Morphological and molecular identification of medically important questing Dermacentor species collected from some recreational areas of Peninsular Malaysia.

Questing is a situation when a tick is seeking to get closer or ambush its potential host. However, information on questing tick species in Malaysia is still lacking, thus the association with tick-borne diseases (TBD) is not completely understood. The aim of this study was to investigate the tick species from five most frequently visited recreational areas in Pahang and Terengganu states, which were recorded to have high potential of TBD cases. By implementing handpick method, a total of 18 males and 15 females belonging to five Dermacentor Koch, 1844 species, were collected, namely D. compactus Neumann 1901, D. tricuspis (Schulze, 1933), D. auratus Supino 1897, D. steini (Schulze, 1933), and D. falsosteini Apanaskevich, Apanaskevich & Nooma respectively. The specimens were collected and identified based on morphological characters prior to obtaining the molecular data of COI and 16S rDNA. The D. compactus was the most abundant species collected in this study, while D. falsosteini was the least. All species were distinctly separated on the Neighbor Joining and Maximum Parsimony tree topologies and supported with high bootstrap values. Furthermore, a low intraspecific variation (0.00 - 0.01) was observed amongst the individuals of the same species in both genes. Meanwhile, each Dermacentor species was genetically different, with interspecific values ranging from 0.13-0.19 and 0.11-0.20 for COI and 16S rDNA. These findings had successfully recorded the tick species that were potentially associated with TBD, and which might be circulated among humans and animals. This study also has some implications on the diversity and geographical extension of Dermacentor ticks, thus should warrant further investigation as a potential vector of tick-borne diseases and public health importance.

Precise identification of different stages of a tick, Ixodes granulatus Supino, 1897 (Acari: Ixodidae)

Objective To identify different stages of Ixodes granulatus (I. granulatus) based on morphological characters prior to molecular identification which is significant for confirming and identifying the nymphal stages of I. granulatus. Methods A total of 14 individuals of adult, engorged and nymphal ticks collected from three different localities were examined morphologically using taxonomic keys, followed by PCR using cytochrome oxidase subunit I (COI). Clustering analysis based on COI sequences was carried out by constructing neighbor-joining and maximum parsimony tree to clarify the genetic variation and diversity of local I. granulatus. Results Based on external morphological characterizations, nine individuals (64.3%) were successfully identified as I. granulatus, while five individuals were recognized only as Ixodes sp. due to lack of morphological characters visible and development during that stage. Molecular analysis of local I. granulatus using COI gene revealed 93%–94% sequence homology from available sequence in GenBank and was in concordance with the morphological identification. Furthermore, a low intraspecific variation was observed among the species of I. granulatus collected from different localities (0%–3.7%). Conclusions These findings demonstrated for the first time the establishment of COI gene for identifying I. granulatus nymphal tick which is of paramount importance to the control of potential tick-borne infections in Malaysia. Moreover, this study provides evidence that a combination of morphology and molecular data was corroborated as an accurate tool for tick identification.

Effect of germicidal UV-C light(254 nm) on eggs and adult of house dustmites, Dermatophagoides pteronyssinus and Dermatophagoides farinae (Astigmata: Pyroglyhidae)

<p><b>OBJECTIVE</b>To examined the immediate and 24 hours post- irradiation germicidal effects of UV-C lamp on eggs and adults of house dust mites Dermatophagoides pteronyssinus (D. pteronyssinus) and Dermatophagoides farinae (D. farinae).</p><p><b>METHODS</b>This study investigated the immediate and 24 hours post irradiation mortalities of adult mites exposed to UV-C at different exposure times (5 mins, 10 mins, 15 mins, 20 mins, 30 mins and 60 mins) and distances (10 cm, 25 cm, 35 cm, 45 cm and 55 cm). Fresh eggs of the 2 dust mites were also irradiated at 10, 35 and 55 cm for 0.5, 1, 2, 3, and 5 minutes, and observed daily post- irradiation for up to 7 days.</p><p><b>RESULTS</b>Highest immediate mortality of 100% occurred with direct irradiation at 10 cm distance from UV-C lamp and for 60 mins, for both species of mites. The post 24 hours mean mortality rates were (58.4±17.4)% for D. pteronyssinus and (27.7±9.7)% for D. farinae when irradiated for 1 hour at 55 cm distance under UV-C lamp. When mites were irradiated in the presence of culture media, the highest mortality rates were lower compared to the direct irradiation; at 10 cm distance and 60 mins exposure, the mean mortality was (74.0±6.8)% for D. pteronyssinus and (70.3±6.7)% for D. farinae. Egg hatchability for both species of mites was also notably reduced by greater than 50% following irradiation.</p><p><b>CONCLUSIONS</b>Ultraviolet C irradiation is lethal to an array of organisms by damaging their nucleic acids (DNA and RNA). This study demonstrates the increasing mite mortalities with increasing exposure times and decreasing distances.</p>