Revisiting detachment techniques in human-biting ticks.

BACKGROUND: Early and complete removal of ticks using the right technique is important to reduce tick-transmitted diseases. Several chemical and mechanical detachment techniques have been described previously. OBJECTIVE: We aimed to compare the performance of 4 tick-detachment techniques that are widely used in human beings and to determine the optimal method from these techniques. METHODS: A cross-sectional study was conducted on 160 patients between April and June 2010. Patients with reported tick bite were reviewed retrospectively and divided into the following 4 groups according to the tick-detachment technique used: card detachment, lassoing, freezing, and tweezers. Performance of each technique was evaluated according to the number of fully detached, nondetached, and crushed ticks and the duration of application. RESULTS: Of the 160 tick-bite cases assessed, we found the following efficacy rates: 82.5% (33/40), technique using tweezers; 47.5% (19/40), lassoing technique; 7.5% (3/40), card detachment; and 0% (0/40), freezing technique. The efficacy rate of the technique using tweezers was significantly higher than that of the other 3 techniques (P < .05). LIMITATIONS: This was a relatively small sample size and not designed as a randomized clinical trial. CONCLUSION: Tick detachment using tweezers, performed in an appropriate manner, is the easiest and most effective technique.

External morphological anomalies in ixodid ticks from Thrace, Turkey.

Of 18,667 ticks examined, 33 specimens from species identified as Haemaphysalis parva, Hyalomma marginatum, Hy. scupense, Rhipicephalus bursa and Rh. turanicus were found to have external morphological anomalies. Anomalous Ha. parva, Hy. scupence and Rh. turanicus were reported in this study for the first time. General anomalies manifested as asymmetry and deformations of the idiosoma, whereas local anomalies occurred in legs, exoskeleton, spiracular, adanal, subanal and accessory plates, mouthparts and capitulum. With this study describing a gynandromorphic Hy. marginatum, the number of gynandromorphic tick cases has been raised to two in Turkey.

[Evaluation of 10 cases of Lyme disease presenting with erythema migrans in Istanbul, Turkey]. / Istanbul'da eritema migrans ile basvuran 10 Lyme olgusunun degerlendirilmesi.

Lyme disease (LD) is a tick-borne, multisystemic infection caused by Borrelia burgdorferi. Although variable rates of seropositivity for B.burgdorferi have been reported between 2% to 44% in Turkey, its actual prevalence is not well-understood. The aim of this retrospective study was to evaluate the characteristics of 10 cases of LD presenting as erythema migrans (EM) between 2009 and 2013 from Istanbul which is one of the metropolitan cities of Turkey. Of the patients, five were male and five were female, ages between 9-51 years (mean age: 34.5 years). Five of the patients were admitted in June, three in October, one in November and two in December and all have the history of tick bite in last 1-2 weeks. There were no clinical symptoms for systemic infection among the patients with normal level routine laboratory test (whole blood count and biochemical tests) results. Five of the cases had EM lesions in the trunk, three in the upper extremities, and two in the lower extremities. Four patients presented with annular, three with solitary macular, and three with target-like EM lesions. In all cases, the biopsy specimens were positive for B.burgdorferi sensu lato DNA with polymerase chain reaction and all were also positive in terms of B.burgdorferi IgM antibodies with ELISA. Nine patients were treated with oral doxycycline, 100 mg twice daily and one child patient was treated with oral amoxicillin 500 mg twice daily for 21 days. EM lesions disappeared within 2-4 weeks in all patients. There was no clinical evidence for systemic involvement in any of the patients like neurologic, cardiac, and joint involvement at the follow-ups on the third, sixth and 12(th) months. To our best knowledge, 10 patients in this study are the largest EM series reported from Turkey. The increase in the number of LD cases may be associated with increased tick bite and increased awareness due to the emergence of concurrent Crimean-Congo hemorrhagic fever epidemic in Turkey. As a result, when enlarged erythematous lesions on the skin were observed, LH must also be considered in differential diagnosis, history of tick bite should be questioned and etiological diagnostic test should be performed.

The first clinical case due to AP92 like strain of Crimean-Congo Hemorrhagic Fever virus and a field survey.

BACKGROUND: Crimean-Congo Hemorrhagic Fever (CCHF) is a fatal infection, but no clinical case due to AP92 strain was reported. We described the first clinical case due to AP92 like CCHFV. METHODS: A case infected by a AP92 like CCHFV was detected in Balkanian part of Turkey. Diagnosis was confirmed by RT-PCR and sequencing. A human serologic and tick survey studies were performed in the region, where the case detected. RESULTS: Thirty eight individuals out of 741 were found to be anti CCHFV IgM positive. The attack rate for overall CCHFV was calculated as 5.2%. In univariate analyses, CCHFV IgM positivity was found to be associated with the age (p < 0.001), male gender (p = 0.001), agricultural activity (p = 0.036), and history of tick bite (p = 0.014). In multivariate analysis, older age (OR: 1.03, CI:1.01-1.05, p < 0.001), male gender were found to be the risk factors (OR: 2.5, CI:1.15-5.63, p = 0.020) for CCHFV infection. CONCLUSION: This is the first human case with AP92 like CCHFV infection. Furthermore, this is the first report of AP92 like strain in Turkey. In the region, elderly males carry the highest risk for CCHFV infection.

Interactions of Human Dermal Dendritic Cells and Langerhans Cells Treated with Hyalomma Tick Saliva with Crimean-Congo Hemorrhagic Fever Virus.

Crimean-Congo hemorrhagic fever virus is one the most important and wide spread tick-borne viruses. Very little is known about the transmission from the tick and the early aspects of pathogenesis. Here, we generate human cutaneous antigen presenting cells-dermal dendritic cells and Langerhans cells-from umbilical cord progenitor cells. In order to mimic the environment created during tick feeding, tick salivary gland extract was generated from semi-engorged Hyalomma marginatum ticks. Our findings indicate that human dermal dendritic cells and Langerhans cells are susceptible and permissive to Crimean-Congo hemorrhagic fever virus infection, however, to different degrees. Infection leads to cell activation and cytokine/chemokine secretion, although these responses vary between the different cell types. Hyalomma marginatum salivary gland extract had minimal effect on cell responses, with some synergy with viral infection with respect to cytokine secretion. However, salivary gland extract appeared to inhibit antigen presenting cells (APCs) migration. Based on the findings here we hypothesize that human dermal dendritic cells and Langerhans cells serve as early target cells. Rather affecting Crimean-Congo hemorrhagic fever virus replication, tick saliva likely immunomodulates and inhibits migration of these APCs from the feeding site.

Modeling the spatial distribution of crimean-congo hemorrhagic fever outbreaks in Turkey.

The more than 1100 human cases of Crimean-Congo Hemorrhagic Fever (CCHF) reported in a continuing series of outbreaks that started in 2003 in Turkey constitute the largest epidemic of the disease since CCHF was first recognized in 1944. The spatial distribution of CCHF case reporting rates in Turkey was studied to look for clusters of disease. We applied the spatial scan analysis to test the hypothesis of whether there were areas with a higher than expected number of CCHF cases. The analysis was conducted through windows of 10, 20, 40 and 80 km in diameter[SR1] to determine whether clustering of cases was dependent on the size of the scanning window. At the largest window size, consistent patterns of significantly higher than expected numbers of CCHF cases were found in a total of 40 administrative districts. A predictive model to map the habitat suitability for the vector tick was developed from satellite-based climate data and high-resolution features of the vegetation from Landsat images covering the whole country. It was found that areas of higher risk (higher CCHF reporting) were correlated (p<0.05) with zones of high climate suitability for the tick together with a high rate of fragmentation of agricultural land interspersed between forest and shrub-type vegetation.

Imported Crimean-Congo hemorrhagic fever cases in Istanbul.

We described a series of imported cases of Crimean-Congo Hemorrhagic Fever (CCHF) in Istanbul and investigated the genetic diversity of the virus. All the suspected cases of CCHF, who were applied to the health centers in Istanbul, were screened for CCHF virus (CCHFv) infection by using semi-nested Polymerase Chain Reaction (PCR) following RT-PCR. Simultaneous blood samples were also sent to the national reference laboratory in Ankara for serologic investigation. In 10 out of 91 patients, CCHFv was detected by PCR, and among 9 out of 10, anti-CCHFv IgM antibodies were also positive. Clinical features were characterized by fever, myalgia, and hemorrhage. The levels of liver enzymes, creatinine phosphokinase, and lactate dehydrogenase were elevated, and bleeding markers were prolonged. All the cases were treated with ribavirin. There was no fatal case. All the strains clustered within the same group as other Europe/Turkey isolates.

The role of ticks in the maintenance and transmission of Crimean-Congo hemorrhagic fever virus: A review of published field and laboratory studies.

This manuscript is part of a series of reviews that aim to cover published research on Crimean-Congo hemorrhagic fever (CCHF) and its etiological agent, CCHF virus (CCHFV). The virus is maintained and transmitted in a vertical and horizontal transmission cycle involving a variety of wild and domestic vertebrate species that act as amplification hosts, without showing signs of illness. These vertebrates have traditionally been considered reservoirs of CCHFV, but in fact they develop only a transient viremia, while the virus can persist in ticks for their entire lifespan, and can also be transmitted vertically to the next generation. As a result, ticks are now considered to be both the vector and the reservoir for the virus. CCHFV has been detected in a wide range of tick species, but only a few have been proven to be vectors and reservoirs, mainly because most published studies have been performed under a broad variety of conditions, precluding definitive characterization. This article reviews the published literature, summarizes current knowledge of the role of ticks in CCHFV maintenance and transmission and provides guidance for how to fill the knowledge gaps. Special focus is given to existing data on tick species in which vertical passage has been demonstrated under natural or experimental conditions. At the same time, we identify earlier reports that used unreliable methods and perceptions to ascribe a vector role to some species of ticks, and have contributed to confusion regarding viral transmission. We also examine epidemiological pathways of CCHFV circulation and discuss priority areas for future research.

Inhibition of inducible nitric oxide synthase in murine visceral larva migrans: effects on lung and liver damage.

The roles of nitric oxide production and oxidative process were studied in mice infected with Toxocara canis and treated with aminoguanidine which is a specific inhibitor of inducible nitric oxide synthase (iNOS). Relations of nitric oxide synthase inhibition and tissue pathology were assessed by biochemical, histological and immunohistochemical methods. In experiments, Balb/c albino mice were inoculated with T. canis eggs either with or without aminoguanidine treatment or alone, at 24th, 48th hours and on 7th days. LPx and SOD values in liver tissue and plasma were measured. Liver and lung tissues were evaluated for the pathological lesions. The expression of eNOS and iNOS in both tissues were studied with immunohistochemistry in the same intervals. We observed significant differences between T. canis infected and aminoguanidine treated animals. Larval toxocarosis led to oxidative stress elevation in plasma. Microscopic examination of the liver histological sections revealed pathological lesions in the hepatic parenchyma in infected mice. In the mice received T. canis eggs plus aminoguanidine, the sinusoidal areas were enlarged. Histological lesions were more severe at 48 hours after infection. Numbers of eNOS and iNOS expressing epithelial cells were increased in the T. canis infected mice. The activities of eNOS and iNOS were also observed in the body of the larvae which have migrated to lung and liver. As a result, we have demonstrated that in vivo production of eNO and iNO during T. canis infection cause direct host damages and it is strongly related to the oxidative stress. We propose that larval NO can also be effective in larval migration, but it needs further investigation on distribution of NO in larvae.

Transstadial Transmission and Long-term Association of Crimean-Congo Hemorrhagic Fever Virus in Ticks Shapes Genome Plasticity.

The trade-off hypothesis, the current paradigm of arbovirus evolution, proposes that cycling between vertebrate and invertebrate hosts presents significant constraints on genetic change of arboviruses. Studying these constraints in mosquito-borne viruses has led to a new understanding of epizootics. The trade-off hypothesis is assumed to be applicable to tick-borne viruses too, although studies are lacking. Tick-borne Crimean-Congo hemorrhagic fever virus (CCHFV), a member of the family Bunyaviridae, is a major cause of severe human disease worldwide and shows an extraordinary amount of genetic diversity compared to other arboviruses, which has been linked to increased virulence and emergence in new environments. Using a transmission model for CCHFV, utilizing the main vector tick species and mice plus next generation sequencing, we detected a substantial number of consensus-level mutations in CCHFV recovered from ticks after only a single transstadial transmission, whereas none were detected in CCHFV obtained from the mammalian host. Furthermore, greater viral intra-host diversity was detected in the tick compared to the vertebrate host. Long-term association of CCHFV with its tick host for 1 year demonstrated mutations in the viral genome become fixed over time. These findings suggest that the trade-off hypothesis may not be accurate for all arboviruses.

In vivo inhibition of inducible nitric oxide and evaluation of the brain tissue damage induced by Toxocara canis larvae in experimentally infected mice.

Nitric oxide (NO) is known to be produced by macrophages, endothelial cells and neurons and synthesized by an enzyme called nitric oxide synthase (NOS). Various effector mechanisms and infections can affect the NO production. Excessive amount of NO will lead to biochemical reactions, which cause toxic effects. In this study the role of NO has been evaluated in larval toxocarosis, which is a systemic parasite infection caused by T. canis larvae. Infection was established in the Balb/c mice with or without inducible NOS (iNOS) inhibition and the effects of infection and NOS inhibition were observed according to the results of SOD and LPx measurements in brain tissue and NADPH-diaphorase (NADP-d) histochemistry. Results of NADPH-d histochemistry indicate that iNOS inhibition has protective effect on the brains of infected mice and that larval T. canis infection could be related to oxidative stress, and NO production and iNOS inhibition can protect the tissue from damage in this infection.

Influence of laboratory animal hosts on the life cycle of Hyalomma marginatum and implications for an in vivo transmission model for Crimean-Congo hemorrhagic fever virus.

Crimean-Congo hemorrhagic fever virus (CCHFV) is one of the most geographically widespread arboviruses and causes a severe hemorrhagic syndrome in humans. The virus circulates in nature in a vertebrate-tick cycle and ticks of the genus Hyalomma are the main vectors and reservoirs. Although the tick vector plays a central role in the maintenance and transmission of CCHFV in nature, comparatively little is known of CCHFV-tick interactions. This is mostly due to the fact that establishing tick colonies is laborious, and working with CCHFV requires a biosafety level 4 laboratory (BSL4) in many countries. Nonetheless, an in vivo transmission model is essential to understand the epidemiology of the transmission cycle of CCHFV. In addition, important parameters such as vectorial capacity of tick species, levels of infection in the host necessary to infect the tick, and aspects of virus transmission by tick bite including the influence of tick saliva, cannot be investigated any other way. Here, we evaluate the influence of different laboratory animal species as hosts supporting the life cycle of Hyalomma marginatum, a two-host tick. Rabbits were considered the host of choice for the maintenance of the uninfected colonies due to high larval attachment rates, shorter larval-nymphal feeding times, higher nymphal molting rates, high egg hatching rates, and higher conversion efficiency index (CEI). Furthermore, we describe the successful establishment of an in vivo transmission model for CCHFV in a BSL4 biocontainment setting using interferon knockout mice. This will give us a new tool to study the transmission and interaction of CCHFV with its tick vector.

Rickettsia species in ticks removed from humans in Istanbul, Turkey.

A total of 167 ticks collected from humans in Istanbul (Turkey) in 2006 were screened for Rickettsia species, and nested PCRs targeting gltA and ompA rickettsial fragment genes were carried out. Rickettsia monacensis (51), R. aeschlimannii (8), R. conorii subsp. conorii (3), R. helvetica (2), R. raoultii (1), R. africae (1), R. felis (1), and other Rickettsia spp. (2), were detected. To our knowledge, these Rickettsia species (except R. conorii) had never been reported in ticks removed from humans in Turkey. The presence of R. africae also had not been previously described, either in Hyalomma ticks or in any European tick species. In addition, R. aeschlimannii and R. felis had not been found associated with Rhipicephalus bursa specimens. The presence of human pathogenic Rickettsia in ticks removed from humans provides information about the risk of tick-borne rickettsioses in Turkey.

Crimean-Congo hemorrhagic fever in European part of Turkey: genetic analysis of the virus strains from ticks and a seroepidemiological study in humans.

A survey of ticks from domestic ruminants, together with a serosurvey in humans was conducted in Thrace (northwestern Turkey) to evaluate the prevalence of Crimean-Congo hemorrhagic fever virus (CCHFV) in ticks and humans. More prevalent ticks were Hyalomma marginatum, Hyalomma aegyptium, Rhipicephalus bursa, and Rhipicephalus (Boophilus) annulatus, with low numbers of Dermacentor marginatus, Rhipicephalus sanguineus group, and Ixodes ricinus. No differences in the tick faunal composition were found among surveyed provinces. CCHFV was detected using specific primers for strains belonging to both Europe 1 and Europe 2 clades in a total of 15 pools of ticks collected in nine localities. The maximum likelihood estimate of infection rate was calculated as 0.72/100 ticks (95% CI = 0.42-1.16). Viral RNA was observed only in H. marginatum, R.(B.) annulatus, and R. bursa with overall maximum likelihood estimate infection rates being 0.93 (95% CI = 0.35-2.05), 0.74 (95% CI = 0.24-1.78), and 1.67 (95% CI = 0.69-3.46), respectively. The surveyed region is the only place where both viral strains are circulating together in nature in Turkey. Results from serosurvey on 193 samples from three localities in the region showed that immunoglobulin M and immunoglobulin G rates are compatible with an epidemiological situation in which the virus has been present for a long time and is not the result of a recent invasive event from the main epidemic center in Anatolia (north-central Turkey). Seropositivity rates cannot be compared against the tick faunal composition, because of the homogeneity in the results about tick surveys. The high rate of seropositivity, and the prevalence of CCHFV in both Europe 1 and 2 clades among the ticks, but few clinical cases suggest that the circulation of both viral strains may confer protection against the CCHFV infection.

A newly identified Crimean-Congo hemorrhagic fever virus strain in Turkey.

Crimean-Congo hemorrhagic fever (CCHF) is a fatal viral disease that occurs in approximately 30 countries. It has the most extensive geographic range among the tick-borne viruses that affect human health. Recently, a 6-year-old boy presented with complaints of fever, fatigue, and loss of appetite. He revealed a history of tick bite in rural Istanbul three days prior to presentation. A hyperemia was detected at the site of the tick bite. Laboratory tests showed that alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and creatine phosphokinase levels were elevated and that the prothrombin time and activated partial thromboplastin time were prolonged. Anti-CCHF virus IgM ELISA and a reverse transcriptase-PCR assay for CCHF RNA were both positive. Phylogenetic studies revealed that the virus was a new AP92-like CCHF strain, which was named KMAG-Hu-07-01 (accession number EU057975). This patient could provide important information on the transmission dynamics of CCHF infection.

An early warning system for Crimean-Congo haemorrhagic fever seasonality in Turkey based on remote sensing technology.

In the last few years, Crimean-Congo haemorrhagic fever (CCHF) has been reported as an emerging tickborne disease in Turkey. This paper deals with the preparation of an early warning system, aimed to predict the beginning of the CCHF season in Turkey based on a clear, simple and repeatable remotely-sensed signal. Decadal (mean of 10 days) values of the normalized difference vegetation index (NDVI) at 1 km resolution were used on a set of 952 confirmed, accurately geo-referenced, clinical cases between 2003 and 2006. A prerequisite is that the signal should be observable between 2 and 3 decadals before a given moment of the season to be of value as early warning. Decadals marking the 10th percentile or the 25th quartile in the frequency distribution of case reporting were selected as markers for the beginning of season of risk. Neither raw nor accumulated decadal NDVI signals were able to predict the onset of this season. However, when we defined the NDVI anomaly (NDVIa) as the positive difference between decadal NDVI values and the average for the previous year, this standardized measure gave a homogeneous overview of the changes in the NDVI signal producing a NDVIa slope for the decadals 10 to 13 that was always greater than 0. We conclude that observing this slope over time can be used as an early-warning system as it would predict the build-up of the number of cases 20 days in advance with an accuracy of 82% (10th percentile) or 98% (25th quartile).