Comparative laboratory bioassays of Tolypocladium cylindrosporum, California strain (Kal) was conducted against third instar larvae of four species of mosquito, viz. Aedes aegypti, Anopheles balabacensis, Culex quinquefasciatus and Mansonia uniformis in Malaysia. Of the four mosquito species tested, Ma. uniformis was found to be the most susceptible, followed by Cx. quinquefasciatus, An. balabacensis and Ae. aegypti, in a decreasing order. The LC50 values for Ma. uniformis, Cx. quinquefasciatus, An. balabacensis and Ae. aegypti after four days of exposure were 1.18 X 10(4), 2.02 X 10(5), 4.76 X 10(5) and 1.84 X 10(7) spores per ml test media, respectively. The high sensitivity of Ma. uniformis and its longer life cycle seems to indicate that T. cylindrosporum Kal has good potential as a biocontrol agent for this species of mosquito. But, for Ae. aegypti, this fungus appears to be less effective.
Matched MeSH terms: Pest Control, Biological/methods*
The pathogenicity of Bacillus sphaericus strain 1593 was tested against laboratory-reared larvae of four local species of mosquitoes of public health importance in Malaysia; Aedes aegypti, Anopheles balabacensis, Mansonia uniformis and Culex quinquefasciatus. The bacteria was shake-cultured at 28 +/- 1 degrees C for three days, using Glucose-Yeast Extract Salts medium. After which, the spores and vegetative cells were harvested and stored at 4 degrees C before use. Conditions for bioassays were mean temperature of 25 +/- 1 degrees C and relative humidity 65 +/- 5.0. Twenty third-instar larvae of each species were assayed in 90 ml of diluted spore solution. Each concentration and a control were replicated three times for each bioassay. Larval mortalities at 24 hours and 48 hours were taken and analyzed through Probit Analysis using a computer (IBM 370). LC50 values after 48 hours of exposure showed an increasing order of larval susceptibility as follows: Ae. aegypti (417.70 x 10(4)), An. balabacensis (45.84 x 10(4)), Ma. uniformis (18.23 x 10(4)) and Cx. quinquefasciatus (4.14 x 10(4) spores/ml). With the ability to kill 90% of the Cx. quinquefasciatus larvae tested with just a concentration of 10(5) spores/ml, B. sphaericus (strain 1593) has shown good potential as a biocontrol agent for this species of mosquito.
Five strains of Ma. uniformis from Malaysia were tested for their susceptibility to infection with subperiodic B. malayi. All were found to be susceptible with infection rates ranging from 62% to 100%. The susceptibility rates were directly related to the microfilarial densities of the cat at the time of feeding. Statistical analysis showed no significant difference (p greater than 0.05) among the means of the indices of experimental infection as well as the percentage of infective mosquitoes of the five strains and an old laboratory colony. They were all equally susceptible to subperiodic B. malayi.
There was high mortality in late larval instars of Aedes albopictus (Skuse) from laboratory and field populations in the 24 h after application of three Bactimos formulations of Bacillus thuringiensis H-14. Mortalities were higher and residual effects longer in field populations than in laboratory ones. Briquets were the most effective formulation (mortality 96-100% after five weeks; 76-92% after eight weeks). Culex quinquefasciatus Say larvae were tested only against the briquet formulation. In the laboratory, 100% mortality of late instars persisted for six weeks and dropped to 48-88% after eight weeks. In the field, late instars were reduced by 62-87% after 24 h and 69-72% after one week compared to increases in an untreated population of 160% and 176% respectively.
A nationwide screening program searching for microbial control agents of mosquitos was initiated in Malaysia in 1986. A total of 725 samples were collected and 2,394 bacterial colonies were isolated and screened for larvicidal activity. From such screening, 20 Bacillus thuringiensis, 6 B. sphaericus, 1 Clostridium bifermentans and 2 Pseudomonas pseudomallei larvicidal isolates were obtained. Of these, a new B. thuringiensis named as subspecies malaysianensis was found, while the C. bifermentans was also a new anaerobe individualized as serovar malaysia. It was concluded that this screening program was highly successful.
Matched MeSH terms: Pest Control, Biological/methods*
Nine species of parasitoids were found parasitizing the pupae of filth flies breeding in refuse dumps and poultry farms throughout peninsular Malaysia. Spalangia were most common, consisting of Spalangia endius Walker, S. cameroni Perkins, S. gemina Boucek, S. nigroaenea Curtis, and two undescribed species. Other parasitoids collected were Pachycrepoideus vindemmiae Rondani, Dirhinus himalayanus Westwood, and an unidentified Hymenoptera. The parasitized fly hosts included Musca domestica L., Chrysomya megacephala (F.), Fannia sp., and Ophyra sp. S. endius was the most common parasitoid attacking M. domestica and C. megacephala at refuse dumps and poultry farms D. himalayanus were found to parasitize only M. domestica pupae collected at poultry farms.
The measurement of the ultimate effects of the microbial insecticides on mosquito density is best obtained by assessment of adult populations. The main aims of this study are: (1) to assess the effect of Bacillus thuringiensis israelensis (Bti) FC Skeetal and Bactimos briquettes on the emergence rate of Mansonia bonneae developed from the introduced first-instar stage larvae and (2) to measure the effect of these two formulations of insecticides on Mansonia adult populations emerging from the natural breeding plots. Bti Skeetal and Bactimos briquettes at the lower applied dosages of 2.3 kg/ha and 1 briquette case/20 m2 respectively achieved 39-40% pupation rates and 31.5-34.2% adult emergence rates. At these low applied dosages, there was little or no direct effect on pupation from the surviving larvae and thereafter on the emergence of adults from the pupae. A two-fold increase in dosage, however, produced a drastic decline in the pupation rate and adult emergence rate. The rates dropped to 6.5% (pupation) and 4.3% (adult emergence) of the total larvae for Bactimos briquettes and to merely 1.5% (pupation) and 1.3% (adult emergence) of the total larvae for Skeetal. In studying the effect of Bti on the field populations of Mansonia mosquitos, two plots each were treated with Bactimos at 1 briquette case/10 m2 and Skeetal at 4.6 kg/ha. A wooden pyramid-shaped screened cage was placed on a cluster of host plants for a period of 2 weeks to trap the emerging adult mosquitoes. There were a total of 24 clusters of host plants in each plot.(ABSTRACT TRUNCATED AT 250 WORDS)
A screening program searching for indigenous microbial control agents of mosquitos in Malaysia is initiated since 1987 and to date at least 20 isolates of mosquitocidal Bacillus thuringiensis serotypes have been obtained. Preliminary field evaluation of several isolates indicated that they are highly effective in the control of medically important mosquito species. For operational purposes, there is an urgent need to produce this agent utilizing cheap and locally available wastes through fermentation biotechnology. Fermentation studies in shake-flasks containing standard nutrient broth and soya bean waste, respectively, indicate that it takes about 37 hours for a Malaysian isolate of B. thuringiensis serotype H-14 to mature. In the grated coconut waste, fishmeal and rice bran, the bacteria took 28 hours, 26 hours and 126 hours respectively to mature. The endotoxin was harvested from the standard nutrient broth at 55 hours and at 50 hours from soybean, grated coconut waste and fishmeal. The endotoxin could only be harvested 150 hours after inoculation from rice bran medium. However, no bacterial growth was detected in palm oil effluent. In terms of endotoxin and biomass production, fishmeal appears to be a suitable medium. Variations in the pH of the fermenting media were also noted.
Matched MeSH terms: Pest Control, Biological/methods*
Five formulations of Bacillus sphaericus (strain 2362) including aqueous suspension BSP 1, BSP 2, technical powder ABG 6184, corncob granules ABG 6185 (potencies 2 x 10(10), 2 x 10(7), 9.5 x 10(10), 5 x 10(10), 5 x 10(10) spore/g, respectively) and wettable powder ABG 6232 (1,000 BS ITU/mg) were tested against laboratory-cultured late third/early fourth instar larvae of Mansonia uniformis in floating screened cages in small plots at swampy ditches on Penang Island, Malaysia. Mean dosage/response values at 90% mortality levels were 6.93, 95.32, 1.45, 11.92 and 2.86 liters or kg per ha, respectively, for the formulations tested. There were practically no residual effects for the formulations tested with larvae introduced at 48, 96, and 168 h post-treatment. In trials of BSP 1, ABG 6184 and ABG 6185 (1 liter or 1 kg per ha) against immature Mansonia spp. in impounded paddy field ditches, improved efficacy and residual effects were obtained with mean reductions of 93.1, 91.9 and 80.4% at days 3, 7 and 14 posttreatment, respectively.
A study on population patterns of the parasitoid Spalangia endius Walker at a dumping ground near Kuala Lumpur city showed that the percentage of S. endius adult emergence varied seasonally. During the relatively heavy rainfall months of August and November 1988, and January, March, and April 1989, the population of S. endius adult emergence were low (0-14.2%) compared to the less rainy months of July, September, and December 1988, and May 1989 (29.3-39.6%). This information could be useful in formulating strategies to reduce house fly population at the refuse dumping ground through integrated pest management programs.
The susceptibility of three strains of Aedes aegypti (L.) to four strains of Ascogregarina culicis were studied under laboratory conditions. The parasite was found to be pathogenic to the mosquito; although pathogenicity varied among geographical strains of A. culicis and susceptibility varied among geographical strains of Ae. aegypti. In addition to affecting the survival of the mosquito larvae, infection with the parasite was found to shorten larval development time. Selected strains of A. culicis may be useful as a biological control agent against Ae. aegypti.
Clostridium bifermentans serovar. malaysia (C.b.m.) is toxic to mosquito larvae. In this study, we quantified its toxicity to the mosquitoes, Aedes aegypti, Ae. albopictus, Ae. caspius, Ae. detritus, Anopheles stephensi, An. gambiae, Culex pipiens and Cx. quinquefasciatus. Anopheles larvae are the most susceptible, followed by Ae. detritus and Ae. caspius, then Culex and other Aedes larvae. According to mosquito species, the LC50 varies from 7 x 10(3) to 1.3 x 10(6) cells/ml. Three concentrations (10(7), 10(6) and 10(5) cells/ml) of C.b.m., Bacillus thuringiensis var. israelensis (B.t.i.) and Bacillus sphaericus were tested on Ae. aegypti, An. stephensi and Cx. pipiens larvae in order to determine the time necessary for each concentration to kill 50 and 90% of the population. Ninety percent of the 3 mosquito populations are killed within 4-15 h by the C.b.m. concentrations. Whatever the concentrations, C.b.m. kills at least 10 times less rapidly than B.t.i. but always quicker than B. sphaericus. Bioassays of C.b.m. bacterial cells or final whole culture were not toxic to Musca domestica and Drosophila melanogaster (Diptera) as well as to Phaedon cochleariae (Coleoptera) and Spodoptera littoralis (Lepidoptera).
Eggs of temperate Aedes albopictus populations are cold hardy and can diapause, but tropical populations are not cold hardy and cannot diapause. Heterozygotes possess intermediate diapause and cold hardiness. Males of a tropical strain from Malaysia with a distinctive genetic marker were released into an existing temperate population in East St. Louis, Illinois. Subsequent egg samples from the release site had genetic marker frequency of up to 24%. Reduced cold hardiness and decreased diapause incidence were also observed in the release site population. No such changes occurred at a nearby control site. The rank order of overwintering survival of eggs at the release site was: Aedes triseriatus > temperate Ae. albopictus > hybrid temperate/tropical Ae. albopictus > tropical Ae. albopictus. Eggs collected from the release population the next summer showed total absence of the genetic marker; presumably carriers were removed by the winter.
Matched MeSH terms: Pest Control, Biological/methods*
Clostridium bifermentans serovar malaysia (C.b.m.) is highly toxic to mosquito larvae. In this study, the following aquatic nontarget invertebrates were treated with high C.b.m. concentrations (up to 1,600-fold the toxic concentration for Anopheles stephensi) to study their susceptibility towards the bacterial toxin: Planorbis planorbis (Pulmonata); Asellus aquaticus (Isopoda); Daphnia pulex (Cladocera); Cloeon dipterum (Ephemeroptera); Plea leachi (Heteroptera); and Eristalis sp., Chaoborus crystallinus, Chironomus thummi, and Psychoda alternata (Diptera). In addition, bioassays were performed with mosquito larvae (Aedes aegypti, Anopheles stephensi, and Culex pipiens). Psychoda alternata larvae were very susceptible, with LC50/LC90 values comparable to those of mosquito larvae (about 10(3)-10(5) spores/ml). The tests with Chaoborus crystallinus larvae showed significant mortality rates at high concentrations, but generally not before 4 or 5 days after treatment. The remaining nontarget organisms did not show any susceptibility. The investigation confirms the specificity of C.b.m. to nematocerous Diptera.
Matched MeSH terms: Pest Control, Biological/methods
A new mosquitocidal Bacillus thuringiensis subsp., jegathesan, has recently been isolated from Malaysia. Parasporal crystal inclusions were purified from this strain and bioassayed against fourth-instar larvae of Culex quinquefasciatus, Aedes aegypti, Aedes togoi, Aedes albopictus, Anopheles maculatus, and Mansonia uniformis. The 50% lethal concentration of crystal inclusions for each species was 0.34, 8.08, 0.34, 17.59, 3.91, and 120 ng/ml, respectively. These values show that parasporal inclusions from this new subspecies have mosquitocidal toxicity comparable to that of inclusions isolated from B. thuringiensis subsp. israelensis. Solubilized and chymotrypsin-activated parasporal inclusions possessed low-level hemolytic activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the crystals were composed of polypeptides of 77, 74, 72, 68, 55, 38, 35, 27, and 23 kDa. Analysis by Western blotting (immunoblotting) with polyclonal antisera raised against toxins purified from B. thuringiensis subsp. israelensis reveals that proteins in parasporal inclusions of subsp. jegathesan are distinct, because little cross-reactivity was shown. Analysis of the plasmid content of B. thuringiensis subsp. jegathesan indicates that the genes for toxin production may be located on 105- to 120-kb plasmids. Cry- clones that have been cured of these plasmids are nontoxic. Southern blot analysis of plasmid and chromosomal DNA from subsp. jegathesan showed little or low homology to the genes coding for CryIVA, CryIVB, and CryIVD from B. thuringiensis subsp. israelensis.