The 'Dextran' and the 'Buffy-coat' methods for isolation of human leucocytes for DNA extraction were compared on the basis of DNA yield from the same amounts (10 ml) of blood. Human leucocytes from a total of 11 samples were isolated using both methods for each sample after which DNA was extracted. Extracted DNA samples were treated with ribonucleases and proteinase K after which the yields were quantitated by measuring absorbance at 260 nm. The 'Buffy-coat' method yielded a mean concentration of DNA of 476.7 micrograms/ml (range: 212 to 700 micrograms/ml) while the 'Dextran' method yielded 188.4 micrograms/ml (range: 64 to 340 micrograms/ml). The difference was confirmed by subjecting the extracted DNA samples to agarose gel electrophoresis.
The Programme on Prevention and Management of Obesity in Preschool Children, aged three to six years, was implemented in 17 Primary Health Clinics in November 1991. The study sample comprised 1128 preschool children who qualified to enter the obesity register, using the defined criteria for obesity of 2 standard deviations above the normal weight for height and age. This group was divided into three categories, namely, mildly overweight (120% to below 140%), moderately overweight (140% to below 160%), and severely overweight (above 160%). The severely overweight category was referred to dietitians for follow-up management, while the other two categories were managed by the clinic staff through pre-planned nurse-conducted counselling sessions. In this paper, we analysed the first 1128 preschool children aged three to six years on the Programme with respect to their demographic characteristics; medical conditions; family history of obesity, diabetes, hypertension and ischaemic heart disease; number of siblings and parents' educational and occupational status. The Malay children showed significantly more severe grades of obesity compared to the Chinese and Indian children. A family history of obesity and hypertension among the three groups were significant (P < 0.001). After one year of follow-up with the intervention programme, the following were found: 40.4% (456) of the children improved in their obesity status and 20.2% (228) reached normal status. The severe, moderate and mild categories reduced from 6.3% to 5.9%, 29.3% to 23.2% and 64.4% to 50.7% respectively and was found to be statistically significant (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
Mosquito adults and larvae were collected from dengue high risk areas and transported to the laboratory for identification. Identified mosquitos were pooled according to the species, date and locality and stored at -70 degrees C. A total of 1,385 pools of Aedes albopictus and 267 pools of Ae. Aegypti were collected from major towns in 12 states in Peninsular Malaysia. Virus isolation was carried out using cell culture (C6/36 clone) of Ae. albopictus and detection of dengue virus by the peroxidase anti-peroxidase staining. All positive isolations were further re-confirmed by the reverse transcriptase-polymerase chain reaction (RT-PCR). Most of the pools were negative with PAP staining and RT-PCR. However, 11 mosquito pools were positive with PAP staining. On the other hand, samples from Terengganu, Pulau Pinang and Johor were positive using both methods.
The diversity of monogeneans from Southeast Asia was examined using information from the literature to show their diversity at different taxonomic (subclass, family, genera, species) levels. Knowledge of monogeneans is still incomplete in Southeast Asia and the present numbers of monogeneans are likely an underestimate of what is present on/in aquatic organisms in the region, since so few hosts have been examined. An estimate of the possible numbers of monogeneans that could be present on/in fishes and turtles in Peninsular Malaysia indicates that only 8% of the monogeneans are presently known. Analysis of the available data on monogenean diversity (or species richness) at different taxonomic levels will provide useful information on their distribution patterns. There is an uneven distribution of investigations on this topic and Malayan fauna is considered to be representative of the Southeast Asian fauna. Southeast Asian (Sundaland) monogeneans are related (at the generic level) to the monogenean fauna of South China, India and Africa.
Neopolystoma liewi sp. n. is described from the conjunctival cavity of the Malayan box turtle Cuora amboinensis (Daudin, 1802), in Peninsular Malaysia. This is the first record of Neopolystoma in Malaysia and the fourth polystomatid species described from C. amboinensis. Of the 27 Malayan box turtles examined, 8 were found to be infected. A maximum of 2 parasites per eye and 4 individuals per host was recorded. N. liewi sp. n. differs from all other members of the genus by possessing few and short genital spines and small marginal hooks. The oncomiracidium has 64 ciliated cells arranged symmetrically about the sagittal axis.
Sundapolystoma chalconotae. n. g., n. sp. (Polystomatidae, Polystomatinae) is proposed for a new polystomatid from the urinary bladder of Rana chalconota (Schlegel) in Peninsular Malaysia. This is the first species of polystomatid to be described from the amphibians of Peninsular Malaysia and the second for the Southeast Asian region. This new genus, as exemplified by S. chalconotae, differs from other polystomatids, and in particular Parapolystoma Ozaki, 1935 (P. bulliense (Johnston, 1912) Ozaki, 1935 and P. johnstoni Pichelin, 1995), in having a tubular uterus and a single diffuse testis. P. crooki Vande Vusse, 1976 is similar to S. chalconotae in having a similar type of uterus and testis, and is re-assigned as Sundapolystoma crooki (Vande Vusse, 1976) n. comb. S. chalconotae differs from S. crooki in having anchors with a longer outer root rather than a longer inner root and 7-8 genital spines compared to 9-13 in S. crooki.
This is a catalogue and discussion of the known dactylogyridean monogenean genera of siluriform fishes of the Old World. Of a total of 38 nominal genera, only 19 are considered valid. Seventeen of these 19 genera are currently in the Ancyrocephalidae (containing the Ancyrocephalinae and Ancylodiscoidinae), whilst the other two (Neocalceostoma and Neocalceostomoides) are in the Neocalceostomatidae. The 17 genera are Anchylodiscus, Ancylodiscoides, Bagrobdella, Bifurcohaptor, Bychowskyella, Chauhanellus, Cornudiscoides, Hamatopeduncularia, Mizelleus, Paraquadriacanthus, Pseudancylodiscoides, Protoancylodiscoides, Quadriacanthus, Schilbetrema, Schilbetrematoides, Synodontella and Thaparocleidus. Clariotrema Long, 1981 and Neobychowskyella Ma, Wang & Li, 1983 are considered synonyms of Bychowskyella Akhmerov, 1952, Anacornuatus Dubey, Gupta & Agarwal, 1992 is considered a synonym of Quadriacanthus Paperna, 1961, Mizellebychowskia Gupta & Sachdeva, 1990 is considered a synonym of Neocalceostoma Tripathi, 1959 and Hargitrema Tripathi, 1959 is treated as a synonym of Hamatopeduncularia Yamaguti, 1953. It is proposed that the Ancylodiscoidinae be raised to family status within the order Dactylogyridea to accommodate these 17 'ancyrocephalid' genera from siluriforms, together with Malayanodiscoides and Notopterodiscoides from notopterids. A key and the diagnostic characteristics of the 19 recognised dactylogyridean genera from catfishes plus two from notopterids, together with a list of species and synonyms, are included. New combinations made in this work are Thaparocleidus avicularia (Chen, 1987) n. comb., T. calyciflorus (Chen, 1987) n. comb., T. choanovagina (Luo & Lang, 1981) n. comb., T. dissimilis (Chen, 1988) n. comb., T. leiocassis (Reichenbach-Klinke, 1959) n. comb., T. meticulosa (Chen, 1987) n. comb., T. parasoti (Zhao & Ma, 1999) n. comb., T. persculpus (Chen, 1987) n. comb., T. valga (Chen, 1987) n. comb. and T. wulingensis (Yao & Wang, 1997) n. comb. [all from Silurodiscoides] and Bychowskyella glyptothoraci (Ma, Wang & Li, 1983) n. comb. [from Neobychowskyella].
Intestinal parasitic infection among five interior communities at Bakun Valley, upper Rejang River, Sarawak, Malaysia, was investigated as part of a public health impact assessment of the proposed US$ 3 billion Bakun Hydroelectric Project. Coproparasitological examination of 355 stool samples from 7 of 16 villages representing 5 of 7 tribes in the area revealed infection rate of 41%. A higher infection rate was found among the settled Kayans (56%) than the seminomadic Penans (29%). Infection rate was high (68%) among children less than 14 years old. Trichuris trichiura accounted for more than 90% of all infections; less common were Ascaris lumbricoides, hookworms and Strongyloides stercoralis. Polyparasitism was found in 8% of the individuals surveyed with dual infection due to T. trichiura and A. lumbricoides being more common than dual infection with T. trichiura and hookworm. Women had higher infection rates (57%) than men (33%).
Four new species of Calydiscoides Young, 1969 are described from three species of nemipterids caught off Kemaman, Terengganu, on the eastern coast of Peninsular Malaysia: C. monogrammae n. sp. from Scolopsis monogramma; C. conus n. sp. from S. magaritifer; C. scolopsidis n. sp. from S. margaritifer and S. monogramma; and C. kemamanensis n. sp. from Pentapodus setosus. The present investigation reveals that the squamodiscs (lamellodiscs) are composed of 10-12 short, complete, interlocking and concentric tubular lamellae. The innermost lamella is attached to a pair of adductor muscles.
VectoBac DT, a tablet formulation of Bacillus thuringiensis israelensis (Bti) was evaluated for the potential control of dengue vectors in various types of potable water containers. On introduction to containers, the tablet sinks to the bottom and the Bti toxins are found concentrated at the sides and the base, while the treated water column is free of Bti toxins within 24 hours after tablet introduction. In a simulated study, earthen, HDPE and plastic containers were kept covered and laboratory-bred larvae were introduced to determine the control by the tablet. The efficacy and persistence of the tablet, with a control of > 90%, was significantly longer in earthen containers in comparison to the HDPE and plastic containers. Efficacy and persistence were observed in earthen containers for a minimum period of 5.5 months (166 days) both without water replenishment and with weekly, 50% water volume, replenishment, and for a maximum period of 2.2 months (66 days) with daily, 50% water volume, replenishment. In plastic and HDPE containers, the tablet activity had a persistence of 2.1 months (63 days) without water replenishment and 1.8 months (54 days) with weekly water replenishment. The efficacy and persistence of the VectoBac DT was significantly longer in the earthen containers, with or without regularly treated water exchange, due to the Bti toxins being embedded in the porous earthen container surfaces, which protects them from rapid degradation. Lesser toxin amounts are removed from the water column during water exchange. The efficacy of VectoBac DT was also evaluated for the control of natural infestation of Aedes larvae which were resistant to temephos at the WHO diagnostic dosage of 0.012 mg/l. The tablet significantly reduced the pupal density by 8 fold in earthen containers for 67 days and 5 fold in HDPE containers for 55 days in comparison to untreated containers (p < 0.05). However, the tablet was effective for a shorter period of 25 days post-tablet-introduction due to fungal infestation in the treated plastic containers. There is a need to determine the capacity of the VectoBac DT to reduce the dengue vector population to a threshold which will prevent dengue outbreaks in dengue endemic areas.
A study of the anterior adhesive apparatus (head organs) of Bravohollisia gussevi Lim, 1995 was carried out using light and electron microscopy. The anterior adhesive apparatus or head organs in B. gussevi comprise 6 circular openings or apertures in the antero-lateral region, associated pits lined with specialized microvillous tegument that differ from the general body tegument, a bundle of ducts, and uninucleate gland cells located lateral to the pharynx. The uninucleate glands of the anterior adhesive apparatus (head organs) comprise 2 types of cells, one kind of cell producing rod-like bodies (S1) and the other oval bodies (S2). The S1 bodies are filled with numerous, less electron-dense vesicles in an electron-dense matrix, while S2 bodies have no vesicles but contain a more homogeneous electron-dense matrix. Interlinking band-like structures were observed between S1 bodies. Similar band-like structures were found between S2 bodies. The formation of S1 bodies was followed by transmission electron microscopy. However, the formation of S2 bodies was unclear and could not be resolved. Uniciliated structures were also observed around the openings of the anterior adhesive apparatus. Each uniciliated structure is usually associated with an opening of a gland cell producing granular, electron-dense, secretory bodies, which differ from the secretions produced by the lateral gland cells of the anterior adhesive apparatus.
Hard structures of helminths have often been used for taxonomic identification but are usually not clearly defined when treated with conventional methods such as ammonium picrate-glycerin for monogeneans and glycerin for nematodes. The present study reports a rapid and simple technique to better resolve the hard parts of selected monogeneans and nematodes using 5-10% alkaline sodium dodecyl sulphate (SDS). In comparison with established methods, SDS-treated worms become more transparent. In monogeneans treated with SDS, clear details of the hooks, hook filaments, anchors, bars and the sclerotized copulatory organs could be observed. In SDS-treated nematodes, spicules and ornamentations of the buccal capsules could be clearly seen.
Two new and two previously described species of diplectanid monogeneans (Heteroplectanum flabelliforme n. sp., Diplectanum sumpit n. sp., D. jaculator Mizelle & Kritsky, 1969 and D. toxotes Mizelle & Kritsky, 1969) were collected from archerfish Toxotes jaculatrix off the Island of Langkawi, Kedah and off Perak, Malaysia. The reproductive systems and squamodiscs of D. jaculator and D. toxotes are described for the first time. D. sumpit n. sp. differs from D. toxotes and D. jaculator in a having a small curved copulatory tube with a distinct accessory piece, compared to the long, tubular copulatory tube of D. jaculator and the slender tube of D. toxotes. D. sumpit n. sp. also differs from D. toxotes in having a larger ventral bar and larger squamodiscs. H. flabelliforme n. sp. differs from all known Heteroplectanum species in the shape and size of the squamodiscs, the arrangement of the sclerites in the squamodiscs, the extremely large ventral bar and the short, curved, non-spinous copulatory tube.
The bioefficacy of indoor residual-sprayed deltamethrin wettable granule (WG) formulation at 25 mg a.i./m2 and 20 mg a.i./m2 for the control of malaria was compared with the current dose of 20 mg/m2 deltamethrin wettable powder (WP) in aboriginal settlements in Kuala Lipis, Pahang, Malaysia. The malaria vector has been previously identified as Anopheles maculatus. The assessment period for the 20 mg/m2 dosage was six months, but for the 25 mg/m2 dosage, the period was 9 months. Collections of mosquitoes using the bare-leg techniques were carried out indoors and outdoors from 7:00 PM to 7:00 AM. All mosquitoes were dissected for sporozoites and parity. Larval collections were carried out at various locations to assess the extent and distribution of breeding of vectors. A high incidence of human feeds was detected during May 2005 and a low incidence during January 2005 for all the study areas. Our study showed that deltamethrin WG at 25 mg/m2 suppressed An. maculatus biting activity. More An. maculatus were caught in outdoor landing catches than indoor landing catches for all the study areas. The results indicate that 25 mg/m2 WG is good for controlling malaria for up to 9 months. Where residual spraying is envisaged, the usual two spraying cycles per year with 20 mg/m2 deltamethrin may be replaced with 25 mg/m2 deltamethrin WG every 9 months.
New conjugate vaccine for Streptococcus pneumoniae has been introduced in Malaysia recently. Information on infection due to S. pneumoniae in Malaysian children is scarce. We conducted a retrospective chart review of childhood invasive pneumococcal disease (IPD) presented to a single centre in Malaysia.
Two known and two new species of Diplectanocotyla Yamaguti, 1953 (D. gracilis Yamaguti, 1953, D. megalopis Rakotofiringa & Oliver, 1987, D. langkawiensis n. sp. and D. parva n. sp.) were collected from Megalops cyprinoides (Megalopidae) off Langkawi, Kedah and Matang, Perak, Peninsular Malaysia. All four species possess similar types of sclerotised male and female reproductive structures and similar soft anatomical features. The squamodisc sclerites of all four species have spine-like projections with varying degrees of visibility and shapes (sharp-pointed to triangular). In D. megalopis and D. langkawiensis n. sp. the spines are sharp-pointed and distinct on sclerites from rows 5-6 onwards. In D. gracilis and D. parva n. sp. the sclerite spines are triangular, lightly sclerotised and occur on almost all of the sclerites. D. parva n. sp. has comparatively the smallest set of anchors, bars, squamodiscs and squamodisc suckers. The anchors and bars of the other three species are almost similar in overall size, and the main distinguishing feature is the relative lengths of the inner and outer roots of the ventral anchors. In D. gracilis the outer root is very much smaller than the inner root and they are disposed almost at a right angle to each other. In D. megalopis the outer root is usually about half the length of the inner root and the roots are inclined at c.60 degrees to each other. In D. langkawiensis n. sp. the roots are inclined at c.40 degrees degrees and the outer root is of a similar length or only slightly shorter than the inner root. The openings of the two squamodisc suckers of all four Diplectanocotyla species are surrounded by tiny scale-like spines. Bifid tegumental spines are found in the posterior region of all four species, differing only in their extent: in D. parva n. sp. the tegumental spines are only distributed in the peduncular region and not beyond, whilst in the other three species the tegumental spines extend from the posterior level of the testis to the end of the peduncle. An amended diagnosis of Diplectanocotyla and a key to its species are appended.
Sixteen labrid species, including four Bodianus spp., were examined in New Caledonia (South Pacific) and monogeneans were found only on Bodianus perditio (Quoy et Gaimard). This species, Haliotrema banana sp. n., is the second Haliotrema species to be described from the labrids, the first being Haliotrema bodiani Yamaguti, 1968 from Bodianus albotaeniatus (Valenciennes), previously designated as B. bilunulatus (Lacépède). The new species is similar to H. bodiani in soft reproductive parts but differs from it in the morphologies of the hard haptoral parts, mainly in the shape of the dorsal bar (bar-shaped vs V-shaped in H. bodiani) and ventral bar. It is similar to Haliotrema spirale Yamaguti, 1968 and Haliotrema minutospirale Yamaguti, 1968 in the shape of the anchors and bars but differs from them in the detailed structures of the copulatory organ and vaginal system.
Numerous specimens of Ancyrocephaloides triacanthi Yamaguti, 1938 and A. chauhani Bychowsky & Nagibina, 1975 were collected from two triacanthid fishes, Triacanthus biaculeatus and Tripodichthys blochii, off Peninsular Malaysia. The two monogenean species are redescribed and considered to be the only valid species of Ancyrocephaloides Yamaguti, 1938. Examinations of these worms revealed new features, e.g. the presence of exudates (both net-like and bundle-like) and superficial grooves in the anchors in both species, which necessitated re-descriptions of the two species and amendments to the generic diagnosis. Both species have relatively small anchors with two lateral superficial grooves along the shaft and point, peduncular glands and four large, pyriform secretory reservoirs in the peduncular-haptoral region, each with a single tubular extension to an associated anchor, and net-like structures (exudate) attached to the anchors. The net-like structures are one of the external manifestations of the secretion produced in the peduncular glands and stored in the pyriform secretory reservoirs. When released within the gill-tissue of the host, the exudate is in the form of bundles which extend within the gill-filament. The small anchors convey secretions from the secretory reservoirs via lateral superficial grooves into the gills as the anchors pierce the host tissue for attachment. The secretion coagulates as left and right thread-like bundles of exudate within the gill tissues and is only apparent as nets when it is released into the surrounding water. The recurved point of the anchor and position of the point of exudation allow the nets to remain attached to the anchor point, even after the detachment of the anchors from the gill tissue. This exudate possibly acts somewhat like a 'belay device' or 'safety belt', preventing the parasite from being washed away by the respiratory current during the onset of its leech-like locomotion, as well as assist the relatively small anchors in attachment.