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This study details the novel application of predacious copepods, genus Mesocyclops, for control of Ochlerotatus tremulus (Theobald) group and Aedes aegypti (L.) mosquito larvae in subterranean habitats in north Queensland, Australia. During June 1997, 50 Mesocyclops sp. 1 were inoculated into one service manhole in South Townsville. Wet season rainfall and flooding in both 1998 and 2000 was responsible for the dispersal of copepods via the underground pipe system to 29 of 35 manholes over an area of 1.33 km2. Significant reductions in Aedes and Ochlerotatus larvae ensued. In these habitats, Mesocyclops and Metacyclops were able to survive dry periods, when substrate moisture content ranged from 13.8 to 79.9%. At the semiarid inland towns of Hughenden and Richmond, cracking clay soil prevents drainage of water from shallow service pits where Oc. tremulus immatures numbered from 292-18,460 per pit. Introduction of Mesocyclops copepods into these sites during May 1999 resulted in 100% control of Oc. tremulus for 18 mo. One uninoculated pit subsequently became positive for Mesocyclops with resultant control of mosquito larvae.

The objective of this study wasto determine the epidemiological significance of subterranean mosquito breeding sites to the 1993 outbreak of dengue fever (type 2) in the northern Queensland town of Charters Towers, Australia. In recent studies on subterranean mosquito breeding, containers such as wells and service manholes have been shown to be important breeding sites to Australia's only dengue vector, Aedes aegypti (L.). This study demonstrates a direct epidemiological association between subterranean breeding sites and dengue virus infection. The mean distance between residents seropositive for dengue 2 and the nearest subterranean container (113 m) was significantly less than for a randomly selected control (191 m), (F = 81.9; df = 1, 478; P < 0.001). Residents positive for dengue 2 antibodies was 2.47 (95% confidence interval 1.88-3.24) times higher for those living within 160 m of a well or service manhole, compared with those residing further away. These findings emphasize the importance of incluuding subterranean water containers in Ae. aegypti surveillance and control programs.

The ability of 2 freshwater fishes, eastern rainbow fish Melanotaenia splendida splendida and fly-specked hardyhead Craterocephalus stercusmuscarum stercusmuscarum, native to North Queensland to prey on immature Aedes aegypti was evaluated under laboratory conditions. The predation efficiency of the 2 species was compared to the exotic guppy, Poecilia reticulata, which is commonly used as a biological control agent of mosquito larvae. Of the 3 fish species tested, M. s. splendida was shown to be the most promising agent for the biological control of Ae. aegypti that breed in wells. Melanotaenia s. splendida consumed significantly greater numbers of immature Ae. aegypti than P. reticulata, irrespective of developmental stage or light conditions. Unlike C s. stercusmuscarum, M. s. splendida could be handled, transported, and kept in captivity for extended periods with negligible mortality. However, M. s. splendida was also an efficient predator of Litoria caerulea tadpoles, a species of native frog found in wells during the dry season. This result may limit the usefulness of M. s. splendida as a biological control agent of well-breeding Ae. aegypti and suggests that predacious copepods, Mesocyclops spp., are more suitable. However, the use of M. s. splendida as a mosquito control agent in containers that are unlikely to support frog populations (e.g., aquaculture tanks and drinking troughs) should be given serious consideration.

The effect of a protracted dry season on the viability of Ae. aegypti (L.) eggs was examined in Townsville, northern Queensland, Australia. Eggs were placed in several different surface and subterranean larval habitats; and after four dry season months, only 1-10% of eggs remained viable in the surface and subterranean sites, respectively. Low humidity and predation by Periplaneta americana (L.) were the major causes of egg mortality in eggs in surface sites. P. americana was the most significant cause of egg predation in subterranean breeding sites but fungi, especially Penicillium citrinum Thom, covered egg batches within 15 d. Mycotoxins produced by the spores of P. citrinum are believed to have killed embryonating eggs. The high mortality rate of Ae. aegypti eggs during the dry season suggests that this survival strategy is unlikely to contribute to rapid and successful recolonization of surface sites at the end of the wet season.

In north Queensland, 14 localities were surveyed for mosquito larvae (third and fourth instar) during summer/autumn and winter from 1996 to 1999. Absolute population numbers in subterranean habitats, mainly service manholes and pits (97%) but also some wells, septic tanks, storm drains, and sumps, were expressed as a proportion of total numbers in these sites plus surface sites within a 100-m radius. When correction factors were applied to subterranean samples, the 472,477 larvae mainly of Aedes tremulus (Theobald) group, Aedes notoscriptus (Skuse), and Aedes aegypti (L.) comprised 78% of the total population. In relation to the proportion of the overall immature mosquito population from subterranean habitats (propsub), linear regression coefficients for minimum temperature, relative humidity, and Mesocyclops copepod prevalence were significant for winter data; but for summer, only relative humidity was significant. Linear regression coefficients for Mesocyclops prevalence approached significance (P = 0.061) in summer. When multiple linear regression was used to model propsub, 68% of the variation was accounted for by relative humidity and the prevalence of Mesocyclops. In the drier and cooler towns, increased use of subterranean sites during winter was caused by reduced availability of surface oviposition sites because of the dry season. In the wetter coastal towns, no such restrictions applied and ambient conditions remained more equitable all year round. Mesocyclops were surprisingly common, particularly in these coastal towns. Release of known numbers of Mesocyclops indicated that 3-sweep netting in service manholes was sensitive down to densities of one Mesocyclops per 10 liters, and overall recovery varied from 1 to 4%. In relation to control, service manholes represent a stable habitat for mosquito (7% positive overall) and Mesocyclops populations. If they remained wet, service manholes positive for mosquito immatures or Mesocyclops during summer/autumn had 96% and 85% chance, respectively, of being positive the following winter. Even allowing for the effect of drying, a mosquito-positive manhole had a 79% chance of remaining positive the following winter. In view of the importance of these sites as refuges from adverse ambient conditions, it is proposed that a winter control strategy using Mesocyclops presents a cost-effective control option to reduce the recolonization of surface sites when conditions become more suitable.

A survey of the dengue vector mosquito Aedes aegypti was undertaken using funnel traps to detect immature stages (larvae and pupae) in flooded disused mine shafts and wells in Charters Towers, Queensland, Northern Australia. The town has a history of dengue fever since 1885 when goldminers were the first recorded victims. During the latest dengue epidemic in 1993, 2% of the population had laboratory-confirmed dengue virus Type 2, despite source reduction of Ae.aegypti breeding-sites at ground level or above. This led to suspicions that dengue vector Ae.aegypti breeding-sites might be below ground level. When surveyed in March 1994, Ae.aegypti immatures were found in 9/10 wells and 1/6 mine shafts. The water in wells and mines had similar characteristics-except that turbidity was higher in the mines, which more often contained predators of mosquito immatures. The copepod Mesocyclops aspericornis was collected from water in 1/10 wells and 2/6 mine shafts. Laboratory predation trials resulted in 95.5-100% predation by 25 copepods/l on Ae.aegypti first-instar larvae up to 200 larvae/l. Five wells containing Ae.aegypti in the survey were inoculated with fifty indigenous M.aspericornis, and five wells (one positive and four negative in the survey) were left untreated as controls. Nine months later, in December 1994, Ae.aegypti had been eliminated from all five treated wells but all untreated control wells contained Ae.aegypti, except for one well that contained a natural population of M.aspericornis. The role of wells and mines as winter/ dry season refuges of Ae.aegypti in northern Australia is reviewed, and we recommend the use of M.aspericornis as a cost-effective, environmentally acceptable and persistent agent for the sustainable control of Ae.aegypti, especially in inaccessible breeding sites.