A Global Perspective on Oral Vaccination of Wildlife against Rabies.

The long-term mitigation of human-domestic animal-wildlife conflicts is complex and difficult. Over the last 50 yr, the primary biomedical concepts and actualized collaborative global field applications of oral rabies vaccination to wildlife serve as one dramatic example that revolutionized the field of infectious disease management of free-ranging animals. Oral vaccination of wildlife occurred in diverse locales within Africa, Eurasia, the Middle East, and North America. Although rabies is not a candidate for eradication, over a billion doses of vaccine-laden baits distributed strategically by hand, at baiting stations, or via aircraft, resulted in widespread disease prevention, control, or local disease elimination among mesocarnivores. Pure, potent, safe, and efficacious vaccines consisted of either modified-live, highly attenuated, or recombinant viruses contained within attractive, edible baits. Since the late 1970s, major free-ranging target species have included coyotes (Canis latrans), foxes (Urocyon cinereoargenteus; Vulpes vulpes), jackals (Canis aureus; Lupulella mesomelas), raccoons (Procyon lotor), raccoon dogs (Nyctereutes procyonoides), and skunks (Mephitis mephitis). Operational progress has occurred in all but the latter species. Programmatic evaluations of oral rabies vaccination success have included: demonstration of biomarkers incorporated within vaccine-laden baits in target species as representative of bait contact; serological measurement of the induction of specific rabies virus neutralizing antibodies, indicative of an immune response to vaccine; and most importantly, the decreasing detection of rabies virus antigens in the brains of collected animals via enhanced laboratory-based surveillance, as evidence of management impact. Although often conceived mistakenly as a panacea, such cost-effective technology applied to free-ranging wildlife represents a real-world, One Health application benefiting agriculture, conservation biology, and public health. Based upon lessons learned with oral rabies vaccination of mesocarnivores, opportunities for future extension to other taxa and additional diseases will have far-reaching, transdisciplinary benefits.

Borrelia persica infection in wild carnivores in Israel: molecular characterization and new potential reservoirs.

BACKGROUND: Borrelia persica causes tick-borne relapsing fever in Israel, the eastern Mediterranean basin, and Asia. Relapsing fever is associated with severe illness and potentially death in humans and animals. Since B. persica infection has rarely been described in wild animals, the aim of this study was to evaluate the prevalence of infection with B. persica in wild carnivores in Israel. METHODS: Spleen and blood clot samples from wild carnivores, which underwent necropsy, were tested for the presence of Borrelia DNA by real-time polymerase chain reaction (PCR). PCR products were sequenced, and the spirochete loads were quantified using a specific quantitative PCR (qPCR). RESULTS: A total of 140 samples from 74 wild carnivores were analyzed for the presence of Borrelia DNA. Six out of the 74 (8.1%) animals were found positive for B. persica by PCR and sequencing of the flagellin B gene, of which 4/74 (5.4%) were also positive by PCR for the glycerophosphodiester phosphodiesterase (glpQ) gene. Positive samples were obtained from three European badgers, and one striped hyena, golden jackal, and red fox each. All B. persica-positive animals were young males (P < 0.0001). Quantifiable results were obtained from 3/5 spleen and 4/5 blood samples. The spirochete loads in the blood were significantly higher than those found in the spleen (P = 0.034). CONCLUSIONS: The prevalence of B. persica infection found in wild carnivores brought for necropsy was unexpectedly high, suggesting that this infection is widespread in some wild animal species in Israel. This is the first report of B. persica infection in the European badger and striped hyena. These carnivores have a wide geographical range of activity, and the results of this survey raise the possibility that they may serve as reservoir hosts for B. persica.

Fecal microbiota of the synanthropic golden jackal (Canis aureus).

The golden jackal (Canis aureus), is a medium canid carnivore widespread throughout the Mediterranean region and expanding into Europe. This species thrives near human settlements and is implicated in zoonoses such as rabies. This study explores for the first time, the golden jackal fecal microbiota. We analyzed 111 fecal samples of wild golden jackals using 16S rRNA amplicon sequencing the connection of the microbiome to animal characteristics, burden of pathogens and geographic and climate characteristics. We further compared the fecal microbiota of the golden jackal to the black-backed jackal and domestic dog. We found that the golden jackal fecal microbiota is dominated by the phyla Bacteroidota, Fusobacteriota and Firmicutes. The golden jackal fecal microbiota was associated with different variables, including geographic region, age-class, exposure to rabies oral vaccine, fecal parasites and toxoplasmosis. A remarkable variation in the relative abundance of different taxa was also found associated with different variables, such as age-class. Linear discriminant analysis effect size (LEfSe) analysis found abundance of specific taxons in each region, Megasphaera genus in group 1, Megamonas genus in group 2 and Bacteroides coprocola species in group 3. We also found a different composition between the fecal microbiota of the golden jackal, blacked-backed jackal and the domestic dog. Furthermore, LEfSe analysis found abundance of Fusobacterium and Bacteroides genera in the golden jackal, Clostridia class in blacked-backed jackal and Megamonas genus in domestic dog. The golden jackal fecal microbiota is influenced by multiple factors including host traits and pathogen burden. The characterization of the microbiota of this thriving species may aid in mapping its spread and proximity to human settlements. Moreover, understanding the jackal microbiota could inform the study of potential animal and human health risks and inform control measures.

Identification, Isolation, and Molecular Characterization of Betacoronavirus in Oryx leucoryx.

Coronaviruses (CoVs) are enveloped viruses with a large RNA genome (26 to 32 kb) and are classified into four genera: Alphacoronavirus, Betacoronavirus, Gammacoronavirus, and Deltacoronavirus. CoV infections cause respiratory, enteric, and neurologic disorders in mammalian and avian species. In 2019, Oryx leucoryx animals suffered from severe hemorrhagic diarrhea and high morbidity rates. Upon initial diagnosis, we found that the infected animals were positive for coronavirus by pancoronavirus reverse transcriptase RT-PCR. Next, we detected the presence of CoV particles in these samples by electron microscopy and immunohistochemistry. CoV was isolated and propagated on the HRT-18G cell line, and its full genome was sequenced. Full-genome characterization and amino acid comparisons of this viral agent demonstrated that this virus is an evolutionarily distinct Betacoronavirus belonging to the subgenus Embecovirus and the Betacoronavirus 1 species. Furthermore, we found that it is most similar to the subspecies dromedary camel coronavirus HKU23 by phylogenetic analysis. Here, we present the first report of isolation and characterization of Betacoronavirus associated with enteric disease in Oryx leucoryx. IMPORTANCE CoVs cause enteric and respiratory infections in humans and animal hosts. The ability of CoVs to cross interspecies barriers is well recognized, as emphasized by the ongoing pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The identification of novel CoV strains and surveillance of CoVs in both humans and animals are relevant and important to global health. In this study, we isolated and characterized a newly identified Betacoronavirus that causes enteric disease in a wild animal, Oryx leucoryx (the Arabian oryx). This work is the first report describing CoV infection in Oryx leucoryx and provides insights into its origin.

Plasma cholinesterase activity: A benchmark for rapid detection of pesticide poisoning in an avian scavenger.

Poisoning due to exposure to organophosphate and carbamate pesticides is a common threat for many wildlife species, especially for scavengers such as vultures. The Griffon vulture population (Gyps fulvus), for instance, is deteriorating in the Eastern Mediterranean, and is considered to be critically endangered in Israel, where 48 out of 107 (45 %) known injury/mortality cases in 2010-2021 were caused by poisoning. Lack of specific clinical indications, together with levels of organophosphate or carbamate pesticides too low to detect, challenge the ability to diagnose and treat such poisoning events. The activity of cholinesterase (ChE) in plasma has the potential to serve as an effective biomarker for monitoring exposure to anticholinesterase pesticides in live vultures. Yet, the applicability of this approach has been limited by intra- and inter-species variations in ChE basal levels. The present study aims to provide a benchmark for ChE activity levels in healthy Griffons and their intra-species variation. Blood samples from free-roaming (n = 231) and captive (n = 63) Griffons were collected during routine monitoring, and ChE levels were determined using a colorimetric method. We established that the ChE in the plasma of Griffons reflects mostly acetylcholinesterase as the dominant form. ChE levels in healthy Griffons are 0.601 ± 0.011 U/ml (mean ± SE), while Griffons with suspected or confirmed pesticide poisoning display much lower levels of ChE activity (typically <0.3 U/ml). We also characterized the age dependence of ChE activity, as well as differences among groups from different locations or origins. Our study provides a rapid diagnostic tool for the detection of exposure to organophosphate and carbamate pesticides that should facilitate the lifesaving treatment and the conservation of this species. Moreover, our protocols can be adapted to other species and geographical areas, addressing pesticide poisoning worldwide and contributing to the protection of endangered species and their ecological functions (e.g. sanitation by scavengers).

Advancing understanding of the taxonomy and diversity of the genus Contracaecum in the great white pelican (Pelecanus onocrotalus).

Despite the wide distribution and health importance of anisakids of the genus Contracaecum, epidemiological data on their occurrence in definitive bird hosts are scarce, particularly from certain parts of the world that represent important wintering sites or migration stopovers for different bird species. In the present study, Contracaecum spp. infecting six great white pelicans (Pelecanus onocrotalus) in Israel were identified using light and scanning electron microscopy and phylogenetic analyses of nuclear internal transcribed spacer (ITS) and mitochondrial cytochrome c oxidase II (cox2). A PCR-RFLP method was also developed and applied to screen large numbers of Contracaecum parasites. Most (415/455) worms recovered were C. micropapillatum, followed by C. gibsoni (31/455), C. quadripapillatum (8/455), and C. multipapillatum E (1/455). Contracaecum micropapillatum from Israel and C. bancrofti from Australia are distinguishable by cox2 but less well resolved with ITS sequences, and could not be distinguished morphologically. Worms with cox2 matching C. gibsoni had ITS matching specimens identified as C. multipapillatum A. To the authors' knowledge, this represents the first of such studies in Israel and provides useful data on the ecology and distribution of different Contracaecum species of health and economic interest.

First detection of avian influenza subtype H4N6 in Israel in a wild mallard (Anas platyrhynchos).

Avian influenza viruses (AIV) are a worldwide threat to animal and human health. As wild waterfowl circulate and spread these viruses around the world, investigations of AIV prevalence in wild populations are critical for understanding pathogen transmission, as well as predicting disease outbreaks in domestic animals and humans. Surveillance efforts in this study have isolated H4N6 for the first time in Israel from a faecal sample of a wild mallard (Anas platyrhynchos). Phylogenetic analyses of the HA and NA genes revealed that this strain is closely related to isolates from Europe and Asia. This Eurasian origin, together with Israel serving as an important migratory bottleneck of the mid Palearctic-African flyway, suggests a potential introduction of this strain by migratory birds. Additional phylogenetic analysis of the isolate's internal genes (PB1, PB2, PA, NP, M and NS) revealed high levels of phylogenetic relatedness with other AIV subtypes, indicating previous reassortment events. High reassortment rates are characteristic for H4N6 viruses, which, together with this subtype's ability to infect pigs and adaptability to the human receptor binding domain, raises the concern that it would potentially become zoonotic in the future. These results emphasize the importance of continuous AIV monitoring in migratory birds.

The association between natural drinking water sources and the emergence of zoonotic leptospirosis among grazing beef cattle herds during a human outbreak.

Leptospirosis is a zoonotic bacterial disease associated with water abundance in tropical and temperate climate zones. Bacterial spread may also occur in dry and warm weather conditions when humans and animals are forced to share depleted water sources. In such settings, farm animals such as beef cattle, which may be present in large numbers in natural water sources, can play a major role in disease spread. However, the risk factors for their infection and the potential control measures to prevent the disease spread have not been adequately studied. In the face of an emerging human leptospirosis outbreak in the dry and warm Israeli 2018 summer, we tested seropositivity to Leptospira serovar Pomona in grazing beef cattle and wild boars located in proximity to the contaminated streams. Additionally, we used the natural setting of the outbreak to identify risk factors for seropositivity in beef cattle. We found high seropositivity to serovar Pomona in grazing beef cattle (233/845), and in wild boars (7/13). Seropositivity was significantly associated with beef cattle drinking from natural water sources compared to beef cattle drinking from water troughs with fresh water supply (Multivariable logistic regression; odds ratio = 18.6, 95% confidence interval = 3-116, p-value<0.01). One Health approach is necessary for mitigating zoonotic Leptospira infections, in which interactions between humans, animals, and the environment play a major role. As the global warming crisis results in severe climate changes, dry and warm weather conditions may become more common worldwide. Under such conditions, reducing inter-species interactions in contaminated natural water sources is essential for protecting public health. Our study demonstrates the role of natural water as a source for beef cattle infection and disease spread. Furthermore, we suggest using water troughs with freshwater supply for preventing future outbreaks in animals and humans in such settings.

Short Term Safety, Immunogenicity, and Reproductive Effects of Combined Vaccination With Anti-GnRH (Gonacon) and Rabies Vaccines in Female Feral Cats.

Overpopulation of free-roaming cats is a major problem leading to negative impacts on animal health and welfare, public nuisance, transmission of zoonotic diseases, and well-documented harm to wildlife. Surgical sterilization had failed to provide a practical solution to free-roaming cats' overpopulation under field conditions; therefore, efficient and safe non-surgical immunocontraception methods are aspired. Rabies is a deadly virus that may infect people and animals. However, the safety and efficacy of combined vaccination with anti-GnRH and rabies vaccines in feral cats, which often suffer from disrupted health conditions and experienced high stress level, has never been studied. Therefore, our objective was to examine the short-term safety and efficacy of anti-GnRH vaccine (Gonacon), in combination with rabies vaccine in female feral cats. Mature feral female cats were captured and divided into the following groups: (I) GonaconX1-Rabies: queens vaccinated with both Gonacon and rabies (n = 5); (II) GonaconX2-Rabies: queens vaccinated twice with Gonacon (3 weeks apart) and with Rabies (n = 4); (III) OVx-Rabies: queens ovariohysterectomized and vaccinated with rabies (n = 4); (IV) Intact-Rabies: queens vaccinated against rabies and remained intact (n = 3). Comprehensive veterinary examinations and blood tests were performed every 2 weeks for 14 weeks. Data were analyzed by Repeated-Measures-ANOVA or Fisher-Exact-Test. There were neither systemic nor local adverse reactions at the vaccination sites. Blood count (PCV, TS, RBC, HGB, HCT, WBC) and chemistry (Total protein, Total globulin, Albumin, Urea, Creatinine, Creatine kinase, Bilirubin, GGT, ALT, AST) analyses revealed no differences among groups. There were no differences in serum rabies antibodies titers among groups, and queens kept a protective titer (>0.5 IU/mL) starting at 2-4 weeks after vaccination. Anti-GnRH antibodies were detected in all Gonacon-vaccinated queens, excluding one queen (GonaconX2-Rabies group). Anti-müllerian hormone serum concentrations reduced significantly after ovariohysterectomy, as well as gradually following vaccination with Gonacon, but it remained high in intact queens. Evaluation of vaginal cytology and ovarian histology suggested that reproductive cyclicity was suppressed in Gonacon-vaccinated queens. Our results support the conclusion that in the short term, the combined vaccination with Gonacon and rabies is safe and effective in female feral cats. However, further long-term studies are warranted to test this immunologic regimen in feral cats.

Transmission of the Human Relapsing Fever Spirochete Borrelia persica by the Argasid Tick Ornithodoros tholozani Involves Blood Meals from Wildlife Animal Reservoirs and Mainly Transstadial Transfer.

Borrelia persica, transmitted by the argasid tick Ornithodoros tholozani, causes human tick-borne relapsing fever in the Middle East and Central Asia. Infection is acquired often when visiting tick-infested caves and reported to be transmitted mainly transovarially between ticks, occasionally infecting humans. To study the epidemiology of this infection, ticks were trapped in 24 caves in 12 geographic zones covering all of Israel and identified morphologically. DNA was extracted from larvae, nymphs, and adult stages from each location and PCR followed by DNA sequencing was performed to identify Borrelia infection, tick species, and tick blood meal sources. We collected 51,472 argasid ticks from 16 of 24 caves surveyed. We analyzed 2,774 O. tholozani ticks, and 72 (2.6%) from nine caves were PCR positive for B. persica Infection rates in male, female, and nymphal ticks (4.4%, 3%, and 3.2%, respectively) were higher than in larva (P < 0.001), with only 3 (0.04%) positive larvae. Presence of blood meal was associated with B. persica infection in ticks (P = 0.003), and blood meals of golden jackals, red foxes, and Cairo spiny mouse were associated with infection (P ≤ 0.043). PCR survey of 402 wild mammals revealed B. persica infection with the highest rates in social voles (22%), red foxes (16%), golden jackals (8%), and Cairo spiny mice (3%). In conclusion, although transovarial tick transmission of B. persica occurs at low levels, ticks apparently acquire infection mainly from wildlife canid and rodents and may eventually transmit relapsing fever borreliosis to humans who enter their habitat.IMPORTANCEBorrelia persica is a spirochete that causes tick-borne relapsing fever in humans in an area that spans from India to the Mediterranean. Until now, it was thought that the soft tick vector of this infection, Ornithodoros tholozani, is also its main reservoir and it transmits B. persica mostly transovarially between tick generations. This study showed that tick infection with B. persica is associated with feeding blood from wild jackals, foxes, and rodents and that transovarial transmission is minimal. Since O. tholozani ticks are found in isolated caves and ruins, it is assumed that wild canids who migrate over long distances have a major role in the transmission of B. persica between remote tick populations, and it is then maintained locally also by rodents and eventually transferred to humans during tick bites. Prevention of human infection could be achieved by restricting entrance of canines and humans to habitats with O. tholozani populations.

First isolation of foot and mouth disease virus from wild boars in the Middle East.

Domestic pigs are susceptible to foot and mouth disease virus (FMDV) infection and suffer from severe clinical disease. Our knowledge on the clinical manifestations of FMD in and its transmission by wild boars (Sus scrofa) is very limited. During an FMD outbreak in the Golan Heights in 2018, wild boars grazing in close proximity to cattle were observed showing lameness. Infectious FMDV was isolated from throat and heart tissues of two young wild boars with FMD clinical signs. This is the first report of wild boars clinically infected with FMDV in the Middle East.

Genetic characteristics of Theileria equi in zebras, wild and domestic donkeys in Israel and the Palestinian Authority.

Equine piroplasmosis (EP) is an important tick-borne disease of equids, caused by Theileria equi and Babesia caballi. It is endemic in most parts of the world, including Israel, and has clinical and economic consequences. This study was set to evaluate the presence of EP parasites in domestic donkeys and in wild equids in Israel and the Palestinian Authority (PA). To assess subclinical EP infection in 98 domestic donkeys (Equus africanus asinus), 9 Asiatic wild donkeys (Equus hemionus), 8 zebras (Equus quagga), 7 African wild donkeys (Equus africanus) and 5 mules, were tested using PCR and qPCR. Positive samples were characterized by amplification and sequencing of a 1600 bp fragment of the 18S rRNA gene. Babesia caballi was not detected in any of the animals. Theileria equi was detected in 32% of the donkeys, 89% of Asiatic wild donkeys, 57% of African wild donkeys, 62% of zebras and none of the mules. Parasitemia was low in all of the positive samples. Risk factors associated with infection in donkeys included one farm (Kiryat Gat) and animal sex (male). The sequences of the 18S rRNA gene from domestic donkeys were all similar, and belonged to the T. equi genotype D, similar to the genotype sequenced from horses in the same area, while sequences from wild donkeys were unique and belonged to the T. equi genotype A. Verification of the T. equi genotype in zebras could not be concluded, suggesting major genetic variation.

Immobilization of captive Persian fallow deer (Dama dama mesopotamica) using medetomidine-ketamine or medetomidine-midazolam.

OBJECTIVE: To establish and compare the effectiveness of two medetomidine-based immobilization protocols in Persian fallow deer (Dama dama mesopotamica). STUDY DESIGN: Prospective, randomized, blinded clinical study. ANIMALS: A group of 31 captive Persian fallow deer. METHODS: Deer scheduled for translocation were immobilized with a combination of medetomidine (76 ± 11 µg kg-1) and ketamine (1.0 ± 0.2 mg kg-1) (MK; n = 15) or medetomidine (77 ± 11 µg kg-1) and midazolam (0.10 ± 0.01 mg kg-1) (MM; n = 16) administered intramuscularly. An observer unaware of group assignments recorded times to immobilization and recovery, monitored physiologic variables and scored the quality of induction, immobilization and recovery (scale 1-5: 1, poor; 5, excellent). Atipamezole was administered for reversal. Data analysis was performed using the t test, the Mann-Whitney U test, the chi-square test and the Fisher's exact test. Significance was set at p < 0.05. RESULTS: Data are presented as mean ± standard deviation or median (range). Time to induce immobilization was 9 ± 4 and 10 ± 4 minutes in the MK and MM groups, respectively. Immobilization quality score was 5 (1-5) following both combinations. Hemoglobin oxygen saturation (SpO2) was significantly lower in the MK (80 ± 8%) than in the MM group (87 ± 8%) although respiratory frequency did not differ between MK and MM (11 ± 5 and 10 ± 2 breaths minute-1, respectively). Recovery times were 13 ± 6 (MK) and 14 ± 7 minutes (MM) and did not differ between groups. No morbidities or mortalities were recorded during 1 month after immobilization. CONCLUSIONS AND CLINICAL RELEVANCE: The MK and MM combinations produced sufficient immobilization in captive Persian fallow deer for short nonpainful procedures. Based on the SpO2 values, the MM combination may be associated with less respiratory depression; nevertheless, both combinations may result in a decrease in SpO2.

Molecular characterization of six Hyalomma species using mitochondrial markers.

Hyalomma species (Acari: Ixodidae) are vectors of several human and animal pathogens. However, due to their similar morphological properties, classification of related Hyalomma species is often challenging. Here we describe a combined approach for molecular characterization of six Hyalomma species: H. aegyptium, H. dromedarii, H. excavatum, H. impeltatum, H. marginatum and H. turanicum. This procedure was developed using a combination of PCR amplification of four molecular markers, followed by sequencing and species-specific restriction analysis. Segments from the following genes were used as markers: 12S rRNA, 16S rRNA, Cytochrome C oxidase subunit 1 (COX1), and Cytochrome B (CytB). Phylogenetic analysis based on the amplified sequences was consistent with the morphology-based classification. It revealed relative close proximity of H. excavatum, H. marginatum and H. turanicum, and close proximity of H. aegyptium and H. dromedarii to each other. H. impeltatum was examined using the COX1 and CytB markers, and in both cases was located on a separate clade from the other five species. Digestion of the amplified products using specific restriction enzymes enabled clear distinction between the six species. This report is the first to describe CytB marker sequences of the studied species, and the first to describe COX1 marker sequences of H. aegyptium, H. excavatum, H. impeltatum and H. turanicum. The information obtained in this study may therefore be useful for future combined morphological-molecular Hyalomma characterization.

Exposure to Neospora spp. and Besnoitia spp. in wildlife from Israel.

Neosporosis and besnoitiosis, caused by cyst-forming protozoa Neospora caninum and Besnoitia besnoiti, respectively, are parasitic infestations of livestock in Israel. These parasites cause significant economic losses in cattle due to reproductive and productive disorders. Both parasites have been detected in several wild ruminant species throughout other regions of the world, while the existence of a sylvatic life cycle in Israel remains uncertain. Thus, a wide panel of 871 sera from two wild carnivores and nine wild ruminant species were tested. All sera were first analysed by MAT for an initial screening and positive samples were confirmed a posteriori by Western blot. Additionally, a complementary IFAT was used for the detection of antibodies against N. caninum. Neospora antibodies were present in six out of the 11 species investigated, whereas Besnoitia antibodies were undetected. Golden jackal, red fox, addax, Arabian oryx, Persian fallow deer, mouflon, mountain gazelle, Nubian ibex, scimitar horned oryx and water buffalo were seropositive against N. caninum infection by IFAT and/or MAT. Moreover, the presence of Neospora spp.-specific antibodies was confirmed by Western blot in golden jackal (6/189; 3.2%), red fox (1/75; 1.3%), Persian fallow deer (13/232; 5.6%), mouflon (1/15; 16.7%), Nubian ibex (22/55; 40%) and water buffalo (12/18; 66.7%). Addax (1/49) and water buffalo (1/18) were MAT-seropositive against B. besnoiti but were seronegative by Western blot. Hence, Neospora sylvatic cycle is present in Israel and may cross over to a domestic life cycle. In contrast, wildlife species investigated are unlikely to present a risk of transmitting Besnoitia to livestock in Israel.

Outbreak of human leptospirosis linked to contaminated water bodies in Northern Israel, June to August 2018.

We report preliminary findings of a large outbreak of human leptospirosis with 36 confirmed/probable and 583 suspected cases from June-August 2018, linked to contaminated water bodies in Northern Israel. There was a travel-associated case in Germany; additional cases are being investigated in other countries. The presumed chain of transmission, implicating wild boar and cattle, raises multiple challenges for risk assessment, risk management and risk communication currently being addressed by a public health response team.

Detection of Toxoplasma gondii in three common bottlenose dolphins (Tursiops truncatus); A first description from the Eastern Mediterranean Sea.

Toxoplasma gondii has been described in several marine mammals around the world including numerous species of cetaceans, yet infection and transmission mechanisms in the marine environment are not clearly defined. The Israel Marine Mammal Research and Assistance Center has been collating a database of all marine mammal stranding events along the country's national coastlines since 1993. In this study, we describe the molecular detection and characterisation of T. gondii in three common bottlenose dolphins (Tursiops truncatus) including one case of coinfection with herpesvirus. The animals were found stranded on the Mediterranean coast of Israel in May and November 2013. In one of the three cases, the dolphin was found alive and admitted to intensive care. To our knowledge, this is the first report of T. gondii infection of marine mammals in the Eastern Mediterranean Sea. As this parasite acts as an indicator for marine pollution and marine mammal health, we believe these findings add important information regarding the state of the environment in this region.

A survey of Babesia spp. and Hepatozoon spp. in wild canids in Israel.

BACKGROUND: Babesia spp. and Hepatozoon spp. are apicomplexan parasites that infect a variety of animals, including canids. Their life-cycle includes an invertebrate hematophagous vector as a definitive host and vertebrates as intermediate hosts. The aims of this study were to investigate the prevalence and risk factors for Babesia spp. and Hepatozoon spp. infections in wild golden jackals (Canis aureus) and red foxes (Vulpes vulpes) in Israel and to compare spleen with blood sample polymerase chain reaction (PCR) for the detection of infection. RESULTS: Blood and spleen samples from 109 golden jackals and 21 red foxes were tested by PCR for the detection of Babesia spp. and Hepatozoon spp. using primers for the 18S ribosomal (r) RNA gene. Hepatozoon canis was detected in 50/109 (46%) of the jackals and 9/21 (43%) of the foxes. "Babesia vulpes" (the Babesia microti-like piroplasm) was detected in 4/21 (19%) of the foxes and in none of the jackals. A previously unknown genotype termed Babesia sp. MML related to Babesia lengau (96-97% identity) was detected in 1/109 (1%) of the jackals and 4/21 (19%) of the foxes. Further characterization of this genotype carried out by PCR of the rRNA internal transcribed spacer 2 (ITS2) indicated that it had only 87% identity with the B. lengau ITS2. Sex (male or female), age (juvenile or adult) and geographic zone (North, Central or South Israel) were not found to be significant risk factors for these protozoan infections. The prevalence of "B. vulpes" and Babesia sp. MML infections was significantly higher in foxes compared to jackals (χ2 = 15.65, df = 1, P < 0.005), while there was no statistically significant difference in the rate of H. canis infection between these two canid species. A fair agreement beyond chance between identification in the blood and spleen of H. canis was found in 21 animals from which both blood and spleen samples were available (k = 0.33). CONCLUSIONS: This study describes a high prevalence of H. canis infection in foxes and jackals and is the first report of "B. vulpes" infection in Israel, an area where Ixodes spp. are rare. It describes infection with a previously unknown genotype of Babesia related to B. lengau from Africa.

Borrelia persica infection in rock hyraxes.

Tick-borne relapsing fever (TBRF) is an acute infectious disease caused by arthropod-borne spirochetes of the genus Borrelia and characterized by recurrent episodes of fever. Borrelia persica, the causative agent of this disease in Israel, is transmitted by the argasid tick Ornithodoros tholozani. There is little information about the maintenance and possible vertebrate reservoirs of B. persica in nature, but the tick O. tholozani is known to feed on animals that enter its habitat in caves, rock crevices and shady environments. The rock hyrax (Procavia capensis) is commonly found in such habitats and may therefore serve as a reservoir host for O. tholozani. Blood and spleen samples from rock hyraxes were collected from twelve locations in Israel and the West Bank during 2009-2014 to test if these animals may be infected with B. persica. Real-time PCR targeting a segment of the flagellin (flaB) gene was initially used to detect B. persica. Positive samples were further analyzed by PCR, using a segment of the glycerophosphodiester phosphodiesterase (GlpQ) gene for additional confirmation. Borrelia species were identified by nucleotide sequence analysis and the copy number of Borrelia was quantified in blood and spleen samples based on the number of Borrelia 16S rRNA gene copies. A total of 112 hyraxes were examined, with both blood and spleen samples tested from 108 animals. Nine hyraxes were infected with B. persica, with a prevalence of 8%. Of these, two animals were positive for both blood and spleen samples, three only for blood and four only for the spleen. The number of DNA copies of Borrelia 16S rRNA was significantly higher in blood (5 × 106 to 9.2 × 108/ml blood) compared to spleen (2.1 × 104 to 1.0 × 106/ml). We conclude that rock hyraxes are possible reservoirs for B. persica because they have long lifespans and gregarious habits, share habitats with vector ticks, and are naturally infected with this spirochete. Further studies should be conducted in the future to evaluate the competence of hyraxes as reservoirs for B. persica infection.

REFRACTOMETRIC URINE SPECIFIC GRAVITY OF FREE-LIVING EGYPTIAN FRUIT BATS (ROUSETTUS AEGYPTIACUS).

In both human and veterinary medicine, urine specific gravity (USG) is commonly measured by refractometry to indirectly reflect the osmolality of urine to thereby evaluate the kidney's ability to concentrate or dilute urine according to physiologic need and certain disease conditions. However, for accurate interpretation of the significance of any value, knowledge of the expected USG for the healthy species in question is required. It is generally believed that fruit bats, and Egyptian fruit bats (Rousettus aegyptiacus) in particular, are unable to highly concentrate their urine. In this study, the USG was determined using a handheld urine refractometer in 43 free-living Egyptian fruit bats of both sexes. The calculated nonparametric 90% confidence interval for Egyptian fruit bats in this study was 1.006-1.050, with no association with capture site, sex, weight, or packed cell volume and total solids. Results suggest that free-living Egyptian fruit bats are able to highly concentrate their urine.