Occurrence of Thelazia lacrymalis (Nematoda, Spirurida, Thelaziidae) in native horses in Abruzzo region (central eastern Italy)

A survey on the prevalence of Thelazia spp. in Abruzzo region (Italy) in slaughtered native horses was conducted from August 29, 1997 to August 28, 1998. Both eyes from 128 eight-month to 11 year-old native animals were examined. 50 horses (39.06%) were found parasitized by Thelazia lacrymalis. 502 specimens (371 females, 88 males and 12 larvae) were collected. In the infected horses the numbers of T. lacrymalis ranged from 1 to 48, with a mean count burden of 3.92 per head (SD = 7.79). T. lacrymalis specimens were mainly in the excretory ducts of the Harderian gland, and also in the ducts of the lacrimal glands, free in the conjunctiva and behind the nictitancte. Gross examination showed a conjunctivitis, more frequently a follicular conjunctivitis, in the 58% of the infected horses.     

Thelazia lacrymalis in horses in Kentucky and observations on the face fly (Musca autumnalis) as a probable intermediate host.

Eyes from 114 (30.3%) of 376 dead horses, examined from 3 April 1975 to 3 April 1976, were naturally infected with adult Thelazia lacrymalis; 1 horse was also infected with 1 male Thelazia skrijabini. Adult T. lacrymalis from dead horses were successfully transferred mechanically to the eyes of 3 of 4 Shetland ponies raised helminth-free. Larvae from gravid female T. lacrymalis underwent development in experimentally infected, laboratory-raised face flies (Musca autumnalis) and third-stage larvae ranging from 1.82 to 2.94 mm in total length were recovered at 12 to 15 days postexposure. A total of 866 naturally occurring face flies were collected from the head region of horses. Twelve of the face flies harbored larval stages of Thelazia spp. One of the larvae resembled third-stage T. lacrymalis that were recovered from the experimentally infected, laboratory-raised face flies. Introduction of 3 third-stage larvae from 1 face fly onto the cornea of a pony raised helminth-free resulted in the recovery of 1 male T. skrjabini 242 days later. In addition to the eyeworm larvae, other parasites recovered from the face flies included Heterotylenchus autumnalis, hypopi of astigmatid mites and a first instar beetle (Coleoptera: Rhipiphoridae). Data from these investigations indicate the likelihood that face flies are an intermediate host for T. lacrymalis and probably other species of Thelazia in this part of the country.     

Thelazia gulosa Railliet & Henry, 1910 and T. skrjabini Erschow, 1928 infection in southern Europe (Italy)

The occurrence of Thelazia gulosa and T. skrjabini in cattle in Italy, together with an update of the prevalence of Thelazia spp. in the southern Italy, is reported. On 764 slaughtered native cattle, fifty-five (7.2%) bovines were infected by Thelazia spp: T. rhodesi was found in 44 (80%) animals, T. gulosa was found in 19 (34.5%) cattle and only one (1.8%) cattle harboured T. skrjabini. This is the first report of T. gulosa and T. skrjabini in Southern Europe.     

Molecular characterization of the first internal transcribed spacer of ribosomal DNA of the most common species of eyeworms (Thelazioidea: Thelazia)

Spirurid nematodes of the genus Thelazia cause ocular infection in several mammals and are transmitted by dipteran flies. Of the 10 described species, T. gulosa, T. rhodesi, T. skrjabini (affecting cattle), T. lacrymalis (affecting horses), and T. callipaeda (affecting carnivores and humans) are the most common. The aim of this study was to characterize the first internal transcribed spacer (ITS1) ribosomal DNA sequences of these 5 species. The length of the ITS1 sequences ranged from 357 bp (T. lacrymalis) to 905 bp (T. callipaeda). Interspecific differences ranged from 35 to 77%. Polymorphic sites were detected in each species. Intraspecific variation varied from 0.3 to 2.5%. The characteristics of the ITS1 of Thelazia spp. show similarities to those of other Spirurida. The findings of this present study show that the ITS1 represent a powerful genetic marker for the molecular identification of eyeworms and a useful tool for developing molecular epidemiological techniques to study Thelazia spp. transmission patterns and prevalence in definitive and intermediate hosts.     

Ecology of Thelazia spp. in cattle and their vectors in Italy

The genus Thelazia (Spirurida, Thelaziidae) includes a cosmopolitan group of eyeworm spirurids responsible for ocular infections in domestic and wild animals and transmitted by different species of muscids. Bovine thelaziosis is caused by Thelazia rhodesi Desmarest 1828, Thelazia gulosa Railliet & Henry 1910, and Thelazia skrjabini Erschow 1928, which occur in many countries; T. gulosa and T. skrjabini have been reported mainly in the New World, while T. rhodesi is particularly common in the Old World. In Italy, T. rhodesi was reported in southern regions a long time ago and, recently, T. gulosa and T. skrjabini have been identified in autochthonous cattle first in Apulia and then in Sardinia. Thirteen species of Musca are listed as intermediate hosts of eyeworms, but only Musca autumnalis and Musca larvipara have been demonstrated to act as vectors of Thelazia in the ex-URSS, North America, ex-Czechoslovakia and more recently in Sweden. In Italy, after the reports of T. gulosa and T. skrjabini in southern regions, the intermediate hosts of bovine eyeworms were initially only suspected as the predominant secretophagous Muscidae collected from the periocular region of cattle with thelaziosis were the face flies, M. autumnalis and M. larvipara, followed by Musca osiris, Musca tempestiva and Musca domestica. The well-known constraints in the identification of immature eyeworms to species by fly dissection and also the time-consuming techniques used constitute important obstacles to epidemiological field studies (i.e. vector identification and/or role, prevalence and pattern of infection in flies, etc.). Molecular studies have recently permitted to further investigations into this area. A PCR-RFLP analysis of the ribosomal ITS-1 sequence was developed to differentiate the 3 species of Thelazia (i.e. T. gulosa, T. rhodesi and T. skrjabini) found in Italy, then a molecular epidemiological survey has recently been carried out in field conditions throughout five seasons of fly activity and has identified the role of M. autumnalis, M. larvipara, M. osiris and M. domestica as vectors of T. gulosa and of M. autumnalis and M. larvipara of T. rhodesi. Moreover, M. osiris was described, for the first time, to act as a vector of T. gulosa and M. larvipara of T. gulosa and T. rhodesi. The mean prevalence in the fly population examined was found to be 2.86%. The molecular techniques have opened new perspectives for further research on the ecology and epidemiology not only of Thelazia in cattle but also of other autochthonous species of Thelazia which have been also recorded in Italy, such as Thelazia callipaeda, which is responsible for human and canid ocular infection and Thelazia lacrymalis, the horse eyeworm whose epidemiological molecular studies are in progress.     

Occurrence of Gasterophilus intestinalis and some Parasitic Nematodes of Horses in Sweden

A survey was performed on the occurrence of some internal parasites in 461 horses (1–30 years old) slaughtered from October 1992 to September 1993 at the Linköping abattoir in central Sweden. Macroscopical examination was carried out specifically for parasites of the tear ducts and conjunctival sacs of the eyes, and of selected parts of the alimentary tract and car-dio-vascular system. The following parasites were found in selected parts of the large intestine: encapsulated cyathostome larvae (in 35.6% of the horses), and mature strongyle worms (17.4%); in the stomach: Gasterophilus intestinalis (12.3% during October-June) and Habronema muscae (1.1%); and in the conjunctival sac: Thelazia lacrymalis (3.1%). Significantly more encapsulated cyathostome larvae were found during January to June than during other times of the year, and horses aged 1–5 years harboured significantly more larvae than older horses. Severe damage to the cranial mesenteric artery and its main branches was noticed in 16 (6.1%) out of 263 horses specifically examined. However, the 4th stage larva of Strongylus vulgaris was only recovered in 6 (2.3%) of the horses. Quantitative and qualitative faecal egg counts were done on 412 and 384 of the horses, respectively. Eggs of strongyles, Parascaris equorum and Strongyloides westeri were found in 78.1%, 1.9% and 0.2% of the faecal samples, respectively. Highest prevalence of strongyle eggs was found during July to September. Third stage larvae of the following nematodes were recovered from faecal cultures: subfamily Cyathostomum sensu lato (78.1%) Triodontophorus spp. (6.5%), Strongylus vulgaris (3.6%), Trichostrongylus axei (1.3%), while Gyalocephalus sp., Oesophagodontus sp., Poteriostomum sp. and Strongylus edentatus each comprised less than 0.5%.     

Embryogenesis and the first-stage larva of Thelazia lacrymalis

The female reproductive system of Thelazia lacrymalis (Nematoda: Thelaziidae) was investigated by light and scanning electron microscopy (SEM) with regard to the developmental stages and the stage deposited by the gravid nematode. Female T. lacrymalis have a didelphic and opisthodelphic type of reproductive system with paired ovaries, oviducts and uteri and a single vagina and vulva. Round and spindle-shaped primary oocytes are documented within the ovaries and oviducts, respectively. The distal part of each uterus provides a fertilization chamber filled with spermatozoa, followed by a sphincter-like part. Further anterior, the uteri broaden gradually containing dividing zygotes, small and large morulae, tadpole-stage embryos and horseshoe-shaped embryos which increase in length and become slimmer forming pretzel-stage embryos and larvae rolled up. The larvae stretch gradually and finally lie straight but still covered with their egg membrane in the vagina. The egg membrane encloses the whole larva and is enlarged at the pointed tail of the larva forming a bulb. At the SEM level, the first-stage larva is shown to have a terminal mouth and three hooks directed posteriorly and a striated cuticle. As morphologically identical larvae were also found in lavages of the conjunctival sac of horses infected with T. lacrymalis, this nematode species can be described as ovoviviparous.     

Bovine thelaziasis in Iowa

The prevalence of developing Thelazia nematodes in face flies (Musca autumnalis) was studied for 7 yr at a beef farm in central Iowa. Juvenile nematodes were not found among flies in reproductive diapause in autumn, nor among nulliparous, overwintered flies in spring, but only among actively reproducing insects. Thus Thelazia probably do not overwinter in face flies. A mean prevalence of 2.0% infected was recorded among flies in 7 fly breeding seasons. No heterogeneity in Thelazia prevalence was detected within fly breeding seasons. The frequency distribution of Thelazia among face flies by year of occurrence was homogeneous, with a mean of 2.75 larval nematodes per infected fly. Thelazia gulosa and T. skrjabini were recovered in necropsy from the eyes of bovines from central Iowa. Prevalence among fat cattle 18-27 mo old was 15%; among cows 3-15 yr old, prevalence was only 3%. Thelazia skrjabini was found in 29 eyes and T. gulosa in 4 eyes. Infections were randomly distributed among the eyes of subject cattle. The mean worm burden was 2.3 T. skrjabini and 12 T. gulosa per infected eye. Thelaziasis is clearly enzootic in Iowa.     

Seroprevalence and isolation of Toxoplasma gondii from free-range chickens in Ghana, Indonesia, Italy, Poland, and Vietnam

The prevalence of Toxoplasma gondii in free-ranging chickens (Gallus domesticus) is a good indicator of the prevalence of the parasite's oocysts in soil because chicken feed from the ground. The prevalence of T. gondii in free-range chickens from Ghana, Indonesia, Italy, Poland, and Vietnam was determined using the modified agglutination test (MAT). Antibodies to T. gondii were found in 41 (64%) of 64 chickens from Ghana, 24 (24.4%) of 98 chickens from Indonesia, 10 (12.5%) of 80 chickens from Italy, 6 (30%) of 20 chickens from Poland, and 81 (24.2%) of 330 chickens from Vietnam. Hearts and brains of chickens were bioassayed for T. gondii. Viable T. gondii was isolated from 2 chickens from Ghana, 1 chicken from Indonesia, 3 chickens from Italy, 2 chickens from Poland, and 1 chicken from Vietnam. Toxoplasma gondii isolates from 9 chickens were genotyped using 10 PCR-RFLP markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. A total of 7 genotypes was identified; the 3 isolates from chickens from Italy were clonal type II, and the others were nonclonal. This is the first report of genetic characterization of T. gondii isolates from animals from these countries.     

Presence of Thelazia callipaeda Railliet & Henry, 1910, in Piedmont, Italy

A sample of 314 dogs from a zone of 80,000 ha in Piedmont, Italy, were examined for the presence of thelaziae under the nictitating membrane. Sixteen animals from eight districts were infected, the foothills being the main location of the parasitosis. All 44 nematodes collected were identified as Thelazia callipaeda.     

Thelazia callipaeda (Spirurida, Thelaziidae) of carnivores and humans: morphological study by light and scanning electron microscopy

Thelazia nematodes cause ocular infection in several mammals and are transmitted by dipteran flies. Thelazia callipaeda Railliet and Henry 1910 (Spirurida, Thelaziidae) is common in the Far East, where it causes eyeworm infection in carnivores and humans. In the past few years, it has also been reported in dogs, cats and foxes in Northern and Southern Italy. Although a great deal of work has been carried out in China and other Eastern countries, knowledge of the morphology of T. callipaeda is scanty. Eighty-three nematodes collected from the eyes of naturally infected dogs from the Basilicata region (southern Italy) were examined by light and scanning electron microscopy, and the most important features of the nematodes were described. The morphological characteristics useful for the identification of T. callipaeda are discussed.     

A study on the prevalence and intensity of occurrence of Thelazia skrjabini (Nematoda: Thelazioidea) in cattle in central Alberta, Canada

The distribution of Thelazia skrjabini in the eyes of cattle was examined from necropsies on 297 animals. Although the overall prevalence in beef cattle (21.5%) was similar to that in dairy cattle (25.7%), in general worms were more abundant in beef than in dairy animals. The worm prevalence was lowest in the months from March to June and highest in September among beef and dairy animals. Thelazia skrjabini was found in 76 (16.8%) eyes from beef, and 23 (16.4%) eyes from dairy cattle. Infections were not randomly distributed among the eyes of cattle. Significantly more dairy and beef cattle contained T. skrjabini in both eyes than expected based on the observed prevalences. Based on chi-square probabilities, significantly more cattle more than 2 yr of age and less than 10 mo of age were infected with T. skrjabini. Although all age groups were infected, more worms were observed in cattle more than 2 yr of age. More female worms than males were collected from cattle. The male-to-female ratio of 1:2 was consistent regardless of cattle age, season, or type.     

Development of the nematode eyeworm, Thelazia skrjabini (Nematoda: Thelazioidea), in experimentally infected face flies, Musca autumnalis (Diptera: Muscidae)

The development of Thelazia skrjabini Erschow, 1928, was studied in experimentally infected laboratory-reared Musca autumnalis De Geer. Thelazia skrjabini developed to the infective third stage in a minimum of 9 days in M. autumnalis maintained at 27 +/- 2 C. First-stage larvae were not observed postinoculation, but second-stage larvae were first observed 3 days postinoculation. Development was asynchronous. Second- and third-stage larvae occur in capsules, occasionally in the head but primarily in the abdomen attached to fat bodies. First-stage larvae have anteriorly 1 ventral and 2 dorsal hooks, directed posteriorly. Second-stage larvae have 4 submedian cephalic papillae and faint annular striations. Third-stage larvae have 6 labial papillae, 4 submedian cephalic papillae and pronounced annulations. Morphometric studies of each larval stage were performed with specimens in glycerine.     

Transfection of Escherichia coli Spheroplasts II. Relative Infectivity of Native, Denatured, and Renatured Lambda, T7, T5, T4, and P22 Bacteriophage DNAs

The change of infectivity of phage DNAs after heat and alkali denaturation (and renaturation) was measured. T7 phage DNA infectivity increased 4- to 20-fold after denaturation and decreased to the native level after renaturation. Both the heavy and the light single strand of T7 phage DNA were about five times as infective as native T7 DNA. T4 and P22 phage DNA infectivity increased 4- to 20-fold after denaturation and increased another 10- to 20-fold after renaturation. These data, combined with other authors' results on the relative infectivity of various forms of phiX174 and lambda DNAs give the following consistent pattern of relative infectivity. Covalently closed circular double-stranded DNA, nicked circular double-stranded DNA, and double-stranded DNA with cohesive ends are all equally infective and also most highly infectious for Escherichia coli lysozyme-EDTA spheroplasts; linear or circular single-stranded DNAs are about 1/5 to 1/20 as infective; double-stranded DNAs are only 1/100 as infective. Two exceptions to this pattern were noted: lambda phage DNA lost more than 99% of its infectivity after alkaline denaturation; this infectivity could be fully recovered after renaturation. This behavior can be explained by the special role of the cohesive ends of the phage DNA. T5 phage DNA sometimes showed a transient increase in infectivity at temperatures below the completion of the hyperchròmic shift; at higher temperatures, the infectivity was completely destroyed. T5 DNA denatured in alkali lost more than 99.9% of its infectivity; upon renaturation, infectivity was sometimes recovered. This behavior is interpreted in terms of the model of T5 phage DNA structure proposed by Bujard (1969). The results of the denaturation and renaturation experiments show higher efficiencies of transfection for the following phage DNAs (free of single-strand breaks): T4 renatured DNA at 10(-3) instead of 10(-5) for native DNA; renatured P22 DNA at 3 x 10(-7) instead of 3 x 10(-9) for native DNA; and denatured T7 DNA at 3 x 10(-6) instead of 3 x 10(-7) for native DNA.     

Renifer aniarum (Digenea: Reniferidae), an introduced North American parasite in grass snakes Natrix natrix in Calabria, southern Italy

Over the past decades, as a result of various human activities involving intentional or unintentional movement of animals, many helminth species have been introduced to new regions with several ecological and epidemiological implications for the native species. A high prevalence of infection with an introduced digenean Renifer aniarum, previously known only from North America, was found in the grass snake Natrix natrix in the Calabria region, southern Italy. Morphological and molecular comparison with North American R. aniarum has confirmed the identity of the Italian specimens. A total of 41 grass snakes were studied for R. aniarum infection. Of 24 snakes sampled between 2009 and 2010, 22 were positive for this parasite. In contrast, all 17 snakes sampled from museum collections between 1983 and 1994 were negative. Our results support the hypothesis that R. aniarum was perhaps introduced into this area during the 1990s by the translocation of the American bullfrog Lithobates (Rana) catesbeianus, a normal second intermediate host of the digenean in its native range in North America. Although the life cycle of R. aniarum is complex and includes 3 host stages, this parasite has found suitable first and second intermediate hosts as well as definitive hosts in Italy. Renifer aniarum was second only to the very common grass snake tapeworm Ophiotaenia europaea in both prevalence and abundance among 9 species of helminths recovered in our study.     

Serological,culture and molecular survey of <Emphasis Type="Italic">Mycobacterium avium paratuberculosis</Emphasis> in a goat flock in Tuscany

Mycobacterium avium paratuberculosis (Map) is a pathogen which causes a chronic progressive granulomatous enteritis known as paratuberculosis or Johne’s disease and it primarily affects wild and domestic ruminants. The aim of this research was to examine a flock which consisted of 294 goats and was located in Garfagnana district (Tuscany, Italy) performing ELISA tests, culture and IS900 PCR assay; direct diagnostic methods were carried out not only on bulk tank milk and cheese samples but also on individual milk and tissue specimens collected from nine subjects positive to ELISA tests. Out of 294 animals, 20 goats (6.8%) were positive to ELISA surveys. Bulk tank milk samples were negative to culture and to PCR assay carried out on the DNA extracted directly from them, while, with respect to cheese, Map was detected by culture in 2/12 (16.66%) cheeses ripened for 3–7 days, and by PCR in 2/12 (16.66%) cheeses ripened for 3–7 days and in 3/12 (25%) cheeses ripened for 45 days. Regarding individual milk samples, Map was detected by culture in 2/9 (22.22%) specimens and by PCR in 5/9 (55.55%) samples. Furthermore, Map was isolated from the intestine in 9/9 (100%) animals, from the mesenteric lymph nodes in 8/9 (88.88%) subjects, from the liver in 4/9 (44.44%) goats, from the spleen in 5/9 (55.55%) animals, while Map DNA was found in all the tissue samples analyzed.The results demonstrated the presence of paratuberculosis in a goat flock located in Garfagnana district (Tuscany, Italy).     

First report of Thelazia callipaeda (Spirurida, Thelaziidae) in wolves in Italy

Thelazia callipaeda (Spirurida, Thelaziidae) infects the eyes of humans and rabbits as well of domestic and wild carnivores (i.e., dogs, cats, and foxes). The first three cases of infection by T. callipaeda eyeworms in wolves (Canis lupus) are here described with up to 96 nematodes collected from a single animal. The host competence of wolf was demonstrated by the retrieval of adult worms in the eyes of all examined animals. All nematodes collected were evaluated by morphologic and molecular techniques. Sequence analysis of the partial mitochondrial cytochrome c oxidase subunit 1 gene (cox1) confirmed that only one haplotype of T. callipaeda was present. The competence of wolves as a definitive host for T. callipaeda is discussed in view of the relevance of wild fauna in maintaining and spreading eyeworm infection in humans and domestic animals.     

Exposure of free-ranging wild carnivores, horses and domestic dogs to Leptospira spp in the northern Pantanal, Brazil

Leptospirosis is a zoonotic disease affecting most mammals and is distributed throughout the world. Several species of domestic and wild animals may act as reservoirs for this disease. The purpose of this study was to assess the exposure of free-ranging wild carnivores, horses and domestic dogs on a private reserve located in the northern Pantanal (Brazil) and the surrounding areas to Leptospira spp from 2002-2006, 75 free-ranging wild carnivores were captured in the Pantanal and serum samples were collected. In addition, samples from 103 domestic dogs and 23 horses in the region were collected. Serum samples were tested for the presence of Leptospira antibodies using the microscopic agglutination test. Thirty-two wild carnivores (42.7%) were considered positive with titres ≥ 100, and 18 domestic dogs (17.5%) and 20 horses (74.1%) were also found to be positive. Our study showed that horses, dogs and several species of free-ranging wild carnivores have been exposed to Leptospira spp in the Pantanal, suggesting that the peculiar characteristics of this biome, such as high temperatures and an extended period of flooding, may favour bacterial persistence and transmission. In this region, wild carnivores and horses seem to be important hosts for the epidemiology of Leptospira species.     

Genetic variation in Przewalski's horses, with special focus on the last wild caught mare, 231 Orlitza III

In our continuing efforts to document genetic diversity in Przewalski's horses and relatedness with domestic horses, we report genetic variation at 22 loci of blood group and protein polymorphisms and 29 loci of DNA (microsatellite) polymorphisms. The loci have been assigned by linkage or synteny mapping to 20 autosomes and the X chromosome of the domestic horse (plus four loci unassigned to a chromosome). With cumulative data from tests of 568 Przewalski's horses using blood, hair or tooth samples, no species-defining markers were identified, however a few markers were present in the wild species but not in domestic horses. Inheritance patterns and linkage relationships reported in domestic horses appeared to be conserved in Przewalski's horses. A derived type for the last wild caught mare 231 Orlitza III provided evidence for markers apparently not found in (or not currently available by descent from) the other species founders that were captured at the end of the nineteenth century. This information has been critical to the development of parentage analyses in the studbook population of Przewalski's horses at Askania Nova, at one time the largest herd of captive animals and the source of stock for reintroduction efforts. Some horses in the study showed genetic incompatibilities with their sire or dam, contradicting published studbook information. In many cases alternative parentage could be assigned from living animals. To assist in identification of correct parentage, DNA marker types for deceased horses were established from archived materials (teeth) or derived from offspring. Genetic markers were present in pedigreed animals whose origin could not be accounted for from founders. Genetic distance analysis of erythrocyte protein, electrophoretic and microsatellite markers in Przewlaski's horses and ten breeds of domestic horse place the Przewalski's horse as an outgroup to domestic horses, introgression events from domestic horses not withstanding.     

Equine piroplasmoses at the reintroduction site of the Przewalski's horse (Equus ferus przewalskii) in Mongolia

Piroplasmosis has been identified as a possible cause of mortality in reintroduced Przewalski's horses (Equus ferus przewalskii) in the Dsungarian Gobi (Mongolia). A cross-sectional and a longitudinal study were conducted in a representative sample (n = 141) of the resident domestic horse population and in 23 Przewalski's horses to assess the prevalence of Theileria equi and Babesia caballi. Piroplasms were detected in blood by light microscopy in 6.7% (95% confidence interval [CI]: 3.6-12.2%) of the domestic horse samples. Antibody prevalence was 88.6% (95% CI: 82.4-92.9%) for T. equi and 75.2% (95% CI: 67.4-81.6%) for B. caballi. Antibody prevalence did not change over time, but antibody prevalence for both piroplasms were significantly lower in animals less than 1 yr of age. For both piroplasms, the prevalence of presumably maternal antibodies (falling titers) in foals was 100%. Only one of 16 foals seroconverted against T. equi during the study period, despite that piroplasms were found in two other individuals. The incidence density (ID) of T. equi in foals was therefore 0.0012 seroconversions per horse day (95% CI: 0.00029-0.0057). In contrast, yearlings had an ID of 0.0080 (95% CI: 0.0049-0.010) for T. equi and 0.0064 (95% CI: 0.0036-0.0093) for B. caballi, and in seven individuals piroplasms were detected. The seroprevalence of both piroplasms rose from 20% in spring to 100% in autumn. Comparison of domestic and Przewalski's horses resulted in a standardized prevalence ratio (SPR) of 0.98 (95% CI: 0.80-1.24, not significant) for B. caballi; in contrast, the prevalence of T. equi in Przewalski's horses was significantly lower than expected (SPR = 0.51, 95% CI: 0.50-0.64).