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.     

Long-term occurrence of Hemolivia cf. mauritanica (Apicomplexa: Adeleina: Haemogregarinidae) in captive Testudo marginata (Reptilia: Testudinidae): evidence for cyclic merogony?

Blood smears from wild-caught, long-term captive tortoises, Testudo marginata, revealed the presence of gametocytes of a Hemolivia mauritanica-like hemogregarine in the erythrocytes of 72% tortoises examined. Significant parasitemia was also found in animals living several years in captivity. Experimentally infected tortoises showed no evidence of a decrease in parasitemia level more than 15 mo after infection. Morphologically, stages found in tortoises' erythrocytes were indistinguishable from those referred to by previous workers as H. mauritanica from Testudo graeca. Moreover, successful experimental transmission to Hyalomma aegyptium confirms the conspecificity with H. mauritanica. The occurrence of H. mauritanica gametocytes in tortoise living up to 8 yr in captivity is suggested to result from continuous, long-lasting cyclic merogony in tortoises' parenchymatous organs, which is an unknown phenomenon in the life cycle of Hemolivia spp.     

Holocene vegetation and climate history of the northern Golan heights (Near East)

In this paper we present and discuss palynological results based on a composite profile from Birkat Ram crater lake (Northern Golan, Near East) in order to reconstruct the environmental history, including human impact, of the last 6500 years. Furthermore we apply a newly-developed botanical climatological transfer function to reconstruct climate variations in the northern Golan Heights based on this pollen data-set. The Birkat Ram record is strongly influenced by anthropogenic indicators in the pollen diagram with high quantities during the Chalcolithic period/Early Bronze Age, during the Hellenistic-Roman-Byzantine periods, during the Crusader period and finally during modern times. The palaeoclimate reconstruction method used is based on a Bayesian approach and is robust in avoiding the influence of these strong anthropogenic signals on the reconstruction results. The area has always had Mediterranean climate conditions and no distinctive climate changes can be identified during the past 6500 years. Because of the orography of the Mt. Hermon region the particular geographical position of the northern Golan Heights is obviously capable of buffering large-scale fluctuations in precipitation, which have otherwise been documented for several regions in the Near East. Electronic Supplementary Material Supplementary material is available for this article at An erratum to this article can be found at     

Natural and experimental infection of the lizard Ameiva ameiva with Hemolivia stellata (Adeleina: Haemogregarinidae) of the toad Bufo marinus

Developmental stages of a haemogregarine in erythrocytes of the lizard Ameiva ameiva (Teiidae), from Pará State, north Brazil, were shown to be those of Hemolivia by the nature of the parasite's sporogonic cycle in the tick Amblyomma rotondatum. The type species, Hemolivia stellata Petit et al., 1990 was described in the giant toad Bufo marinus and the tick Amblyomma rotondatum, also from Pará State, and in view of the fact that A. ameiva and Bufo marinus share the same habitat and are both commonly infested by A. rotondatum, the possibility that the parasite of A. ameiva is H. stellata had to be considered. Uninfected lizards fed with material from infected ticks taken from B. marinus, and others fed with liver of toads containing tissue-cysts of H. stellata, were shown to subsequently develop typical Hemolivia infections, with all stages of the development similar to those seen in the naturally infected lizards. Conversely, a juvenile, uninfected toad became infected when fed with sporocysts of Hemolivia in a macerated tick that had fed on an infected A. ameiva and pieces of liver containing tissuecysts from the same lizard. The remarkable lack of host specificity shown by H. stellata, in hosts so widely separated as an amphibian and a reptile, is discussed.     

Risk for the introduction of exotic ticks and pathogens into Italy through the illegal importation of tortoises,Testudo graeca

In April 2008, 585 tortoises illegally imported into Italy from North Africa were examined for the presence of ticks. Of these, 221 tortoises (37.8%) were infested with a mean intensity of 3.9 ± 3.1 ticks (range 1–17 ticks). A total of 798 ticks (672 males, 125 females and one nymph) were collected and identified as Hyalomma aegyptium (L.) (Acari: Ixodidae). The overall male : female ratio observed was 5 : 1. The prevalence and mean intensity [±standard deviation (SD)] of ticks were higher among male (67.4%, 4.0 ± 3.2) than female (55.6%, 3.8 ± 3.1) tortoises, although this difference was not significant. By contrast, the prevalence and mean intensity of ticks were significantly higher on tortoises weighing >100 g (61.5%, 4.0 ± 3.2) compared with tortoises weighing <100 g (12.1%, 2.1 ± 1.1). Of the infested tortoises, 89.8% had ticks on their hind limbs, 21.0% on forelimbs, 18.6% on the tail and pre‐anal area, and 4.8% on the head; thus the hind limbs were evidently the preferred attachment site. The present report highlights the need to develop surveillance systems to prevent the introduction and spreading of exotic ticks and tick‐borne pathogens in Italy and other European countries.     

Identification of the etiological agent for necrotizing scute disease in the Texas tortoise

Epidermal lamellae (scutes) of the Texas tortoise, Gopherus berlandieri, from southern Texas (USA) were observed to be in various stages of necrosis, ranging from localized whitish blemishes to complete degradation of the external portion of the scute. Fusarium semitectum was consistently isolated from slivers of infected scute from tortoises. The fungus was not isolated from tortoises exhibiting no lesions. Confocal microscopy confirmed the presence of septate mycelia inside the scutes, and isolates of F. semitectum grown in the laboratory were successfully transferred to non-infected tortoises. Twenty-four tortoises maintained by two rehabilitators in southern Texas exhibited lesions; however, only one of 27 tortoises from Dimmit and Zavala counties was infected.     

A nonsense mutation in the LDL receptor gene leads to familial hypercholesterolemia in the Druze sect.

Familial hypercholesterolemia (FH) is an autosomal dominant disease caused by mutations in the LDL receptor gene. Here we characterize an LDL receptor mutation that is associated with a distinct haplotype and causes FH in the Druze, a small Middle Eastern Islamic sect with a high degree of inbreeding. The mutation was found in FH families from two distinct Druze villages from the Golan Heights (northern Israel). It was not found neither in another Druze FH family residing in a different geographical area nor in eight Arab and four Jewish FH heterozygote index cases whose hypercholesterolemia cosegregates with an identical LDL receptor gene haplotype. The mutation, a single-base substitution, results in a termination codon in exon 4 of the LDL receptor gene that encodes for the fourth repeat of the binding domain of the mature receptor. It can be diagnosed by allele-specific oligonucleotide hybridization of PCR-amplified DNA from FH patients.     

Reconstructing Mid-Pleistocene paleovegetation and paleoclimate in the Golan Heights using the δ(13)C values of modern vegetation and soil organic carbon of paleosols

The Golan Heights borders the Upper Jordan Valley on its eastern side and likely served as a prime foraging area for hominin groups that inhabited the Upper Jordan Valley during the Mid-Pleistocene. This study tests the hypothesis that Mid-Pleistocene climate in the Golan region was similar to that of the present day. Carbon isotope composition of present day plant communities and soil organic carbon from the Golan were compared to those of paleosols from Nahal Orvim to reconstruct Mid-Pleistocene paleoclimatic conditions. After correcting the paleosol values for recent changes in atmospheric carbon isotope values and potential biodegradation, the isotopic results show a strong similarity to those of present day local plants and soils. These results indicate that during the Mid-Pleistocene, the Golan was dominated by C₃ vegetation, shared similar climatic conditions with the present day, and displayed long-term environmental stability. The span of time of paleosol formation is unknown and might cover multiple climatic episodes; thus, although short climatic fluctuations may have occurred, their impact was not substantial enough to be detected in the Nahal Orvim paleosols. This study concludes that the Golan slopes provided hominins and large grazers with a reliable and highly nutritious foraging area that complemented the Jordan Valley riparian ecosystem.     

Evidence of Cowdria ruminantium infection (heartwater) in Amblyomma sparsum ticks found on tortoises imported into Florida

Amblyomma marmoreum and A. sparsum ticks were collected from tortoises imported into Florida from Africa and were tested for Cowdria ruminantium infection using a C. ruminantium-specific pCS20 polymerase chain reaction assay. In I shipment imported from Zambia, 15 of the 38 A. sparsum male ticks collected from the leopard tortoises (Geochelone pardalis) were found to be positive for infection with C. ruminantium. In contrast, all 148 A. marmoreum tested were negative for C. ruminantium infection. This is the first reported evidence of the introduction of heartwater-infected ticks into the United States, but there were no opportunities to confirm isolation of C. ruminantium from the ticks by either culture or transmission studies.     

Transstadial Transmission of Borrelia turcica in Hyalomma aegyptium Ticks

Borrelia turcica comprises the third major group of arthropod-transmitted borreliae and is phylogenetically divergent from other Borrelia groups. The novel group of Borrelia was initially isolated from Hyalomma aegyptium ticks in Turkey and it was recently found in blood and multiple organs of tortoises exported from Jordan to Japan. However, the ecology of these spirochetes and their development in ticks or the vertebrate hosts were not investigated in detail; our aims were to isolate the pathogen and to evaluate the possibility of transstadial transmission of Borrelia turcica by H. aegyptium ticks. Ticks were collected from Testudo graeca tortoises during the summer of 2013 from southeastern Romania. Engorged nymphs were successfully molted to the adult stage. Alive B. turcica was isolated from molted ticks by using Barbour-Stoenner-Kelly (BSK) II medium. Four pure cultures of spirochetes were obtained and analyzed by PCR and sequencing. Sequence analysis of glpQ, gyrB and flaB revealed 98%–100% similarities with B. turcica. H. aegyptium ticks collected from T. graeca tortoises were able to pass the infection with B. turcica via transstadial route, suggesting its vectorial capacity.     

Isolation and characterization of a novel Borrelia group of tick-borne borreliae from imported reptiles and their associated ticks

The members of the genus Borrelia are transmitted by arthropods and known to be infectious to vertebrates. Here we found isolates and DNAs belonging to the Borrelia turcica and unknown Borrelia species from imported reptiles and their ectoparasites. The Borrelia strains were isolated from blood and multiple organs of exotic tortoises, and were experimentally infectious to captive-bred tortoises. These findings suggest that these tortoises may be a candidate as the reservoir host of the Borrelia species. In this study, the Borrelia strains were also isolated from and/or detected in hard-bodied ticks, Amblyomma ticks and Hyalomma ticks. In some of these ticks, immunofluorescence imaging analysis revealed that the Borrelia had also invaded into the tick salivary glands. Accordingly, these ticks were expected to be a potential vector of the Borrelia species. Sequencing analyses of both housekeeping genes ( flaB gene, gyrB gene and 16S rDNA gene) and 23S rRNA gene-16S rRNA gene intergenic spacer region revealed that these Borrelia strains formed a monophyletic group that was independent from two other Borrelia groups, Lyme disease Borrelia and relapsing fever Borrelia . From these results, the novel group of Borrelia comprises the third major group of arthropod-transmitted borreliae identified to date.     

Monitoring phytoplasma-bearing leafhoppers/planthoppers in vineyards in the Golan Heights, Israel

In the course of identifying the vector(s) of the grapevine yellows (GY) and western-X (WX) phytoplasmas in the Golan Heights, leafhopper and planthopper species were examined. The planthopper, Hyalesthes obsoletus , was trapped on yellow sticky traps or collected on weeds; there was a relatively small peak during two weeks in June and during four weeks starting mid-September. The leafhoppers, Neoaliturus spp. and Circulifer sp. (of the haematoceps complex) were both trapped early or very late in summer, but could be collected on weeds throughout the summer. The two remaining leafhopper species, Macrosteles quadripunctulatus and Orosius orientalis (= albicinctus ) were only rarely caught on sticky traps, but were found on weeds throughout the summer. Phytoplasmas were found within the body of these five species; all are historically known to be efficient vectors of various phytoplasmas and were therefore chosen for further investigations.     

Tick prevalence and species diversity on Aldabran giant tortoises (Dipsochelys dussumieri) in relation to host range and host size in a restored ecosystem, Kenya

Tick species density and diversity on Aldabran tortoises was investigated in relation to the habitat range and size of each tortoise. Identification of tick infestation patterns forms an important aspect of effective tick control. Ten Aldabran tortoises were de‐ticked and monitored over the course of 2 months. Tick species found were Amblyomma sparsum, Amblyomma nuttalli, Amblyomma hebraeum and Boophilus decoloratus, with the most prevalent species found being A. sparsum. Tick loads varied considerably from 20 to 214 ticks per tortoise, with most ticks collected from the head/neck region. Tortoises ranging outside Haller Park had higher tick loads (70–214) compared with tortoises ranging within Haller Park (20–99). Tick load was not correlated with tortoise size. Results indicate that tick loads are related to the habitat range of the tortoises and may indirectly also be related to food preference and host food availability. Implications of the findings and appropriate tick control measures are discussed.     

Acquired immunity to larvae of Amblyomma marmoreum and Ahebraeum by tortoises, guinea-pigs and guinea-fowl

A study of acquired resistance in guinea-pigs, guinea-fowl and tortoises to larvae of the ticks Amblyomma hebraeum Koch and A.marmoreum Koch (Acari: Ixodidae) showed that repeated infestations of the laboratory host resulted in a 29.3-49.4% reduction respectively in weight of engorged larvae. No resistance was induced in guinea-fowl, the natural host for larvae of both species. A small decline in the mean weight of engorged larvae was observed for both species of ticks after three successive infestations on tortoises. The reduction in weight was more pronounced in A.hebraeum (14.5%) which does not naturally feed on tortoises than for A.marmoreum (8.4%), a common tick on this host. The occurrence of acquired resistance in natural tick host relationships as opposed to unnatural tick host relationships is discussed.     

Amblyomma marmoreum on tortoises of southern province, Zambia

Reptiles from Southern Province, Zambia were captured and inspected for ticks. Seven of 18 tortoises (Geochelone pardalis) were infested by small numbers of adults and nymphs of Amblyomma marmoreum. Under such circumstances, the tick had little apparent effect on the host. None of the 13 specimens representing four species of snakes examined were infested with ticks.     

Hyalomma aegyptium as dominant tick in tortoises of the genus Testudo in Balkan countries, with notes on its host preferences

Collection of 1327 ticks sampled throughout Greece, Bulgaria, Romania and Croatia, from 211 tortoises belonging to three species, Testudo marginata Schoepff, T. graeca Linnaeus, and T. hermanni Gmelin, revealed the presence of four species of ixodid ticks, namely Hyalomma aegyptium (Linnaeus), Haemaphysalis sulcata Canestrini and Fanzago, H. inermis Birula and Rhipicephalus sanguineus (Latreille). Study confirmed the strong dominance of all life stages of H. aegyptium among ticks parasitizing west Palaearctic tortoises of genus Testudo Linnaeus. Furthermore, a considerable portion of ticks collected from tortoises in southwestern Bulgaria represent larvae and nymphs of H. sulcata. At the same area we collected as exception one larva and one nymph of H. inermis from a single specimen of T. hermanni. Our findings of four adults of R. sanguineus is the first record of this species from reptilian host. According to our results achieved on localities with syntopic occurrence of two tortoise species, T. marginata and T. graeca represent in the Balkans the principal hosts of H. aegyptium, whereas T. hermanni serves only as an alternative host in the areas close to range of either T. marginata or T. graeca.     

Dynamics of Borrelia burgdorferi infection in nymphal Ixodes ricinus ticks during feeding

We report the sequential developmental events of Borrelia burgdorferi in histological sections of Ixodes ricinus nymphs before, during and after feeding. During the blood meal a decrease of approximately 50% in the number of infected ticks was recorded (eight out of 76, 11%) in comparison with the infection rate of unfed ticks (12 out of 56, 21%). Spirochetes were detected in tick salivary glands only after 2 days of attachment. From day 3 until drop-off, the number of infected ticks increased to 31% (15 out of 49). A quadratic logistic regression analysis showed that the variation in the number of infected ticks was significant, but only during the blood meal. The drop in the percentage of infected ticks during the first hours following attachment to the host is explained by our observation of spirochetes in the faeces of the ticks. The increase in the infection rate of replete ticks may be due to an uptake of spirochetes from the host skin at the feeding site.     

Polymorphism of the 12S rRNA gene and phylogeography of the Central Asian tortoises Agrionemys horsfieldii gray, 1844

Based on intraspecific polymorphism of 12S rRNA gene, genetic variation of isolated populations of the Central Asian tortoise, Agrionemys horsfieldii, was for the first time investigated on a large part of the species distribution range, encompassing Uzbekistan, southern Kazakhstan, and northern and eastern Iran. In 59 tortoises, four haplotypes were discovered, including two (AH1 and AH2), described earlier. Haplotype AH1 was detected in 52 tortoises, inhabiting southern Kazakhstan and Uzbekistan. Haplotype AH2 was found in four tortoises from the border territory between Uzbekistan, Tajikistan, and Afghanistan. Two novel haplotypes, AH3 and AH4, were detected in the three tortoises from Iran. Based on nucleotide substitutions in the 12S rDNA sequence, the possible divergence time between the tortoises from different parts of the range was estimated. Possible pathways of the formation of modern intraspecific groups of A. horsfieldii are discussed.     

Survey of the vectorial competence of ticks in an endemic area of spotted fever group rickettsioses in Fukui Prefecture, Japan

The prevalence of SFGR in ixodid ticks in the Mt. Arashima-dake area in the northern part of Fukui Prefecture was surveyed, because of strong suspicions that the first case identified in this Prefecture had become infected with R. helvetica in this region. The ticks identified consisted of three genera and six species; I.ovatus, I. persulcatus, I. monospinosus, H. flava, H. japonica and D. taiwanensis. Of all 222 ticks collected, only I. monospinosus ticks (8 of 32 examined) were positive for SFGR isolates, which were genetically identified as R. helvetica. Ticks (157 of all 222) positive for SFGR-DNA fragments consisted of I. monospinosus (14 of 32), I. persulcatus (11 of 55), I. ovatus (3 of 38), H. flava (5 of 21) and H. japonica (2 of 9). Of these, thirteen I. monospinosus, eight I. persulcatus, three I. ovatus, two H. flava and one H. japonica were identified by nucleotide sequences as positive for R. helvetica. DNA fragments from three H. flava and one H. japonica showed greater homology to R. japonica than to R. helvetica or R. asiatica. The present results indicate that it is most likely that the vector tick of R. helvetica infection in Fukui Prefecture is I. monospinosus.     

Development of infectivity in Hyalomma detritum (Schulze, 1919) ticks infected with Theileria annulata (Dchunkowsky and Luhs, 1904).

The time-course for the development of infectivity was studied in Hyalomma detritum ticks fed as pre-imagoes on calves infected with Theileria annulata. Unfed adults derived from infected nymphs were non-infectious when inoculated into susceptible calves, whereas ticks of both sexes that had fed for 2-3 days or longer on calves or rabbits were always infectious. Some adults fed for only 1-2 days were infectious, whereas others were not. As few as 2 ticks were capable of infecting susceptible calves.