Prevalence of ixodid ticks on goats and risk factors for tick-borne diseases in humans in the Kyasanur forest disease endemic area of Kerala, India.

In India, tick-borne diseases are prevalent in many states due to the presence of tick vectors. However, information on disease-causing ticks from domestic animals and the associated risk factors for residents living in the disease-endemic area is lacking. Therefore, we conducted this study to identify ixodid ticks in goats and human risk factors in three villages of the Wayanad district of Kerala. We examined 202 goats and collected 741 ticks, of which 69.8% were ticks belonging to the genus Haemaphysalis. The maximum number of ticks was collected from Thirunelli (81.3%), followed by Noolpuzha (76.27%) and Pulpally (45.6%). Overall, H. bispinosa Neumann, 1897 (54.6%), was the most common species, followed by H. turturis Nuttall and Warburton, 1915 (38.0%), H. spinigera Neumann, 1897 (5.4%), and H. intermedia Warburton and Nuttall, 1909 (1.8%). We included 428 participants (men and women) in this study. The average age of the respondents was 43 years. We found significant associations between accessing the forest for cattle grazing and other activities and tick-borne diseases (χ2 = 9.5, p = 0.002), between workers who were bitten by ticks and tick-borne diseases (χ2 = 3.8, p = 0.05), and between number of tick bites per day > 6 and tick-borne diseases (χ2 = 12.1, p = 0.001). The high frequency of Haemaphysalis spp. found in goats highlighted the risk of tick exposure and tick-borne diseases, such as Kyasanur forest disease in humans, and the need for the development and implementation of effective measures to control ticks.

Study of the clinical manifestations and risk factors in people affected during the first Kyasanur Forest Disease outbreak in Goa, India: A mixed method study.

BACKGROUND OBJECTIVES: Kyasanur Forest Disease (KFD) is a tick-borne, zoonotic viral hemorrhagic fever, previously known to be endemic to the state of Karnataka, India. The first outbreak of KFD in Goa state was reported in the Sattari taluka, in North Goa in 2015. This study aimed to investigate the outbreak and report the clinical manifestations and risk factors in people diagnosed with KFD. METHODS: A mixed methods approach was used, which included a case series report and 19 in-depth interviews (IDIs) conducted with people diagnosed with KFD. The recorded IDIs were transcribed and translated and themes were coded for the analysis. RESULTS: There were 73 suspected cases of which 30 were confirmed to have KFD using RT-PCR. There were four suspected deaths of which two were confirmed by RT-PCR. Most of the affected individuals were found to be dependent on the forest for their livelihood. Most of the people in the region were engaged in cashew plantations and had to travel to the forest to fetch firewood and cashew, hence were at a higher risk. They lived near the forest. The lack of hemorrhagic manifestation was noteworthy in the current outbreak. INTERPRETATION CONCLUSION: The 'One Health' approach should be implemented to control KFD. Tick bite prevention measures coupled with vaccination of high-risk groups and intensive health education should be carried out, especially before the transmission season. There is a need to have high clinical suspicion for KFD in the region bearing in mind the non-hemorrhagic manifestation in this outbreak.

Sofosbuvir and its tri-phosphate metabolite inhibit the RNA-dependent RNA polymerase activity of non-structural protein 5 from the Kyasanur forest disease virus.

Kyasanur forest disease is a neglected zoonotic disease caused by a single-stranded RNA-based flavivirus, the incidence of which was first recorded in 1957 in the Southern part of India. Kyasanur forest disease virus is transmitted to monkeys and humans through the infected tick bite of Haemophysalis spinigera. Kyasanur forest disease is a febrile illness, which in severe cases, results in neurological complications leading to mortality. The current treatment regimens are symptomatic and supportive, and no targeted therapies are available for this disease. In this study, we evaluated the ability of FDA-approved drugs sofosbuvir (and its active metabolite) and Dasabuvir to inhibit the RNA-dependent RNA polymerase activity of NS5 protein from the Kyasanur forest disease virus. NS5 protein containing the N-terminal methyl transferase domain and C-terminal RNA-dependent RNA polymerase domain was expressed in Escherichia coli, and RNA-dependent RNA polymerase activity was demonstrated with the purified protein. The RNA-dependent RNA polymerase assay conditions were optimized, followed by the determination of apparent Km,ATP to validate the enzyme preparation. Half maximal-inhibitory concentrations against RNA-dependent RNA polymerase activity were determined for Sofosbuvir and its active metabolite. Dasabuvir did not show detectable inhibition with the tested conditions. This is the first demonstration of the inhibition of RNA-dependent RNA polymerase activity of NS5 protein from the Kyasanur forest disease virus with small molecule inhibitors. These initial findings can potentially facilitate the discovery and development of targeted therapies for treating Kyasanur forest disease.

Species distribution and ecological suitability analysis for potential tick vectors of Kyasanur forest disease in the Western Ghats of Kerala.

Kyasanur forest disease (KFD) is a tick-borne zoonotic viral disease with an estimated case fatality rate of 3% to 5% in humans. The KFD virus is transmitted to both humans and animals by infected ticks, primarily the Haemaphysalis species. Our study was undertaken following reports of five confirmed cases and three deaths due to KFD in Malappuram district of Kerala in 2014 to determine the diversity of Ixodidae ticks on vegetation and their possible infection with the KFD virus. Overall, 3502 hard ticks belonging to three genera and eight species were collected from two forest divisions. Haemaphysalis was the predominant tick species (92.72%). Tick positivity for the KFD virus was 4 (5.33%) of 75 pools of ticks tested. Kyasanur forest disease viral RNA was detected from the genera Haemaphysalis and Amblyomma. The KFD virus was detected in 2 of 35 pools (5.71%) of Haemaphysalis spinigera, 1 of 30 pools (3.33%) of Haemaphysalis turturis, and 1 of 3 pools (3.33%) of Amblyomma integrum from the south forest division. The ticks reached their peak density between December and February and then decreased from the end of May. The temperature in the area ranges from 28°C to 30°C, which is suitable for tick survival.

Kyasanur forest disease and climatic attributes in India.

BACKGROUND & OBJECTIVES: In India, Kyasanur Forest Disease has been reported from the states of Karnataka, Kerala, Goa, and Maharashtra. The relationship between climatic factors and transmission of KFD remains untouched, therefore, the present study was undertaken. METHODS: Based on the occurrence of cases, Shivamogga district (Karnataka) and Wayanad district in Kerala and northern Goa (Goa state) were selected for the study. Data on the incidence of KFD and climate factors were collected from concerned authorities. To determine the relationship between dependent and independent variables, spearman's correlation was calculated for monthly as well as with lag months. RESULTS: KFD cases and temperature (°C) were found significantly correlated up to 1 months' lag period (p<0.05) while with precipitation relationship was found negatively significant for 0-3 months' lag. The range of suitable temperature for KFD in Shivamogga, Goa and Wayanad was found as 20-31°C, 25-29°C and 27-31°C respectively. The cumulative precipitation during transmission months (November-May) ranged from <150-500mm, while in non-transmission months (June-October) from >1100-2400mm. INTERPRETATION & CONCLUSION: The analysis of three sites revealed that with the increase in temperature, the intensity of KFD transmission decreases as corroborated by the seasonal fluctuations in Shivamogga, Goa and Wayanad. High precipitation from June to October rovides suitable ecology to tick vector and sets in transmission season from November to May when cumulative precipitation is <500 mm.

Effectiveness of One Health approach for control of Kyasanur Forest Disease in Wayanad, Kerala, India.

BACKGROUND & OBJECTIVES: Kyasanur Forest Disease (KFD) is a vector borne haemorrhagic fever that is endemic in the Wayanad region located in Northern part of Kerala, India. The region is managing the outbreak well ever since the major epidemic of 2015. This was because of the successful implementation of One Health (OH) initiative concentrating on multisectoral collaboration between regional institutions involved in public, animal and environmental health domains. The article presents how OH was implemented for the first time in the district in the year 2015 and evaluates the degree OH-ness of the Initiative. METHODS: The OH approach involved trans-disciplinary stakeholder meetings and reviews, outbreak management and integrated surveillance targeting ticks, monkeys and humans. The degree of OH-ness used for addressing KFD during the year 2015 was evaluated following the protocol developed by the Network for Evaluation of One Health (NEOH). In detail, we (i) described the OH initiative and its system (Aim, stakeholders, action strategy) and (ii) scored different aspects of this initiative (i.e., OH-thinking, -planning, -working, -sharing, -learning, -organization), with values from 0 (=no OH approach) to 1 (=perfect OH approach). RESULTS: We obtained a median score for each aspect evaluated. We reached high scores for OH systemic organization (1.0), OH thinking (0.83) and OH working (0.83). Lower scores were attributed to OH planning (0.58), OH sharing (0.50) and OH learning (0.33). The OH index was 0.36 and OH ratio was 0.95, indicating a balance between the OH operations and supporting infrastructures. INTERPRETATION & CONCLUSION: With this we could high-light some critical issues related to communication on sharing data as well as learning gaps for consideration to control future outbreaks. The strengths and weaknesses detected may be used to refine the initiative, aiming to provide a basis for the development of shared recommendations in a more OH-oriented perspective. This model of evaluation criteria will serve to create a database of OH success stories in India that will in turn help to institutionalize the approach at ministerial level. Future India is moving towards implementing a One Health, hence, this study data will provide an ideal opportunity for all sectors to control any vector borne diseases.

Identification of bio-climatic determinants and potential risk areas for Kyasanur forest disease in Southern India using MaxEnt modelling approach.

BACKGROUND: Kyasanur forest disease (KFD), known as monkey fever, was for the first time reported in 1957 from the Shivamogga district of Karnataka. But since 2011, it has been spreading to the neighbouring state of Kerala, Goa, Maharashtra, and Tamil Nadu. The disease is transmitted to humans, monkeys and by the infected bite of ticks Haemaphysalis spinigera. It is known that deforestation and ecological changes are the main reasons for KFD emergence, but the bio-climatic understanding and emerging pathways remain unknown. METHODS: The present study aims to understand the bio-climatic determinants of distribution of tick vector of KFD in southern India using the Maximum Entropy (MaxEnt) model. The analysis was done using 34 locations of Haemaphysalis spinigera occurrence and nineteen bio-climatic variables from WorldClim. Climatic variables contribution was assessed using the Jackknife test and mean AUC 0.859, indicating the model performs with very high accuracy. RESULTS: Most influential variables affecting the spatial distribution of Haemaphysalis spinigera were the average temperature of the warmest quarter (bio10, contributed 32.5%), average diurnal temperature range (bio2, contributed 21%), precipitation of wettest period (bio13, contributed 17.6%), and annual precipitation (bio12, contributed 11.1%). The highest probability of Haemaphysalis spinigera presence was found when the mean warmest quarter temperature ranged between 25.4 and 30 °C. The risk of availability of the tick increased noticeably when the mean diurnal temperature ranged between 8 and 10 °C. The tick also preferred habitat having an annual mean temperature (bio1) between 23 and 26.2 °C, mean temperature of the driest quarter (bio9) between 20 and 28 °C, and mean temperature of the wettest quarter (bio8) between 22.5 and 25 °C. CONCLUSIONS: The results have established the relationship between bioclimatic variables and KFD tick distribution and mapped the potential areas for KFD in adjacent areas wherein surveillance for the disease is warranted for early preparedness before the occurrence of outbreaks etc. The modelling approach helps link bio-climatic variables with the present and predicted distribution of Haemaphysalis spinigera tick.

The species distribution of ticks & the prevalence of Kyasanur forest disease virus in questing nymphal ticks from Western Ghats of Kerala, South India.

Background & objectives: Kyasanur forest disease (KFD) is a zoonotic tick-borne disease across the Western Ghats of India. With the discovery of a cluster of human KFD cases in the Wayanad district of Kerala, the present study was focused on detecting KFD virus (KFDV) in tick populations. To manage this disease, it is necessary to understand the diversity of the tick species and factors influencing the distribution, abundance and prevalence of infected ticks in Wayanad district. Methods: Surveys were conducted from November 2016 to May 2018 in four forest ranges of Wayanad district. Ticks were collected by the dragging method and were identified to species level and assayed for virus detection using real-time polymerase chain reaction. Results: A total of 25,169 ticks were collected from 64 sites. Of the identified species, Haemaphysalis spinigera was the most abundant (56.64%), followed by H. turturis 9047 (35.94%), H. bispinosa 999 (3.96%), Amblyomma integrum 691 (2.74%), H. kyasanurensis (0.55%), Rhipicephalus sanguineus (0.08%), Hyalomma marginatum (0.02%), H. cuspidata (0.01%), R.microplus (0.01%) and Dermacentor auratus (0.003%). The nymphal stage was predominant from December to February having peak activity in January. A total of 572 pools were screened for the presence of KFDV, of which 21 pools were positive. The infection rates in H. spinigera and H. turturis tick were 2.62 and 1.04 per cent, respectively. Interpretation & conclusions: The circulation of KFDV was detected and its correlation with the prevalence in ticks near the fragmented forest and teak plantation areas of Wayanad district. Residents and visitors of these regions may become vulnerable to tick bites and to an increased risk of KFD as the distribution of established, infected tick populations continues to expand.

Experiences of Indian Council of Medical Research with tick-borne zoonotic infections: Kyasanur Forest disease & Crimean-Congo haemorrhagic fever in India with One Health focus.

Emergence and re-emergence of several pathogens have been witnessed by this century in the form of outbreaks, epidemics and pandemics. In India, the influencing factor that promotes dissemination of emerging and re-emerging viral infections is the biogeographical zones: a megadiverse country, characterized by varied geographical, climatic conditions and ever-changing socio-economical and geopolitical issues. These influence the movement of humans and animals and add layers of complexity for the identification and timely management of infectious diseases. This review focuses on two tick-borne infections: Crimean-Congo haemorrhagic fever (CCHF) and Kyasanur forest disease (KFD). In the last two decades, these viruses have emerged and caused outbreaks in different parts of India. KFD virus was initially identified in 1957 and was known to be endemic in Karnataka State while CCHF virus was first identified during 2010 in Gujarat State, India. These viruses have managed to emerge in new areas within the last decade. With changing epidemiology of these arboviruses, there is a probability of the emergence of these viruses from new areas in future. The investigations on these two diseases under the One Health focus involved early detection, quickly developing diagnostic tools, identifying stakeholders, capacity building by developing collaboration with major stakeholders to understand the epidemiology and geographical spread in domestic animal reservoirs and tick vectors in the affected areas, developing laboratory network, providing diagnostic reagents and biosafety and laboratory diagnosis training to the network laboratories to control these diseases.

Update on prevalence and distribution pattern of tick-borne diseases among humans in India: a review.

In the present scenario, tick-borne diseases (TBDs) are well known for their negative impacts on humans as well as animal health in India. The reason lies in their increased incidences due to global warming, environmental and ecological changes, and availability of suitable habitats. On a global basis, they are now considered a serious threat to human as well as livestock health. The major tick-borne diseases in India include Kyasanur forest disease (KFD), Crimean-congo hemorrhagic fever (CCHF), Lyme disease (LD), Q fever (also known as coxiellosis), and Rickettsial infections. In recent years, other tick-borne diseases such as Babesiosis, Ganjam virus (GANV), and Bhanja virus (BHAV) infections have also been reported in India. The purpose of this paper is to review the history and the current state of knowledge of tick-borne diseases in the country. The conclusion of this review is extending the requirement of greater efforts in research and government management for the diagnosis and treatment and as well as prevention of these diseases so that tick-borne disease burden should be minimizing in India.

Spatial distribution of Haemaphysalis species ticks and human Kyasanur Forest Disease cases along the Western Ghats of India, 2017-2018.

Kyasanur Forest Disease (KFD) is a viral haemorrhagic fever, transmitted to humans and other hosts by a tick vector of genus Haemaphysalis. It affects 400-500 people annually in the Western Ghats region of India through spring to summer season. To understand the species composition, distribution, and abundance of Haemaphysalis ticks in endemic taluks (sub-districts) of India, a surveillance for ticks was conducted between October 2017 and January 2018. In total 105 sites were selected based on grid sampling from five taluks representing five KFD endemic states in south India. A sum of 8373 ticks were collected by using standard flagging method. The study showed a wide distribution of host seeking tick species among the selected taluks, wherein Haemaphysalis spinigera was predominant in 3/5 taluks, Haemaphysalis bispinosa in 1/5 taluks, and both the species in 1/5 taluks. Further, the H. spinigera abundance was categorised and compared with the incidence of human cases during the same season. The grids with very high and high H. spinigera abundance had 70% of the 205 human cases reported. This method of tick surveillance could be efficiently used as a standard model for KFD transmission risk assessment and prediction of impending outbreaks.

Clinical & epidemiological significance of Kyasanur forest disease.

Kyasanur forest disease (KFD) is a known viral haemorrhagic fever in India, for the last 60 years. However, in recent years, the change in epidemiological profile of the disease has suggested that it is now time to consider KFD as an emerging tropical disease in India. The preference should be to educate not only the villagers where it is being reported or detected but also to public health experts, veterinarians, forest officials and medical professionals to pay attention while seeing a patient overlapping with endemic diseases such as Japanese encephalitis, West Nile, dengue, chikungunya, malaria and tuberculosis. Although the existence of KFD is known for a long time, updated understanding of its clinical profile in humans is still limited. This article describes in detail the clinical presentation of KFD reported till date. It also highlights geographical distribution of the disease, risk factors for virus transmission, biochemical/haematological findings and control measures. There is an urgent need for research on KFD, particularly for understanding biphasic nature of illness, development of cost-effective diagnostic tools, utility of non-invasive samples for diagnosis and development of new vaccines.

Prevalence and spatial distribution of Ixodid tick populations in the forest fringes of Western Ghats reported with human cases of Kyasanur forest disease and monkey deaths in South India.

Kyasanur forest disease (KFD) is a major tick-borne viral haemorrhagic fever caused by KFD virus (KFDV) (Flaviviridae). The disease was reported to be confined to five districts of Karnataka state India until 2011. During 2012-2016, emergence of KFD has been reported in newer areas of Karnataka and adjoining states. Therefore, survey of tick vectors was carried out in these new areas of Karnataka and adjoining states reported with monkey deaths and human cases of KFD. In all selected sites, ticks from the forest floor were collected by lint clothes using flagging method. Tick samples were tested for KFDV nucleic acid by real-time RT-PCR. A total of 4772 ticks, comprising eight species of genus Haemaphysalis and one species each of genus Amblyomma, Ixodes and Rhipicephalus was collected. Haemaphysalis spinigera, the principal vector of KFDV was the predominant tick species (59.5%) collected followed by H. turturis (8.6%). The abundance of H. spinigera ranged from 9.2 to 33.9 per man-hour in the six districts surveyed. Of 214 (4418 tick samples) pools screened by real-time RT-PCR, two pools of H. spinigera were positive for KFDV. High abundance of Haemaphysalis vectors in the six districts indicated that the districts are receptive for KFD outbreaks. KFDV was detected in the tick vectors in the new foci of the KFD. Data on tick distribution will be useful in creating KFD risk map for strengthening the ongoing preventive measures such as vaccination and supply of insect repellents to the high risk groups and intensive health education.

Preliminary risk maps for transmission of kyasanur forest disease in Southern India.

Kyasanur forest disease is known to be transmitted across forested regions of Southern India. The disease appears to be hosted in wild mammals and transmitted by tick vectors although the diversity and identity of host and vector species remain unclear. The area across which risk exists of contracting the disease through transfer from the hosts or vectors, however, has never been mapped in detail, such that the area that surveillance, education, and investment in diagnostic facilities should cover remains unknown. This contribution uses known occurrences of the disease from the year 2000 till date to create and test a correlational ecological niche model that translates into preliminary transmission risk maps, which are summarized in terms of risk presented in each district in the region, as well as across peninsular India.

Kyasanur Forest disease outbreak and vaccination strategy,Shimoga District, India, 2013-2014.

We investigated a Kyasanur Forest disease outbreak in Karnataka, India during December 2013-April 2014. Surveillance and retrospective study indicated low vaccine coverage, low vaccine effectiveness, and spread of disease to areas beyond those selected for vaccination and to age groups not targeted for vaccination. To control disease, vaccination strategies need to be reviewed.

Clinical, epidemiological, and molecular investigation of Kyasanur forest disease from Karnataka state, India during 2018-2019.

BACKGROUND: In this study, we carried out an investigation of Kyasanur Forest Disease (KFD) suspected human cases reported in Karnataka state, India from December 2018 to June 2019. METHODS: The clinical samples of KFD suspected cases (n = 1955) from 14 districts of Karnataka were tested for KFD using real-time RT-PCR and IgM ELISA. Further, the KFD-negative samples were tested for IgM antibodies against dengue and chikungunya viruses. Monkey samples (n = 276) and tick pools (n = 11582) were also screened using real-time RT-PCR. KFD-positive samples were further analysed using next-generation sequencing along with clinico-epidemiological analysis. RESULTS: Of all, 173 (8.8%) cases tested positive for KFD either by real-time RT-PCR (n = 124), IgM ELISA (n = 53) or both tests (n = 4) from seven districts. Among KFD-negative cases, IgM antibody positivity was observed for dengue (2.6%), chikungunya (5.8%), dengue and chikungunya coinfection (3.7%). KFD cases peaked in January 2019 with fever, conjunctivitis, and myalgia as the predominant symptoms and a mortality of 4.6%. Among confirmed cases, 41% received a single dose and 20% received two doses of the KFD vaccine. Of the seven districts with KFDV positivity, Shivamogga and Hassan districts reported KFD viral RNA positivity in humans, monkeys, and ticks. Sequencing analysis of 2019 cases demonstrated a difference of less than 1.5% amino acid compared to prototype KFDV. CONCLUSION: Although the KFD has been endemic in many districts of Karnataka state, our study confirms the presence of KFDV for the first time in two new districts, i.e. Hassan and Mysore. A comparative analysis of KFDV infection among the KFD-vaccinated and non-vaccinated populations demonstrated an insignificant difference.

Knowledge, Attitudes, and Practices Regarding Ticks, Tick-Borne Diseases, and Ethnomedicine Among an at-Risk Population in Kerala.

Tick and tick-borne diseases (TBDs) are increasing annually, and the study of ticks has gained importance after the outbreak of Kyasanur Forest disease (KFD) in the South Western Ghats of India. Wayanad district of Kerala, with the highest tribal population in South India, is a KFD endemic state, owing to the lack of knowledge, attitude, and practice studies on TBDs and ethnomedicines against ticks. This study was carried out to assess their baseline knowledge, attitude, and ethnomedicinal practice against ticks. A structured questionnaire was used to conduct a survey of 499 tribal members living in forest fringe areas. Logistic regression analysis was performed to identify the factors that influence the knowledge, attitude, and practice of tribes on TBDs. More than 70% of the population visit the forests on a regular basis, with 65.7% of the population regularly exposed to tick bites; however, only 47.7% were aware of TBDs. About 47.4% of the respondents took precautions like therapeutics and natural remedies to avoid tick bites. Ten species of medicinal plant belonging to eight different families have been identified from the survey. The tribal population use these plants to repel ticks as well as treat tick bites. From the study, we concluded that the limited in-depth knowledge displayed by the tribes can be strengthened by conducting community programs such as awareness classes on TBD and its control measures. The ethnobotanicals identified can be used to formulate novel tick repellents in the future.

Kyasanur Forest disease, India, 2011-2012.

To determine the cause of the recent upsurge in Kyasanur Forest disease, we investigated the outbreak that occurred during December 2011-March 2012 in India. Male patients >14 years of age were most commonly affected. Although vaccination is the key strategy for preventing disease, vaccine for boosters was unavailable during 2011, which might be a reason for the increased cases.