Hemolivia Mauritanica Research Articles

Pathogens in ticks collected in Israel: I. Bacteria and protozoa in Hyalomma aegyptium and Hyalomma dromedarii collected from tortoises and camels

Ticks were collected from 30 Greek tortoise (Testudo graeca), and 10 Arabian camels (dromedary) (Camelus dromedarius) in Israel. All those collected from Greek tortoises belonged to Hyalomma aegyptium, while all specimens collected from the camels belonged to Hyalomma dromedarii. Out of 84 specimens of H. aegyptium, 31 pools were examined by PCR, while from 75 H. dromedarii specimens nine pools were studied. Out of 31 pools of H. aegyptium 26 were positive for pathogens or endosymbiont; 14 for one, 11 for two and one for three pathogens. Out of nine pools prepared from H. dromedarii, seven were positive for pathogens (two for C. burnetii and five for Leishmania infantum). In H. aegyptium, Rickettsia africae, Rickettsia aeschlimannii, Rickettsia endosymbiont, Coxiella burnetii, Hemolivia mauritanica, Babesia microti, Theileria sp., and Leishmania infantum was detected, while in H. dromedarii C. burnetii and L. infantum were found. None of the ticks were positive for Anaplasma/Ehrlichia, Listeria monocytogenes, Bartonella spp., Hepatozoon spp. and Toxoplasma gondii. H Rickettsia endosymbionts, C. burnetii, B. microti, Theileria sp. and L. infantum are reported for the first time in H. aegyptium, and C. burnetii and L. infantum for the first time in H. dromedarii.

Diversity of microorganisms in Hyalomma aegyptium collected from spur-thighed tortoise (Testudo graeca) in North Africa and Anatolia.

Ticks carry a diverse community of microorganisms including non-pathogenic symbionts, commensals, and pathogens, such as viruses, bacteria, protozoans, and fungi. The assessment of tick-borne microorganisms (TBM) in tortoises and their ticks is essential to understand their eco-epidemiology, and to map and monitor potential pathogens to humans and other animals. The aim of this study was to characterize the diversity of microorganisms found in ticks collected from the spur-thighed tortoise (Testudo graeca) in North Africa and Anatolia. Ticks feeding on wild T. graeca were collected, and pathogens were screened by polymerase chain reaction using group-specific primers. In total, 131 adult Hyalomma aegyptium ticks were collected from 92 T. graeca in Morocco (n=48), Tunisia (n=2), Algeria (n=70), and Turkey (n=11). Bacteria and protozoa detected included Hemolivia mauritanica (22.9%), Midichloria mitochondrii (11.4%), relapsing-fever borreliae (8.4%), Ehrlichia spp. (7.6%), Rickettsia spp. (3.4%), Borrelia burgdorferi s.l. (0.9%), Francisella spp. (0.9%), and Wolbachia spp. (0.8%). The characterization of Rickettsia included R. sibirica mongolitimonae (Algeria), R. aeschlimannii (Turkey), and R.africae (Morocco). Hemolivia mauritanica and Ehrlichia spp. prevalence varied significantly with the sampling region/country. We did not detect significant associations in microorganism presence within ticks, nor between microorganism presence and tick mitochondrial DNA haplogroups. This is the first report of Francisella persica-like, relapsing fever borreliae, M. mitochondrii, and Wolbachia spp. in H. aegyptium ticks collected from wild hosts from the South and Eastern Mediterranean region, and of R. sibirica mongolitimonae and R. africae in H. aegyptium from Algeria and Morocco, respectively. Given that T. graeca is a common species in commercial and non-commercial pet trade, the evaluation of the role of this species and its ticks as hosts for TBM is particularly relevant for public health.

Molecular Evidence of Hemolivia mauritanica, Ehrlichia spp. and the Endosymbiont Candidatus Midichloria Mitochondrii in Hyalomma aegyptium Infesting Testudo graeca Tortoises from Doha, Qatar

Simple SummaryDue to the veterinary and medical importance of pathogens transmitted by Hyalomma aegyptium, we tested ticks removed from Testudo graeca tortoises for the presence of Anaplasma, Ehrlichia, Hemolivia, Babesia and Hepatozoon. Forty-three percent of the examined adult ticks were infected with at least one agent. The most prevalent agent identified was Hemolivia mauritanica (28.6%), followed by Candidatus Midichloria mitochondrii (9.5%) and Ehrlichia spp. (4.7%). Our study reported for the first time H. mauritanica, Ehrlichia spp. and Candidatus M. mitochondrii in H. aegyptium ticks collected from pet spur-thighed tortoises, in Qatar, providing data that adds to the geographical extension of these agents.Tick-borne agents constitute a growing concern for human and animal health worldwide. Hyalomma aegyptium is a hard tick with a three-host life cycle, whose main hosts for adults are Palearctic tortoises of genus Testudo. Nevertheless, immature ticks can feed on a variety of hosts, representing an important eco-epidemiological issue regarding H. aegyptium pathogens circulation. Hyalomma aegyptium ticks are vectors and/or reservoirs of various pathogenic agents, such as Ehrlichia, Anaplasma, Babesia and Hepatozoon/Hemolivia. Ehrlichia and Anaplasma are emergent tick-borne bacteria with a worldwide distribution and zoonotic potential, responsible for diseases that cause clinical manifestations that grade from acute febrile illness to a fulminant disease characterized by multi-organ system failure, depending on the species. Babesia and Hepatozoon/Hemolivia are tick-borne parasites with increasing importance in multiple species. Testudo graeca tortoises acquired in a large animal market in Doha, Qatar, were screened for a panel of tick-borne pathogens by conventional PCR followed by bidirectional sequencing. The most prevalent agent identified in ticks was Hemolivia mauritanica (28.6%), followed by Candidatus Midichloria mitochondrii (9.5%) and Ehrlichia spp. (4.7%). All samples were negative for Babesia spp. and Hepatozoon spp. Overall, 43% of the examined adult ticks were infected with at least one agent. Only 4.7% of the ticks appeared to be simultaneously infected with two agents, i.e., Ehrlichia spp. and H. mauritanica. This is the first detection of H. mauritanica, Ehrlichia spp. and Candidatus M. mitochondrii in H. aegyptium ticks collected from pet spur-thighed tortoises, in Qatar, a fact which adds to the geographical extension of these agents. The international trade of Testudo tortoises carrying ticks infected with pathogens of veterinary and medical importance deserves strict control, in order to reduce potential exotic diseases.

Absence of Hemolivia mauritanica (Apicomplexa: Haemogregarinidae) in natural populations of Testudo graeca in Morocco.

During spring 2018, we captured 101 spur-thighed tortoises, Testudo graeca, from seven localities in central Morocco. All tortoises were examined for the presence of blood parasites Hemolivia mauritanica and Hyalomma aegyptium ticks, the known vectors. We looked for H. mauritanica infection by examination of blood smears and by genetic analysis with PCR using extractions from both tortoises and ticks. On all tortoises collected, 71.29% were infested with at least one tick, with a mean infestation intensity of 7.12 ticks/tortoise and maximum of 15.55 ticks/tortoises in Had Draa locality (Essaouira region). Although the definitive host is present and abundant in all tortoise populations, all blood samples were Hemolivia-negative. Our results support and confirm the finding of studies previously conducted in other populations of Morocco and indicate that H. mauritanica has a narrower distribution range than its tick vector.

Identification of tick-borne pathogens in ticks collected from wild animals in Turkey.

Many enzootic life cycles involving wild animals and non-nidicolous ixodids are still unknown. Therefore, the aim of this study was to report the identified tick species collected from seven different animal species (red deer, brown bear, gray wolf, Eurasian lynx, red fox, European hare, and Mediterranean spur-thighed tortoise) living in the wild in Turkey and to investigate the presence of a wide range of tick-borne microorganisms in the tick samples obtained from these animals. The collected ticks (n = 98) were identified as Dermacentor reticulatus, Haemaphysalis parva, Hyalomma aegyptium, Hyalomma excavatum, Hyalomma marginatum, Ixodes ricinus, and Rhipicephalus turanicus. All engorged ticks collected from the wild animals and unfed larvae (n = 30) obtained from a single Rh. turanicus female were also analyzed individually for tick-borne bacterial and protozoan agents via PCR-sequencing. The molecular analyses revealed the presence of Babesia sp. tavsan2, Theileria capreoli, four Hepatozoon spp. (Hep. ursi, Hep. canis, Hep. felis, and Hepatozoon sp.), Hemolivia mauritanica, and three SFG rickettsiae (Candidatus Rickettsia barbariae, Ca. R. goldwasserii, and Rickettsia hoogstraalii) in the collected ticks. This represents the first report of Th. capreoli, Hep. ursi, and Ca. R. barbariae in ticks from Turkey. The evolutionary relationships of microbes in the different host and tick species are also discussed. Multiple novel tick-host associations in the tick life cycle were also revealed.

Bacterial and protozoan agents found in Hyalomma aegyptium (L., 1758) (Ixodida: Ixodidae) collected from Testudo graeca L., 1758 (Reptilia: Testudines) in Corum Province of Turkey

Hyalomma aegyptium (L., 1758) (Ixodida: Ixodidae) is a hard tick and the main host for adults are Palearctic tortoises of the genus Testudo, while larvae and nymphs are less host-specific and nymphs also attach to humans. In the present study, a total of 261 H. aegyptium ticks were removed from 26 Testudo graeca L., 1758 in Corum Province of Turkey. The most prevalent pathogens identified molecularly in the ticks were Hemolivia mauritanica (51.9 %), followed by Rickettsia aeschlimannii (32.6 %), Ehrlichia spp. (30.2 %), and Bartonella bovis (0.8 %). All samples were negative for Coxiella burnetii, Francisella tularensis, Anaplasma phagocytophilum, Babesia spp., Hepatozoon spp. and Theileria spp. Overall, 97.4 % of the examined adult ticks and 26.3 % of nymphs were infected with at least one pathogen, while 40.9 % of all ticks were infected with only one pathogen, 27.4 % with two pathogens, and 9.9 % with three pathogens, concomitantly. Overall, 80.8 % of the examined blood smears of tortoises were H. mauritanica-positive, and the mean intensity of parasitemia was 4.8 % (1–21). As a conclusion, since the examined tortoises were sampled in gardens and vineyards close to human habitation, and as a relatively large percentage of them were infested with ticks carrying pathogenic agents affecting also humans, the importance of tortoises, their ticks and pathogens in terms of the public health should be farther examined.

Turkey tick news: A molecular investigation into the presence of tick-borne pathogens in host-seeking ticks in Anatolia; Initial evidence of putative vectors and pathogens, and footsteps of a secretly rising vector tick, Haemaphysalis parva

This Turkey-based study investigated the presence of various tick-borne microorganisms in a broad-range of host-seeking ticks (n = 1019) that exhibit both hunter and ambusher characteristics. All collected ticks were analyzed individually via PCR-sequencing, resulting in the identification of 18 different microorganisms: six Babesia spp., including one putative novel species (Ba. occultans, Ba. crassa, Ba. rossi, Babesia sp. tavsan1, Babesia sp. tavsan2, and Babesia sp. nov.); six SFG rickettsiae (Ri. aeschlimannii, Ri. s. mongolitimonae, Ri. slovaca, Ri. raoultii, Ri. monacensis, and Ri. hoogstraalii); two Borrelia burgdorferi sensu lato spp. (Bo. afzelii and Bo. lusitaniae); two unnamed Hepatozoon spp.; Theileria annulata; and Hemolivia mauritanica. This provided evidence for the natural transstadial survival of these tick-borne microorganisms in adult ticks (in addition a nymph) of Turkey. Surprisingly, this study determined the presence of five different microorganisms (Ba. crassa, Ba. rossi, Babesia sp. Ucbas, Hepatozoon sp., and Ri. hoogstraalii) in host-seeking Haemaphysalis parva adults, for which poor data exist on its vectorial competence. Therefore, this study provides important data indicating the potential vectorial capacity of Ha. parva. This study also revealed the presence of the close ecological and evolutionary relationships between two important vector ticks, Hyalomma marginatum and Hy. aegyptium and determined genetic variations (distinct phylogenetic divergences inside the main clades) in some pathogenic SFG rickettsiae that are found in these ticks. Additionally, the presence of two Babesia species described very recently in hares with unknown vectors, namely Babesia sp. tavsan1 and Babesia sp. tavsan2, were detected for the first time in ticks. Finally, two unnamed Hepatozoon spp. were detected in Haemaphysalis ticks and their phylogenetic positions were demonstrated. Consequently, this study provides important data on the diversity of tick-borne microorganisms in host-seeking ticks and on potentially novel microorganisms (Babesia and Hepatozoon species) and their possible vectors (Ha. parva, Ha. sulcata, Hy. aegyptium, Hy. marginatum, and Rh. turanicus).

Molecular detection of a novel Babesia sp. and pathogenic spotted fever group rickettsiae in ticks collected from hedgehogs in Turkey: Haemaphysalis erinacei, a novel candidate vector for the genus Babesia

In this study, a total of 319 ticks were obtained from hedgehogs (Erinaceus concolor). All ticks were pooled into groups and screened by PCR for tick-borne pathogens (TBPs). PCR and sequence analyses identified the presence of a novel Babesia sp. in adult Haemaphysalis erinacei. In addition, the presence of natural transovarial transmission of this novel Babesia sp. was detected in Ha. erinacei. According to the 18S rRNA (nearly complete) and partial rRNA locus (ITS-1/5.8S/ITS-2) phylogeny, it was determined that this new species is located within the Babesia sensu stricto clade and is closely related to Babesia spp. found in carnivores. Furthermore, the presence of three pathogenic spotted fever group (SFG) rickettsiae was determined in 65.8% of the tick pools: Rickettsia sibirica subsp. mongolitimonae in Hyalomma aegyptium (adult), Hyalomma spp. (larvae), Rhipicephalus turanicus (adult), and Ha. erinacei (adult); Rickettsia aeschlimannii in H. aegyptium (adult); Rickettsia slovaca in Hyalomma spp. (larvae and nymphs) and H. aegyptium (adult). To our knowledge, this is the first report of R. sibirica mongolitimonae in H. aegyptium, Ha. erinacei, and Rh. turanicus, and the first report of R. slovaca in H. aegyptium. In addition, the presence of a single Hemolivia mauritanica haplotype was detected in H. aegyptium adults. Consequently, the presence of a novel Babesia sp. has been identified in a new candidate vector tick species in this study. Additionally, three SFG rickettsiae that cause infections in humans were identified in ticks collected from hedgehogs. Therefore, environmental wildlife monitoring for hedgehogs should be carried out for ticks and tick-borne pathogens in the region. Additionally, studies regarding the reservoir status of hedgehogs for the aforementioned pathogens must be carried out.

A cross-sectional screening by next-generation sequencing reveals Rickettsia, Coxiella, Francisella, Borrelia, Babesia, Theileria and Hemolivia species in ticks from Anatolia

BackgroundTicks participate as arthropod vectors in the transmission of pathogenic microorganisms to humans. Several tick-borne infections have reemerged, along with newly described agents of unexplored pathogenicity. In an attempt to expand current information on tick-associated bacteria and protozoans, we performed a cross-sectional screening of ticks, using next-generation sequencing. Ticks seeking hosts and infesting domestic animals were collected in four provinces across the Aegean, Mediterranean and Central Anatolia regions of Turkey and analyzed by commonly used procedures and platforms.ResultsTwo hundred and eighty ticks comprising 10 species were evaluated in 40 pools. Contigs from tick-associated microorganisms were detected in 22 (55%) questing and 4 feeding (10%) tick pools, with multiple microorganisms identified in 12 pools. Rickettsia 16S ribosomal RNA gene, gltA, sca1 and ompA sequences were present in 7 pools (17.5%), comprising feeding Haemaphysalis parva and questing/hunting Rhipicephalus bursa, Rhipicephalus sanguineus (sensu lato) and Hyalomma marginatum specimens. A near-complete genome and conjugative plasmid of a Rickettsia hoogstraalii strain could be characterized in questing Ha. parva. Coxiella-like endosymbionts were identified in pools of questing (12/40) as well as feeding (4/40) ticks of the genera Rhipicephalus, Haemaphysalis and Hyalomma. Francisella-like endosymbionts were also detected in 22.5% (9/40) of the pools that comprise hunting Hyalomma ticks in 8 pools. Coxiella-like and Francisella-like endosymbionts formed phylogenetically distinct clusters associated with their tick hosts. Borrelia turcica was characterized in 5% (2/40) of the pools, comprising hunting Hyalomma aegyptium ticks. Co-infection of Coxiella-like endosymbiont and Babesia was noted in a questing R. sanguineus (s.l.) specimen. Furthermore, protozoan 18S rRNA gene sequences were detected in 4 pools of questing/hunting ticks (10%) and identified as Babesia ovis, Hemolivia mauritanica, Babesia and Theileria spp.ConclusionsOur metagenomic approach enabled identification of diverse pathogenic and non-pathogenic microorganisms in questing and feeding ticks in Anatolia.

Bacterial and protozoal pathogens found in ticks collected from humans in Corum province of Turkey.

BackgroundTick-borne diseases are increasing all over the word, including Turkey. The aim of this study was to determine the bacterial and protozoan vector-borne pathogens in ticks infesting humans in the Corum province of Turkey.Methodology/Principal findingsFrom March to November 2014 a total of 322 ticks were collected from patients who attended the local hospitals with tick bites. Ticks were screened by real time-PCR and PCR, and obtained amplicons were sequenced. The dedected tick was belonging to the genus Hyalomma, Haemaphysalis, Rhipicephalus, Dermacentor and Ixodes. A total of 17 microorganism species were identified in ticks. The most prevalent Rickettsia spp. were: R. aeschlimannii (19.5%), R. slovaca (4.5%), R. raoultii (2.2%), R. hoogstraalii (1.9%), R. sibirica subsp. mongolitimonae (1.2%), R. monacensis (0.31%), and Rickettsia spp. (1.2%). In addition, the following pathogens were identified: Borrelia afzelii (0.31%), Anaplasma spp. (0.31%), Ehrlichia spp. (0.93%), Babesia microti (0.93%), Babesia ovis (0.31%), Babesia occultans (3.4%), Theileria spp. (1.6%), Hepatozoon felis (0.31%), Hepatozoon canis (0.31%), and Hemolivia mauritanica (2.1%). All samples were negative for Francisella tularensis, Coxiella burnetii, Bartonella spp., Toxoplasma gondii and Leishmania spp.Conclusions/SignificanceTicks in Corum carry a large variety of human and zoonotic pathogens that were detected not only in known vectors, but showed a wider vector diversity. There is an increase in the prevalence of ticks infected with the spotted fever group and lymphangitis-associated rickettsiosis, while Ehrlichia spp. and Anaplasma spp. were reported for the first time from this region. B. microti was detected for the first time in Hyalomma marginatum infesting humans. The detection of B. occultans, B. ovis, Hepatozoon spp., Theileria spp. and Hemolivia mauritanica indicate the importance of these ticks as vectors of pathogens of veterinary importance, therefore patients with a tick infestation should be followed for a variety of pathogens with medical importance.

First Hemolivia from southern Africa: reassigning chelonian Haemogregarina parvula Dias, 1953 (Adeleorina: Haemogregarinidae) to Hemolivia (Adeleorina: Karyolysidae)

To date, only a single species of Hemolivia, Hemolivia mauritanica (Sergent & Sergent, 1904), has been described from African terrestrial tortoises. Although various haemogregarines have been described from southern African terrapins and tortoises, including species from the genus Haemogregarina and one from the genus Hepatozoon, no species of Hemolivia have been identified previously from southern Africa. Since its morphological redescription, the taxonomic placement of one of these species, Haemogregarina parvula Dias, 1953, was in doubt. Hence, research was undertaken to resolve the true taxonomic position of this haemogregarine. Blood smears from nine wild tortoises of two species, Stigmochelys pardalis and Kinixys zombensis, from South Africa were screened, with the focus on H. parvula. Parasite DNA was extracted from ethanol-preserved blood samples, and fragments of the 18S rDNA gene were amplified by PCR using the primer sets HepF300/HepR900 and 4558/2733. Maximum likelihood and maximum parsimony analyses of 18S rDNA sequences showed that Haemogregarina parvula formed a monophyletic clade with species of Hemolivia within the Hepatozoon clade, not with species of Haemogregarina. It is thus recommended that this haemogregarine be reassigned to the genus Hemolivia, rendering Hemolivia parvula (Dias, 1953) the first species of the genus recorded from southern African tortoises.

Distribution and abundance of Hemolivia mauritanica (Apicomplexa: Haemogregarinidae) and its vector Hyalomma aegyptium in tortoises of Iran

AbstractDistribution pattern, prevalence and intensity of parasitaemia of heteroxenous apicomplexan blood parasite Hemolivia mauritanica and its vector tick Hyalomma aegyptium have been studied in 264 tortoises (212 Testudo graeca and 52 T. horsfieldii) throughout the Iranian territory. In T. graeca the highest prevalence and intensity of parasitaemia for H. mauritanica were recorded in the temperate mid and northern parts of the Iranian plateau, while the lowest values were found in the arid central part of Iran. No Hemolivia-positive samples were found in T. horsfieldii. Hyalomma aegyptium ticks were diagnosed in both tortoise species. The highest prevalence and intensity of infestation with H. aegyptium were recorded in western part of Iran, where climatic conditions are not so arid compared to central and eastern parts. No significant differences were found in intensity of parasitaemia of H. mauritanica between males and females. Similarly, no differences in intensity of parasitaemia were detected between different age groups (adults; more than 10 years, young; 5-10 years and juveniles; less that 5 years). Prevalence of H. mauritanica and H. aegyptium in adults was higher than in young and in juveniles. While intensity of infestation with H. aegyptium was significantly higher in adult tortoises compare to juveniles, no significant difference were found between intensity of H. aegyptium infestation in male and female tortoises.

Molecular Detection ofHemolivia(Apicomplexa: Haemogregarinidae) from Ticks of North AfricanTestudo graeca(Testudines: Testudinidae) and an Estimation of Their Phylogenetic Relationships Using 18S rRNA Sequences

ABSTRACT: Hemolivia mauritanica is an apicomplexan parasite infecting the tortoises Testudo graeca and Testudo marginata as the intermediate vertebrate host, and the tick Hyalomma aegyptium as the definitive invertebrate host. Here, we used molecular techniques to detect an apparent H. mauritanica in the ticks H. aegyptium, attached to Testudo graeca from North Africa. Sequences from 18S rRNA were used to place the new sequences, and others retrieved from GenBank, in a phylogenetic framework. A single Hemolivia haplotype was recovered from 16 ticks. Prevalence was estimated at 22.6% of hosts having ticks infected with H. mauritanica.

A redescription of Haemogregarina fitzsimonsi Dias, 1953 and some comments on Haemogregarina parvula Dias, 1953 (Adeleorina: Haemogregarinidae) from southern African tortoises (Cryptodira: Testudinidae), with new host data and distribution records

Blood films were examined from 154 wild and captive tortoises from four provinces of South Africa, including Gauteng, Kwazulu-Natal, North West and Western Cape. The five species ofchelonians studied were Chersina angulata (Schweigger), Kinixys belliana belliana (Gray), K. lobatsiana Power, K. natalensis Hewitt, and Stigmochelys pardalis (Bell). Two species of haemogregarines, previously reported from Mozambique, were identified in blood films, namely Haemogregarina fitzsimonsi Dias, 1953 and Haemogregarina parvula Dias, 1953. Additional stages of development (trophozoites and probable meronts, merozoites and immature gamonts) in blood preparations from South Africa warranted the redescription of H. fitzsimonsi. A variety of hosts and broad host distribution range were observed for this haemogregarine, with all five species of tortoises parasitized, wild and captive, from all four provinces, in all seasons. In contrast, only two individuals of K. b. belliana and one S. pardalis, all three captive in Kwazulu-Natal, contained H. parvula with encapsulated stages resembling those of Hemolivia mauritanica (Sergent et Sergent, 1904). For H. fitzsimonsi, parasite prevalences, but not parasitaemias, were significantly higher in captive than wild S. pardalis; captive female S. pardalis also showed a significantly greater prevalence of infection than males, but younger, lighter hosts were not significantly more heavily parasitized than older, heavier individuals. The ticks, Amblyomma marmoreum Koch, 1844 and A. sylvaticum (De Geer, 1778), found attached to some tortoises, may prove to be definitive hosts for the two species of haemogregarines observed.

Co-distribution Pattern of a Haemogregarine Hemolivia mauritanica (Apicomplexa: Haemogregarinidae) and Its Vector Hyalomma aegyptium (Metastigmata: Ixodidae)

Hyalomma aegyptium ticks were collected from tortoises, Testudo graeca, at localities in northern Africa, the Balkans, and the Near and Middle East. The intensity of infestation ranged from 1-37 ticks per tortoise. The sex ratio of feeding ticks was male-biased in all tested populations. Larger tortoises carried more ticks than did the smaller tortoises. The juveniles were either not infested, or carried only a poor tick load. Hyalomma aegyptium was absent in the western Souss Valley and Ourika Valley in Morocco, the Cyrenaica Peninsula in Libya, Jordan, and the Antilebanon Mountains in Syria. Hemolivia mauritanica, a heteroxenous apicomplexan cycling between T. graeca and H. aegyptium, was confirmed in Algeria, Romania, Turkey, Syria, Lebanon, and Iran. Its prevalence ranged from 84% in Romania (n = 45), 82% in eastern Turkey (n = 28), and 82% in the area of northwestern Syria with adjacent Turkish borderland (n = 90), to 38% in Lebanon (n = 8) and in only 1 of 16 sampled tortoises in Algeria. The intensity of parasitemia in the studied areas ranged from 0.01% up to 28.17%. The percentage of Hemolivia-infected erythrocytes was significantly higher in adults. All tortoises from Hyalomma-free areas were Hemolivia-negative. Remarkably, all 29 T. graeca from Jabal Duruz (southwestern Syria) and 36 T. graeca from the area north of Middle Atlas (Morocco) were Hemolivia-negative, despite the fact that ticks parasitized all adult tortoises in these localities. Identical host preferences of H. aegyptium and H. mauritanica suggest the occurrence of co-evolution within the Testudo-Hyalomma-Hemolivia host-parasite complex.

Endogenous development of Hemolivia mauritanica (Apicomplexa: Adeleina: Haemogregarinidae) in the marginated tortoise Testudo marginata (Reptilia: Testudinidae): evidence from experimental infection

Six young tortoises Testudo marginata Schoepff, 1792 were experimentally infected with Hemolivia mauritanica (Sergent et Sergent, 1904). The prepatent period ranged from 6 to 8 weeks. Young, smaller, club-like forms (6-9 x 3-6 Am) of gametocytes appeared in the peripheral blood first, whereas mature, elongated, cylindrical forms (9-12 x 5-7 Am) were detected after 1-2 weeks and predominated during later patency. Three of the infected tortoises were euthanized and dissected to study the endogenous stages. Meronts occurred in the cells of the reticulo-endothelial system and in the erythrocytes; these were observed mostly in parenchymatous organs. Mature forms measured 14.2 x 9.3 microm and contained 7-12 merozoites. Cysts with two (exceptionally one) cystozoites were also found predominantly in parenchymatous organs and measured 14.8 x 7.9 microm. Pathological changes attributable to Hemolivia were mild and limited to liver and kidneys. The role of individual developmental stages of haemogregarines is discussed with respect to evolution of heteroxenous life cycle and long-term persistence of parasites in their intermediate hosts.

Prevalence of Hemolivia mauritanica (Apicomplexa: Adeleina: Haemogregarinidae) in natural populations of tortoises of the genus Testudo in the east Mediterranean region

Hemolivia mauritanica (Sergent et Sergent, 1904) infections were found in 14% (n= 14) of Testudo graeca Linnaeus, 1758 tortoises in Bulgaria and in 92% (n = 26) of those in Turkey. Hemolivia mauritanica-like gametocytes were found in 81% (n = 47) of wild Testudo marginata Schoepff, 1792 tortoises in Greece. Parasitaemia intensity (the percentage of infected red blood cells found in approximately 104 cells) was 0.03-22.4% in T. graeca and 0.06-12.27% in T. marginata. On the other hand, all blood samples from 40 Bulgarian, 38 Greek, and 18 Croatian specimens of Testudo hermanni Gmelin, 1789 were negative.