve antioxidant status. Although our results did not support the in vivo anticoccidial activity of CINN, it had a promising potential to improve antioxidant status and body weight in the chukar partridge.

Protozoan and Helminth Infections of Aviary-Reared Alectoris rufa (Galliformes: Phasianidae) before Releasing for Hunting in the Czech Republic: Infection Dynamics and Potential Risks.

Age-related dynamics of protozoan and helminth infections in the red-legged partridge, Alectoris rufa, were studied in an intensive breeding aviary in the Czech Republic before releasing birds for hunting purposes. Pooled fecal samples (n = 900) were examined over 3 rearing seasons (2012-2014). A total of 4 protozoan species, Cryptosporidium baileyi, Cryptosporidium meleagridis, Eimeria kofoidi, and Eimeria legionensis, and 4 helminths, Capillaria phasianina, Eucoleus perforans, Heterakis gallinarum, and Syngamus trachea, was found. The most common parasite was S. trachea (prevalence 20-26%) in dead birds (n = 99), which represents a high risk for breeders. Co-occurrence of protozoans and helminths indicated similarities in infection dynamics throughout the 3 breeding seasons. Mixed infections of Cryptosporidium baileyi and C. meleagridis with other parasitic species are reported for the first time. Our findings provide new insights into breeding of A. rufa and may help to improve the efficacy of disease control strategies and prevention, especially with the potential for spreading of parasitic infections to wildlife through released birds into open areas.

Correlates of pathological lesions associated with respiratory cryptosporidiosis prevalence in shot red grouse Lagopus lagopus scotica from moors in northern England.

Infection of wild red grouse Lagopus lagopus scotica by Cryptosporidium baileyi was first diagnosed in 2010. Within three years, signs of infection were reported from grouse on half of all grouse moors in northern England, bringing severe concerns of economic losses to grouse shooting. A total of 45,914 red grouse shot from 10 moors in northern England between 2013 and 2018 were visually screened for signs of respiratory cryptosporidiosis. Prevalence varied with age, being twice as high in juveniles (4.5%) as in adults (2.4%). It also varied nine-fold between moors and three-fold between years. Prevalence was highest in grouse shot later in the shooting season. Our results are consistent with the concept that disease incidence is highest in naïve juveniles that have previously not been exposed to infection, with prevalence dropping as birds develop immunity. We found no evidence of increased prevalence over time, and fears of escalated disease prevalence, bringing with it increased mortality and lowered productivity, that may have significant impacts on the economic viability of shoots, have not yet been realized. We recommend continued annual screening for clinical signs amongst shot birds, better hygiene associated with potential reservoirs of infection, and practices that both improve the detection and selective culling of diseased individuals and generally reduce overall grouse densities.

Parasitological and molecular survey of scattered parasitism by trichomonads in some avian species in Iran.

Outbreaks of avian trichomonosis are being reported worldwide; meanwhile, the genetic and virulence variations are under investigation. In this study, the occurrence and genetic variability of oral or faecal trichomonads among various avian species were investigated. Samples obtained from either the oropharyngeal cavity, crop/oesophagus, droppings/cloaca, or conjunctival swabs of avian species were inspected for flagellates. Phylogenetic analysis of partial ITS1-5.8s rRNA-ITS2 sequences from selected samples was performed to investigate the genetic diversity of the isolates. Investigation of 737 birds revealed an infection rate of 15.7% in the upper gastrointestinal tract, 7.3% in the faecal samples, and 0.7% involvement of the conjunctiva. Phylogenetic analysis of partial ITS1-5.8s rRNA-ITS2 sequences from selected samples, identified genotypes A and B of Trichomonas gallinae and genogroups A-C and E of Tetratrichomonas gallinarum. A novel ITS genotype of intestinal trichomonads was also detected in hooded crow (Corvus cornix) and common mynah (Acridotheres tristis). In the present study, in addition to Columbiformes and Falconiformes, trichomonads were detected in Passeriformes and Galliformes with the involvement of organs other than the gastrointestinal tract. Genotype A T. gallinae was detected in domestic pigeons (Columba livia domestica), a laughing dove (Spilopelia senegalensis), a common kestrel (Falco tinnunculus), a budgerigar (Melopsittacus undulates), and a canary (Serinus canaria). Distinct genotype B was detected in a common mynah and a budgerigar. Genogroups A-C of T. gallinarum were also demonstrated in Galliformes and Anseriformes. Furthermore, two novel trichomonad ITS genotypes were detected in hooded crows and a common mynah warranting detailed multi-locus molecular analysis.RESEARCH HIGHLIGHTSITS diversity of trichomonads was shown in various avian species.Diversity of the parasites' target organ and clinical manifestations was demonstrated.Two novel ITS genotype trichomonads from common mynah and hooded crow were identified.

Occurrence of a Cryptosporidium xiaoi-like genotype in peafowl (Pavo cristatus) in China.

The aim of this study was to survey the Cryptosporidium species in peafowls (Pavo cristatus) in Henan Province, China. A total of 143 fecal specimens collected from a breeding farm were tested for Cryptosporidium by nested PCR targeting the small subunit rRNA (SSU rRNA), 70-kDa heat shock protein (HSP70), and actin genes of Cryptosporidium followed by sequence analysis. Only one isolate from an asymptomatic host was obtained, and the isolate differed from a new C. xiaoi-like genotype by one nucleotide and from C. xiaoi or C. bovis at the SSU rRNA locus by six nucleotides. Likewise, the actin gene shared 99% identity with the C. xiaoi-like genotype, accompanied by four nucleotide mutations. A complete sequence of the HSP70 gene was obtained, and exhibited 96% similarity with that from C. xiaoi and differed by one nucleotide from that with the C. xiaoi-like genotype. Phylogenetic analysis of the current isolate revealed genetic relatedness to the C. xiaoi-like genotype and distinction from C. xiaoi and C. bovis. Therefore, our results provided the first documentation of avian infection with a C. xiaoi-like genotype in China and further insight into the diversity of Cryptosporidium spp. in avians.

Heterakis isolonche Linstow, 1906 ­ a new nematode species found in ornamental pheasants in Poland

Studies were carried out on 25 pheasants belonging to 13 Phasianinae species of the Phasianidae family. The research material was collected from private breeders ­ both Polish and from abroad. The results confirm that the most frequent nematode is Heterakis gallinarum, while for the first time in the country ­ in two of the examined birds of Polish flocks ­ the occurrence of Heterakis isolonche was found.