Marginatum Rufipes Research Articles

Diversity of Hard Ticks (Acari, Ixodidae) Infestion in Arabian Camel in Iraq

A total of 1346 hard ticks (863♂ and 483 ♀) infested 104 camels, 60 alive camels with 93.33% infestation rate and 44 carcasses of camels had 79.54% infestation rate The total infestation rate was 87.5 %. The current study results revealed ten species of hard ticks family Ixodidae Koch, 1844 related to genus Hyalomma as following: H. dromedarii Koch, 1844, H. schulzii Morel, 1969, H. turanicum Pomerantsev, 1946, H. excavatum Koch, 1844, H. truncatum Koch, 1844, H. scupense Schulzii, 1919, H. marginatum Koch, 1844, H. anatolicum Koch, 1844, H. rufipes Koch, 1844, H. impeltatum Schulze & Schlottke, 1930 from camel Camelus dromedarius Linnaeus, 1758 collected from 21 regions belonging to six provinces in middle, west and south of Iraq where camels were bred in abundance. According to the current results, camels are considered a new host for three species of genus Hyalomma: H. truncatum, H. marginatum rufipes. These results may be of more importance as being the available data for risk topic about camels infested with hard ticks.

Diversity and Phylogeny of Cattle Ixodid Ticks and Associated Spotted Fever Group Rickettsia spp. in Tunisia.

Tick-borne rickettsioses are mainly caused by obligate intracellular bacteria belonging to the spotted fever group (SFG) of the Rickettsia genus. So far, the causative agents of SFG rickettsioses have not been detected in cattle ticks from Tunisia. Therefore, the aim of this study was to investigate the diversity and phylogeny of ticks associated with cattle from northern Tunisia and their associated Rickettsia species. Adult ticks (n = 338) were collected from cattle in northern Tunisia. The obtained ticks were identified as Hyalomma excavatum (n = 129), Rhipicephalus sanguineus sensu lato (n = 111), Hyalomma marginatum (n = 84), Hyalomma scupense (n = 12) and Hyalomma rufipes (n = 2). After DNA extraction from the ticks, 83 PCR products based on the mitochondrial 16S rRNA gene were sequenced and a total of four genotypes for Rh. sanguineus s.l., two for Hy. marginatum and Hy. excavatum and only one for Hy. scupense and Hy. rufipes were recorded, with the occurrence of one, two and three novel genotypes, respectively, for Hy. marginatum, Hy. excavatum and Rh. sanguineus s.l. mitochondrial 16S rRNA partial sequences. The tick DNA was tested for the presence of Rickettsia spp. by using PCR measurements and sequencing targeting three different genes (ompB, ompA and gltA). Of the 338 analyzed ticks, 90 (26.6%), including 38 (34.2%) Rh. sanguineus s.l., 26 (20.1%) Hy. excavatum, 25 (29.8%) Hy. marginatum and one (50%) Hy. rufipes tick, were positive for Rickettsia spp. Based on 104 partial sequences of the three analyzed genes, the BLAST analysis and phylogenetic study showed the infection of Hy. excavatum, Hy. marginatum and Rh. sanguineus s.l. tick specimens with R. massiliae, R. aeschlimannii and R. sibirica subsp. mongolitimonae and one Hy. rufipes tick specimen with R. aeschlimannii. In addition, coinfection with R. massiliae and R. aeschlimannii was reported in one Hy. marginatum and one Rh. sanguineus s.l. tick specimen, while a coinfection with R. massiliae and R. sibirica subsp. mongolitimonae was recorded in one Rh. sanguineus s.l. tick specimen. In conclusion, our study reports, for the first time in Tunisia, the infection of cattle ticks belonging to Hyalomma and Rhipicephalus genera with zoonotic Rickettsia species belonging to the SFG group.

The climate niche of the invasive tick species Hyalomma marginatum and Hyalomma rufipes (Ixodidae) with recommendations for modeling exercises

The finding of immature stages of some Hyalomma spp. feeding on migratory birds in Europe is unexceptional. The reports of adults of Hyalomma in Europe (incl. the British Isles) after successful molting from immatures have increased in recent years. It has been claimed that the warming of the target territory could favor the populations of these invasive ticks. Although evaluations of the impact on health or measures of adaptation are on their way, the climate niches of these species remain undefined, preventing preventive policies. This study delineates such niches for both Hyalomma marginatum (2,729 collection points) and Hyalomma rufipes (2,573 collections) in their distribution area, together with 11,669 points in Europe where Hyalomma spp. are believed to be absent in field surveys. Niche is defined from daily data of temperature, evapotranspiration, soil humidity and air saturation deficit (years 1970–2006). A set of eight variables (annual/seasonal accumulated temperature and vapor deficit) has the maximum discriminatory power separating the niches of both Hyalomma and a negative dataset, with an accuracy near 100%. The sites supporting H. marginatum or H. rufipes seem to be controlled by the joint action of the amount of water in the air (accounting for mortality) and the accumulated temperature (regulating development). The use of accumulated annual temperature as the only variable for predictive purposes of colonization of Hyalomma spp. looks unreliable, as far as values of water in air are excluded.

A new report of adult Hyalomma marginatum and Hyalomma rufipes in the Czech Republic

Hyalomma marginatum and Hyalomma rufipes are important vectors of Crimean-Congo Hemorrhagic Fever Virus (CCHFV) in North Africa and Southern Europe. They are occasionally also reported from Central and Western Europe where they are likely introduced from their natural range by migratory birds. In this study, we report findings and molecular identification of adults and one nymph of H. marginatum and H. rufipes, primarily from horses from different regions of the Czech Republic. While the number of the reported ticks is small, this is likely to be an underrepresentation of the actual number. Due to their vector competence for CCHFV and potential expansion into new areas with a changing climate, surveillance programs in Europe are warranted.

Molecular Detection of Theileria annulata among Dairy Cattle and Vector Ticks in the Herat Area, Afghanistan.

Theileriosis is one of the most important diseases in tropical and subtropical regions and leads to annual economic losses, such as the reduction of dairy products and casualties. Although the clinical form of bovine theileriosis has been observed in Afghanistan, to the best of our knowledge, no comprehensive study has been conducted on this issue. This molecular survey was performed to identify Theileria annulata and tick vectors in dairy cattle in the Herat area, Afghanistan, from June 2015-September 2016. A total of 100 dairy cattle were clinically examined and their blood smears, EDTA blood samples, and ixodid ticks were collected. The blood samples were transported to the laboratory, followed by the preparation of the blood smears and staining with the Giemsa method. The collected ticks were identified at the species (spp) level using the identification key and were then separated into 70 tick pools according to their species. Subsequently, the salivary glands were dissected out in 0.85% saline under a stereomicroscope. The DNA of blood and salivary glands was extracted using a commercial kit and analyzed by polymerase chain reaction (PCR). The ring form of Theileria spp infection was observed in 22 (22%) of blood smears, while 74% of blood samples were T. annulata positive using PCR. Among the collected ticks, the numbers of male and female ticks were obtained at 219 and 130 ticks, respectively. The frequency of tick spp was rated in descending order as Hyalomma annatolicum (73.9%), Hyalomma excavatum (22.3%), Hyalomma nymph spp (12%), Hyalomma marginatum (1.7%), Hyalomma asiaticum (1.1%), and Hyalomma rufipes (0.75%). The PCR results showed that seven pools belonging to salivary glands of H. anatolicum were infected with T. annulata. Based on the obtained results, it can be concluded that T. annulata had a high frequency in dairy cattle and H. anatoloicum was also identified, such as the vectors of T. annulata in the Herat area, Afghanistan.

Distribution and Seasonal Dynamics of Tick Species Infesting Cattle in Nocturnal and Daytime Systems of Livestock in the Far North Region, Cameroon

Diseases transmitted by ticks constitute a threat to cattle in Cameroon, Far North Region. The objective of this study was to identify ticks and to determine the structure and seasonal variation of tick communities infesting cattle in two types of farming in the Far North Region of Cameroon. Investigation on cattle were conducted from August 2018 to July 2019. Thus, ticks were collected monthly from 108 cattle in three districts (Bogo, Pétté, Kalfou). A total of 13151 adult ticks were collected. Identification revealed the presence of six tick species belonging to three genera whose relative abundance varied according to the different tick species: <i>Amblyomma variegatum</i> (39.88%), <i>Hyalomma marginatum rufipes</i> (33.43%), <i>Hyalomma truncatum</i> (14.70%); <i>Hyalomma impeltatum</i> (9.88%); <i>Boophilus decoloratus</i> (1.58%); <i>Rhipicephalus sanguineus</i> (0.51%). Ticks were most abundant at the beginning of the rainy season (June-July) and least abundant during the dry season (October-April). There was no significant difference between the abundance of different tick species in the two farming systems. Older animals and females were the most infested with ticks. The most infested parts by ticks were udders and testicles (40.08%), anal region (18.04%), legs and armpits (13.27%) and chest (13.96%). This study can help in improving of appropriate strategies for the control of ticks infestation in cattle.

First record of hard tick species, Hyalomma marginatum marginatum and H. marginatum rufipes (Acari: Ixodidae), as probable vectors of Crimean-Congo hemorrhagic fever virus, from the spur-thighed tortoise, Testudo graeca (Reptilia: Testudinidae), SE Iran

Ticks (Acari: Ixodidae) are one of the most successful and widespread vectors of zoonotic pathogens of medical interest. Most ticks use a wide spectrum of vertebrate hosts, including tortoises. They are obligatory ectoparasitic blood sucking arthropods on egg laying (ovipositing) and/or mammalian hosts. The present investigation was part of a wider survey to study the fauna of wild ticks on primarily selective domesticated mammals and subsequently the likely tick infection with Crimean-Congo hemorrhagic fever (CCHF) virus RNA. Ticks were morphologically identified to species level using a valid regional taxonomic key. They were subsequently processed for the presence of CCHF virus RNA using reverse transcription polymerase chain reaction (RT-PCR). During this survey, two natural tortoises were found infested with ticks at highland. Hard tick species of Hyalomma marginatum marginatum, Hy. marginatum rufipes and Hy. marginatum aegyptium were found in attachment to two Eastern spur-thighed tortoise species of Testudo graeca (Reptilia: Testudinidae) for the first time in southeast Iran. These host tortoises were discovered at high altitude region of about 2800-2900m above sea level in the vicinity of a village in Jiroft county, Kerman province, Iran. None of these tortoise-associated ticks were infected with CCHF virus RNA. Since herd breeding nomads often climb to highland to improve animal grazing in open shrub land, the importance of infestation with wild ticks potentially infected with various pathogens is described.

Imported Hyalomma ticks in Germany in 2018

BackgroundHyalomma marginatum and Hyalomma rufipes are two-host tick species, which are mainly distributed in southern Europe, Africa and middle-eastern Asia. They are well-known vectors of Crimean Congo hemorrhagic fever (CCHF) virus and other viruses as well as Rickettsia aeschlimannii. In recent years, these tick species have been found sporadically in Germany, but they do not belong to the autochthonous tick fauna in Germany.MethodsTicks with unusual morphology were collected and sent from private persons or public health offices to involve institutions for morphological identification and further testing. All ticks identified as Hyalomma spp. were tested using molecular detection methods for CCHF virus, Rickettsia spp., Coxiella burnetii and Coxiella-like organisms, Babesia spp. and Theileria spp.ResultsThirty-five ticks with an unusual appearance or behaviour were reported to us during summer-autumn 2018. For 17 of them, the description or photos implied that they belong to the hard tick genus Hyalomma. The remaining 18 ticks were sent to us and were identified as adult Hyalomma marginatum (10 specimens) or adult Hyalomma rufipes (8 specimens). All ticks tested negative for CCHF virus, Coxiella burnetii, Coxiella-like organisms, Babesia spp. and Theileria spp. The screening for rickettsiae gave positive results in 9 specimens . The Rickettsia species in all cases was identified as R. aeschlimannii.ConclusionsThese results show that exotic tick species imported into Germany were able to develop from the nymphal to the adult stage under appropriate weather conditions. Fifty percent of the ticks carried R. aeschlimannii, a human pathogen, while CCHF virus or other pathogens were not detected. Imported Hyalomma ticks may be the source of exotic diseases acquired in Germany.

PREVALENCE OF TICK INFESTATION ON SMALL RUMINANTS IN AND AROUND DIRE DAWA, EASTERN ETHIOPIA

Across section, study of tick was conducted to identify types of tick species and determine the Prevalence of tick infestation in small ruminants in and around Dire Dawa Administrative council, Eastern Ethiopia. Collection and identification of the ticks were undertaken from November 2014 to May 2015. The overall prevalence of tick infestation in the study area was 278 (72.39% /384). All visible individual adult ticks were collected from the body of 196 goats and 188 sheep. The prevalence of tick infestation in goats and sheep was found to be 73.9% and 70.7%, respectively. In this study, eight species of ticks, which grouped under four genera, were identified. The most abundant species found in this study were Rhipicephalus pulchellus (34.1%), Amblyomma variegatum (24.5%), Rhipicephalus evertsievertsi (22.1%), Hyalomma truncatum (15.6%), Hyalomma marginatum rufipes (12.2%) and Amblyomma gemma (10.9 according to predominance. Hyalomma dromedari (4.2%) and Boophilus decoloratus was the minor species observed on both goats and sheep in the study area. The difference in prevalence of tick infestation was found statistically significant variation (P &lt; 0.05) between the sex, age exception of Boophilus decoloratus and Hyalomma dromedari in all cases male ticks dominated females.

STUDY ON THE STATUS OF BOVINE TICK INFESTATION, IN GUBA-KORICHA DISTRICT IN WEST HARARGHE ZONE, EAST - ETHIOPIA

A cross-sectional study was conducted from November 2010 to April 2011 to determining the prevalence of tick infestation, to identify the common tick species in cattle and to assess the major risk factors associated with the occurrence of tick species in Guba koricha district West Hararghe zone of Oromia regional state, southeast Ethiopia. Ticks were collected by searching and half body of animals using forceps on different regions of the animals’ body. A total sample of 384 cattle, 234 were found to be infested by one or more tick species an overall prevalence of 60.9%. The most predominant isolated ticks species in this study were R. pullchelus with isolation rate of 49.4% followed A. Varigatum with isolation rate of 30.3%, H. marginatum rufipes, the third predominant with isolation rate 11.8%. However, A. gemma, H. trucatum and R. Evertsi-evertsi was the least isolate which accounts for 3.6%, 2.4% and 2.35% respectively. Age, sex and body conditions scoring were found to be important risk factors associated with tick infestation. The prevalence of tick infestation between age and sex was statistically significant (X2=32.3075, CI=0.1323358-0265075, P=0.000 andX2=5.117, CI=0.1953184-0.0061713, P=0.037) respectively. However, breed and body condition were not statistically significant (P&gt;0.05). Hereof, Ticks are obligate, blood feeding ecto-parasites that cause severe damage to the hides and skins of domestic cattle due to this reduce the foreign exchange of the country; and transmit tick borne diseases. Therefore, effective tick control programs should be formulated and implemented at national or regional level.

STUDIES ON ROLE OF TICKS IN TRANSMISSION OF SOME BLOOD PARASITES IN CAMEL

This study was carried out in the imported camels that enter Egypt coming from Sudan those were passed on Abu-Simbuel region (Abu-Simbuel Abatoir) of Aswan Governorate, southern Egypt in the period from November, 2015 to October, 2016, to identify tick species infesting camels, study the prevalence of ticks infestation in camels all over the year andthe prevalence of Trypanosoma and microfilaria infection of camels, also the role of ticks in transmission of Trypanosoma species. The overall prevalence in the tick infestation was recorded as (60.96 %) (1463/2400) .The higher infestation were recorded in November, December (2015) and January (2016) and lower infestation were recorded in June.A total of (960) ticks were collected from the infested camels and identified. Hyalomma dromedarii was the predominant tick species and comprised (83.54 %) of the collected ticks. Other tick species found in very low numbers were Rhipicephalus praetextatus(4.27 %), Hyalomma anatolicum anatolicum(3.85%),Hyalomma impeltatum (2.92%), Rhipicephalus sanguineus (2.29 %),Hyalomma anatolicum excavatum(1.35 %), Hyalomma marginatum rufipes (1.12 %), and Hyalomma truncatum (0.63 %).(99) blood samples from (4800 )collected blood samples was infected by Trypanosoma evansi and hence constitute a prevalence rate of (2.0625%),with highest prevalence rate infection was (8.5 %) in October, alsofounded that (2) blood samples from( 4800) samples that infected by microfilaria in February and December (winter) with prevalence rate (0.042 %)(2/4800).We founded two samples in gut of male ticks contain developed stages of Trypanosoma in (March and June).

Tick repellents and acaricides of botanical origin: a green roadmap to control tick-borne diseases?

Arthropods are dangerous vectors of agents of deadly diseases, which may hit as epidemics or pandemics in the increasing world population of humans and animals. Among them, ticks transmit more pathogen species than any other group of blood-feeding arthropods worldwide. Thus, the effective and eco-friendly control of tick vectors in a constantly changing environment is a crucial challenge. A number of novel routes have been attempted to prevent and control tick-borne diseases, including the development of (i) vaccines against viruses vectored by ticks; (ii) pheromone-based control tools, with special reference to the "lure and kill" techniques; (iii) biological control programmes relying on ticks' natural enemies and pathogens; and (iv) the integrated pest management practices aimed at reducing tick interactions with livestock. However, the extensive employment of acaricides and tick repellents still remains the two most effective and ready-to-use strategies. Unfortunately, the first one is limited by the rapid development of resistance in ticks, as well as by serious environmental concerns. On the other hand, the exploitation of plants as sources of effective tick repellents is often promising. Here, we reviewed current knowledge concerning the effectiveness of plant extracts as acaricides or repellents against tick vectors of public health importance, with special reference to Ixodes ricinus, Ixodes persulcatus, Amblyomma cajennense, Haemaphysalis bispinosa, Haemaphysalis longicornis, Hyalomma anatolicum, Hyalomma marginatum rufipes, Rhipicephalus appendiculatus, Rhipicephalus (Boophilus) microplus, Rhipicephalus pulchellus, Rhipicephalus sanguineus and Rhipicephalus turanicus. Eighty-three plant species from 35 botanical families were selected. The most frequent botanical families exploited as sources of acaricides and repellents against ticks were Asteraceae (15% of the selected studies), Fabaceae (9%), Lamiaceae (10%), Meliaceae (5%), Solanaceae (6%) and Verbenaceae (5%). Regression equation analyses showed that the literature grew by approximately 20% per year (period: 2005-2015). Lastly, in the final section, insights for future research are discussed. We focused on some caveats for future data collection and analysis. Current critical points mainly deal with (a) not uniform methods used, which prevent proper comparison of the results; (b) inaccurate tested concentrations, frequently 100% concentration corresponded to the gross extract, where the exact amounts of extracted substances are unknown; and (c) not homogeneous size of tested tick instars and species. Overall, the knowledge summarized in this review may be helpful for comparative screening among extensive numbers of plant-borne preparations, in order to develop newer and safer tick control tools.

Multiple Pathogens Including Potential New Species in Tick Vectors in Côte d'Ivoire.

BackgroundOur study aimed to assess the presence of different pathogens in ticks collected in two regions in Côte d’Ivoire.Methodology/Principal FindingsReal-time PCR and standard PCR assays coupled to sequencing were used. Three hundred and seventy eight (378) ticks (170 Amblyomma variegatum, 161 Rhipicepalus microplus, 3 Rhipicephalus senegalensis, 27 Hyalomma truncatum, 16 Hyalomma marginatum rufipes, and 1 Hyalomma impressum) were identified and analyzed. We identified as pathogenic bacteria, Rickettsia africae in Am. variegatum (90%), Rh. microplus (10%) and Hyalomma spp. (9%), Rickettsia aeschlimannii in Hyalomma spp. (23%), Rickettsia massiliae in Rh. senegalensis (33%) as well as Coxiella burnetii in 0.2%, Borrelia sp. in 0.2%, Anaplasma centrale in 0.2%, Anaplasma marginale in 0.5%, and Ehrlichia ruminantium in 0.5% of all ticks. Potential new species of Borrelia, Anaplasma, and Wolbachia were detected. Candidatus Borrelia africana and Candidatus Borrelia ivorensis (detected in three ticks) are phylogenetically distant from both the relapsing fever group and Lyme disease group borreliae; both were detected in Am. variegatum. Four new genotypes of bacteria from the Anaplasmataceae family were identified, namely Candidatus Anaplasma ivorensis (detected in three ticks), Candidatus Ehrlichia urmitei (in nine ticks), Candidatus Ehrlichia rustica (in four ticks), and Candidatus Wolbachia ivorensis (in one tick).Conclusions/SignificanceFor the first time, we demonstrate the presence of different pathogens such as R. aeschlimannii, C. burnetii, Borrelia sp., A. centrale, A. marginale, and E. ruminantium in ticks in Côte d’Ivoire as well as potential new species of unknown pathogenicity.

Occurrence and Genotyping of Coxiella burnetii in Ixodid Ticks in Oromia, Ethiopia

This study was conducted from September 2011 to March 2014 to address the occurrence and genotypes of Coxiella burnetii using molecular methods in ticks collected from domestic animals in Ethiopia. Ticks were tested for C. burnetii by quantitative real-time polymerase chain reaction (qPCR) targeting two different genes followed by multispacer sequence typing (MST). An overall prevalence of 6.4% (54/842) of C. burnetii was recorded. C. burnetii was detected in 28.6% (14/49) of Amblyomma gemma, 25% (31/124) of Rhipicephalus pulchellus, 7.1% (1/14) of Hyalomma marginatum rufipes, 3.2% (2/62) of Am. variegatum, 3.1% (4/128) of Am. cohaerens, 1.6% (1/63) of Rh. praetextatus, and 0.6% (1/153) of Rhipicephalus (Boophilus) decoloratus. Significantly higher overall frequencies of C. burnetii DNA were observed in Am. gemma and Rh. pulchellus than in other tick species (Mantel-Haenszel [MH], P < 0.0001). The overall frequency of C. burnetii was significantly higher (MH, P < 0.0001) in ticks from southeastern districts (Arero, Moyale, and Yabelo) than that from other districts. This study demonstrated the presence of C. burnetii genotype MST 18 in ticks in southeastern districts and genotype MST 20 in ticks in central districts. This study highlights the importance of ticks in the epidemiology of C. burnetii in Ethiopia.

Identification and Mechanistic Analysis of a Novel Tick-Derived Inhibitor of Thrombin.

A group of peptides from the salivary gland of the tick Hyalomma marginatum rufipes, a vector of Crimean Congo hemorrhagic fever show weak similarity to the madanins, a group of thrombin-inhibitory peptides from a second tick species, Haemaphysalis longicornis. We have evaluated the anti-serine protease activity of one of these H. marginatum peptides that has been given the name hyalomin-1. Hyalomin-1 was found to be a selective inhibitor of thrombin, blocking coagulation of plasma and inhibiting S2238 hydrolysis in a competitive manner with an inhibition constant (Ki) of 12 nM at an ionic strength of 150 mM. It also blocks the thrombin-mediated activation of coagulation factor XI, thrombin-mediated platelet aggregation, and the activation of coagulation factor V by thrombin. Hyalomin-1 is cleaved at a canonical thrombin cleavage site but the cleaved products do not inhibit coagulation. However, the C-terminal cleavage product showed non-competitive inhibition of S2238 hydrolysis. A peptide combining the N-terminal parts of the molecule with the cleavage region did not interact strongly with thrombin, but a 24-residue fragment containing the cleavage region and the C-terminal fragment inhibited the enzyme in a competitive manner and also inhibited coagulation of plasma. These results suggest that the peptide acts by binding to the active site as well as exosite I or the autolysis loop of thrombin. Injection of 2.5 mg/kg of hyalomin-1 increased arterial occlusion time in a mouse model of thrombosis, suggesting this peptide could be a candidate for clinical use as an antithrombotic.

An additional records of Hyalomma marginatum rufipes Koch, 1844 (Acari: Ixodidae) in southwestern and southern Iran with a molecular evidence

Hyalomma marginatum rufipes (Acari: Ixodidae) along with three closely related subspecies is considered as marginatum group. The subspecies had proven as main vector of Crimean Congo hemorrhagic fever virus, the cause of human death in Asia, Africa and Europe. This tick is also a vector of parasitic protozoan Theileria annulata, agent of tropical theileriosis in cattle. Nonetheless, taxonomical status of this tick not recognized or confirmed in tick’s fauna of Iran, then we decided to show most taxonomic characteristics and confirm the presence of this subspecies in Iran by molecular methods. Tick specimens were collected from cattle in Manujan township, Kerman province, southern Iran. Specimens were identified morphologically using suitable taxonomical identification keys. The morphologically identified specimens were subjected to molecular studies. Morphological and COI gene analysis clearly confirmed the occurrence of H. m. rufipes in Iran, however, according to ITS2 fragment H. m. rufipes can be the same H. m. marginatum. Thus, it seems that based on most popular molecular markers, H. m. rufipes and its relative H. m. marginatum really should be assigned as a polymorphic species H. marginatum.

Identification of tick species and disseminate pathogen using hemolymph by MALDI-TOF MS

BackgroundMatrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) is increasingly emerging tool for identification of arthropods including tick vectors using whole or body part of specimens. The challenges of the present study were to assess MALDI-TOF MS profiling for the both identification of tick species and Rickettsia spp. in infected ticks using hemolymph as protein mixture. MethodsFirstly, hemolymph protein mixture from legs of 5 tick species, Rhipicephalus sanguineus, Rhipicephalus bursa, Dermacentor marginatus, Hyalomma marginatum rufipes and Amblyomma variegatum infected by Rickettsia africae were submitted to MALDI-TOF MS to assess tick species identification ability. Secondly, hemolymph MS spectra from Rh. sanguineus infected or not by Rickettsia c. conorii were compared to detect protein profiles changes. Finally, leg hemolymph MS spectra from new specimens of the 5 tick species were tested blindly including ticks infected by R. c. conorii. Discriminating mass peaks distinguishing the R. c. conorii infected and non-infected Rh sanguineus were determined. ResultsConsistent and reproducible MS profiles were obtained into each tick species. Comparison of MS spectra revealed distinct hemolymph protein profiles according to tick species. MS spectra changes were observed between hemolymphs from R. c. conorii-infected and non-infected Rh. sanguineus specimens, revealing 17 discriminating mass peaks. Clustering analysis based on MS protein profiles highlighted that hemolymph samples were grouped according to tick species. All tick hemolymph samples blindly tested against our home-made arthropod MS reference database were correctly identified at the species distinguishing also R. c. conorii-infected from Rickettsia-free Rh. sanguineus specimens. ConclusionThe present study demonstrated the use of hemolymph MS profiles for dual identification of tick species and associated pathogens. This concomitant identification could be helpful for tick entomological diagnosis, notably for specimens removed directly on patients.

Phenotypic and Genotypic Identification of Ticks Sampled from Wildlife Species in Selected Conservation Sites of Kenya

Hard ticks are blood feeding ectoparasites that infest humans and animals and are vectors of pathogenic microorganism that cause severe infectious diseases. Morphological identification has been the main approach of identifying ticks but the technique is considered inaccurate and difficult. Molecular techniques have recently been considered to be appropriate approaches for accurate and rapid identification and Internal Transcribed Spacer 2 (ITS 2) has been shown to differentiate genus of hard ticks. Currently, genetic identification of ticks using ITS 2 has not been carried out in Kenya. In this study, 80 tick samples were collected from Lake Nakuru and Tsavo National Parks and were identified morphologically using appropriate identification keys. DNA was extracted from the appendages using DNA extraction kit followed by partial amplification of ITS 2 gene. The PCR products were then analyzed by gel electrophoresis and positive PCR products were sequenced. Of the tick samples four genera were identified morphologically; Amblyomma, Hyalomma, Rhipicephalus and Dermacentor. Of the tick samples identified and compared with the sequences in the GenBank, six and seven samples showed 98-100% homology with A. variegatum and R. pulchellus respectively and they clustered in their respective monophyletic group in the phylogeny tree with a bootstrap of 99%. Two samples showed 92% homology with H. dromedarii and the study sequences clustered with the reference sequence with a bootstrap of 99% while six samples showed 95% homology with H. marginatum rufipes, however, only four of these samples clustered together with the reference sequence in the phylogeny with a bootstrap of 95%. One sample showed 91% homology with A. humerali and did not cluster together in the phylogeny tree. Congruency between both techniques was high with a correlation coefficient of 0.941. This is the first report of phenotypic and genotypic traits of tick species in Kenya and the findings will add value to the existing knowledge in identification of ticks.

Spotted fever group rickettsiae in ixodid ticks in Oromia, Ethiopia

In Ethiopia, information on the transmission of human zoonotic pathogens through ixodid ticks remains scarce. To address the occurrence and molecular identity of spotted fever group rickettsiae using molecular tools, a total of 767 ixodid ticks belonging to thirteen different species were collected from domestic animals from September 2011 to March 2014. Rickettsia africae DNA was detected in 30.2% (16/53) Amblyommma variegatum, 28.6% (12/42) Am. gemma, 0.8% (1/119) Am. cohaerens, 18.2% (4/22) Amblyomma larvae, 6.7% (2/60) Amblyomma nymphs, 0.7% (1/139) Rhipicephalus (Boophilus) decoloratus and 25% (1/4) nymphs of Rh. (Bo.) decoloratus. A markedly low prevalence of R. africae was recorded in both Am. cohaerens and Rh. (Bo.) decoloratus (p<0.0001) compared with that in Am. variegatum and Am. gemma. The prevalence of R. africae was markedly low in the western districts (Gachi and Abdela) (p<0.0001); however, the prevalence of R. africae was relatively high in the central (Ada’a, Wolmara and Arsi) and eastern (Arero, Moyale and Yabelo) districts, where Am. variegatum and Am. gemma were predominantly associated with R. africae, respectively. R. aeschlimannii DNA was detected in 45.4% (5/11) Hyalomma marginatum rufipes and 2.2% (1/46) Hy. truncatum. Moreover, the first report of R. massiliae DNA in 1.9% (1/52) Rhipicephalus praetextatus ticks in Ethiopia is presented herein. Altogether, these results suggest that the transmission of spotted fever group rickettsiae through ixodid ticks is a potential risk for human health in different parts of Ethiopia. Clinicians in this country should consider these pathogens as a potential cause of febrile illness in patients.

Ixodid ticks infesting cattle in three agroecological zones in central Oromia: species composition, seasonal variation, and control practices

A cross-sectional study was conducted to determine the species, prevalence, and seasonal variation of ticks on cattle in central Oromia. In addition, questionnaire survey was used to assess the control practices of ticks in central Oromia. During the study period from August 2008 through April 2009, a total of 10,440 adult ixodid ticks were collected from a total of 918 cattle (153 cattle during dry and wet seasons from each of three agroecologies) in three agroecological zones during wet and dry seasons. A total of seven species of ixodid ticks belonging to four genera were identified during the wet period, whereas similar numbers of genera but six species of ixodid ticks were recorded during the dry season. Amblyomma, Boophilus, Rhipicephalus, and Hyalomma tick genera were identified during the two seasons. Five different species of ticks were identified from the highland altitude. Significantly (p < 0.05) higher proportion of Boophilus decoloratus (88.2 %) was identified during the wet season, Hyalomma truncatum (1.5 %) was with the least proportion. Rhipicephalus evertsi evertsi (50.8 %) was the most abundant during the dry period, whereas Hyalomma marginatum rufipes (5 %) was with smallest proportion. In the midland, a total of six species of ticks were identified. Significantly (p < 0.05) higher proportion of Amblyomma variegatum (76.6 %) than the other species was recorded during the wet season in the midland. But Rhipicephalus pulchellus (0.05 %) was collected with the lowest proportion. R. e. evertsi (46.3 %) was the most abundant tick species during the dry season, whereas R. pulchellus (0 %) was with lowest proportion. In the lowland, a total of seven species of ticks during the wet and six during the dry season were identified. A significantly (p < 0.05) higher proportion of A. variegatum (30.3 %) was recorded during the wet season, whereas R. pulchellus (1.4 %) was identified with the lowest proportion in the lowland. R. e. evertsi (24.8 %) was the most abundant tick species during the dry season in the lowland, but R. pulchellus (0 %) was with the lowest proportion during the same season in the lowland. Results of the study revealed that the highest mean total tick burden was recorded during the early rainy season than the dry season from the end of March to April. And the lowest mean tick count was recorded during the early dry and the late dry season during December and February, respectively. The overall tick count across seasons showed significant variation (F = 114 and p = 0.000). The questionnaire survey on the tick control practices has revealed that (90 %) diazinon and (57.3 %) cypermethrin were the commonly used acaricides with a frequency of one to three times per year during the peak of tick infestation. In conclusion, this study demonstrated that ixodid ticks occur with high prevalence and burden and still play major roles in reducing productivity of cattle and cause health problems of cattle in central Oromia. Further studies on the role of ticks in the transmission of pathogens to cattle, other animals, and zoonotic pathogens to man in Oromia are urgently needed.