Evidence of natural infection of Orientia tsutsugamushi in vectors and animal hosts � Risk of scrub typhus transmission to humans in Puducherry, South India

Background: Scrub typhus infection is endemic in India and reported to be the major cause for acute encephalitis syndrome (AES) in humans. Periodic occurrence of scrub typhus cases and presence of pathogen in rodents were also reported in areas with human cases of scrub typhus in Puducherry. Objectives: This study was carried out to screen Orientia tsutsugamushi in rodent/shrew reservoirs and vectors in villages of Puducherry with no reports of human scrub typhus cases. Methods: This study was conducted during October 2017 to January 2018 in ten randomly selected villages in Puducherry. Rodents/shrews in the peridomestic area were trapped using Sherman traps. Screening of O. tsutsugamushi in rodents/shrews and mite vectors was done by polymerase chain reaction (PCR). Weil-Felix test was done to screen antibodies against O. tsutsugamushi in rodent serum samples. Results: Among the 54 rodents trapped, Suncus murinus was the major small animal and Leptotrombidium deliense was the major mite species retrieved. PCR screening revealed pathogen positivity in 8 rodent blood and 3 pooled mite samples. Phylogenetic analysis has shown that Kato was the circulating serotype of O. tsutsugamushi. None of the rodent serum samples was tested positive for antibodies against O. tsutsugamushi by Weil-Felix test. Conclusions: The presence of pathogen in both vectors and reservoir animal hosts imposes a risk for scrub typhus transmission to the inhabitants; hence, initiation of vector control measures before the start of winter is recommended in the study area. It is also recommended to screen scrub typhus in patients with undifferentiated acute febrile illness and AES.

Research Progress on Leptotrombidium deliense

This article reviews Leptotrombidium deliense, including its discovery and nomenclature, morphological features and identification, life cycle, ecology, relationship with diseases, chromosomes and artificial cultivation. The first record of L. deliense was early in 1922 by Walch. Under the genus Leptotrombidium, there are many sibling species similar to L. deliense, which makes it difficult to differentiate L. deliense from another sibling chigger mites, for example, L. rubellum. The life cycle of the mite (L. deliense) includes 7 stages: egg, deutovum (or prelarva), larva, nymphochrysalis, nymph, imagochrysalis and adult. The mite has a wide geographical distribution with low host specificity, and it often appears in different regions and habitats and on many species of hosts. As a vector species of chigger mite, L. deliense is of great importance in transmitting scrub typhus (tsutsugamushi disease) in many parts of the world, especially in tropical regions of Southeast Asia. The seasonal fluctuation of the mite population varies in different geographical regions. The mite has been successfully cultured in the laboratory, facilitating research on its chromosomes, biochemistry and molecular biology.

Chigger Mite (Acari: Trombiculidae) Survey of Rodents in Shandong Province, Northern China

Chigger mites are parasites of rodents and other vertebrates, invertebrates, and other arthropods, and are the only vectors of scrub typhus, in addition to other zoonoses. Therefore, investigating their distribution, diversity, and seasonal abundance is important for public health. Rodent surveillance was conducted at 6 districts in Shandong Province, northern China (114–112°E, 34–38°N), from January to December 2011. Overall, 225/286 (78.7%) rodents captured were infested with chigger mites. A total of 451 chigger mites were identified as belonging to 5 most commonly collected species and 3 genera in 1 family. Leptotrombidium scutellare and Leptotrombidium intermedia were the most commonly collected chigger mites. L. scutellare (66.2%, 36.7%, and 49.0%) was the most frequently collected chigger mite from Apodemus agrarius, Rattus norvegicus, and Microtus fortis, respectively, whereas L. intermedia (61.5% and 63.2%) was the most frequently collected chigger mite from Cricetulus triton and Mus musculus, respectively. This study demonstrated a relatively high prevalence of chigger mites that varied seasonally in Shandong Province, China.

Catenotaenia dendritica (Cestoda: Catenotaeniidae) and Three Ectoparasite Species in the Red Squirrel, Sciurus vulgaris, from Cheongju, Korea

Parasites are recorded from the red squirrel, Sciurus vulgaris, from Cheongju, the Republic of Korea. A total of 5 road-killed squirrels were thoroughly examined for internal and external parasites from November 2011 to May 2014. Total 4 parasite species, including 1 tapeworm and 3 ectoparasite species were recovered. They were morphologically identified as Catenotaenia dendritica (Cestoda: Catenotaeniidae), Hirstionyssus sciurinus, Leptotrombidium pallidum, and Ceratophyllus (Monopsyllus) indages. Among them, C. dendritica and H. sciurinus are recorded for the first time in the Korean parasite fauna. In addition, the possibility that the red squirrel could act as a reservoir host for a zoonotic disease like tsutsugamushi disease with L. pallidum as its vector has been raised.

Seroepidemiological Survey of Zoonotic Diseases in Small Mammals with PCR Detection of Orientia tsutsugamushi in Chiggers, Gwangju, Korea

Serosurveillance for zoonotic diseases in small mammals and detection of chiggers, the vector of Orientia tsutsugamushi, were conducted from September 2014 to August 2015 in Gwangju Metropolitan Area. Apodemus agrarius was the most commonly collected small mammals (158; 91.8%), followed by Myodes regulus (8; 4.6%), and Crocidura lasiura (6; 3.5%). The highest seroprevalence of small mammals for O. tsutsugamushi (41; 26.3%) was followed by hantaviruses (24; 15.4%), Rickettsia spp. (22; 14.1%), and Leptospira (2; 1.3%). A total of 3,194 chiggers were collected from small mammals, and 1,236 of 3,194 chiggers were identified with 7 species of 3 genera: Leptotrombidium scutellare was the most commonly collected species (585; 47.3%), followed by L. orientale (422; 34.1%), Euchoengastia koreaensis (99; 8.0%), L. palpale (58; 4.7%), L. pallidum (36; 2.9%), Neotrombicula gardellai (28; 2.3%), and L. zetum (8; 0.6%). L. scutellare was the predominant species. Three of 1,236 chigger mites were positive for O. tsutsugamushi by PCR. As a result of phylogenetic analysis, the O. tsutsugamushi strain of chigger mites had sequence homology of 90.1-98.2% with Boryong. This study provides baseline data on the distribution of zoonotic diseases and potential vectors for the development of prevention strategies of vector borne diseases in Gwangju metropolitan area.

Larval Chigger Mites Collected from Small Mammals in 3 Provinces, Korea

A total of 9,281 larval chigger mites were collected from small mammals captured at Hwaseong-gun, Gyeonggi-do (Province) (2,754 mites from 30 small mammals), Asan city, Chungcheongnam-do (3,358 mites from 48 mammals), and Jangseong-gun, Jeollanam-do (3,169 for 62 mammals) from April-November 2009 in the Republic of Korea (= Korea) and were identified to species. Leptotrombidium pallidum was the predominant species in Hwaseong (95.8%) and Asan (61.2%), while Leptotrombidium scutellare was the predominant species collected from Jangseong (80.1%). Overall, larval chigger mite indices decreased from April (27.3) to June (4.9), then increased in September (95.2) and to a high level in November (169.3). These data suggest that L. pallidum and L. scutellare are the primary vectors of scrub typhus throughout their range in Korea. While other species of larval chigger mites were also collected with some implications in the transmission of Orientia tsutsugamushi, they only accounted for 11.2% of all larval chigger mites collected from small mammals.

Outbreak of scrub typhus in the North East Himalayan region-Sikkim: An emerging threat.

Scrub typhus is an acute febrile illness that is known to be endemic in the South East Asian countries and the Western Pacific region. We here report an outbreak in the tiny Himalayan state of Sikkim. Patients with pyrexia of unknown origin were evaluated. They were screened by Weil-Felix test and the rapid immunochromatographic method. Samples that were positive by either Weil-Felix agglutination test or by rapid immunochromatography were confirmed by IgM enzyme-linked immunosorbent assay (ELISA). A total 204 samples were screened. Sixty-three patients were confirmed positive among which 42 were male and 21 were female. Effective management and early administration of antibiotics will help prevent the complications and mortality associated with scrub typhus.

Geographical distribution and host selection of Leptotrombidium rubellum in some parts of Yunnan province / 中华流行病学杂志

Objective To investigate the geographical distribution and host selection of Leptotrombidium rubellum among different small mammal hosts in some areas of Yunnan province,China.Methods Field survey was carried out in 23 counties of Yunnan province between 2001 and 2011.Small mammal hosts were captured with mouse cages and traps with baits.Chigger mites on the surface of two auricles were scraped off by a bistoury,and then preserved in 70％ ethanol.Every specimen of the chigger mites on the slides was finally identified into species under a microscope.Some conventional statistical methods were adopted to calculate all the collected chigger mite species and the constituent ratios of L,.rubellum in different areas and on different hosts,with prevalence (P),mean intensity (MI) and mean abundance (MA) on different hosts calculated.Linear regression was used to analyze the relationship among P,MI and MA.Patch index (m*/m) was used to measure the spatial patterns of L.rubellum among different individuals of related small mammal hosts.Results A total of 108 480 chigger mites were collected from the body surface of all the captured small mammal hosts.All the collected chigger mites were identified as 3 subfamilies,24 genera and 234 species.Of the 234 species of chigger mites,654 individuals of L.rubellum were collected,only in 4 counties.The collected individuals of L.rubellum accounted for 0.603％ of the total mites (108 480 individuals).96.637％ of L.rubellum came from flatland areas and habitats while only 3.363％ of the L.rubellum were from the mountainous regions.The orderings of the hosts appeared as Rodentia and Insectivora harbored 96.296％ and 2.469％ respectively,of the collected while Lagomorpha and other orders there was no L rubellum found.Of 67 species (in 34 genera and 12 families of 5 orders),Rattus tanezumi (in genus Rattus and family Muridae of Rodentia)harbored 96.788％ of the collected L.rubellum with relatively low prevalence (P=3.776％) or mean intensity (MI=8.792 mites/per host),followed by Bandicota indica.Data from the patch index showed that L rubellum had an aggregated distribution pattern among different individuals of their hosts.Conclusion L.rubellum existed in Yunnan province with relatively rare numbers of the individuals.In Yunnan province,L.rubellum was mainly found in the flatland regions and habitats with relatively low altitude.L.rubellum could parasitize several different species of hosts with low host specificity,but it prefered to choose Rattus tanezumi and some other house-dwelling small mammals as its main hosts.

Monthly Occurrence of Vectors and Reservoir Rodents of Scrub Typhus in an Endemic Area of Jeollanam-do, Korea

Monthly surveys were conducted to investigate the occurrence of chigger mites and seroprevalence of scrub typhus among small mammals in Jeollanam-do, the southwestern part of Korea, from November 2006 through October 2007. Fifty-eight small mammals, including 57 Apodemus agrarius (98.3%) and 1 Crocidura lasiura (1.7%), were captured, and a total of 4,675 chigger mites representing 4 genera and 8 species were collected from them. The chigger infestation rate among small mammals was 69.0%. The most predominant species in A. agrarius was Leptotrombidium scutellare (54.0%), followed by Leptotrombidium pallidum (39.4%), Leptotrombidium orientale (4.4%), Leptotrombidium palpale (1.1%), Neotrombicula tamiyai (0.6%), Eushoengastia koreaensis (0.3%), Neotrombicula gardellai (0.3%), and Cheladonta ikaoensis (<0.1%). The chigger index of A. agrarius was the highest in October (740.0), followed by November (242.0), September (134.6), March (98.3), February (38.2), January (35.3), December (34.5), April (30.8), and May (1.7). The average antibody positive rate of scrub typhus in wild rodents was 50.0%. The seropositive rates were high in October (100.0%) and November (83.3%), whereas those in other months were relatively low (28.6-57.1%). The chigger index of L. scutellare rapidly increased in September to form an acuminate peak in October, followed by a gradual decline. These results suggest that the outbreak of scrub typhus in the southwestern part of Korean peninsula is mostly due to L. scutellare.

Emergence of Schoengastiella ligula as the vector of scrub typhus outbreak in Darjeeling: Has Leptotrombidium deliense been replaced.

Background: Following a suspected outbreak of scrub typhus in Kurseong, Darjeeling, the Armed Forces Medical College, Pune was requested by the National Institute of Epidemiology and the State authorities to undertake investigation of the ongoing scrub typhus outbreak and suggest containment measures. Materials and Methods:The epidemic team undertook clinical, entomological and serological studies to understand the local disease pattern and delineate high risk areas, host diversity by rodent trapping using Sherman traps, mite fauna diversity, abundance and vector species identification by phase contrast microscopy for preparation of electronic database and rodent and human serological studies by Weil Felix and PCR. Results:The results indicate no association of scrub typhus with age and sex (P=0.37 and 0.74 respectively). The maximum cases occurred amongst the tea garden workers (73%) in the age group of 25-44 years. The predominant clinical presentation was fever (100%) with headache (75%), lymphadenopathy (45%) and presence of eschar (76.7%). The dominant host species (50% of trapped rodents) was shrew Suncus murinus, the index animal for scrub typhus, which contributed maximally to the vector abundance (52.96%) with a chigger index of 61.56. The trombiculid mite Schoengastiella ligula was the vector species much against the expected mite vector Leptotrombidium deliense, in the area. The study found the presence and abundance of vector species which corroborated well with the occurrence of cases in the various localities within the subdivisions. Conclusion:The study thus establishes Schoengastiella ligula as the vector of scrub typhus outbreak in Kurseong, Darjeeling. Preventive and containment measures with emphasis on reduction of man - vector contact were suggested to the state authorities to contain the outbreak.

Preliminary survey on the distribution of Leptotrombidium deliense in some areas of Yunnan province / 中华流行病学杂志

Objective To investigate the distribution of Leptotrombidium deliense among different small mammal hosts in some areas of Yunnan province. Methods A field survey was carried out in some counties of Yunnan province and the small mammal hosts were captured, using mouse cages and traps with baits. Chigger mites on the surface of two auricles were scraped off by a bistoury, and then preserved in 70% ethanol. Every specimen of the chigger mites on the slides was finally identified into species under a microscope. Some conventional statistical methods were adopted to calculate all the collected chigger mite species and the constituent ratios of Leptotrombidium deliense in different areas and on different hosts, together with its prevalence and mean abundance on different hosts. Results A total of 10 222 small mammal hosts were captured from 19 counties and identified as 11 families, 34 genera and 62 species in 5 orders, and 92 990 individuals of chigger mites were collected from the body surface of these small mammal hosts. All the collected chigger mites were identified as 3 subfamilies, 22 genera, and 225 species. Meanwhile, Leptotrombidium deliensee only accounted for 1.659% of the total. The host specificity of Leptotrombidium deliense was very low and 1544 individuals of Leptotrombidium deliense collected from 8518 small mammal hosts belonged to 6 families, 13 genera and 19 species in 3 orders. Our results showed that Leptotrombidium delienses were mainly collected from Insectivora and Rodentia. Leptotrombidium deliense had long been considered as the dominant species of chigger mites and the main vector of tsutsugamushi disease in Yunnan province of China, but our results seemed not thoroughly supporting this point of view. Conclusion Traditionally, Leptotrombidium deliense was the dominant species and the main vectors of scrub typhus in Yunnan province. However, based on our results, the above view might be true in some local places and the composition of chigger mites and the main vector of tautsugamushi disease might be different in regions and habitats in Yunnan province.

Serosurveillance of Scrub Typhus in Small Mammals Collected from Military Training Sites near the DMZ, Northern Gyeonggi-do, Korea, and Analysis of the Relative Abundance of Chiggers from Mammals Examined

Comprehensive quarterly serosurveillance on scrub typhus in small mammals collected from military training sites located near the Demilitarized Zone (DMZ), northern Gyeonggi-do (Province), ROK was conducted to determine the potential rodent-borne and associated ectoparasite disease risks to military personnel. A total of 1,196 rodents and insectivores representing 8 species, Apodemus agrarius (87.3%, n = 1,044), Mus musculus (5.4%, n = 65), Crocidura lasiura (3.3%, n = 40), Microtus fortis (2.6%, n = 31), Micromys minutus (0.3%, n = 4), Tscherskia triton (0.3%, n = 4), Rattus norvegicus (0.3%, n = 4), and Myodes regulus (0.3%, n = 4) were assayed for the presence of antibodies to Orientia tsutsugamushi. O. tsutsugamushi antibodies were detected in 6 of 8 species and seroprevalence determined; A. agrarius (45.6%), M. musculus (23.1%), M. fortis (48.4%), M. minutus (50.0%), T. triton (50.0%), and R. norvegicus (25.0%). A total of 31,184 chigger mites collected from 508 rodents and insectivores were slide-mounted and 10 species belonging to 4 genera were identified. Leptotrombidium pallidum (53.4%) was the most frequently collected, followed by L. palpale (15.7%), Neotrombicula tamiyai (14.3%), L. orientale (10.7%), L. zetum (3.1%), Walchia fragilis (2.1%), and L. gemiticulum (0.8%), while the remaining 3 species, L. subintermedium, N. gardellai, and Euschoengastia koreaensis were rarely observed (prevalence < 10%). In contrast to previous surveys, higher chigger indices of the primary scrub typhus vectors, L. pallidum (165.4), L. orientale (45.0), and L. palpale (21.4), were observed during the spring season.

Geographical Distribution and Relative Abundance of Vectors of Scrub Typhus in the Republic of Korea

A survey to determine the geographical distribution and relative abundance of potential vectors of scrub typhus was conducted from October to November 2006 at 13 localities throughout the Republic of Korea. Apodemus agrarius accounted for 97.6% (80/82) of all rodents, while only 2 Myodes regulus (2/82) were collected. A total of 10,860 chiggers were collected from A. agrarius belonging to 4 genera and 8 species, while only Walchia fragilis (40) was collected from Myodes regulus. Leptotrombidium pallidum (8,137; 74.9%), a vector of scrub typhus, was the predominant species collected from A. agrarius followed by Leptotrombidium scutellare (2,057, 18.9%), Leptotrombidium palpale (279; 2.7%), Leptotrombidium orientale (232; 2.1%), and Leptotrombidium zetum (79; 0.7%), Neotrombicula tamiyai (58; 0.5%), Euschoengastica koreaensis (16; 0.1%), and Cheladonta ikaoensis (2; < 0.1%). L. pallidum was the predominant chigger collected at collection sites in Gangwon (100%), Gyeonggi (87.2%), Chungnam (100%), Chungbuk (100%), Jeonbuk (73.9%), Jeonnam (77.0%), and Gyeongbuk (66.1%) provinces, whereas L. scutellare was the predominant chigger collected in Gyeongnam province (77.9%) and Jeju Island (62.3%). Data suggest a correlation between chigger population abundance and human cases of scrub typhus in Korea.

A Study on the Epidemiologic Characteristics of Scrub Typhus in Gyeongsangbuk-do, 1999-2001 / 한국역학회지

ONJECTIVE: This study was carried out to investigate the epidemiologic characteristics and magnitudes of scrub typhus (tsutsugamushi disease) in Gyeongsangbuk-do, Korea. METHODS: Resources of data collected were epidemiologic surveillance report to public health centers on each disease occurrence. The authors reviewed 344 epidemiologic surveillance reports for cases of scrub typhus developed in Gyeongsangbuk-do, Korea from Jan 1, 1999 to Dec 31, 2001. RESULTS: The scrub typhus cases were 376 during three year period, and the incidence was significantly higher in the county area than in the city area. Incidence of scrub typhus was increasing each year. In 344 epidemiologic surveillance reports, the job of the cases was farmer (67.2%), housewife (4.7%), clerk (3.5%). Two hundred ninety eight cases (86.6%) have experienced outdoor activities before development of the scrub typhus. The outdoor places of exposure were dry field (64.8%), rice field (51.7%), hill (12.8), and grass field (9.7%). The symptoms reported were fever (93.3%), headache (83.7%), chill (69.5%), and rash (54.4%). CONCLUSIONS: Incidence of scrub typhus was increasing in Gyeongsangbuk-do, Korea, this phenomenon may be related to change of climate, the reporting rate of disease and outing activities. Good surveillance and adequate analysis of epidemiologic data is a essential for the prevention of the disease. With this result, high risk population was identified and effective intervention should be done to prevent the disease.

The Value of PCR Method for Diagnosing Tsutsugamushi Disease / 대한피부과학회지

BACKGROUND: Tsutsugamushi disease, is an acute febrile illness caused by the infection of Orientia(O.) tsutsugamushi which is transmitted by mites such as Leptotrombidium(L.) pallidum. Because the antibody titer against O. tsutsugamushi has not risen enough to be detected at the initial stage of tsutsugamushi disease, the early serologic diagnosis of this disease has some difficulties. OBJECTIVE: The aim of this study was to show that the recently introduced technique of polymerase chain reaction(PCR) is excellent for the detection of rickettsial DNA in the patient's blood at the acute phase of illness. METHODS: Nested PCR with Boryong specific 56kDa protein antigen was done with sera from twenty one patients with tsutsugamushi disease and with two groups of mites(L. pallidum). The presence of O. tsutsugamushi DNA in the peripheral blood mononuclear cells of patients with tsutsugamushi disease was also determined by nested PCR during the antibiotic treatment. RESULTS: 1. Positive results with nested PCR with Boryong specific 56kDa protein antigen were found in eleven out of twenty one patients with tsutsugamushi disease and two samples of L. pallidum extracts. 2. After the initiation of antibiotic treatment, all samples(11 patients) were positive on the second day, and seven samples were positive on the seventh day, showing a slow action of the drugs against the rickettsia within the cells. CONCLUSION: Nested PCR is an ideal method for the detection of O. tsutsugamushi DNA during the early phase of tsutsugamushi disease.