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.

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.

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.

Predicting Kyasanur forest disease in resource-limited settings using event-based surveillance and transfer learning.

In recent years, the reports of Kyasanur forest disease (KFD) breaking endemic barriers by spreading to new regions and crossing state boundaries is alarming. Effective disease surveillance and reporting systems are lacking for this emerging zoonosis, hence hindering control and prevention efforts. We compared time-series models using weather data with and without Event-Based Surveillance (EBS) information, i.e., news media reports and internet search trends, to predict monthly KFD cases in humans. We fitted Extreme Gradient Boosting (XGB) and Long Short Term Memory models at the national and regional levels. We utilized the rich epidemiological data from endemic regions by applying Transfer Learning (TL) techniques to predict KFD cases in new outbreak regions where disease surveillance information was scarce. Overall, the inclusion of EBS data, in addition to the weather data, substantially increased the prediction performance across all models. The XGB method produced the best predictions at the national and regional levels. The TL techniques outperformed baseline models in predicting KFD in new outbreak regions. Novel sources of data and advanced machine-learning approaches, e.g., EBS and TL, show great potential towards increasing disease prediction capabilities in data-scarce scenarios and/or resource-limited settings, for better-informed decisions in the face of emerging zoonotic threats.

Simplified visual detection of Kyasanur Forest Disease virus employing Reverse Transcriptase-Polymerase Spiral Reaction (RT-PSR).

Among recently prevalent tick-borne infections in India, Kyasanur Forest Virus Disease (KFD) is an important public health concern. During last decade the emergence of cases apart from endemic zone raised concern about case positivity. Early diagnosis is therefore very important in disease management and primary containment. This study, aimed to develop a simplified viral RNA extraction in combination to dry down format of novel isothermal assay for (Reverse Transcription- Polymerase Spiral reaction) specific and rapid identification of Kyasanur Forest Disease Virus targeting viral envelope gene. The one step method was optimized by magnetic bead based viral RNA extraction followed by isothermal RT-PSR assay in heat bath at 63°C for 60 minutes. Further, visual results interpretation was done by color change of Hydroxy Naphthol Blue dye. The detection limit of the assay was found 10 RNA copies/rxn with comparable to silica column based viral RNA combined real time qPCR. No cross reactivity was observed with other closely related flaviviruses. The assay was evaluated with clinical samples has shown >99% concordance between two methods. This is the first report of sample extraction coupled isothermal detection of KFD in a simplified manner without a need of any hi-end equipment. The assay developed here has potential to use as an alternate for field-based detection in resource limited settings for KFD.

Using mechanistic models to highlight research priorities for tick-borne zoonotic diseases: Improving our understanding of the ecology and maintenance of Kyasanur Forest Disease in India.

The risk of spillover of zoonotic diseases to humans is changing in response to multiple environmental and societal drivers, particularly in tropical regions where the burden of neglected zoonotic diseases is highest and land use change and forest conversion is occurring most rapidly. Neglected zoonotic diseases can have significant impacts on poor and marginalised populations in low-resource settings but ultimately receive less attention and funding for research and interventions. As such, effective control measures and interventions are often hindered by a limited ecological evidence base, which results in a limited understanding of epidemiologically relevant hosts or vectors and the processes that contribute to the maintenance of pathogens and spillover to humans. Here, we develop a generalisable next generation matrix modelling framework to better understand the transmission processes and hosts that have the greatest contribution to the maintenance of tick-borne diseases with the aim of improving the ecological evidence base and framing future research priorities for tick-borne diseases. Using this model we explore the relative contribution of different host groups and transmission routes to the maintenance of a neglected zoonotic tick-borne disease, Kyasanur Forest Disease Virus (KFD), in multiple habitat types. The results highlight the potential importance of transovarial transmission and small mammals and birds in maintaining this disease. This contradicts previous hypotheses that primates play an important role influencing the distribution of infected ticks. There is also a suggestion that risk could vary across different habitat types but currently more research is needed to evaluate this relationship. In light of these results, we outline the key knowledge gaps for this system and future research priorities that could inform effective interventions and control measures.

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.

Development and application of a recombinant Envelope Domain III protein based indirect human IgM ELISA for Kyasanur forest disease virus.

Kyasanur forest virus disease (KFD) is a major public health concern in India. Its etiology KFD virus causes haemorrhagic fever with severe sequelae in humans. Due to continuous spatiotemporal expansion of KFD in last decade, the incidences of positive cases have been increasing in both humans and primates. Early diagnosis is of prime importance for disease management and epidemiological containment. In the present study, the highly immunogenic Envelope Domain III (EDIII) antigen was produced using prokaryotic expression system with an yield of 8 mg/L. The protein was purified using affinity chromatography and confirmed for its immuno-reactivity by western blot and UPLCMS/MS analysis. The recombinant EDIII was used as an antigen for the standardization of ELISA to detect anti KFD IgM antibodies in humans. The ROC curve was prepared to set the optimum cut-off OD for the assay. The comparative evaluation of the assay with a reference MAC ELISA revealed 86.96% concordance, 82.22% sensitivity and 91.48% specificity. Inter-rater agreement was performed with kappa index revealing significant agreement between the assays. This is the first study using safe recombinant protein antigen-based detection of anti KFDV antibodies in humans. This simple and scalable ELISA assay will be applicable for large scale screening of samples for combating the emerging threats of KFD in newer territories.

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.

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.

Impact of Plantation Induced Forest Degradation on the Outbreak of Emerging Infectious Diseases-Wayanad District, Kerala, India.

The world has been facing a pandemic owing to COVID-19. We have also seen the geographic expansion and outbreaks of other emerging infectious diseases (EID) in recent years. This paper investigates the direct and indirect effects of land use land cover change (LULCC) on EID outbreaks in the context of Wayanad District of Kerala, India. Wayanad is in the vulnerable tropical forested region, and it is named as one of the four environmental change hotspots. The focus of this project is mainly three EIDs prevalent in this region: Kyasanur forest disease (KFD), Dengue and Leptospirosis. Our results, based on topographical map, remote sensing and extensive field work, show that the natural forest in Wayanad was replaced with agriculture and forest plantation during 1950-2018. This paper further suggests that encroachment of forest by forest plantation causes the human-animal conflict resulting in the outbreak of KFD cases. Our analysis reveals that a high number of Dengue cases is found in the forested regions of the district and over the adjacent human-made agriculture plantation areas. High and medium number of Leptospirosis cases contain a high portion of land area devoted to paddy cultivation and agricultural plantation. In summary, the results clearly show the linkage between the outbreak of above mentioned EIDs and LULCC in the context of Wayanad district, Kerala. We also discuss in detail the causal pathway involving human-environmental dynamics through which plantation leads to the outbreak of KFD. Replacing forests with plantations poses an alarming threat of disease outbreak in the community.

Kinetics of human infection with Kyasanur Forest Disease Virus.

BACKGROUND: Kyasanur Forest disease (KFD) was first reported in 1957 and became an emerging tick-borne viral disease of public health importance in India. However, very little is known about the host-virus interaction and pathogenesis of KFD in humans. This study described the presence, duration, and kinetics of KFDV RNA in body fluids in infected human cases. METHODOLOGY: We enrolled 76 laboratory-confirmed KFD individuals and followed them up in the study. We obtained serial samples of blood, throat swabs in viral transport medium (VTM), urine, stool, and semen during the acute and convalescent phase of KFD illness. In addition, specimens were inactivated, and nucleic acid was extracted and tested for KFDV real-time reverse transcriptase -PCR. Clinical data was also obtained from the subjects. RESULT: The study provides evidence of KFD virus RNA in different biological body fluids of humans. The percentage positivity of KFDV RNA in blood was 100% during the first four days of illness. PCR became negative in most cases by 7-8 days; a subset of cases (14%) had prolonged viremia for up to 15 days post-onset of illness. Relatively low copies of KFDV RNA were also detected in throat swabs and urine in the first week of illness. In addition, we detected KFDV RNA in stool samples of cases of those who had diarrhea at an early stage of infection. CONCLUSION: The study provides evidence of KFDV RNA in different biological body fluids, which will help understand the pathogenesis, transmission pattern and develop diagnostic algorithms of KFDV in humans. In Kyasanur Forest disease infection, the blood has more RNA copies/ml than other body fluids, and viremia may last up to two weeks post-infection.

Neurological manifestations of Kyasanur Forest disease: a retrospective cohort study from South India.

BACKGROUND: Kyasanur Forest disease (KFD) is a viral zoonotic disease where patients present with febrile illness and haemorrhagic manifestations in the first phase. In a small fraction of patients, the fever may be biphasic. This study aimed to describe the neurological manifestations of patients with KFD in the first and second phases of the illness. METHODS: This is a retrospective cohort study of 297 patients admitted with a molecular diagnosis of KFD from December 2018 to December 2020. The case records of these patients were reviewed for evidence of neurological involvement. RESULTS: A total of 34 (11.5%) patients in the first phase and 16 (36.4%) patients in the second phase had neurological involvement. Altered sensorium, seizures and focal infarcts were common in the first phase, while cerebellar signs and leptomeningeal enhancement were common in the second phase. CONCLUSIONS: Neurological involvement is seen in both phases of KFD. While in the first phase it is a result of possible encephalitis/encephalopathy, the second phase involvement is possibly due to postinfectious cerebellitis or meningitis.

Kyasanur forest disease: a state-of-the-art review.

Kyasanur forest disease (KFD) virus is a flavivirus that can be transmitted to humans from monkeys or other mammals through hard ticks (Haemaphysalis spinigera). The disease is endemic to 16 districts in 5 states of Southern India and is reported in the dry season, most commonly in humans travelling to the forests in these areas. The aim of this systematic review is to raise awareness of the clinical and laboratory manifestation of KFD among physicians and travel medicine practitioners. A total of 153 articles were screened of which 16 articles that met the inclusion and exclusion criteria were included for qualitative analysis. KFD is an acute haemorrhagic fever with a biphasic component in some individuals. The second phase is usually marked by neurological symptoms. Leucopoenia, thrombocytopenia and elevated transaminases are the hallmarks of the first phase of KFD. The diagnostic modality of choice in the first few days of illness is polymerase chain reaction assay, whereas serology is used in the late phase. In the absence of a specific antiviral treatment, the clinical management of patients is limited to supportive care. Avoidance of exposure and vaccination is recommended to prevent this infection.

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.

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.

Kyasanur Forest Disease, is our surveillance system healthy to prevent a larger outbreak? A mixed-method study, Shivamogga, Karnataka, India: 2019.

BACKGROUND: Kyasanur Forest disease (KFD) is a tick-borne zoonosis that is endemic in Karnataka. Against the backdrop of the recent geographical expansion of KFD, indicating the inadequacy of policy and surveillance systems, the present study was performed to evaluate the KFD surveillance system in Shivamogga. METHODS: US Centers for Disease Control and Prevention guidelines for surveillance system evaluation were followed. Nine attributes of the system towards its objectives were evaluated in a mixed study in Shivamogga. RESULTS: Two of three medical officers and four of six health staff at the institutions visited were found to be untrained in KFD surveillance. Integrated disease surveillance formats did not capture KFD data. Surveillance (tick, monkey, human) was mostly driven by the Health Department. Some of the critical findings of the evaluations were the absence of an animal and entomological surveillance line list, non-standardized reporting formats for human suspects, varying delays in the time-to-test across laboratories (2-16 days), and a lack of systematic data-sharing practices. Significant issues that emerged in the interview were deforestation with a change in ecosystem dynamics, limited diagnostic capacity, non-availability of point-of-care tests, outdated surveillance guidelines, a confusing surveillance perimeter (5 km), non-existing co-ownership among stakeholders, limited vaccine production capacity, and inadequate operational research. CONCLUSIONS: The system should consider integrating a One Health approach with defined ownership of activities among stakeholders. Revision of the guidelines is mandatory.

Point of care real-time polymerase chain reaction-based diagnostic for Kyasanur forest disease.

OBJECTIVES: Due to the remote forest area locations of sporadic cases and outbreaks of Kyasanur forest disease (KFD), rapid diagnosis poses a significant challenge. This study aimed to evaluate the diagnostic performance of Truenat KFD, a simple, rapid and user-friendly point-of-care test for detection of KFD and compare diagnostic accuracy with conventional real-time reverse transcription-polymerase chain reaction (RT-PCR) testing. Truenat KFD can be deployed in a field laboratory setting. METHODS: The study involved 145 clinical specimens, including human serum, monkey necropsy tissues and tick pool, to validate Truenat KFD (Molbio Diagnostics Pvt.Ltd.) for KFD diagnosis. RESULTS: We have optimized and validated the microchip-based Truenat KFD (Molbio Diagnostics Pvt.Ltd.) for KFD diagnosis. Point-of-care testing was highly sensitive and specific, with a detection limit of up to 10 copies of KFD viral RNA. Results were comparable with the gold-standard TaqMan and commercially available Altona RealStar AHFV / KFDV real-time RT-PCR assays. Screening results for human, monkey and tick specimens were 100% concordant across the assays. CONCLUSION: Truenat KFD(Molbio Diagnostics Pvt.Ltd.) was found to be highly sensitive and specific with a significant limit of detection. This point-of-care test would be useful in rapid diagnosis of KFD in remote and/or field settings, quick patient management and control of virus spread.