First Detection of Anti-Besnoitia spp. Antibodies in Equids in Israel and the Palestinian Authority.

Besnoitia is a tissue cyst forming coccidia, which affects multiple host species worldwide. Equine besnoitiosis is characterized mainly by generalized skin lesions and cysts in the scleral conjunctiva. Recent reports revealed exposure to Besnoitia in equines in Europe and the United States. However, the exposure to Besnoitia spp. in the Israeli equine population was never investigated. The aim of this study was to evaluate the seroprevalence and associated risk factors for besnoitiosis in equids in Israel. A cross-sectional serosurvey was performed using serum samples of apparently healthy horses (n = 347), donkeys (n = 98), and mules (n = 6), and exposure to Besnoitia spp. was determined by an immunofluorescent antibody test (IFAT). Anti-Besnoitia spp. antibodies were detected in 17.7% equids, 6.9% horses, 33.3% mules, and 55.1% donkeys. The seroprevalence in donkeys was significantly higher than in horses (p < 0.001). A significant association between the geographic location and seropositivity was found both in horses and donkeys, which was significantly higher (p = 0.004) in horses sampled in southern Israel, and donkeys sampled in Israel versus the Palestinian Authority (p < 0.001). This is the first serosurvey of Besnoitia infection in equines in Israel, and the results are consistent with reports from Europe. The clinical significance of equine besnoitiosis should be further investigated.

Spinal cord protothecosis causing paraparesis in a dog.

Protothecosis, an infectious disease caused by the green algae Prototheca zopfii and P. wickerhamii, occurs sporadically in domestic animals and humans. Diagnosis of CNS protothecosis is based on neurologic signs that indicate multifocal nervous system lesions and that follow a period of chronic diarrhea and weight loss, cytologic observation of algae in fecal culture or histopathology, and detection of the agent by PCR assay of infected tissues. Here, we report a case of a paraparetic dog with CNS protothecosis that was diagnosed definitively antemortem using CSF cytology, PCR, and DNA sequencing. A 4-y-old mixed-breed dog developed progressive paraparesis that followed weight loss and diarrhea. CSF analysis revealed marked eosinophilic pleocytosis. Prototheca organisms were detected by microscopic examination of the CSF, and speciated as P. zopfii by CSF PCR and DNA sequencing. Other possible causes of paraparesis were ruled out using computed tomography, serology, and CSF PCR. The dog's condition deteriorated despite treatment, developing forebrain and central vestibular system clinical signs, and it was euthanized at the owner's request. Postmortem examination was declined. Our findings indicate that when CNS protothecosis is suspected, antemortem diagnosis can be made using CSF analysis and a PCR assay.

Cryptosporidium parvum subtypes from diarrheic dairy calves in Israel.

Cryptosporidium are protozoan parasites with worldwide distribution, infecting a wide range of terrestrial and aquatic animals, as well as humans. Cryptosporidium parvum is the most important zoonotic species and is the primary cause of cryptosporidiosis in preweaned calves, a highly prevalent, economically important disease. Extensive subtyping of C. parvum from infected humans and animals has expanded current understanding of the parasites' epidemiology. Israel has a highly developed dairy sector with intensive, zero-grazing operations. While C. parvum has been found in dairy calves throughout the country, and subtype data from human patients have also been published, subtype data from animals, and in particular preweaned ruminants, are lacking. We carried out an initial study of Cryptosporidium species and subtypes from preweaned diarrheic calves. Cryptosporidium species were determined in 71 fecal samples from 43 different dairy farms using 18S rRNA PCR, and subtyping of C. parvum based on the 60-kDa glycoprotein (gp60) sequences was done on one sample per farm. C. parvum was the only species found, with eight different subtypes belonging to the zoonotic IIa and IId families. Subtype IIaA15G2R1 was the most prevalent and widespread, found in 50% of the farms over a wide geographical distribution. Our results confirm the presence of subtypes IIaA15G2R1 and IIdA20G1, which were previously found in human patients in Israel, also in Israeli calves. In addition, subtype IIaA12G1R1 is reported here for the first time in an animal. These findings demonstrate the value of monitoring C. parvum subtypes in animal samples, and suggest that the role of calves as well as other young ruminants in the transmission of zoonotic cryptosporidiosis in Israel should be further studied.

Genomic Detection of Schmallenberg Virus, Israel.

We discuss genomic detection of Schmallenberg virus in both Culicoides midges and affected ruminants during June 2018-December 2019, demonstrating its circulation in Israel. This region is a geographic bridge between 3 continents and may serve as an epidemiologic bridge for potential Schmallenberg virus spread into Asia.

Detection of Neospora caninum Infection in Aborted Equine Fetuses in Israel.

In horses, Neospora caninum and Neospora hughesi have been associated with fetal loss, and neurological disease, respectively. This study investigated the role of Neospora spp. infection in equine abortion in Israel. The presence of anti-Neospora spp. antibodies was evaluated in 31 aborting mares by indirect fluorescent antibody test (IFAT) and the presence of parasite DNA in their aborted fetuses was evaluated by polymerase chain reaction (PCR), using two target loci (ITS1 and Nc5). The seroprevalence found in aborting mares was 70.9% and the prevalence by DNA detection in the aborted fetuses was 41.9%. Transplacental transmission from positive mares to their fetuses was 45.4% (10/22), while 33.3% (3/9) of fetuses of seronegative mares also tested positive for Neospora. The use of two PCR targets improved the sensitivity of parasite detection, and positive samples were identified by sequence analyses as N. caninum. These finding suggest that N. caninum could be a significant cause of abortion in horses, and that transplacental transmission in horses is an important way of transmission of N.caninum. The results presented here demonstrated the necessity to use several tests concurrently, including serological and molecular assays in order to confirm the involvement of Neospora in mare abortions.

Leishmania infection in cats and dogs housed together in an animal shelter reveals a higher parasite load in infected dogs despite a greater seroprevalence among cats.

BACKGROUND: An outbreak of leishmaniosis was studied in cats and dogs housed together with no separation in an animal shelter in Israel. METHODS: The study included recording of clinical signs, serology for Leishmania infection by ELISA, PCR of blood for Leishmania DNA by ITS1 HRM and kDNA PCR, parasite quantification, and trapping of sand flies around the shelter. RESULTS: Thirty-seven % (22/60) of the dogs and 75% (50/67) of the cats were seropositive to L. infantum with a significantly higher seropositivity rate in the cat population (χ2 = 42.160, P < 0.0001). Twenty-five percent (15/60) of the dogs were positive for Leishmania by blood PCR, 12% by the Leishmania ITS1 HRM PCR and 22% by kDNA PCR. Of the cats, 16% (11/67) were positive by kDNA PCR and none by ITS1 HRM PCR. All the PCR-positive animals were infected by L. infantum verified by DNA sequencing and there was no significant difference between the PCR-positivity in the dog and cat populations. Altogether, 43% (26/60) of the dogs and 79% (53/67) of the cats were positive by serology or PCR for L. infantum. The average Leishmania parasite load in the blood of PCR-positive dogs (42,967 parasites/ml) was significantly higher than in PCR-positive cats (1259 parasites/ml) (t(12) = 2.33, P = 0.037). Dogs that were positive by the Leishmania ITS1 HRM PCR and kDNA PCR had significantly higher parasite loads than dogs positive only by the kDNA PCR (t(11) = - 3.186580, P < 0.009). No significant effect was found for FIV seropositivity on Leishmania infection in the cats (χ2 = 0.506, P = 0.777). A higher percentage of Leishmania-positive dogs showed clinical signs compatible with leishmaniosis compared to Leishmania-positive cats (100 vs 52.8%, χ2 =15.242, P < 0.0001). Phlebotomus perfiliewi, a proven vector of L. infantum, comprised 92% of trapped sand flies. CONCLUSIONS: Comparisons of populations of cats and dogs exposed to sand flies and L. infantum under the same conditions indicated that although a high rate of exposure was detected in cats as manifested by a significantly greater degree of seropositivity, dogs had significantly higher blood parasite loads, and were likely to be more infectious to sand flies than cats.

Genetic Characterization of Piroplasms in Donkeys and Horses from Nigeria.

Equine piroplasmosis (EP) is a tick-borne disease of equids, caused by the two haemoprotozoal parasites: Theileria equi and Babesia caballi. Nigeria constitutes a major crossroads of animal transport in West Africa and may serve as a factor in EP dissemination in the region. The study aim was to characterize EP parasites in donkeys and horses in northern Nigeria using a molecular approach. Blood was collected from 57 donkeys and 47 horses. EP infection was detected and characterized by polymerase chain reaction (PCR). Twenty five donkeys (43.8%) were infected with T. equi, five (8.8%) with B. caballi, three (5.3%) with dual infections. Four horses (8.5%) were infected by T. equi and none by B. caballi. Four of the five known T. equi 18S rRNA genotypes (A, B, C and D) were identified. Theileria equi ema-1 and ema-2 genes were amplified in only 2 and 10 samples, respectively, showing no genetic variation. All B. caballi isolates were classified as rap-1 genotype A1. Twenty-two (42.3%) of the donkeys were positive for anti-T. equi antibodies and 29 (55.8%) were positive for anti-B. caballi antibodies, using immunofluorescence antibody test (IFAT). The study results demonstrate high genetic variation within T. equi parasites, suggesting that donkeys may be reservoirs of EP parasites in West Africa.