Allergens of Dermatophagoides and Blomia species are well-characterized but not for other species. This study was conducted to determine the prevalence of allergic sensitization to house dust (HDM) and storage mites (SM). One hundred adult subjects (aged ≥ 18) were recruited. The mite specific IgE of all allergic subjects were higher compared with healthy subjetcs despite being not statistically significant except for D. farinae and G. malaysiensis. The mean serum IgE levels against HDM and SM for allergic subjects were significantly higher compared with those in healthy subjects. They were mainly sensitized to Dermatophagoides farinae (35%) and Glycycometus malaysiensis (37%). Immunoblots revealed not all allergic subjects showed positive immuno-reactivity against the mites tested. Single or multiple bands were observed for different species. The subjects were commonly sensitized to Group 2 (9-12 kDa), 10 (38 kDa) and 18 (40-48 kDa) allergens. Twenty-one out of 60 allergic subjects were sensitized to either one or more species. The majority of them (71%) were sensitized to single species. The allergic subjects were mainly sensitized to D. pteronyssinus, followed by Tyrophagus putrecentiae and Aleuroglyphus ovatus. Seven were solely sensitized to HDM while 10 were solely sensitized to SM. Four subjects were sensitized to both. Pre-adsorption study revealed no cross-reactivity. There was difference between the prevalence and reactivity to allergens of HDM and SM in these subjects. Both ELISA and immunoblot did not correlate well but can complement each other in improving the detection of mite allergens to the species level.
Scanning electron microscope (SEM) images of dust mites, Suidasia pontifica, is presented to provide an improved visualization of the taxonomic characters of these mites. Suidasia pontifica can easily be identified by its scale-like cuticle, presence of external vertical setae (ve), longer external scapular setae (sce) compared to internal scapular setae (sci) and 3 ventral spines on apex of tarsus I. The differences in morphology of male and female S. pontifica are also discussed.
House dust mites and storage mites are well-known causes for allergenic diseases. The aim of this study was to investigate the immunogenic sites of Blomia tropicalis, Aleurogyphus ovatus and Glycycometus malaysiensis. The mites were maintained in a culture medium at 25ºC and 75% relative humidity. Mites were harvested either with heat escape or floatation method, purified, homogenized, quantified and used for the production of polyclonal antibody and immunostaining. For each species of mites, five male mice and five male rats were randomly selected and immunized intraperitoneally with respective crude mite extract at two-weekly intervals. Blomia tropicalis, A. ovatus or G. malaysiensis whole mites and paraffin-embedded mite sections were immunostained with the respective polyclonal antibody. The faecal pellets of mites were intensely stained for all the three species in the present study. The legs of sectioned A. ovatus were not immunogenic as compared with those of G. malaysiensis and B. tropicalis. The outer layer (cuticle) of whole mites and the eggs for these species were very immunogenic. Hence, the polyclonal antibodies obtained in this study may serve as potential tools in detecting the eggs and immature mites in environmental samples. Future studies should focus on the antigenic components of eggs since they were relatively abundant in dust and highly antigenic as seen in the present study.
Many finer taxonomic characters of Blomia tropicalis are not distinctly visible under conventional light microscopy. Scanning electron micrographs of this mite are therefore presented in this paper for better appreciation of the inconspicuous features of the morphology of the species. The differences in morphology of male and female B. tropicalis are also briefly discussed.
Scanning electron microscope (SEM) images of two dust mites, Sturnophagoides brasiliensis and Sturnophagoides halterophilus, are presented to provide an improved visualization of the taxonomic characters of these mites. Sturnophagoides halterophilus can be differentiated from S. brasiliensis by their expanded genu and femur of leg I. The differences in morphology of male and female S. brasiliensis are also discussed.
Tick-borne encephalitis (TBE) is a viral infection of the central nervous system and is caused by tick bites, usually after travel to rural or forested areas. The disease is prevalent in Scandinavia, Western Europe, Central Europe and the former Soviet Union and East Asia including Japan. In Malaysia, so far there are no reported cases of TBE. In the present time, many illnesses have been attributed to traveling to other parts of the world. Thus it is important to carry out TBE prevalence study to determine whether the virus is present among Malaysian population. Samples (sera and CSF) from patients admitted to major MOH hospitals in Peninsular Malaysia and Sabah with a clinical diagnosis of encephalitis but is IgM negative for JE, were tested for TBEV IgM ELISA and TBEV IgG ELISA (DRG, Germany). Out of the 600 samples screened for TBEV IgG, all were non-reactive. In addition, out of the 100 samples screened for TBEV IgM, all the samples were also non-reactive. Our results indicate that currently TBE is not present in the Malaysian population. Among the reasons for this could be lack of the infection agent, absence of the suitable vector or subjects selected for the study did not fit the criteria of possible exposure to TBE infections. Hence we recommend that for any future study, the selection of subjects should include those who returned from tick-infested forested areas.
A serosurvey was conducted in 1995-97 among 1596 febrile patients from 8 health centres in Malaysia for antibodies against Orientia tsutsugamushi (OT), Rickettsia typhi (RT) and TT118 spotted fever group rickettsiae (SFGR) by using an indirect immunoperoxidase assay. A total of 51.4% patients had antibody against at least 1 of those rickettsiae. Antibody to SFGR was most prevalent (42.5%), followed by RT (28.1%) and OT (24.9%). The seroprevalences of antibodies to SFGR, RT or OT alone were 12.4, 3.6 and 4.3%, respectively. Antibodies against more than 1 species of rickettsiae were presence in 31.1% of the patients, suggesting the possibility of co-infection, previous exposures or serological cross-reactivities. Seroprevalence of the various rickettsiae varied according to locality, with SFGR antibodies being the most prevalent in most areas. There was no significant association of prevalence of rickettsial antibody with gender. The seroprevalence of OT, SFGR and RT increased with patient age but an increase of antibody titre with age was not significant. Those working in the agricultural sectors had significantly higher seroprevalence of OT, SFGR and RT than those not related with agricultural activities. Scrub typhus remains a public health problem with an estimated annual attack rate of 18.5%. Tick typhus and murine typhus as shown in this serosurvey appear much more widespread than scrub typhus in this country.
Dermatophagoides pteronyssinus and Dermatophagoides farinae mites are commonly found in house dust, and are important sources of allergens affecting humans. Various approaches to killing the mites have been examined. This study investigated the mortalities of adult mites exposed to 2,450 MHz microwave radiation produced by 3 ovens at various exposure times and power settings. The ovens all had 3 power settings. The average maximum water temperatures generated at high, medium and low power settings were 99.4 +/- 0.2, 84.1 +/- 0.4 and 44.8 +/- 0.9 degrees C, respectively. At high and medium settings, there was 100.0% mortality in both species when exposed for 300 seconds. The mean mortality rates at low power were 10.8 +/- 0.7% for D. pteronyssinus and 9.7 +/- 2.6% for D. farinae. When mites were exposed in the presence of culture media, the mortality rates decreased with increasing weight of media. The mean mortality with the largest amount of media tested at high power setting was 61.4%.
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.
A survey of ticks and other ectoparasites was carried out during a national biodiversity scientific expedition at Ulu Muda Forest Reserve, Kedah, Malaysia from 23-29 March 2003. A total of 161 animals comprising 20 species of birds, 16 species of bats, six species of non-volant small mammals and 12 species of reptiles were examined for ticks and other ectoparasites. From these animals, nine species in five genera of ticks, 10 species in two families of Mesostigmatid mites and five species of chiggers were collected. Three of the ectoparasitic species found, Dermacentor auratus, Ixodes granulatus and Leptotrombidium deliense are of known public health importance. This survey produced the first list of ticks and other ectoparasites in the forest reserve and the third study of ectoparasites in Kedah. Fourteen species of these ectoparasites are new locality records.
The residual effect of a commercial spray containing 2.0% benzyl benzoate against Dermatophagoides pteronyssinus was evaluated in the laboratory. Filter papers treated with benzyl benzoate were dried for 2 hours and placed in Petri dishes. Adult mites were then added and mortality was assessed following 24 hour contact and at succeeding 4 weeks intervals up to week 28 post-treatment using the same treated filter papers. There was 100% mortality at 24 hours post-treatment, declining progressively to 71% by week 28. Probit analysis estimated a median lethal time (LT50) of 59 weeks.
A survey of ticks and other ectoparasites was carried out during a national biodiversity scientific expedition at Gunung Stong Forest Reserve, Kelantan, Malaysia from 23-29 May 2003. A total of 272 animals comprised of 12 species of birds, 21 species of bats, 7 species of rodents and 2 species of insects were examined for ticks and other ectoparasites. From these animals, 5 species in 4 genera of ticks; 7 species in 2 families of Mesostigmatid mites and 5 species of chiggers were collected. Among the ectoparasites found were Ixodes granulatus and Leptotrombidium deliense, which are of known medical importance. A tick island consisting of 10 nymphal stages of Dermacentor spp was observed feeding on Rattus tiomanicus.
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.
The pathogenicity of Malaysian isolates of Orientia tsutsugamushi was investigated by a mouse virulence assay. The isolates could be differentiated as low (4 isolates), moderately (3 isolates) and highly virulent (2 isolates) based on the different responses in infected mice. No direct correlation between severity of human scrub typhus infections and virulence of the O. tsutsugamushi in mice was observed. Mice infected with virulent strains of O. tsutsugamushi showed splenomegaly, ascitis accumulation and enlargement of kidneys and livers whereas avirulent O. tsutsugamushi strains were asymptomatic and exhibited ruffled fur for a short period after infection. There was low antibody response in mice infected with isolates of low pathogenicity as compared with those of highly virulent isolates. Upon dissection of the infected mice, enlargement of mouse organs such as spleen, kidney and liver was noted. Presence of rickettsemia in mice was confirmed by the growth of O. tsutsugamushi in the L929 cells when inoculated with blood from infected mice. O. tsutsugamushi was also cultured from the peritoneal exudates of the infected mice. However, DNA of O. tsutsugamushi was only detected in the peritoneal exudates (by PCR) and blood (by cell culture) and not from other tissue samples.
The seroprevalence of various Orientia tsutsugamushi (OT) strains among Malaysian patients with suspected scrub typhus infections was determined using an indirect immunoperoxidase (IIP) assay. IgG against a single OT strain were detected in six sera (3 Karp, 1 Gilliam and 2 TC586), whereas IgM antibodies against a single OT strain (Gilliam) were noted in 3 sera (Gilliam). IgG reactive to all OT strains were present in 33 (47.1%) of the 70 sera and IgM reactive to all OT strains were present in 22 (78.6%) of the 28 sera. The fact that most sera were reactive to multiple OT strains suggests that group-specific antigens are involved in scrub typhus infections, whereas very few were due to strain-specific epitopes present on these strains. Peak IgG and IgM titers were noted more frequently against Gilliam, Karp, and TA763 strains: this suggests that these strains may be the commonest infecting strains among Malaysian patients. Two predominant OT polypeptides consistently reacted with patients' sera were the 70 kDa and 56 kDa proteins.
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.
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.
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.