Iranihindia martellata (Senior-White, 1924) is recorded from peninsular Malaysia for the first time. Male and female specimens in the recent collections of forensically important sarcophagid flies were examined and identified based on morphology and DNA sequencing analysis. Male genitalia offer unambiguous species identification characteristics in the traditional taxonomy of flesh flies but the female flies are very similar to one another in general morphology. Female of I. martellata was determined by DNA sequencing (COI and COII) and PCR-RFLP (COI) analysis. Identified females were carefully examined and compared with the morphologically similar species, Liopygia ruficornis (Fabricius, 1794). Female genitalia are re-described and illustrated in this paper.
We described a new species of cockroach, Periplaneta gajajimana sp. nov., which was collected in Gajajima, Kagoshima-gun Toshimamura, Kagoshima Prefecture, Japan, on November 2012. The new species is characterized by its reddish brown to blackish brown body, smooth surface pronotum, well developed compound eyes, dark brown head apex, dark reddish brown front face and small white ocelli connected to the antennal sockets. In male, the tegmen tip reach the abdomen end or are slightly shorter, while in the female, it does not reach the abdominal end and exposes the abdomen beyond the 7th abdominal plate. We confirmed the validity of this new species by breeding the specimens in our laboratory to demonstrate that the features of the progeny were maintained for several generations. For comparison and easy identification of this new species, the key to species identification of the genus Periplaneta that had been reported in Japan to date are also presented.
Beta toxin (CPB) is a lethal toxin and plays a key role in enterotoxemia of ruminants caused by Clostridium perfringens type C strain. The existing vaccines based on crude CPB need time-consuming detoxification and difficult quality control steps. In this study, we synthesized the rCPBm4 of C. perfringens type C strain and small ubiquitin-like modifier (SUMO)-tag CPBm4 (rSUMO-CPBm4) by introducing four amino acid substitutions: R212E, Y266A, L268G, and W275A. Compared with rCPBm4, rSUMO-CPBm4 was expressed with higher solubility in Escherichia coli BL21 (DE3). Neither rCPBm4 nor rSUMO-CPBm4 was lethal to mice. Although rCPBm4 and rSUMO-CPBm4 were reactogenic with polyclonal antibodies against crude CPB, rabbits vaccinated with rSUMO-CPBm4 developed significant levels of toxin-neutralizing antibody (TNA) titers that conferred protection against crude toxin challenge. These data suggest that genetically detoxified rSUMO-CPBm4 is a promising subunit vaccine candidate for C. perfringens type C beta enterotoxemia.
Three-dimensional (3D) models of the marginal hooks, dorsal and ventral anchors, bars and haptoral reservoirs of a parasite, Sundatrema langkawiense Lim & Gibson, 2009 (Monogenea) were developed using the polygonal modelling method in Autodesk 3ds Max (Version 9) based on two-dimensional (2D) illustrations. Maxscripts were written to rotate the modelled 3D structures. Appropriately orientated 3D haptoral hard-parts were then selected and positioned within the transparent 3D outline of the haptor and grouped together to form a complete 3D haptoral entity. This technique is an inexpensive tool for constructing 3D models from 2D illustrations for 3D visualisation of the spatial relationships between the different structural parts within organisms.
A survey of ectoparasites was carried out during Eco-Zoonoses Expedition in Bukit Aup Jubilee Park (BAJP), Sibu, Sarawak, Malaysian Borneo from 5(th) to 9(th) June 2008. A total of nine individuals comprising two species of rodents were captured. The species of rodents screened for ectoparasites were Sundamys muelleri and Callosciurus notatus. Four genera and six species of ectoparasites were collected, namely, Ixodes granulatus, Ixodes sp., Laelaps sedlaceki, Laelaps nuttalli, Hoplopleura dissicula and Listrophoroides sp. Three species of the ectoparasites are known to have potential health risk. The species were Ixodes granulatus, Laelaps nuttalli and Hoplopleura dissicula. This survey produced the first list of ectoparasites in Bukit Aup Jubilee Park, Sarawak, Malaysia.
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.
This study reported the ant species that were recovered from monkey carcasses in three different ecological habitats in Malaysia. The study was conducted from 9 May - 10 October 2007, 6 May - 6 August 2008 and 26 May - 14 July 2009 in forested area (Gombak, Selangor), coastal area (Tanjong Sepat, Selangor) and highland area (Bukit Cincin, Pahang), respectively. Monkey carcass was used as a model for human decomposition in this study. A total of 4 replicates were used in each of the study sites. Ants were observed to prey on eggs, larvae, pupae and newly emerged flies. This study found that ant species could be found at all stages of decomposition, indicating that ants were not a significant indicator for faunal succession. However, different species of ants were obtained from monkey carcasses placed in different ecological habitats. Cardiocondyla sp. was only found on carcasses placed in the coastal area; while Pheidole longipes, Hypoponera sp. and Pachycondyla sp. were solely found on carcasses placed in the highland area. On the other hand, Pheidologeton diversus and Paratrechina longicornis were found in several ecological habitats. These data suggests that specific ant species can act as geographic indicators for different ecological habitats in forensic entomology cases in Malaysia.
This preliminary study was carried out in a palm oil plantation in Tanjung Sepat, Selangor in 17 May 2007 by using pig (Sus scrofa) as a carcass model in forensic entomological research. A 3 month old pig (8.5 kg) that died of pneumonio was placed in the field to observe the decomposition stages and the fauna succession of forensically important flies. Observation was made for two weeks; two visits per day and all climatological data were recorded. The first visitor to the pig carcass was a muscid fly, seen within a minute, and followed by ants and spiders. Within half an hour, calliphorid flies came over. On the second day (fresh), few calliphorid and sarcophagid flies were found on the carcass. Two different species of moths were trapped in the hanging net. The first larva mass occurred on the third day (bloated) around the mouthpart, with some L1 and L2 found in the eyes. Reduvid bugs and Staphylinidae beetles were recovered on the fourth day (active decay), and new maggot masses occurred in the eyes and anus. L3 larvae could be found beneath the pig carcass on the fourth day. On the fifth day (active decay), new maggot masses were found on neck, thorax, and hind legs. Advance decay occurred on the sixth day with abundant maggots covering all over the body. The main adult fly population was Chrysomya megacephala (day 2 to day 6), but the larvae population was mainly those of Chrysomya rufifacies (day 4 to day 14). The dry stage began on the eighth day. Hermetia illucens adult was caught on day-13, and a larvae mass of Chrysomya rufifacies was seen burrowing under the soil. This forensic entomological research using pig carcass model was the first record in this country.
Vegetative proteins from Malaysian strains of Bacillus thuringiensis israelensis strains (Bt 11, Bt 12, Bt 15, Bt 16, Bt 17, Bt 21 and Bt 22) and Bacillus sphaericus H-25 strains (Bs 1 and Bs 2) were screened for haemolytic, cytotoxic and larvicidal activity. SDS-PAGE profiles of the Bacillus thuringiensis strains studied consistently showed major bands of 33-37 kDa and 47 kDa. Bt 16 also showed two bands of 66 kDa and 45 kDa similar to the previously reported binary vegetative protein, Vip1Ac (66 kDa) and Vip 2Ac (45 kDa). Both the Bacillus sphaericus strains showed a 35 kDa band that was similiar to a previously reported vegetative protein, the Mtx2 protein. Bs 2 also contains a 37 kDa band, similar to another vegetative protein, the Mtx 3 protein. With the exception of Bt 17 and Bt 21, vegetative proteins from all Bacillus thuringiensis and Bacillus sphaericus strains were highly haemolytic to human erythrocytes, causing more than 75% haemolysis at the highest concentration of 200 microg/ml. High haemolytic activity was associated with high cytotoxic activity with most of the haemolytic strains being indiscriminately cytotoxic to both CEM-SS (human T lymphoblastoid) and HeLa (human uterus cervical cancer) cell lines. Interestingly, the less haemolytic vegetative proteins from Bt 17 and Bt 21 demonstrated cytotoxic activity comparable to that of the highly haemolytic vegetative proteins. Bt 21 displayed toxicity towards both cell lines while Bt 17 was more toxic towards CEM-SS cells. Bioassay against Aedes aegypti and Culex quinquefasciatus larvae revealed that vegetative proteins from the Bacillus thuringiensis strains had activity against both species of larvae but vegetative proteins from Bacillus sphaericus were weakly larvicidal towards Cx. quinquefasciatus only.
This study was carried out in an oil palm plantation in Bandar Baharu, Kedah using monkey carcasses and focuses in documenting the decomposition and dipteran colonization sequences in 50 days. This is the first study of Diptera associated with the exploitation of carcasses conducted in the north of peninsular Malaysia during the dry and wet seasons thereat. During the process of decomposition in both seasons, five phases of decay were recognized namely fresh, bloated, active decay, advance decay and dry remain. In this decomposition study, biomass loss of carcass occurred rapidly during the fresh to active decay stage due to the colonization and feeding activity of the Diptera larvae. The duration of the fresh and bloated stages of decay were the same in wet and dry seasons but later stages of decay were markedly shorter during the wet season. Twenty one species of adult Diptera were identified colonizing carcasses in the study period. Among the flies from the family Calliphoridae, Chrysomya megacephala Fabricius and Chrysomya nigripes Aubertin were recognized as the earliest arrivals on the first day of exposure. Adult Ch. nigripes was abundant for approximately two weeks after placement of the carcasses. By comparing the percentages of adults collected during the study period, the calliphorids abundance in percentages in wet season was 50.83%, but in dry season, the abundance was only about 35.2%. In contrast, the percentage of Sphaeroceridae in wet season was only 3.33%, but in the dry season, the abundance was 20.8%. Dipteran in family Phoridae, Piophilidae, Sepsidae, Drosophilidae and Dolichopodidae colonized the carcasses for a long period of time and were categorized as long term colonizers.
Trapping of small mammals was conducted at 5 study sites in Kuala Selangor Nature Park (KSNP) from 20-24 June 2005. A total of 11 animals comprising 2 species of rodents, Maxomys whiteheadi and Rattus exulans were caught from 3 sites, i.e from an area of mixed secondary forest and mangrove swamp; an area of mangrove swamp, and from an area of lalang fringing mangrove swamp. From these animals, the following 7 species of ectoparasites were found: Laelaps echidninus, Laelaps nuttalli, Ascoschoengastia indica, Leptotrombidium deliense, Hoplopleura pectinata, Hoplopleura pacifica and Polyplax spinulosa. One of the ectoparasites found, L. deliense is a known vector of scrub typhus and thus may pose potential health risks to visitors to KSNP.
Shigellosis is a foodborne illness caused by the genus Shigella and is an important global health issue. The development of effective techniques for rapid detection of this pathogen is essential for breaking the chain of transmission. Therefore, we have developed a novel loop-mediated isothermal amplification (LAMP) assay targeting the invasion plasmid antigen H (ipaH) gene to rapidly detect Shigella species. This assay could be performed in 90 min at an optimal temperature of 64ºC, with endpoint results visualized directly. Notably, the method was found to be more sensitive than conventional PCR. Indeed, the detection limit for the LAMP assay on pure bacterial cultures was 5.9 x 10(5) CFU/ml, while PCR displayed a limit of 5.9 x 10(7) CFU/ml. In spiked lettuce samples, the sensitivity of the LAMP assay was 3.6 x 10(4) CFU/g, whereas PCR was 3.6 x 10(5) CFU/g. Overall, the assay accurately identified 32 Shigella spp. with one enteroinvasive Escherichia coli displaying positive reaction while the remaining 32 non-Shigella strains tested were negative.
The pupae of Desmometopa sp. (Diptera: Milichiidae) were collected from a human corpse found indoor in active decay stage together with the larvae of Sarcophagidae, Synthesiomyia nudiseta (Wulp), Chrysomya megacephala (Fabricius) and Chrysomya rufifacies (Macquart). This research note is the first report of the Desmometopa sp. recovered from a human corpse in Malaysia.
Some 2,000 species of mites of the family Trombiculidae are known in the world. The 6-legged larvae are mostly ectoparasites of reptiles, birds, mammals and invertebrates. Their 8-legged active nymphs and adults are free-living predators. In the Asia-Pacific region, a few species in various genera are vectors of scrub typhus and scrub-itch. In this a paper, a very bizarre trombiculid species, Vatacarus ipoides Southcott 1957, endoparasitic in the trachea of the amphibious sea snake, Laticauda colubrina (Schenider) is re-described based mostly on new-born larvae reared in the laboratory. Life history study of the mite produced very novel and interesting results. A brief account of the life-cycle was presented at the first laboratory demonstration of the Malaysian Society of Parasitology and Tropical Medicine Meeting by Nadchatram and Audy (1965). The life history is illustrated and described here in greater detail. The active nymphal, and the akinetic teleiophane stages are bypassed, which is unusual in the life-cycle of the family Trombiculidae. Also, the larva is the only stage in the life-cycle that feeds. The sexes are predetermined in the larval neosomatic stage and give rise to small males and bigger females. Having obtained adults of the species, by rearing, it is deemed unnecessary for the original proposal by Southcott to erect a new family, Vatacaridae, because the adults share all the attributes of the family Trombiculidae. The male and female obtained through laboratory rearing are illustrated for the first time. Relationship of V. ipoides with Laticauda snakes show close host-specificity, in a group of acarines that are generally habitat-specific. Possible explanations for their association are discussed. The unusual morphology and the formation of new structures during an instar is of ontogenetic and evolutionary importance. The hypertrophic larvae are superficially vermiform, rather than typically acarine in shape. This, and other biological features, necessitated the proposal of new morphological terms, and they are discussed here.
This paper reviews the literature on human leptospirosis in Malaysia from its first description in 1925 until the present day. Fletcher diagnosed the first case of human leptospirosis in Malaysia in 1925. Following Fletcher, many investigations on human leptospirosis in Malaysia disclosed a high prevalence of infection. These investigations indicated that the disease was endemic in the country. Examination of 1993 suspected human cases of leptospirosis by Tan indicated 28 % of the cases were positive. In a recent survey, 2190 serum samples from patients with different clinical manifestations in the country disclosed 12.6% were positive for antibodies to leptospires. The risk to leptospiral infection with respect to occupation, location, sex, race and age groups was demonstrated. Both civilians and military personnel were affected. Thirty-seven serovars from thirteen serogroups have been identified in the country. Recent studies on animal leptospirosis showed that the disease was highly endemic in the animal population. It is considered that the majority of leptospirosis cases in humans were due to association of man with animals and disease-infected environment.
Clinical utilization of carbapenems remains under threat with the emergence of acquired carbapenemase-producing bacteria, particularly metallo-β-lactamases (MBL). Rapid detection of MBL-producing Gram-negative bacilli is essential to prevent their widespread dissemination. However, no standardized detection method is available for routine laboratory use. The purpose of the study was to evaluate a chelating-agent based double disk synergic test and disk potentiation test for MBL-producing strain detection and to determine the isolation rate of MBL-producing Pseudomonas aeruginosa and Acinetobacter from clinical samples in our tertiary teaching hospital. A total of 22 and 66 imipenem-resistant P. aeruginosa and Acinetobacter isolates respectively were tested with ceftazidime (CAZ) disk by modified double disk synergic test and disk potentiation test using ethylenediaminetetraacetic acid (EDTA) and 2-mercaptopropionic acid (as chelating agents) to detect MBL production. The tests were compared with EDTA-phenanthroline-imipenem (EPI) microdilution MIC test as gold standard. MBL positive strains were detected in 17 (77.3%) P. aeruginosa and 2 (3.5%) Acinetobacter isolates. The disk potentiation test with 2-mercaptopropionic acid (2-MPA) dilution of 1:12 provided the most acceptable sensitivities and specificities (88.2% sensitivity and 100% specificity in P. aeruginosa; 100% sensitivity and specificity in Acinetobacter) compared to other screening methods used in this study. This study provided useful information on the local prevalence of MBL-producing P. aeruginosa and Acinetobacter in our hospital. Disc potentiation test with CAZ/2-MPA disc appears to be reliable and convenient MBL detection method in the routine clinical laboratory.
Appendicitis has a worldwide prevalence and affects all age groups. The aetiology of acute appendicitis is still much debated, many factors have been implicated. The pathology is likely to be due to obstruction of the lumen of the appendix. Parasites, both helminths and protozoa have been suggested to be the cause of acute appendicitis. Studies have demonstrated that parasites are present in the appendix specimens removed from surgery methods.
Amoebic serodiagnosis at Hospital Universiti Sains Malaysia (HUSM), Kelantan employs an indirect haemagglutination assay (IHA) which detects anti-Entamoeba histolytica antibodies in patients' serum samples. In an amoebiasis endemic area such as Kelantan, interpretation of a positive IHA result can be problematic due to the high background antibody levels. The TechLab E. histolytica II ELISA is a commercial kit for detection of specific Gal/GalNAc lectin antigen in stool samples, and has been reported to be able to detect the antigen in serum samples from patients with amoebic liver abscess (ALA). Thus in this study we investigated the usefulness of TechLab E. histolytica II ELISA for diagnosis of ALA by comparing it with IHA. This is a cross sectional study involving 58 suspected ALA patients who were admitted to the surgical ward, HUSM, Kelantan. The diagnosis of ALA was established based on clinical symptoms and signs, ultrasound and/or CT scan results. The serum specimens obtained from the patients were tested with IHA (Dade Behring Diagnostics, Marburg, Germany) and TechLab E. histolytica II ELISA (Techlab, Blacksburg, Virginia, USA) according to the manufacturers' instructions. Of the 58 patients, 72.4% (42) were positive by IHA and only 8.6% (5) were positive by the TechLab E. histolytica II ELISA. Agreement between the IHA and ELISA was poor (kappa value 0.019, p=0.691). There was also no correlation between ELISA results and IHA antibody titers. The TechLab E. histolytica II ELISA was not sensitive in detecting amoebic antigen in samples from ALA patients. In addition the results of the test did not correlate with the IHA anti-E. histolytica antibody titres. Therefore, the TechLab E. histolytica II ELISA was found not to be useful for serological diagnosis of ALA at HUSM.
A survey of Caseous Lymphadenitis (CLA), a bacterial infection in sheep and goats was conducted on small ruminant farms in two districts in Perak, namely Kinta and Hilir Perak. The objective of this survey is to determine the status of CLA infection in small ruminants. A total of 8 farms were screened, involving a total of 579 animals. Agar Gel Precipitation Test (AGPT) and Enzyme Linked Immuno Absorbent Assay (ELISA) were conducted on serum samples obtained from the animals. Results show that 8.5% of the animals had a positive reaction for AGPT test. It was found that 36 samples (17%) were found positive using both AGPT and ELISA methods, 9 samples (4%) were found positive only using AGPT method, 14 samples (6%)were found positive only using ELISA and 157 samples (73%) were found negative using both methods. Since there is no available data on the prevalence of the disease in the country, further epidemiological studies as well as reliable diagnostic detection methods need to be assessed for aiding in control and eradication programmes for this disease.
This paper reports the occurrence of helminth and protozoan infections on small ruminants from eight farms situated in Kinta and Perak Tengah district, Perak. The results of this survey indicate that helminthiasis and coccidiosis is rampant in sheep and goat farms. Several anthelmintics have been used for the control of helminths. The smallholders depended on health and extention services from the State Veterinary Department. This survey is part of an ongoing programme by the Department of Veterinary Services to upgrade services and report the current status of parasitic diseases in the state.