Mites are known causes of allergic diseases. Currently, identification of mites based on morphology is difficult if only one mite is isolated from a (dust) sample, or when only one gender is found, or when the specimen is not intact especially with the loss of the legs. The purpose of this study was to use polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the ITS2 gene, to complement the morphological data for the identification of mites to the species level. For this, six species were cultured: Dermatophagoides pteronyssinus, D. farinae, Blomia tropicalis, Tyrophagus putrescentiae, Aleuroglyphus ovatus and Glycycometus malaysiensis. Genomic DNA of the mites was extracted, quantified, amplified and digested individually with restriction enzymes. Hinf I and Ple I differentiated the restriction patterns of D. pteronyssinus and D. farinae. Bfa I and Alu I enzymes differentiated B. tropicalis and G. malaysiensis. Ple I enzyme was useful for the differentiation between T. putrescentiae and A. ovatus. Bfa I was useful for the differentiation of G. malaysiensis from the rest of the species. In conclusion, different species of mites can be differentiated using PCR-RFLP of ITS2 region. With the established PCR-RFLP method in this study, identification of these mites to the species level is possible even if complete and intact adult specimens of both sexes are not available. As no study to date has reported PCR-RFLP method for the identification of domestic mites, the established method should be validated for the identification of other species of mites that were not included in this study.
In the present study on the life-cycle of Blomia tropicalis, freshly laid eggs were observed until they developed into adults; the development periods between stages were recorded. The eggs took an average of 22.9 +/- 6.4 days to develop to adults. For longevity experiments, newly emerged adults were kept at 25 degrees C and observed until they died. There was no significant difference in longevities of the different sexes (p = 0.053). Production of eggs by mated females were monitored until egg production stopped and the female died. Mated females and males survived an average of 32.2 +/- 15.4 and 30.9 +/- 17.7 days respectively. The difference in longevity of the mated females, and males was not significant (p = 0.747). Longevity of the mated females was found to be significantly (p < 0.05) shorter than unmated females.
Two commercial repellants were evaluated in the laboratory against Leptotrombidium fletcheri chiggers. The active ingredient in one was DEET and in the other was citrus oil. Excito-toxicity effect was studied and it was determined by the time ("escape time") chiggers took to move off filter papers treated with the repellants. All chiggers exposed on filter papers treated with DEET died and did not move off the treated papers. None of the chiggers that were placed on papers treated with citrus oil were killed. Escape times on papers treated with a 2-sec spray of citrus oil were longer than those for the 4- and 8-sec sprays. The weights of citrus oil deposited increased with increasing spray times. Electron microscopy showed that the repellants had no effect on the texture of the filter papers. It was concluded that the spray containing DEET was more effective; however, both repellants should be further evaluated under field conditions for protection against chigger bites.
Four commercial repellents were evaluated in the laboratory against Leptotrombidium deliense chiggers. Both in vitro and in vivo methods were used to determine repellency of the compounds. The repellents were Kellis (containing citronella oil, jojoba oil and tea tree oil), Kaps (containing citronella oil), BioZ (containing citronella oil, geranium oil and lemon grass oil) and Off (containing DEET). The combination of three active ingredients: citronella oil, geranium oil, lemon grass oil gave the highest repellency (87%) followed by DEET (84%). In vitro repellencies ranged from 73% to 87%. There was no significant difference between the four products. All the repellents had 100% in vivo repellency compared to 41-57% for the controls.
Five pesticides were evaluated against laboratory colonies of Leptotrombidium fletcheri (Womersly and Heaslip) by the Pasteur pipet technique. The pesticides were dieldrin (LC50 = 3.6 ppm, LC99 = 18.2 ppm), bromopropylate (LC50 = 9.2 ppm, LC99 = 239.6 ppm), dicofol (LC50 = 27.8 ppm, LC99 = 118.1 ppm), fenthion (LC50 = 15.4 ppm, LC99 = 29.7 ppm), and malathion (LC50 = 84.7 ppm, LC99 = 313.9 ppm). Dieldrin was the most toxic. Dicofol was recommended for further evaluation in field trials.
Isolation of rickettsiae from patients' blood samples and organ samples of wild rodents from areas with high seroprevalence of rickettsial infections was attempted using cell culture assay and animal passages. L929 mouse fibroblast cells grown in 24 well tissue culture plate were inoculated with buffy coat of febrile patients and examined for the growth of rickettsiae by Giemsa, Gimenez staining and direct immunofluorescence assay. No rickettsiae were isolated from 48 patients' blood samples. No symptomatic infections were noted in mice or guinea pigs infected with 50 organ samples of wild rodents. There was no rickettsial DNA amplified from these samples using various PCR detection systems for Orientia tsutsugamushi, typhus and spotted fever group rickettsiae.
Using cultured mouse fibroblast L929 cells, this study demonstrated the hemolytic and cytotoxic activities and induction of apoptosis in cells infected with Orientia tsutsugamushi. Low levels of hemolytic activity were detected using heavily infected cells. No hemolysin or cytotoxin were detected in the infected culture fluid regardless of the pathogenicity of the O. tsutsugamushi strains in mice. Using propidium iodide uptake assay, acridine orange/ethidium bromide staining and terminal deoxynucleotide transferase-mediated dUTP-digoxigenin nick-end labeling assay, apoptosis was observed in L929 cells infected with Karp and Gilliam strains.
Accidental entry of insects and other arthropods is a common aetiology of aural foreign bodies (FB) presenting to accident and emergency departments. A retrospective study revealed that the FB in almost half (148) of 348 cases of aural FB investigated at the Universiti Sains Malaysia Hospital was an arthropod. The most common arthropod encountered was the cockroach, followed by a cattle tick. The high prevalence of tick infestation of the human ear canal (i.e. human otoacariasis) currently appears to be unique to the Malaysian state of Kelantan. The presentation of patients with intra-aural ticks, the methods used to remove the ticks, the complications encountered, and recommendations for an appropriate course of action in such cases are discussed.
Allergy to house dust mites (HDM) is an important cause of asthma and rhinitis in Malaysia. This study was carried out to evaluate the dust mite fauna in the Klang Valley. Dust samples were collected from 20 houses from March 1994 to February 1995. Thirty-three dust samples from mattresses were examined monthly for the occurrence of HDM. A total of 22 species in 9 families of HDM was identified. The most common and densely populated species was Blomia tropicalis with an average density of 8,934 mites/g of dust. Dermatophagoides pteronyssinus was the next in abundance, followed by Malayoglyphus intermedius. All houses surveyed were found to be infested with HDM and every house had at least 6 species of HDM. B. tropicalis and D. pteronyssinus were found in all mattresses. HDM in the Klang Valley were found to be highly prevalent and present in high densities. In this study, counts of D. pteronyssinus was found to exceed the proposed exposure threshold of 500 mites/g dust, for triggering acute asthma. Although counts of B. tropicalis exceeded D. pteronyssinus, no conclusion could be made because there is currently no exposure threshold for triggering acute asthma, for this species. Monthly distribution of B. tropicalis and D. pteronyssinus showed 2 peaks and 4 peaks, respectively. The major peak for D. pteronysinus was in January 1995 whereas for B. tropicalis, the major peak was more variable and occurred between November 1994 to January 1995. Both the species showed minor peak in April 1994.
A species of house dust mite, Suidasia pontifica, was recently shown to produce allergens affecting man. The species may be as important as other allergen producing mite in sensitization and causing allergic symptoms in Malaysians. Surveys conducted demonstrated that 80% of the houses surveyed were positive for this mite with densities ranged from 2 to 50 mites per gram of dust. Colonies of the species has been successfully established and materials from those colonies have been used to produce extracts for studies on sensitization to the mites. A total of 85 suspected allergic rhinitis patients were tested and 74.1% demonstrated positive reactions. Extract of this mite should be considered for routine diagnostic testing and possible immunotherapy.