Tick-borne pathogens in ticks (Acari: Ixodidae) collected from various domestic and wild hosts in Corsica (France), a Mediterranean island environment.

Corsica is a mountainous French island in the north-west of the Mediterranean Sea presenting a large diversity of natural environments where many interactions between humans, domestic animals and wild fauna occur. Despite this favourable context, tick-borne pathogens (TBPs) have not systematically been investigated. In this study, a large number of TBPs were screened in ticks collected over a period of one year from domestic and wild hosts in Corsica. More than 1,500 ticks belonging to nine species and five genera (Rhipicephalus, Hyalomma, Dermacentor, Ixodes and Haemaphysalis) were analysed individually or pooled (by species, gender, host and locality). A real-time microfluidic PCR was used for high-throughput screening of TBP DNA. This advanced methodology enabled the simultaneous detection of 29 bacterial and 12 parasitic species (including Borrelia, Anaplasma, Ehrlichia, Rickettsia, Bartonella, Candidatus Neoehrlichia, Coxiella, Francisella, Babesia and Theileria). The Crimean-Congo haemorrhagic fever (CCHF) virus was investigated individually in tick species known to be vectors or carriers of this virus. In almost half of the tick pools (48%), DNA from at least one pathogen was detected and eleven species of TBPs from six genera were reported. TBPs were found in ticks from all collected hosts and were present in more than 80% of the investigated area. The detection of DNA of certain species confirmed the previous identification of these pathogens in Corsica, such as Rickettsia aeschlimannii (23% of pools), Rickettsia slovaca (5%), Anaplasma marginale (4%) and Theileria equi (0.4%), but most TBP DNA identified had not previously been reported in Corsican ticks. This included Anaplasma phagocytophilum (16%), Rickettsia helvetica (1%), Borrelia afzelii (0.7%), Borrelia miyamotoi (1%), Bartonella henselae (2%), Babesia bigemina (2%) and Babesia ovis (0.5%). The high tick infection rate and the diversity of TBPs reported in this study highlight the probable role of animals as reservoir hosts of zoonotic pathogens and human exposure to TBPs in Corsica.

Research on Piroplasmorida and other tick-borne agents: Are we going the right way?

The use of molecular diagnostic techniques in the study of ticks and tick-borne agents has increased dramatically in the last few decades, and while conferring the advantages of rapidity, sensitivity and accuracy, they can also be misused by the research community, who have shown an increasing overreliance on the data they produce, often completely abandoning more traditional approaches such as microscopy and serology. In this short review examples are provided of 'findings' that are apparently at odds with well-established eco-epidemiological knowledge, ranging from descriptions of new species of pathogens to their detection in new hosts and ticks, often immediately incriminated as new reservoir hosts and vector ticks, respectively. Most examples have been taken from the Piroplasmorida or "piroplasm" species as they are often called. Researchers are urged to utilize molecular methods with as much awareness as possible of the system or systems they are investigating, and journal editors and their teams of reviewers should be encouraged to adopt a more critical approach to papers that contain data based solely on molecular methods without due regard to the underlying biology. Explanations for such findings should be offered and discussed.

Ticks (Acari: Ixodidae) infesting cattle and some other domestic and wild hosts on the French Mediterranean island of Corsica.

BACKGROUND: Corsica is a mountainous French island in the north-western Mediterranean presenting a large diversity of natural environments where many interactions between domestic animals and wild fauna occur. Despite a favourable context for ticks and tick-borne diseases (TBDs), the tick fauna of Corsica has not systematically been investigated. METHODS: For one year (May 2014-May 2015), a survey of ticks infesting cattle was performed in the three Corsican cattle slaughterhouses. Two of these were visited monthly and one quarterly; the ticks were manually collected, just after flaying. Ticks were identified on their morphology; when necessary, some specimens were also molecularly identified by sequencing mitochondrial cox1 (cytochrome c oxidase subunit 1) and 16S ribosomal RNA genes and ITS2 (internal transcribed spacer 2). During the same period, ticks from other domestic animals (small ruminants, horses, domestic carnivores) and wild animals (wild boars, mouflons, deer) were occasionally collected. RESULTS: A total of 1,938 ticks was collected from 264 of 418 cattle examined, reared in 86 different localities. Eight tick species were found infesting cattle: Rhipicephalus bursa (56.1 %), Hyalomma marginatum (21.5 %), Hy. scupense (8.7 %), Ixodes ricinus (5.7 %), Haemaphysalis punctata (4.8 %), Rh. sanguineus (sensu lato) (2.3 %), Rh. (Boophilus) annulatus (0.7 %) and Dermacentor marginatus (0.2 %). The cattle infestation rate remained high all year (more than 50 %). Several tick species showed seasonal variation of their activity. From other Corsican animals 1,196 ticks were collected. Comparing ticks collected from cattle with those found on other animals, several host preferences were shown. A noteworthy record is that of a few Ha. sulcata on mouflons which were mainly infested by Rh. bursa. CONCLUSION: The Corsican tick fauna is characterized by typical Mediterranean species (Rh. bursa and Hy. marginatum), but the mild climate and diversified environment provide satisfactory habitats both for species usually found in dry areas (Hy. scupense) and species usually collected in humid areas (I. ricinus).

First report of the tick Hyalomma scupense (natural vector of bovine tropical theileriosis) on the French Mediterranean island of Corsica.

Hyalomma scupense (Acari, Ixodidae) is a common tick species found in several areas in North Africa, Asia and South Europe and an efficient natural vector of bovine tropical theileriosis (Theileria annulata), a livestock disease with an important economic impact. For one year, 1938 ticks were collected on cattle in several Corsican slaughterhouses; 168 of them were morphologically identified as H. scupense. This result was confirmed by genetic identification using sequences of mitochondrial cytochrome c oxidase subunit I (COI) and ribosomal internal transcribed spacer 2 (ITS2) genes. The presence of 2 different stages (adults and nymphs), collected in various areas of the island, indicates that a population of H. scupense is established in Corsica. However, bovine tropical theileriosis has not been diagnosed on the island so far.

[A legend of tropical veterinary medicine: Dick Zwart]. / Een icoon in de tropische diergeneeskunde: Dick Zwart.

An overview is given of the achievements of Professor Dr. D. Zwart (1930-2005). Following a brief introduction dealing with his background, his first eleven years of working in tropical countries are reviewed. Initially he was employed as a government veterinarian in Dutch New Guinea, followed by five years as a scientist at the agricultural faculty in Kumasi, Ghana, where he completed his PhD on liver cirrhosis in local pigs. He was seconded by the Department of Tropical Veterinary Medicine at Utrecht University to work as a virologist in Vom, Nigeria. Once back in The Netherlands, he was selected to take over the position of head of the Department due to the unexpected death of Professor S. G. Wilson. He stayed for 20 years at Utrecht University and was for another nine years department head at the Agricultural University of Wageningen. He was an influential man during this long period as department head, not only for his students, but also in the international field. Four aspects of the achievements by Dick are highlighted in detail: as a scientist, as an educator, as a stimulating figure, and as a visionary.

Ticks of the Central African Republic.

A review is presented of the ticks of the Central African Republic (C.A.R.), based on unpublished data and a literature review. We consider as established in the C.A.R. two species of Argasidae (one species of each of the genera Carios and Ornithodoros) and 49 species of Ixodidae: 5 Ixodes, 11 Amblyomma, 2 Dermacentor, 9 Haemaphysalis, 4 Hyalomma, and 18 Rhipicephalus species, 3 of which belong to the subgenus Boophilus. Two of these species, one Amblyomma and one Dermacentor, may by now have disappeared from the country together with their hosts, the black and white rhinoceroses. Moreover, four other species of Ixodidae have been recorded, but are not necessarily established in the country, and 6 species of Ixodidae have apparently been reported incorrectly; the data for the occurrence of three other species of Ixodidae are uncertain. Rhipicephalus species in the C.A.R. present the greatest difficulties in identification, on the one hand because of uncertainties in taxonomy, particularly in the capensis group, on the other hand because of individual variability, which means that individual specimens cannot always be reliably identified to species. Examining the dissected and cleared gonopore structure of females helps, but is very time-consuming where large numbers are involved. Some of the species of the genus Haemaphysalis are also difficult to identify with certainty. There are only a few species of Hyalomma in the country. We revised also some old records of ticks in C.A.R., which we consider as unreliable and should be disregarded because of subsequent changes in taxonomy and nomenclature, unless the material is still available for examination.

[Tropical veterinary medicine and education in The Netherlands]. / Tropische diergeneeskunde en onderwijs in Nederlanda.

in this lecture an overview is given of the development of tropical veterinary medicine and education in The Netherlands after the Dutch colonial period. The starting point is the development of tropical veterinary medicine in general, especially in Europe and Africa. It is pointed out that just now it is very important to have specialists in tropical diseases not only in the tropics but also in the western world since globalization involves the import of a lot of tropical diseases. The speaker is an advocate of a course on tropical veterinary medicine on an European level, but at the same time he is sceptical about it. In the second part he gives an overview of the education programme on tropical veterinary medicine from the foundation of the Institute at Utrecht University in 1948 until its decline.

Polyphasic taxonomy.

Several organisms from a number of prokaryotic and eukaryotic groups have presented problems for systematists for a long time. Both phenotypic and genotypic methods for sorting out these relationships have been employed. There are limitations with each method when taken alone. Since the purpose of systematics is to determine the correct genealogical relationships among biological organisms, it is necessary to use all available means to arrive at consensus associations, and polyphasic taxonomy, which takes into consideration both methods, is a rational approach. In this short article, we provide a number of examples where polyphasic taxonomy is serving as the means of arriving at the desired consensus.

Babesia--a historical overview.

The history of the genus Babesia is briefly outlined. The classical differences with the main other genus of non-pigment-forming hemoparasites, Theileria, are the absence of extra-erythrocytic multiplication (schizogony) in Babesia and the cycle in the vector tick, which includes transovarial transmission in Babesia but only transstadial transmission in Theileria. Also, the multiplication in the red cell of Babesia, by budding, most often results in two daughter cells (merozoites), while that of Theileria gives four merozoites, often as a Maltese cross. In particular this means that what is still commonly called Babesia microti is not a Babesia and that it would be just as logical to speak of human theileriosis as of babesiosis. The small piroplasm of horses, long known as Babesia equi, is already commonly designated as Theileria equi. However, on molecular grounds, it may be necessary to create a new genus for these parasites. The Babesia species of domestic animals are briefly discussed and presented in a table.

A note on Hyalomma nitidum, its distribution and its hosts.

The paper summarises the morphological characteristics, the known geographical distribution and hosts of Hyalomma nitidum, a tick which has never been properly characterised. H. nitidum was first described in Cameroon in 1919, long considered as a synonym of Hyalomma truncatum and recognised as a separate and valid species only since the early 1970s. H. nitidum occurs from Senegal to the Central African Republic in more humid conditions than H. truncatum, roughly to the south of the isohyet of 900 mm. Adults are parasitic on various wild and domestic ungulates, on which immatures have not been found. Nymphs, identified after moulting to adults, have been found on a hare, and tentatively identified larvae and nymphs of this species have also been collected on hare as well as on a zebra mouse. The tick has been found infected with the virus of Crimean-Congo haemorrhagic fever, so far no other disease associations are known.

On molecular taxonomy: what is in a name?

Gene sequences of small portions of the genome are often used for premature detailed taxonomic changes, neglecting polyphasic taxonomy, which should also consider phenotypical characteristics. Three examples are given: (i) Recently, members of the genera Eperythrozoon and Haemobartonella have been moved, correctly so, from the Rickettsiales to the Mycoplasmatales, but were assigned to the genus Mycoplasma, mostly on the basis of 16S rRNA sequence analysis. Not only is the 16S rRNA sequence similarity between 'classical' Mycoplasma and these species of Eperythrozoon and Haemobartonella less than that between some other well-recognised bacterial genera, but their biological differences amply justify their classification in different genera of the Mycoplasmatales. Furthermore, the move creates considerable confusion, as it necessitates new names for some species, with more confusion likely to come when the 16S rRNA sequences of the type species of Eperythrozoon, a name which has priority over Mycoplasma, will be analysed. (ii) In the Rickettsiales, members of the genera Anaplasma, Ehrlichia, Cowdria, Neorickettsia and Wolhbachia are so closely related phylogenetically on the basis of 16S rRNA sequences, and for some also of groESL operon sequences, that they have recently been fused, correctly so, into one family, the Anaplasmataceae, while the tribes Ehrlichieae and Wolbachieae have been abolished. Sequence diversity within the 'classical' genus Ehrlichia has led to classifying E. phagocytophila (including E. equi and the agent of human granulocytic ehrlichiosis), E. platys and E. bovis in the genus Anaplasma, while others have been retained in Ehrlichia, which also includes Cowdria ruminantium. E. sennetsu and E. risticii have been transferred to the genus Neorickettsia. 16S rRNA and GroEL sequences of 'classical' Anaplasma and some members of 'classical' Ehrlichia do show a close relationship, but differences in citrate synthase gene sequences, the GC content of this gene, and sequences of the gene encoding the beta-subunit of RNA polymerase, not to speak of the phenotypical differences, do not justify the fusion into one genus. Because of the phylogenetical diversity in Ehrlichia it is recommended that a new genus name be created for the E. phagocytophila genogroup (and E. platys and E. bovis). (iii) One of the conclusions of studies on the phylogeny of ticks of the subfamilies Rhipicephalinae and Hyalomminae, based on nucleotide sequences from 12S rRNA, cytochrome c oxidase I, the internal transcribed spacer 2, 18S rRNA, as well as morphological characters, is that Boophilus should be considered as a subgenus of Rhipicephalus. While Boophilus and Rhipicephalus are undoubtedly close, the obviously important morphological and biological differences between the genera Rhipicephalus and Boophilus are thus overruled by similarities in the sequences of a number of genes and this leads to considerable confusion. Polyphasic taxonomy amply justifies maintaining Boophilus as a separate genus, phylogenetically near to Rhipicephalus. This note is a plea for a cautious and balanced approach to taxonomy, taking into account molecular genotypical information, as far as is possible from different genes, as well as phenotypical characteristics.