Systemic toxoplasmosis in a cat under cyclosporine therapy.

Toxoplasma gondii, an obligatory intracellular protozoan parasite infecting warm-blooded animals, can cause toxoplasmosis, a major zoonosis. A male neutered, domestic cat was referred to the Hebrew University Veterinary Teaching Hospital due to dyspnea after long term treatment with cyclosporine for obsessive self-grooming and pruritis. After thorough diagnostics, including non - invasive imaging, broncho-alveolar lavage, blood serology, hematology and biochemistry, and evaluation of the aspirated fluid components, a severe pneumonia and abdominal effusion were detected with observation of free tachyzoites under light microscopy from lavage fluids. PCR and DNA sequencing of broncho-alveolar lavage was positive for T. gondii. Despite aggressive treatment with antibiotics, oxygen supplementation and T. gondii specific antimicrobials, the cat died. It is suggested that potential candidates for cyclosporine be screened for T. gondii antibodies, kept entirely indoors and not fed uncooked meat in order to prevent exposure to T. gondii infection.

Cutaneous habronemosis in horses: First molecular characterization of Habronema muscae in Israel.

Draschia megastoma, Habronema microstoma, and Habronema muscae are the etiological agents of cutaneous habronemosis, commonly known as summer sores, an inflammatory cutaneous and ocular parasitic disease of horses and other equids transmitted by flies. Here, we describe a cluster of cutaneous habronemosis in five horses that showed single or multiple typical cutaneous ulcerative wounds located on the face, lower forelegs or hindquarters in Israel with the presence of typical "sulphur granules." All affected animals were confirmed by histopathological and/or molecular methods to be infected by H. muscae. This constitutes the first report of cutaneous habronemosis in Israel in which the causative nematode, H. muscae, was identified by molecular means. Cutaneous habronemosis should be considered as a differential diagnosis in equids with cutaneous ulcerative lesions during the summer months, especially when affected animals are refractive to antibiotic treatment alone.

Troglostrongylus brevior is the dominant lungworm infecting feral cats in Jerusalem.

Feline lungworms infect the respiratory tract of wild and domestic cats, causing infection often associated with clinical disease. Until recently, Aelurostrongylus abstrusus has been considered the most relevant species of lungworm, while Troglostrongylus brevior was considered of less significance. Fecal samples of feral cats from Jerusalem, Israel, collected over a year, were examined for first stage lungworm larvae (L1) using the Baermann method. Positive samples were morphologically identified, and their species identity was molecularly confirmed. Forty of 400 (10.0%) cats were lungworm-positive, of which 38/40 (95.0%) shed Troglostrongylus brevior and 6/40 (15.0%) shed Aelurostrongylus abstrusus. Four cats (10.0%) had mixed infections with both lungworm species. L1 shedding was associated with clinical respiratory signs in 11 (19.0%) T. brevior shedding cats of a total of 58 cats manifesting respiratory signs, while 23/342 (6.7%) cats without respiratory signs were L1-positive (p = 0.006). Non-respiratory clinical signs were also found to be more prevalent in L1 shedders (p = 0.012). A young kitten ≤ 4 weeks of age shed T. brevior L1 larvae. DNA sequences of both lungworm species using the ribosomal internal transcribed spacer 2 (ITS2) locus were > 99% similar to other sequences deposited in GenBank, suggesting that T. brevior and A. abstrusus ITS2 sequences are both highly conserved. In conclusion, L1 shedding in feral cats from Jerusalem were mostly caused by T. brevior with only a small proportion involving A. abstrusus, different from many studies from other geographical regions.

First report of Onchocerca lupi from Israel and confirmation of two genotypes circulating among canine, feline and human hosts.

Onchocerca lupi is a parasitic filarioid and the causative agent of canine ocular onchocercosis, a zoonotic disease of domestic dogs with sporadic reports in humans. A 13-year-old dog with no travel history outside of Israel was presented to an ophthalmology veterinary clinic in Israel with severe right ocular and periocular disease. After surgical exploration, thin helminths were removed from the dorsal sclera of the eye and identified as Onchocerca lupi by polymerase chain reaction according to the cytochrome c oxidase subunit I (cox1), reduced nicotinamide adenine dinucleotide dehydrogenase subunit 5 (nad5) and 12S rRNA genes. Phylogenetic trees and haplotype networks of the cox1 and nad5 genes confirmed the circulation of two genotypes: genotype 1 with worms from dogs, cats and humans from both the Old and New Worlds, and genotype 2 with specimens from Portugal and Spain. The Israeli sequences clustered in genotype 1 and were identical to O. lupi from the USA. Evidence of two genotypes separated geographically sheds light on the phylogeography and evolution of this zoonotic pathogen, and suggests a diverse pathology observed in different regions of the world.

Detection of Intraspinal Spirocerca lupi in Canine Cerebrospinal Fluid by Polymerase Chain Reaction.

Aberrant migration of Spirocerca lupi into the spinal cord is an important cause of severe progressive neurological dysfunction in dogs. Although early diagnosis is essential to prevent deterioration, ante-mortem diagnosis of this condition remains challenging. The aim of this study was to evaluate the detection of the 18S ribosomal DNA (rDNA) S. lupi gene in the cerebrospinal fluid (CSF) of presumptively-affected dogs using polymerase chain reaction (PCR). Dogs with a non-compressive spinal cord lesion, pleocytosis with presence of eosinophils in the CSF and a characteristic clinical presentation were included. CSF samples from eight dogs were available for the study, of which seven were definitively diagnosed with intraspinal spirocercosis by PCR of either the CSF samples (6/7) or tissue samples obtained at necropsy examination (3/7), or both (2/7). Of these seven positive cases, only one dog had a negative CSF PCR, indicating a sensitivity of 86% for detecting nematode DNA in the CSF of infected dogs using this PCR protocol. The nematode DNA sequences obtained from the CSF of six dogs and the spinal cord tissue of three dogs were 98-100% identical to the publicly available sequences of S. lupi, confirming the diagnosis. These findings indicate that PCR targeting the 18S rDNA of S. lupi in CSF is useful for the ante-mortem diagnosis of canine intraspinal spirocercosis.

Urinary incontinence associated with Mesocestoides vogae infection in a dog.

Peritoneal larval cestodiasis caused by Mesocestoides spp. is a rare infection in dogs. A 6-year-old female dog was presented for veterinary care with urinary incontinence which started 1 year earlier. After performing hematology, ultrasound, and computerized tomography, an exploratory laparotomy revealed canine peritoneal larval cestodiasis (CPLC) with the presence of Mesocestoides vogae (syn. Mesocestoides corti) tetrathyridia confirmed by morphological identification and PCR and DNA sequencing. Parasitic cysts were found around the urinary bladder and appeared to inhibit its normal function. An initial treatment with 5 mg/kg praziquantel subcutaneously every 2 weeks for four treatments failed to alleviate the clinical signs, and only treatment with fenbendazole at 100 mg/kg P.O. twice daily for 28 days was associated with the disappearance of ascites and regaining of urinary control. This is the first report of CPLC associated with urinary incontinence in dogs and the first description of this cyclophyllidean cestode in dogs in Israel.

Molecular investigations of cat fleas (Ctenocephalides felis) provide the first evidence of Rickettsia felis in Malta and Candidatus Rickettsia senegalensis in Israel.

Rickettsia felis, the causative agent of flea-borne spotted fever, occurs on all continents except Antarctica, owing to the cosmopolitan distribution of its cat flea vector. In this study, cat fleas were collected in two countries where the occurrence of R. felis was either unknown (Malta) or where accurate prevalence data were lacking (Israel). Altogether 129 fleas were molecularly analysed for the presence of rickettsial DNA. On the basis of three genetic markers, R. felis was identified in 39.5% (15/38) of the cat fleas from Malta. Sequences showed 100% identity to each other and to relevant sequences in GenBank. Among the 91 cat fleas from Israel, two (2.2%) contained the DNA of Candidatus Rickettsia senegalensis. Phylogenetically, the R. felis and Candidatus R. senegalensis identified here clustered separately (with high support) but within one clade, which was a sister group to that formed by the typhus group and spotted fever group rickettsiae. This is the first record of R. felis in Malta and of Candidatus R. senegalensis outside its formerly reported geographical range including Africa, Asia and North America.

Use of acute phase proteins for the clinical assessment and management of canine leishmaniosis: general recommendations.

BACKGROUND: Dogs with canine leishmaniosis (CanL) due to Leishmania infantum can show a wide spectrum of clinical and clinicopathological findings at the time of diagnosis. The aim of this paper is to describe the possible application of acute phase proteins (APPs) for the characterization and management of this disease, based on previously published information on the utility of APPs in CanL and the experience of the authors in using APPs as analytes in the profiling of canine diseases. MAIN BODY: Dogs diagnosed with L. infantum infection by serology, polymerase chain reaction, cytological or histopathological identification, can be divided into three groups based on their clinical condition at physical examination and their APPs concentrations: Group 1: dogs with no clinical signs on physical examination and APPs in reference range; Group 2: dogs with changes in APPs but no clinical signs on physical examination; Group 3: dogs with clinical signs and changes in APPs. This report describes the main characteristics of each group as well as its association with the clinical classification schemes of CanL. CONCLUSION: APPs concentration can be a useful clinical tool to characterize and manage CanL.

Morphological and molecular identification of Rhipicephalus (Boophilus) microplus in Nigeria, West Africa: a threat to livestock health.

The cattle tick Rhipicephalus (Boophilus) microplus was first reported in West Africa in Ivory Coast, in 2007. Since then it has made an aggressive eastward advancement having been reported in four other West African countries: Mali, Burkina Faso, Togo and Benin. We herein report the first molecular identification of this tick species in Nigeria, West Africa, and highlight the threat it poses to livestock health. A nation-wide tick survey was conducted in 12 out of 36 states across the various agro ecological zones of Nigeria over a 1 year period (April 2014-March 2015). In total 1498 ticks belonging to three genera collected from cattle were morphologically identified. Overall, Amblyomma species constituted the highest percentage of sampled ticks, 50.2% (752/1498), followed by Rhipicephalus (including the subgenus Boophilus) species, 29.4% (440/1498) and Hyalomma species, 20.4% (306/1498). The presence of Rh. (B.) microplus was identified morphologically from four out of the 12 states. This finding was confirmed for the first time in Nigeria using a molecular method targeting the ITS-2 region of the ticks in three of the 12 states. This study ascertained the presence of Rh. (B.) microplus in Nigeria in addition to a broad variety of cattle tick species, most of which are of veterinary importance. The implication of this finding is that there may be additional economic burden to livestock farmers due to increased cost of tick control occasioned by the acaricide resistance by this tick species widely reported from different climes. Additionally, there may be a potential upsurge in incidence of hemoparasitic infections in cattle leading to increased morbidity, cost of treatment and mortalities.

Major Parasitic Zoonoses Associated with Dogs and Cats in Europe.

Some of the most important zoonotic infectious diseases are associated with parasites transmitted from companion animals to man. This review describes the main parasitic zoonoses in Europe related to dogs and cats, with particular emphasis on their current epidemiology. Toxoplasmosis, leishmaniosis, giardiosis, echinococcosis, dirofilariosis and toxocariosis are described from the animal, as well as from the human host perspectives, with an emphasis on parasite life cycle, transmission, pathogenicity, prevention and identification of knowledge gaps. In addition, priorities for research and intervention in order to decrease the risks and burden of these diseases are presented. Preventing zoonotic parasitic infections requires an integrated multidisciplinary 'One Health' approach involving collaboration between veterinary and medical scientists, policy makers and public health officials.

Prioritization of Companion Animal Transmissible Diseases for Policy Intervention in Europe.

A number of papers have been published on the prioritization of transmissible diseases in farm animals and wildlife, based either on semiquantitative or truly quantitative methods, but there is no published literature on the prioritization of transmissible diseases in companion animals. In this study, available epidemiological data for diseases transmissible from companion animals to man were analysed with the aim of developing a procedure suitable for their prioritization within a European framework. A new method and its associated questionnaire and scoring system were designed based on methods described by the World Organisation for Animal Health (OIE). Modifications were applied to allow for the paucity of specific information on companion animal transmissible diseases. The OIE method was also adapted to the subject and to the regional scope of the interprofessional network addressing zoonotic diseases transmitted via companion animals in Europe: the Companion Animals multisectoriaL interprofessionaL Interdisciplinary Strategic Think tank On zoonoses (CALLISTO). Adaptations were made based on information collected from expert groups on viral, bacterial and parasitic diseases using a structured questionnaire, in which all questions were closed-ended. The expert groups were asked to select the most appropriate answer for each question taking into account the relevance and reliability of the data available in the scientific literature. Subsequently, the scoring of the answers obtained for each disease covered by the questionnaire was analysed to obtain two final overall scores, one for human health impact and one for agricultural economic impact. The adapted method was then applied to select the 15 most important pathogens (five for each pathogen group: viral, bacterial and parasitic) on the basis of their overall impact on public health and agriculture. The result of the prioritization exercise was a joint priority list (available at www.callistoproject.eu) of relevant pathogens according to these two criteria. As the scope of CALLISTO was comprehensive in terms of geographical area, animal species involved and impact of the diseases, the list of prioritized diseases had to accommodate the realities in different European countries and the differences in biology and animal-human relationships in a wide range of species including cats and dogs, pet pigs and sheep as well as captive reptiles. The methodology presented in this paper can be used to generate accurate priority lists according to narrower and more specific objectives.

Neospora caninum in crows from Israel.

A cross-sectional Neospora caninum seroprevalence study was performed on free ranging crows (Corvus cornix, Corvus monedula and Corvus splendens) from Israel in order to assess their exposure to this pathogen and evaluate their role as potential hosts or as sentinels of infection. Using the modified agglutination test (MAT) with a cutoff titer of 1:100, 30 out of 183 crows (16.4%) were found to be N. caninum seropositive. Positive results were validated and confirmed by the indirect fluorescent antibody test (IFAT). There was 100% agreement between tests when cut-off titers of 1:50 and 1:100 were applied for the IFAT and MAT, respectively. PCR analysis of brain extracts from all crows resulted in the detection of N. caninum DNA for the first time in crows belonging to two species, C. cornix and C. monedula. The high N. caninum seroprevalence in crows suggests that widespread exposure to infection with N. caninum exists especially in central and northern Israel and that crows may act as suitable markers for disease prevalence in the areas in which they are found.

Classification of Babesia canis strains in Europe based on polymorphism of the Bc28.1-gene from the Babesia canis Bc28 multigene family.

The vast majority of clinical babesiosis cases in dogs in Europe is caused by Babesia canis. Although dogs can be vaccinated, the level of protection is highly variable, which might be due to genetic diversity of B. canis strains. One of the major merozoite surface antigens of B. canis is a protein with a Mr of 28 kDa that belongs to the Bc28 multigene family, that comprises at least two genes, Bc28.1 and a homologous Bc28.2 gene. The two genes are relatively conserved but they are very distinct in their 3' ends, enabling the design of specific primers. Sequencing of the Bc28.1 genes from 4 genetically distinct B. canis laboratory strains (A8, B, 34.01 and G) revealed 20 mutations at conserved positions of which three allowed the classification of B. canis strains into three main groups (A, B and 34.01/G) by RFLP. This assay was subsequently used to analyze blood samples of 394 dogs suspected of clinical babesiosis from nine countries in Europe. All blood samples were first analyzed with a previously described assay that allowed detection of the different Babesia species that infect dogs. Sixty one percent of the samples contained detectable levels of Babesia DNA. Of these, 98.3% were positive for B. canis, the remaining cases were positive for B. vogeli. Analysis of the Bc28.1 gene, performed on 178 of the B. canis samples, revealed an overall dominance of genotype B (62.4%), followed by genotypes A (37.1%) and 34 (11.8%). Interestingly, a great variation in the geographical distribution and prevalence of the three B. canis genotypes was observed; in the North-East genotype A predominated (72.1% A against 27.9% B), in contrast to the South-West where genotype B predominated (10.3% A against 89.7% B). In the central part of Europe intermediate levels were found (26.0-42.9% A against 74.0-57.1% B, from West to East). Genotype 34 was only identified in France (26.9% among 78 samples) and mostly as co-infection with genotypes A or B (61.9%). A comparative analysis of the classification of 35 B. canis strains in genotypes A and B using a previously described 18SrDNA-derived PCR-RFLP test revealed a partial but no direct correlation with the classification based on polymorphism of the Bc28.1-gene described here.

Molecular detection of Rickettsia aeschlimannii in Hyalomma spp. ticks from camels (Camelus dromedarius) in Nigeria, West Africa.

Several species of the spotted fever group rickettsiae have been identified as emerging pathogens throughout the world, including in Africa. In this study, 197 Hyalomma ticks (Ixodida: Ixodidae) collected from 51 camels (Camelus dromedarius) in Kano, northern Nigeria, were screened by amplification and sequencing of the citrate synthase (gltA), outer membrane protein A (ompA) and 17-kDa antigen gene fragments. Rickettsia sp. gltA fragments were detected in 43.3% (42/97) of the tick pools tested. Rickettsial ompA gene fragments (189 bp and 630 bp) were detected in 64.3% (n = 27) and 23.8% (n = 10) of the gltA-positive tick pools by real-time and conventional polymerase chain reaction (PCR), respectively. The amplicons were 99-100% identical to Rickettsia aeschlimannii TR/Orkun-H and R. aeschlimannii strain EgyRickHimp-El-Arish in GenBank. Furthermore, 17-kDa antigen gene fragments of 214 bp and 265 bp were detected in 59.5% (n = 25) and 38.1% (n = 16), respectively, of tick pools, and sequences were identical to one another and 99-100% identical to those of the R. aeschlimannii strain Ibadan A1 in GenBank. None of the Hyalomma impressum ticks collected were positive for Rickettsia sp. DNA. Rickettsia sp. gltA fragments (133 bp) were detected in 18.8% of camel blood samples, but all samples were negative for the other genes targeted. This is the first report to describe the molecular detection of R. aeschlimannii in Hyalomma spp. ticks from camels in Nigeria.

Bartonella species in fleas from Palestinian territories: prevalence and genetic diversity.

Bartonellosis is an infectious bacterial disease. The prevalence and genetic characteristics of Bartonella spp. in fleas of wild and domestic animals from Palestinian territories are described. Flea samples (n=289) were collected from 121 cats, 135 dogs, 26 hyraxes and seven rats from northern (n=165), central (n=113), and southern Palestinian territories (n=11). The prevalent flea species were: Ctenocephalides felis (n=119/289; 41.2%), Ctenocephalides canis (n=159/289; 55%), and Xenopsylla sp. (n=7/289; 2.4%). Targeting the Intergenic Transcribed Spacer (ITS) locus, DNA of Bartonella was detected in 22% (64/289) of all fleas. Fifty percent of the C. felis and 57% of the Xenopsylla sp. contained Bartonella DNA. DNA sequencing showed the presence of Bartonella clarridgeiae (50%), Bartonella henselae (27%), and Bartonella koehlerae (3%) in C. felis. Xenopsylla sp. collected from Rattus rattus rats were infected with Bartonella tribocorum, Bartonella elizabethae, and Bartonella rochalimae. Phylogenetic sequence analysis using the 16S ribosomal RNA gene obtained four genetic clusters, B. henselae and B. koehlerae as subcluster 1, B. clarridgeiae as cluster 2, while the rat Bartonella species (B. tribocorum and B. elizabethae) were an outgroup cluster. These findings showed the important role of cat and rat fleas as vectors of zoonotic Bartonella species in Palestinian territories. It is hoped that this publication will raise awareness among physicians, veterinarians, and other health workers of the high prevalence of Bartonella spp. in fleas in Palestinian territories and the potential risk of these pathogens to humans and animals in this region.

Prevalence and molecular characterization of Hepatozoon canis in dogs from urban and rural areas in Southeast Brazil.

The objective of this survey was to investigate the prevalence of Hepatozoon infection in dogs in the rural and urban areas of Uberlândia, Brazil by PCR and molecular characterization. DNA was obtained from blood samples collected from 346 local dogs from both genders and various ages. Seventeen PCR products from positive blood samples of urban dogs and 13 from the rural dogs were sequenced. Partial sequences of the 18S rRNA gene indicated that all 30 dogs were infected with Hepatozoon canis similar in sequence to H. canis from southern Europe. Four local dog sequences were submitted to GenBank (accessions JN835188; KF692038; KF692039; KF692040). This study indicates that H. canis is the cause of canine hepatozoonosis in Uberlândia and that infection is similarly widespread in rural and urban dogs.

Ectoparasites in urban stray cats in Jerusalem, Israel: differences in infestation patterns of fleas, ticks and permanent ectoparasites.

In a period cross-sectional study performed to examine ectoparasites on 340 stray cats in Jerusalem, Israel, 186 (54.7%) were infested with the cat flea, Ctenocephalides felis (Siphonaptera: Pulicidae), 49 (14.4%) with the cat louse, Felicola subrostratus (Phthiraptera: Trichodectidae), 41 (12.0%) with the ear mite, Otodectes cynotis (Astigmata: Psoroptidae), three (0.9%) with the fur mite, Cheyletiella blakei (Trobidiformes: Cheyletidae), two (0.6%) with the itch mite Notoedres cati (Astigmata: Sarcoptidae), and 25 (7.3%) with ticks of the species Rhipicephalus sanguineus sensu lato (Ixodida: Ixodidae), Rhipicephalus turanicus or Haemaphysalis adleri (Ixodida: Ixodidae). A higher number of flea infestations was observed in apparently sick cats (P < 0.05) and in cats aged < 6 months (P < 0.05). The proportion of flea-infested cats (P < 0.01), as well as the number of fleas per infested cat (P < 0.01), was higher in autumn than in other seasons. By contrast with findings in cats with flea infestations, rates of infestation with ticks were higher amongst cats with clinical signs (P < 0.01) and cats aged ≥ 6 months (P < 0.05). The high rates of ectoparasite infestation in the cats studied constitute a risk for the spread of vector-borne infections of zoonotic and veterinary importance.

Serum ferritin and paraoxonase-1 in canine leishmaniosis.

Ferritin and paraoxonase-1 (PON-1) were measured in dogs experimentally infected by Leishmania infantum (during experimental infection and following treatment) and also in naturally-infected dogs which presented different degrees of proteinuria. Experimentally-infected dogs were monitored for 7 months post-infection, then treated for 3 months with allopurinol, and their response to therapy was followed for 11 additional months. Naturally-infected dogs were staged based on the urine protein/creatinine (UPC) ratio into three groups as follows: group 1 (non-proteinuric; UPC ratio: <0.2), group 2 (borderline proteinuric; UPC ratio: 0.2-0.5) and group 3 (proteinuric; UPC ratio>0.5). An increase in serum ferritin values and a decrease in PON-1 activity were observed 2 months after infection. Both analytes returned to preinfection values following treatment. Significantly higher concentrations of ferritin were observed in dogs classified as either borderline or proteinuric when compared with non-proteinuric dogs whereas serum PON-1 activity was decreased only in proteinuric dogs.

Prevalence of Babesia microti-like infection in red foxes (Vulpes vulpes) from Portugal.

The prevalence of piroplasm (order Piroplasmida) infection was assessed in blood and bone marrow samples from 91 red foxes (Vulpes vulpes) from northern, central and southern Portugal by means of molecular methods. PCR for the 18S rRNA gene of Babesia spp. followed by sequencing revealed 63 foxes positive for the Babesia microti-like piroplasm (syn. Theileria annae) (69.2%; 95% confidence interval [CI]: 58.7-78.5%) and one fox positive for Babesia canis (1.1%; 95% CI: 0.0-6.0%). Positivity to the B. microti-like piroplasm or B. canis in 43 blood samples (83.7%) was significantly higher (p<0.001) than in 43 paired bone marrow samples (20.9%). There were no statistically significant differences in the prevalence of infection between genders (p=0.219) or age groups (<2 years vs. ≥ 2 years) (p=1.0). This is the first report of the B. microti-like piroplasm in foxes from Portugal as well as the first report on detection by PCR and genotyping of B. canis in a red fox worldwide. A natural cycle of the B. microti-like piroplasm is suggested in red fox populations based on the high prevalence of the protozoan. Red foxes might be a reservoir of the B. microti-like piroplasm and a source of infection to dogs.

Toxoplasma gondii prevalence in Israeli crows and Griffon vultures.

A cross-sectional Toxoplasma gondii seroprevalence study was performed on free ranging crows (Corvus cornis, Corvus monedula, Corvus splendens) and Griffon vultures (Gyps fulvus) from Israel in order to assess exposure to this pathogen in scavenger birds that feed on animal carcasses and their possible role in the epidemiology of toxoplasmosis. Using the modified agglutination test (MAT) with a cutoff titer of 1:25, 52 of 122 crows (42.6%) and 40 of 101 Griffon vultures (39.6%) were found to be T. gondii seropositive. Crow T. gondii seroprevalence was significantly higher in northern areas of Israel (p=0.007) where annual precipitation is higher and annual summer maximum temperatures are lower than in the drier and warmer south. Seroprevalence in crows was positively associated with higher human population densities possibly related to the increased cat population in these areas. PCR analysis of brain extracts from crows resulted in the detection of T. gondii DNA in 1 seropositive crow from northern Israel. Genetic analysis of DNA from the positive crow brain confirmed infection with T. gondii type 2 using a multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP) of the SAG1, 5-3' SAG2, alt.SAG2, SAG3, BTUB, GRA6, C22-8, c29-2, L358, PK1 and Apico loci. The high T. gondii seroprevalence in these bird species suggests that infected carrion may be responsible for widespread infection of carcass scavenger birds which may further transmit infection to other carnivorous intermediate hosts or feline definitive hosts when consumed post-mortally.