Molecular detection of Orientia tsutsugamushi strains circulating in Nagaland.

The antigenic heterogeneity of Orientia in India is still unknown in many disease endemic areas. The present study aims to characterize the strains of O. tsutsugamushi circulating in Nagaland, Northeast India. Two patients clinically diagnosed with ST and hospitalized in Mon district hospital, Nagaland were identified from whom eschar tissues were collected. Both patients demonstrated antibodies against O. tsutsugamushi along with positive PCR amplification for 56 â€‹kDa gene. The prototype strain TA763 shared 90.4% homology with the sequences. Both the sequences formed a distinctive cluster demonstrating 100% similarity with strains identified from Thailand, Vietnam, China and southern parts of India.

Rickettsiae in fleas infesting domestic pets of eastern Himalayan terrains of India.

BACKGROUND: Flea-borne rickettsioses have been limitedly explored in the Indian sub-Himalayan belt, including the North Eastern Region (NER) of India. This study investigates the presence of rickettsiae hosts and their probable pathogens in the disease-endemic hilly state of the NER of India. METHODS: Entomological surveys were carried out in disease-reporting localities in a hilly state in India. Fleas collected from domesticated animals were processed for detection of a Rickettsia-specific 17-kDa gene. RESULTS: Sequence analysis revealed Rickettsia felis in six flea pools (40%), Candidatus Rickettsia senegalensis in two pools (13.3%) and Rickettsia asembonensis in one pool (6.6%). CONCLUSIONS: Our findings suggest Ctenocephalides felis, Ctenocephalides canis and Pulex irritans as potential carriers of R. felis and R. felis-like organisms in India.

Seroprevalence of Rickettsial Infections in Northeast India: A Population-Based Cross-Sectional Survey.

A cross-sectional survey was undertaken to estimate seroprevalence of immunoglobulin G antibodies against scrub typhus, spotted fever group rickettsiae, and typhus group rickettsiae in randomly selected 48 clusters in 12 districts of 3 Northeast states of India: Assam, Meghalaya, and Tripura. Individuals in 3 age groups (5-8, 9-17, and 18-45 years) were selected from each cluster. Sera (N = 2360) tested were collected as part of a national survey on dengue seroprevalence conducted between September 2017 and February 2018. Overall, seroprevalence of 2.5% was detected against rickettsioses, with highest positivity against spotted fever group rickettsiae, followed by scrub typhus and typhus group rickettsiae. Seroprevalence was highest in Tripura (3.7%), followed by Assam (2.6%) and Meghalaya (1.04%). Adults of 18 to 45 years of age were found to be most affected (3.8%). The study findings indicate the need for increasing testing facilities for active case detection at hospital levels. Efforts on implementing effective preventing strategies are suggested to be targeted in disease-specific endemic foci.

Spotted Fever Group Rickettsiae among Children with Acute Febrile Illness, in Gorakhpur, India.

Seasonal outbreaks of acute encephalitis syndrome have been occurring in Gorakhpur division in the Indian state of Uttar Pradesh during monsoon and post-monsoon months. Orientia tsutsugamushi was identified as the major aetiology of these outbreaks. Orientia tsutsugamushi was also identified as one of the important aetiology of febrile illness among children attending peripheral health facilities. The present study was undertaken to detect antibodies against spotted fever group rickettsiae (SFGR) and typhus group rickettsiae (TGR) among children with acute febrile illness presenting at peripheral health facilities in Gorakhpur district. Of the 224 blood samples tested, SFGR infection was detected in 13 (6%) patients. None of the samples tested positive for TGR.

Evaluation of Th1 and Th2 immune response in clinical and sub-clinical scrub typhus infection.

Scrub typhus (ST), caused by a gram negative intracellular bacteria- Orientia tsutsugamushi, is one among the leading causes of febrile illness across Southeast Asia, including India. Clinical presentation can vary from asymptomatic to severely fatal. Th1-cell mediated immunity has been suggested to play an important role against ST infection in animal models. However, human data on protective immunity are limited. The present study was undertaken to identify host immune correlates that could confer protection in individuals that remain clinically asymptomatic/sub-clinical. Serum cytokine profiling and mRNA expression levels of Th1 (TNF-α, IFN-γ, IL-2) and Th2 (IL-10, IL-6, IL-4) cytokines was studied amongst the clinical and sub-clinical infections. It was observed that a Th1/Th2 pattern is not involved in human ST infection irrespective of being a symptomatic or asymptomatic presentation. However, significant difference was observed in IL-10 serum and gene expression levels. This study suggests a possible role of IL-10 in disease phenotypic presentation. Over-production of IL-10 was found to be a significant factor contributing to the severity of the disease whereas a protective immune mechanism might exist with a low level of IL-10 in ST infection.

Genetic diversity of Orientia tsutsugamushi strains circulating in Northeast India.

Background: Scrub typhus, caused by the intracellular bacteria Orientia tsutsugamushi is widely distributed in Southeast Asian countries with antigenically divergent strains reported across the Asia-Pacific belt. The present study was conducted to characterize the circulating strains of Orientia tsutsugamushi prevailing in two Northeastern states of India-the gateway to most Southeast Asian countries. Methods: A total of 278 (98 clinical and 180 field collected) scrub typhus positive blood samples collected from December 2014 to December 2016 were subjected for amplification of partial 56 KDa, 47 KDa and 16SrRNA genes of Orientia. Results: Highest number of PCR positives were obtained for 56 KDa gene (17.3%); followed by 11.2% for 47 KDa gene and 5.1% for 16S rRNA gene. High degree of genetic diversity was identified among the identified strains, especially within the 56 KDa gene. Different strains of Orientia circulate in the northeastern part of India, with a pre-dominance of Karp-like strains. Independently branched isolates formed distinct clades, suggesting the possibility of a new strain type of Orientia. Conclusion: This study provides insight into the genetic and evolutionary relationship of Orientia strains prevalent in this part of the country. Understanding the regional genetic diversity is crucial for its implications in vaccine developments strategies as well as sero-diagnostics. Accession numbers: KU163366, KY594249, KY594248, KY594251, KU163359, KU163361, KU163363, KU163369, KY594250, KP067915, KU163373, KU163372, KU163370, KU163364, KU163362, KY594247, KY594252, KU163360, KU163365, KU163367, KU163368, KU163371, KX1555826, KY594257, KY594255, KY594256, KX155825, KX155829, KX155827, KX155828, KY594254, KY594253, KY594258, KY583503, KY583499, KY583501, KY583500, KY583502.

Scrub Typhus Leading to Acute Encephalitis Syndrome, Assam, India.

To determine the contribution of Orientia tsutsugamushi, the agent of scrub typhus, as a cause of acute encephalitis syndrome (AES) in Assam, India, we conducted a retrospective study of hospital patients with symptoms of AES during 2013-2015. Our findings suggest that O. tsutsugamushi infection leads to AES and the resulting illness and death.

Seroepidemiology of rickettsial infections in Northeast India.

BACKGROUND: Resurgence of scrub typhus was reported in Northeast India in 2010 after a gap of 67 years since World War II. However, the presence of other rickettsial infections remained unknown from this region. A seroepidemiological investigation was undertaken in the scrub typhus affected areas from 2013-2015 in Assam, Arunachal Pradesh and Nagaland to assess the exposure to other rickettsial diseases besides scrub typhus. METHODS: Samples were collected from people residing in scrub typhus reporting areas. Serology was performed by an indirect ELISA for the three rickettsial agents' viz., scrub typhus group orientiae (STGO), spotted fever group rickettsiae (SFGR) and typhus group rickettsiae (TGR). A sample with total net absorbance ≥1.000 was considered as positive. An entomological survey was also carried out in the affected areas. RESULTS: Overall, 1265 human blood samples were collected, of which 30.8% (n=390), 13.8% (175) and 4.2% (53) had antibodies against STGO, SFGR and TGR respectively. Presence of antibodies against more than one of the rickettsial groups was also detected. Among the arthropods collected, chiggers of Leptotrombidium deleinse, fleas belonging to Ctenocephalides felis and Pulex irritans, ticks belonging to Rhipicephalus microplus, Haemaphysalis spp. were predominant. Candidatus Rickettsia senegalensis was detected in C. felis. CONCLUSIONS: Our findings confirm wide circulation of rickettsial infections and their probable vectors in the northeast region of India.Accession numbers: KU163367, KU163368, KU499847, KU499848.