Coxiella-like bacteria in Haemaphysalis wellingtoni ticks associated with Great Hornbill, Buceros bicornis

@#Birds are known to be the most mobile hosts and are therefore considered to be hosts with potential to contribute to the long-distance spread and transmission of tick-borne pathogens. In the present study, ticks were collected from a hornbill nest at Chaiyaphum Province, Thailand. They were screened for the presence of Coxiella bacteria using conventional PCR. The evolutionary relationships of positive Coxiella-like bacteria (CLB) were analysed based on the gene sequences of 16S rRNA, groEL and rpoB. Among all 22 tested ticks, CLB infections were found in 2 Haemaphysalis wellingtoni individuals. In a phylogenetic analysis, the Coxiella 16S rRNA gene detected in this study formed a separate clade from sequences found in ticks of the same genus. In contrast, the phylogenetic relationships based on groEL and rpoB revealed that these two genes from H. wellingtoni ticks grouped with CLB from the same tick genus (Haemaphysalis). This study is the first to report the presence of CLB in H. wellingtoni ticks associated with the Great Hornbill, Buceros bicornis in Thailand. Three genes of CLB studied herein were grouped separately with Coxiella burnetii (pathogenic strain). The effects of CLB in the ticks and Buceros bicornis require further investigation.

A new record of Rickettsia japonica in ticks infesting a Burmese ferret-badger in Thailand

@#Ticks are important vectors of arthropod-borne diseases and they can transmit a wide variety of zoonotic pathogens to humans, domestic and wild animals. Rickettsia japonica is a member of SFG rickettsiae causing Japanese spotted fever (JSF) and can transmit to humans via infected ticks. In this study, we report the first case of Rickettsia japonica in Haemaphysalis hystricis tick collected from a roadkill Burmese ferret-badger (Melogale personata) in Loei province, northeastern Thailand. According to the DNA sequences and phylogenetic analyses of the outer membrane protein A and B genes (ompA and ompB), the detected R. japonica was identical to those found in JSF patients in Korea, Japan, and China, and closely related to Rickettsia detected by ompA in a tick from Thailand. Further study on the prevalence of R. japonica and diversity of mammalian reservoir hosts will be useful to gain a better understanding of JSF epidemiology.

Cytogenetic, crossing and molecular evidence of two cytological forms of Anopheles argyropus and three cytological forms of Anopheles pursati (Diptera: Culicidae) in Thailand

Nine and 11 isolines of Anopheles argyropus and Anopheles pursati, respectively, were established from individual females collected from cow-baited traps, and the characteristics of metaphase chromosomes were investigated in their F1-progenies. As determined by the different amounts of extra heterochromatin on sex chromosomes, 2 types of X (X1, X2) and Y (Y1,Y2), and 2 types of X (X1, X2) and 3 types of Y (Y1, Y2, Y3) chromosomes were obtained from An. argyropus and An. pursati, respectively. These types of sex chromosomes comprised 2 [Forms A (X1, Y1) and B (X1, X2, Y2)] and 3 [Forms A (X1, X2, Y1), B (X1, X2, Y2) and C (X2, Y3)] karyotypic forms of An. argyropus and An. pursati, respectively. All karyotypic forms acquired from An. pursati are new one that were discovered in this study, of which Forms A, B and C were found generally in Chiang Mai Province, while only 1 isoline of Form B was obtained in Ratchaburi Province. Form A was recovered from An. argyropus only in Ubon Ratchathani Province, whereas Form B from that species was found commonly in both Ubon Rathchathani and Nakhon Si Thammarat Provinces. Crossing experiments among the 2 and 3 isolines representing 2 and 3 karyotypic forms of An. argyropus and An. pursati, respectively, indicated genetic compatibility in yielding viable progenies and synaptic salivary gland polytene chromosomes through F2-generations. The conspecific natures of these karyotypic forms in both species were further supported by very low intraspecific sequence variations (average genetic distance: An. argyropus = 0.003-0.007, An. pursati = 0-0.005) of ribosomal DNA (ITS2) and mitochondrial DNA (COI and COII).

Population cytogenetics of the malaria vector Anopheles leucosphyrus group.

Until recently, very little was known of Anopheles species complexes and their relationships to epidemiology and malaria transmission in Southeast Asia. During the past eight years, extensive studies on the genetics of natural populations of anopheline mosquitoes in this region, involving the interdisciplinary efforts of taxonomists, operational entomologists and biologists, have revealed groups of cryptic species of Anopheles vectors, particularly the An. leucos phyrus group. This species group comprise seventeen species and two subspecies widely distributed in the forested areas of Southeast Asia. Among these species. An. dirus Peyton and Harrison, has been shown by cytogenetic and morphological studies to be a complex of at least seven isomorphic species, provisionally designated species A, B, C, D, E, F and takasagoensis, on the Southeast Asian mainland. Cytological identification of these species is based on distinct banding patterns of salivary gland polytene chromosomes as well as heterochromatin differences in mitotic karyotypes. The five species found in Thailand (A-D, F) exhibit distinct geographic distributions. Species A is widespread throughout Thailand except in the south. Species B had been found in sympatry with species C in southern Thailand and both seem to show north-south clinal geographic variation. Species D is common on the west side of southern Thailand and along the Thai-Burmese border in sympatry with species A. Species F, An. nemophilous Peyton and Ramalingam, has been found in a population at the Thai-Malaysian border in this study although it was known to be common in southern and western Thailand and Peninsular Malaysia. Species E is known only from western India. The five species found in Thailand also exhibit seasonal variation in relative abundance and different nocturnal biting cycles. Chromosomal polymorphisms have been observed in mitotic and polytene chromosomes of An. dirus A and D. Species B and C also show heterochromatin variation in the sex chromosomes, but are monomorphic for the standard sequence in polytene chromosomes. These biological characteristics of the An. dirus complex may have implications for understanding the epidemiology of malaria in Southeast Asia. Recent cytogenetic studies of wild-caught samples of An. leucosphyrus from Sumatra, Kalimantan and southern Thailand have revealed the presence of two distinct species within this taxon. Species A is widely distributed in southern Thailand, East Malaysia and Kalimantan, while species B is confined to Sumatra. The two isomorphic species are vectors of human malaria within their range of distribution.(ABSTRACT TRUNCATED AT 400 WORDS)

Distribution and chromosomal polymorphism of the malaria vector Anopheles dirus species D.

Preliminary results indicate that species D of the Anopheles dirus complex is widespread west of the Thai-Burma border in Burma and Bangladesh. A chromosomal study of An. dirus species D in these areas has revealed that this malaria vector is highly polymorphic for chromosomal rearrangements in salivary gland polytene chromosomes. The data from the limited number of wild-caught samples suggest that different geographically isolated populations may occur with respect to the frequency of inversions 2La, 3Ra and 3La. The distribution of chromosomal polymorphisms may be associated with the geography and epidemiology of human malaria in this region.

Geographic distribution and biting behaviour of four species of the Anopheles dirus complex (Diptera: Culicidae) in Thailand.

A cytogenetic analysis of the species status of members of the Anopheles dirus group, from natural populations, over a six-year period has produced biogeographic and behavioural data which are presented herein. Species A, B, C and D have quite distinct geographic distributions in Thailand. Species A is the only species in most of the mainland but is absent from the southern half of the peninsula. Species B is dominant in the far south of the peninsula giving way to species C on the north-east side and rarely occurring on the west side of the peninsula; it is unknown from the northern half or the rest of the country. Species C is known from the middle eastern side of the peninsula and from a site far to the north along the Burmese border. Species D occurs down the mountains along the Burmese/Thai border and along the north half of the western peninsula. Each species appears to have distinct times of biting during the night. They also appear to have differential seasonal abundance. These data together with the differing geographic distributions suggest some implications for Plasmodium transmission and that care should be taken to identify these genetic species during malariometric studies.

Some observations on sympatric populations of the malaria vectors Anopheles leucosphyrus and Anopheles balabacensis in a village-forest setting in South Kalimantan.

Human bait collections of biting anopheline mosquitoes were made on five consecutive nights during September 1986 in a remote village located in a heavily forested area of South Kalimantan, Indonesia. Anopheles leucosphyrus and An. balabacensis comprised 97.7% of the total number of specimens collected outside houses in the village. Anopheles balabacensis were slightly fewer in total numbers than leucosphyrus. Mosquitoes were collected simultaneously in the village and the forest on two nights. The numbers of leucosphyrus and balabacensis biting in the forest were small in comparison with the populations encountered in the village. Approximately 75% of the specimens were checked individually for sporozoite infections using ELISA for P. falciparum and P. vivax. Sporozoites of P. falciparum were detected in one specimen of leucosphyrus and one of balabacensis. The sporozoite infection rate was 1.0% for leucosphyrus and 1.3% for balabacensis.

Cytogenetic studies of some species complexes of Anopheles in Thailand and Southeast Asia.

Recent studies on cytogenetics, behavioral, geographical and distinct morphological characters on adult, pupal and larval stages have revealed that "balabacensis" is a species complex. Anopheles dirus the mainland species, is distributed widely in Thailand and is renowned for its role as primary vector of human malarial parasites. Further, evidence from cytogenetic and taxonomic studies suggests that "An. dirus" is a species complex comprising at least four distinct species provisionally designated: dirus A, B, C and D. These cryptic species are distinguishable only partially morphologically, but can be separated on the basis of metaphase chromosomes using the Giemsa and Hoechst 33258 staining techniques. Apparently, these siblings show distinct patterns of geographic distribution in Thailand and Peninsular Malaysia. The recognition of dirus as a complex of species in Thailand and Peninsular Malaysia requires a re-evaluation of the role that the individual members of this complex have in the transmission of malaria parasites in this region. Cytological analysis of gene rearrangements in ovarian polytene chromosomes has shown that An. maculatus is a sibling-species complex consisting of at least four species in Thailand provisionally designated: maculatus A, B, C and G. These siblings are sympatric in some populations. Furthermore, species B is so highly polymorphic for chromosome rearrangements that four geographic forms can be recognized. It is not known whether these four forms are subspecies or yet further species within the species B complex. These sibling-species must be differentiated in order to understand any differential capabilities in their transmission of human malaria parasites. Anopheles nivipes was elevated from synonymy under An. philippinensis to full species status by Reid, a decision recently confirmed by cross mating experiments. The Thailand Malaria Division does not differentiate these two species and only identifies An. philippinensis, yet, An. nivipes is by far the most common of the two species in Thailand. Furthermore, preliminary surveys of the ovarian polytene chromosomes of several widely separated populations of An. nivipes in Thailand have revealed at least two distinct chromosomal types of nivipes based on fixed inversions on the X chromosomes.

Genetic response of Aedes aegypti to different environmental temperatures.

Three groups of Aedes aegypti maintained 8 generations in the laboratory at 20 degrees C, 27 +/- 1 degree C (normal control), and at 35 degrees C, respectively were analyzed for allozyme variability. Of the 8 larval loci examined Est alpha-1 and Lap-2 in generation 8 were still variable, while the other allozymes became uniform. The 35 degrees C population had mean level of heterozygosity of about 6% of individual per locus, less than those observed in the 20 degrees C and the control populations which were 14% and 13%, respectively. It is suggested that in the experimental populations, the rate of loss of alleles can be increased by high temperature which must not be higher than the optimum temperature of enzymes.