A study was carried out to investigate correlation between presence of specific microalgal species and Leptoconops biting midge larvae in its breeding habitats. Sand samples containing microalgae were collected from the beach where the midges were most commonly biting and from sand beaches which are potential as breeding habitats of Leptoconops. The survey covered sand beaches from 12 seperate islands. At all sites, the Bacillariophyta constituted the largest representatives of the microalgae community with the majority from the Naviculaceae family. A total of 24 microalgal species were identified from the sand samples collected from the study sites. Sand samples from Kentot Kecil Island had the highest number of algal species (11.0) and the highest algae species diversity ( Shanon-Weiner Diversity Index, H' = 0.884). Besar Island (Johor) had the lowest number of algal species (2.0) whereas Tengol A Island had the lowest algae species diversity (H'=0.234). Highest similarity index was recorded between sand samples collected from Tengol A Island and Tengol B Island (75.0%) followed by Besar Island (Melaka) and Tengol B Island (62.0 %). The variation between other islands were relatively high. Virtually many kinds of algae were found where Leptoconops were breeding but Fragilaria intermedia, Mastigloia minuta and Navicula advena were particularly common.
Ovitrap surveillance was conducted in four dengue endemic areas in Kuala Lumpur and Selangor, Malaysia to determine the distribution and percentage of mixed breeding of both Aedes aegypti and Aedes albopictus. The percentage of mixed breeding in all study sites both indoors and outdoors accounted for 10 to 32 % from the total ovitraps collected. Ae. aegypti was found at a higher frequency than Ae. albopictus in these ovitraps. This study again indicates that ovitrap is a sensitive tool to attract gravid females of more than one mosquito species to oviposit in the container.
The bioefficacy of a commercial formulation of temephos, Creek against Aedes aegypti larvae was studied in the laboratory. Earthen jars were filled with 10 L tap water each. One g of temephos (Creek) sand granule formulation was added into each earthen jar as recommended by the manufacturer. The final test concentration of Creek was 1 mg a.i./L. One earthen jar was filled with 10 L tap water and served as a test control (untreated). Thirty late 3(rd) or early 4(th) instar of lab-bred Ae. aegypti larvae were added into each earthen jar. Mortality of the larvae was recorded after 24 hours and percent mortality was calculated. Test was repeated every week. The results showed that complete larval mortality was achieved after 24 hours. The residual effect lasted 15 weeks (105 days), indicating that Creek is effective at the dosage recommended by the manufacturer which is 1 mg a.i./L.
Forensic entomological specimens received by the Unit of Medical Entomology, IMR., from hospitals and the police in Malaysia in the last 3 decades (1972 - 2002) are reviewed. A total of 448 specimens were received. From these, 538 identifications were made with the following results: Eighteen species of cyclorrphaga flies were identified consisting of Chrysomya megacephala (Fabricius) 215 cases (47.99%), Ch. rufifacies (Masquart) 132 (29.46%), Ch. villeneuvi Patton 10 (2.23%), Ch. nigripes Aubertin 7 (1.56%), Ch. bezziana Villeneuve 4 (0.89%), Ch. pinguis (Walker) 1 (0.22%), Chrysomya sp. 47 (10.49%), Sarcophaga sp. 28 (6.25%), Lucilia sp. 21 (4.69%), Hermetia sp. 15 (3.35%), He. illucens (Linnaeus) 1 (0.22%), Hemipyrellia ligurriens (Wiedemann) 3 (0.67%), Hemipyrellia sp. 2 (0.45%), Ophyra spinigera 1 (0.22%), Ophyra sp. 6 (1.34%), Calliphora sp. 24 (5.36%), Synthesiomyia nudiseta (Wulp) 1 (0.22%) and Eristalis sp. 1 (0.22%). Other non - fly insect specimens are Pthirus pubis (Linnaeus) (Pubic louse) 2 (0.45%) and Coleoptera (Beetles) 1 (0.22%). Ch. megacephala and Ch. rufifacies were the commonest species found in cadavers from different ecological habitats. Sy. nudiseta is an uncommon species, thus far found only on cadavers from indoors. Sy. nudiseta is reported for the second time in Peninsular Malaysia. A total of 329 cases (73.44%) had a single fly infestation, 109 cases (24.33%) had double fly infestation and 10 cases (2.23%) had triple fly infestation. Five cases (1.12%) had eggs and 3 cases (0.67%) had larval stages that were not identifiable. No arthropods were retrieved from cadavers in 8 cases (1.79%). In conclusion, although large number of fly species were found on human cadavers, the predominant species are still those of Chrysomya.
The rapid detection of dengue infection in mosquito vectors is important for early warning to forestall an outbreak. Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) provides a rapid method for dengue detection in man and mosquitoes. An RT-PCR kit developed by the Medical Entomology Unit, Institute for Medical Research to detect dengue infection in mosquitoes, was tested for its shelf life at 3 storage temperatures: room temperature, refrigerator and freezer. Test kits were tested once every 3 days for kits stored at room temperature, and once every week for those stored at refrigerator and freezer temperatures. The results showed that the test kit could only be stored above its recommended storage temperature of -20 degrees C for not more than 3 days. DNA 100 bp markers in the kits appeared to be stable at the tested temperatures and were usable up to the 20th day when stored at 2 degrees C and below.
The susceptibility of Culex quinquefasciatus to chemical insecticides in two field sites in Kuala Lumpur was evaluated using the WHO standard susceptibility test. Less then 7 days old female mosquitos, reared from wild caught females were exposed to discriminating dosages of insecticides at recommended exposure periods. The larval bioassay were conducted using the multiple concentrations and the LC50 value was determined. The results indicated that cyfluthrin is the most effective among all the insecticides tested with LT50 value of 29.95 min and 28.59 min, for the strain from Ampang Hill and Pantai Dalam, respectively. It was surprisingly to note that both these field strains showed 0% mortality when tested against malathion and DDT. The LC50 value indicated that both strains were highly resistant to malathion with resistance ratio of 17,988 folds and 14,053 folds, respectively. This concludes that resistance at larval stages is extremely high compared to adult stages.
In order to control any pest it is essential to study the life cycle, biology and bionomics of the target pest under control. With this respect, we have studied the flight range of the house fly Musca domestica (L.). The flight range of the house fly from two sites i.e, the poultry farm and a stable farm has been studied. The flight range study was conducted using a mark release technique. The approach we used in this study was that the flies collected from the respective farms were marked and released at different distances from the farms. The flies were then re-captured from the poultry farm and the stable farm. Studies conducted elsewhere use the technique of releasing the insect species at one spot and recapturing the insect species with the help of baited traps placed at various locations from the release point. The advantage of the approach used in this study was that the flight range as well as the homing effect was determined. From this study, the flight range of house flies released at the poultry farm was 7 km whereas flight range for flies release from stable farm was 5 km. The recovery rate of house flies at the poultry and stable farm was 0.05% and 0.016%, In this study, marked specimens has been detected up to 8 days in field conditions indicating that under field condition the life expectancy could be in the range of 1-2 weeks.
The hornets are a group of venomous stinging insects that at times cause human death. A fatal case of a child stung by the lesser banded hornet Vespa affinis indosinesis is reported. Though often covered by the mass media, this constitutes the first scientifically reported case.
To determine resistance level and characterize malathion and permethrin resistance in Culex quinquefasciatus, two methods were used namely: WHO procedures of larval bioassay to determine the susceptibility of lethal concentration (LC) and adult bioassay to determine the lethal time (LT) which are resistant to malathion and permethrin. These mosquito strains were bred in the Insectarium, Division of Medical Entomology, IMR. Thousands of late fourth instar larvae which survived the selection pressure to yield 50% mortality of malathion and permethrin were reared and colonies were established from adults that emerged. Larvae from these colonies were then subjected to the subsequent 10 generations in the test undertaken for malathion resistant strain (F61 - F70) and permethrin resistant strain (F54 - F63). Selection pressure at 50% - 70% mortality level was applied to the larvae of each successive generation. The rate of resistance development and resistance ratio (RR) were calculated by LC5 0 for larval bioassay and LT50 value for adult bioassay. The lab bred Cx. quinquefasciatus was used as a susceptible strain for comparison purpose. The adult bioassay test was carried out by using diagnostic dosages of malathion 5.0%, permethrin 0.75% and with propoxur 0.1%. All bioassay results were subjected to probit analysis. The results showed that LC5 0 for both malathion (F61 - F70) and permethrin (F54 - F63) resistant Cx. quinquefasciatus increased steadily to the subsequent 10 generations indicating a marked development of resistance. The adult female malathion resistant strain have developed high resistance level to malathion diagnostic dosage with resistance ratio 9.3 to 9.6 folds of resistance. Permethrin resistance ratio remained as 1.0 folds of resistance at every generation. It was obvious that malathion resistance developing at a higher rate in adult females compared to permethrin. Female adults exposed to 2 hours of exposure period for propoxur 0.1% showed presence of cross-resistance among the both strains of mosquitoes towards propoxur and it was indicated by 70%-100% mortality at 24 hours post-recovery period.
Detail chemical investigations on the stem bark of Mesua daphnifolia gave three triterpenoids and four xanthones. They are friedelin (1), friedelan-1,3-dione (2), lup-20(29)- en-3ss-ol (3), cudraxanthone G (4), ananixanthone (5), 1,3,5-trihydroxy-4-methoxyxanthone (6) and euxanthone (7). These chemical constituents were tested in vitro for their cytotoxic activities against four cell lines, MDA-MB-231 (human estrogen receptor negative breast cancer), HeLa (cervical carcinoma), CEM-SS (T-lymphoblastic leukemia) and CaOV3 (human ovarian cancer). Compound 4 showed a broad spectrum of activity against the MDA-MB-231, HeLa and CEM-SS cell lines with IC5 0 values of 1.3, 4.0 and 6.7 microg/ml respectively. Meanwhile, the other compounds 1, 2, 3, 5, 6 and 7 gave only selective activities against the cell lines.
A study of chikungunya virus was carried out to establish Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR) as a rapid detection technique of the virus. The susceptibility of lab-colonized Aedes aegypti to chikungunya virus was also determined. Artificial membrane feeding technique was used to orally feed the mosquitoes with a human isolate of chikungunya virus. A total of 100 fully engorged female Ae. aegypti were obtained and maintained for 7 days. Seventy of them survived and then pooled at 10 individuals per pool. Total RNA was extracted from the samples and RT-PCR amplifications were carried out. Five out of 7 pools showed positive PCR band at 350-bp, indicating Ae. aegypti is a potential vector of chikungunya virus. The minimum infection rate (MIR) was 71% within these laboratory colonies. RT-PCR is a sensitive technique that is useful in detecting infected mosquitoes in epidemic areas. This technique can de used as a rapid detection method and provide an early virologic surveillance systems of chikungunya virus infected mosquitoes.
Field bioefficacy of residual-sprayed deltamethrin against Aedes vectors was evaluated in an urban residential area in Kuala Lumpur. The trial area consisted of single storey wood-brick houses and a block of flat. The houses were treated with outdoor residual spraying while the flat was used as an untreated control. Initial pre-survey using ovitrap surveillance indicated high Aedes population in the area. Deltamethrin WG was sprayed at a dosage of 25mg/m2 using a compression sprayer. The effectiveness of deltamethrin was determined by wall bioassay and ovitrap surveillance. The residual activity of 25mg/m2 deltamethrin was still effective for 6 weeks after treatment, based on biweekly bioassay results. Bioassay also indicated that both Aedes aegypti and Aedes albopictus were more susceptible on the wooden surfaces than on brick. Aedes aegypti was more susceptible than Ae. albopictus against deltamethrin. Residual spraying of deltamethrin was not very effective against Aedes in this study since the Aedes population in the study area did not reduce as indicated by the total number of larvae collected using the ovitrap (Wilcoxon Sign Test, p> 0.05). Further studies are required to improve the effectiveness of residual spraying against Aedes vectors.
Ticks are vectors of bacteria, protozoa and viruses capable of causing serious and life threatening diseases in humans and animals. Disease transmission occurs through the transfer of pathogen from tick bites to susceptible humans or animals. Most commonly known tick-borne pathogens are obligate intracellular microorganisms but little is known on the prevalence of culturable pathogenic bacteria from ticks capable of growth on artificial nutrient media. One hundred and forty seven ticks originating from dairy cattle, goats and rodents were collected from nine selected sites in Peninsular Malaysia. The culture of surfacesterilized tick homogenates revealed the isolation of various pathogenic bacteria including, Staphylococcus sp., Corynebacterium sp., Rothia sp., Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli and Bacillus sp. and its derived genera. These pathogens are among those that affect humans and animals. Findings from this study suggest that in addition to the regular intracellular pathogens, ticks could also harbor extracellular pathogenic bacteria. Further studies, hence, would be needed to determine if these extracellular pathogens could contribute to human or animal infection.
The mechanism of insecticide resistance is traditionally attributed to detoxification enzymes, target site alteration, decreased penetration of insecticides and behavioural resistance. Other form of mechanisms, such as the role of protein(s) in resistance is unknown. In the present study, the protein profiling of both IMR-PSS strain (permethrin-selected) and IMR-LS strain (laboratory-susceptible) 24 hours post exposure period to permethrin was carried out via 1D-gel electrophoresis and liquid chromatography mass spectrometry (LC-MS/ MS). The bands which appeared in the gel of 1D-electrophoresis revealed an abundance of proteins. The band pattern of both strains looked macroscopically alike and differed only in band intensity. However, LC-MS/MS analysis revealed that the IMR-PSS strain produced extra 388 peptides that were not found in the IMR-LS strain, indicating that IMR-PSS strain reacted differently from IMR-LS strain as a result of persistent exposure to permethrin. Since the complex banding patterns of 1D-gel electrophoresis were difficult to interpret the significance of the protein difference between IMR-PSS and IMR-LS strain, hence LC-MS/MS analysis is ideally suited for better protein resolution and thus will allow more in-depth comparison of the complex pattern. The findings here provide the first preliminary evidence that insecticide resistance in mosquito induces up regulation of proteins that may be protective to mosquitoes against insecticide and proteins could be another mechanism that contributes to development of resistance.
The continued absence of an effective and safe tetravalent dengue vaccine and the lack of specific anti-viral treatment have made mosquito vector control using chemical insecticides as the mainstream for dengue prevention and control. However, the long-term use of chemical insecticides may induce resistance. Hence detection of insecticide resistance in dengue vectors is crucially important in ensuring the insecticide-based intervention in dengue control program is still effective and reliable. In this study, the susceptibility status of Aedes aegypti from five selected dengue hotspots in Klang Valley, Malaysia against pyrethroids was determined by employing the World Health Organization (WHO) protocol of adult bioassay. Four types of pyrethroids were tested against adult female Aedes aegypti to determine the knockdown rate, post 24-h adult mortality and resistance ratio. All field-collected Aedes aegypti strains were resistant to the four pyrethroids tested, except for the Taman Sungai Jelok (TSJ) strain. Permethrin exhibited the lowest knockdown rate against Aedes aegypti, followed by deltamethrin, cyfluthrin and lambda-cyhalothrin. This present study indicated the widespread of pyrethroid resistance in Aedes aegypti in Klang Valley, indicating the needs of implementing alternative measures in vector control program. The data in this study can be utilised as an input for insecticide resistance management of Aedes aegypti in Malaysia.
This study was carried out to determine from bacterial profiling to the bacterial profiles of head lice among the Orang Asli communities. The head lice were collected from Orang Asli community volunteers. The surface sterilized head lice pools were subjected to genomic DNA extraction while next generation sequencing of the 16S rRNA gene was performed using the Illumina MiSeq platform. Six female and three male head lice identified as Pediculus humanus capitis were collected. A total of 111 368 number of NGS sequencing reads were recorded while another 223 bacterial taxa sequences were obtained. Symbiotic bacteria showed the highest number of reads, with Arsenophonus and Rhodococcus sequences being the most abundant genera in the female and male samples, respectively. The female head lice contained a more distinct microbial diversity. Amongst the pathogenic bacterial species sequences noted were the methicillin-resistant Staphylococcus aureus, Streptobacillus moniliformis, Haemophilus influenzae, Bordetella pertussis and Acinetobacter baumannii. The 16S rRNA genome sequencing revealed a number of rare and pathogenic bacterial species within the head lice of the Orang Asli. The socio-economic practices of the community which involved forest foraging and hunting, and their poor living conditions potentially facilitated the transmission of zoonotic bacterial pathogens, including those found within the head lice. Hence, there is the possibility that the head lice could serve as vectors for the transmission of pathogenic bacteria. This study highlighted the diverse microbial community found within the head lice's gut of the Orang Asli, with the detection of multiple rare and pathogenic bacteria capable of causing severe infections.
Susceptibility status of Aedes albopictus (Skuse) sampled from residential areas in Interior, Sandakan and Tawau divisions of Sabah, Malaysia, was evaluated based on the WHOrecommended doses of organochlorine and organophosphate larvicides. To determine susceptibility status, larval bioassays were carried out and post 24-hour mortalities based on WHO resistance classifications were adopted. The results demonstrated that Ae. albopictus larvae were resistant toward 5 out of the 8 larvicides tested. Larvae from all populations were resistant against bromophos, fenitrothion, malathion, temephos and dichlorodiphenyltrichloroethane (DDT), with mortalities ranging from 0.00 to 89.33%. Dieldrin, on the other hand, could induce 100.00% mortalities in all populations, followed by fenthion and chlorpyrifos, with mortalities ranging from 97.33 to 100.00% and 81.33 to 100.00% respectively. Despite most populations exhibiting similitude in their resistance status, larvae from Sandakan exhibited the highest resistance level whereas the lowest level was observed in Keningau. In view of the inadequacy of some larvicides in controlling Ae. albopictus in this study, integrated management such as insecticide rotation or combination of interventions is warranted.
The spraying of insecticide on foliage to provide an insecticidal barrier may serve as part of the vector control measures to combat the increasing threat of Aedes-borne diseases. The effectiveness of insecticide barrier spraying was evaluated by assessing the residual efficacy of deltamethrin sprayed on foliage against Malaysian Ae. aegypti (L.) and Ae. albopictus (Skuse). In this semi-field study, landscape plants grown within the vicinity of the Institute for Medical Research (IMR), Malaysia, were treated with deltamethrin suspension concentrate (SC) with the dosage of 30 mg/m2 and 50 mg/m2 in three rounds of spraying. Deltamethrin residual activity on treated and untreated leaves was investigated using standard WHO cone bioassays. Wild Aedes populations at both deltamethrin-treated and untreated plant clusters were monitored by ovitrap surveillance. Ovitrap monitoring revealed that the mean number of Ae. albopictus larvae at deltamethrin-treated were significantly lower than the mean number of larvae of the same species at the untreated plant cluster. Cone bioassay results showed that the insecticide remained effective for up to 4 weeks (> 80% mortality), but the insecticide residual activity was affected by rainfall. These results suggest that insecticide barrier spraying is a promising tool and may be used along with other mosquito control tools such as indoor residual spray and space spraying to reduce the dengue burden.
Diseases such as malaria, dengue, Zika and chikungunya remain endemic in many countries. Setting and deploying traps to capture the host/vector species are fundamental to understand their density and distributions. Human effort to manage the trap data accurately and timely is an exhaustive endeavour when the study area expands and period prolongs. One stop mobile app to manage and monitor the process of targeted species trapping, from field to laboratory level is still scarce. Toward this end, we developed a new mobile app named "PesTrapp" to acquire the vector density index based on the mobile updates of ovitraps and species information in field and laboratory. This study aimed to highlight the mobile app's development and design, elucidate the practical user experiences of using the app and evaluate the preliminary user assessment of the mobile app. The mobile app was developed using mobile framework and database. User evaluation of the mobile app was based on the adjusted Mobile App Rating Scale and Standardized User Experience Percentile Rank Questionnaire. The process flows of system design and detailed screen layouts were described. The user experiences with and without the app in a project to study Aedes surveillance in six study sites in Selangor, Malaysia were elucidated. The overall mean user evaluation score of the mobile app was 4.0 out of 5 (SD=0.6), reflects its acceptability of the users. The PesTrapp, a one-stop solution, is anticipated to improve the entomological surveillance work processes. This new mobile app can contribute as a tool in the vector control countermeasure strategies.