Effect of water temperature on reproductive development of Benedenia seriolae (Monogenea: Capsalidae) from Seriola lalandi in Australia

The monogenean Benedenia seriolae (Yamaguti, 1934) Meserve, 1938 is a major pathogen of farmed yellowtail kingfish Seriola lalandi in South Australia. To control parasite populations in commercial farms, an understanding of the effect of water temperature on parasite development is vital. This study investigated the effect of water temperature (14, 18, 22 and 26 +/- 0.5 degrees C) on development, growth rates and age at sexual maturity of B. seriolae. Five distinct developmental stages, defined by the progressive development of reproductive organs, were used to describe development of B. seriolae from recently invaded larvae to sexually mature adults. Parasite age at the first sign of sexual maturity (ability to lay eggs) was strongly influenced by water temperature and was attained at 41, 24, 16 and 14 d post infection (p.i.) at 14, 18, 22 and 26 +/- 0.5 degrees C respectively. Four parameters (parasite total length, maximum parasite width, accessory sclerite length and anterior hamulus length) were examined for suitability as an index of parasite age. Growth rates for each parameter increased with water temperature; however, mean anterior hamulus length was identified as the most reliable index of parasite age. Equations derived from these data can be used to estimate parasite age and time to sexual maturity at water temperatures ranging from 14 to 26 degrees C. The equations provide a simple tool to assist implementation of strategic treatment plans for B. seriolae infections in commercial kingfish farms, not only in South Australia but in other localities worldwide where S. lalandi is farmed.     

Efficacy of orally administered praziquantel against Zeuxapta seriolae and Benedenia seriolae (Monogenea) in yellowtail kingfish Seriola lalandi

We investigated the efficacy of praziquantel (PZQ) administered orally to yellowtail kingfish (Seriola lalandi in sea-cage aquaculture in South Australia) against the monogeneans Zeuxapta seriolae and Benedenia seriolae infesting gills and skin, respectively. PZQ was administered to fish by surface-coating feed pellets (Trial 1) or by direct intubation of the stomach (Trial 2). In both trials 4 daily doses were administered: 50 and 75 mg kg(-1) body weight (BW) d(-1) for 6 d, and 100 and 150 mg kg(-1) BW d(-1) for 3 d. Mean parasite intensity was compared between medicated fish and unmedicated control fish. In Trial 1, fish fed lower daily doses of PZQ for 6 d (50 and 75 mg kg(-1) BW d(-1)) had fewer Z. seriolae and B. seriolae than fish fed higher daily doses for 3 d (100 and 150 mg kg(-1) BW d(-1)). Fish rejected feed pellets surface-coated with PZQ, suggesting PZQ affected palatability of feed, and may explain differences in efficacy between treatments. In Trial 2, where PZQ was administered by intubation, there were fewer Z. seriolae and B. seriolae in medicated fish than control fish. Intubated PZQ was also effective against newly recruited Z. seriolae and B. seriolae. PZQ could be developed as a useful treatment for Z. seriolae and B. seriolae parasitising S. lalandi in sea-cage aquaculture if suspected palatability problems are resolved.     

Metazoan parasite assemblages of wild Seriola lalandi (Carangidae) from eastern and southern Australia

Yellowtail kingfish, Seriola lalandi support significant commercial and recreational fisheries as well as aquaculture operations throughout the world. Metazoan parasite infections of S. lalandi are of considerable economic and ecological importance, yet very little is known about wild parasite assemblages. S. lalandi were collected from the east coast and south coast of Australia and examined for metazoan parasites. Forty-three parasite taxa were identified, including 26 new host records. Four of the parasite species recovered have been previously associated with disease or mortality in Seriola aquaculture. Comparisons are made between ectoparasite and endoparasite prevalence and intensity of S. lalandi from New South Wales and Victoria. S. lalandi sampled from the east coast of Australia shared ectoparasites previously documented from this species in New Zealand, providing support that S. lalandi in the Tasman Sea comprise a single stock. Based on previously used criteria to evaluate the suitability of parasites as biological tags, the monogenean Paramicrocotyloides reticularis Rohde and the copepod Parabrachiella seriolae Yamaguti and Yamasu may be potentially useful for stock discrimination.     

Attachment-inducing capacities of fish skin epithelial extracts on oncomiracidia of Benedenia seriolae (Monogenea: Capsalidae)

Attachment-inducing capacities of skin epithelial extracts of yellowtail, Japanese flounder and red sea bream on oncomiracidia of the monogenean Benedenia seriolae were examined. Clear differences were not detected in the capacity among the fish species, although B. seriolae infects only yellowtail and its congeners in Seriola. This suggests that either the capacity is not host specific or host-specific attachment-inducing capacity cannot be detected by the assay method. Further, the attachment-inducing capacities were suppressed by wheat-germ lectin and concanavalin A in skin epithelial extracts of Japanese flounder and yellowtail, respectively. This suggests that some sugar-related chemical substances existing in fish epithelia induce the attachment of B. seriolae oncomiracidia.     

Effects of temperature during chilling and pre-chilling periods on diapause and post-diapause development in a katydid, Eobiana engelhardti subtropica

In Eobiana engelhardti subtropica, early laid eggs reach the diapause stage in early autumn. For long periods before winter, the eggs are exposed to temperatures higher than their theoretical lower threshold for development. In contrast, late-laid eggs cannot reach their diapause stage before winter. Our study showed that E. e. subtropica copes with these difficulties via the thermal response involving embryonic diapause. In this katydid, the almost fully developed embryo undergoes an obligatory diapause. When diapause eggs were maintained at a temperature of 20 degrees C or higher, diapause persisted for a long time. Diapause was effectively terminated by temperatures ranging from 1 to 11 degrees C, and hatching occurred successfully at temperatures from 11 to 15 degrees C. In addition to the chilling temperature, pre-chilling temperature modified diapause intensity and hatching time. Diapause eggs hatched earlier after chilling when the pre-chilling temperature was lower, within a range of 14.5-25 degrees C. Thus, the low-temperature requirement for diapause termination prevents early laid eggs from untimely hatching in autumn, and low temperatures before and during winter decrease diapause intensity and shorten the hatching time in the following spring. When eggs were chilled before diapause, they tolerated chilling and averted diapause. Thus, even if eggs encounter low temperatures before diapause, they can hatch in the following spring.     

Egg-laying patterns and in vivo egg production in the monogenean parasites Heteraxine heterocerca and Benedenia seriolae from Japanese yellowtail Seriola quinqueradiata

Egg-laying patterns and egg production in Heteraxine heterocerca from the gills and Benedenia seriolae from the skin of Japanese yellowtail Seriola quinqueradiata in Japan were investigated in vivo. Eggs were collected every 3 h from sexually mature H. heterocerca and B. seriolae infecting 3 S. quinqueradiata kept individually over 3 consecutive days and exposed to alternating periods of illumination and darkness (LD 12:12; light on 06.00, light off 18.00) and maintained at 23.8 +/- 0.1 degrees C and 35 ppt salinity. A well-defined egg-laying rhythm was demonstrated for H. heterocerca while B. seriolae was shown torelease eggs continuously. A total of 114,000 H. heterocerca eggs was collected and of these, 45.4 (42.5-49.9)% were collected during the first 3 h period following dark at 18.00 h. A total of 662,857 B. seriolae eggs was collected and these were distributed over each 3 h period ranging from 11.1 to 14.1% of the daily egg output. All eggs extracted from the uterus of each H. heterocerca were joined together forming an 'egg-string'. The contrasting egg-laying patterns of H. heterocerca and B. seriolae suggest that each species makes use of a different infection strategy to infect the same host species, S. quinqueradiata.     

Impacts of diclidophorid monogenean infections on fisheries in Japan

Many monogeneans are pathogenic to economically important fish in Japan. However no other monogenean is comparable with the diclidophorids, Heterobothrium okamotoi and Neoheterobothrium hirame, on the scale of impacts they inflict on Japanese fisheries. The shared importance of the two monogenean infections lies in their pathogenicity, fecundity and tolerance to chemical treatment. Heterobothrium okamotoi infects the gills and wall of the branchial cavity of the tiger puffer, Takifugu rubripes (Tetraodontidae), which is widely cultured in western Japan. The main presenting signs of infected fish are anaemia and extensive necrosis caused by adult worms. This monogenean deposits long strings of eggs, which reach lengths of almost 3m. Egg entanglement with the mesh of culture nets increases the chance of hatched larvae encountering susceptible fish. The oncomiracidium maintains infectivity for up to 4 days after hatching. Hydrogen peroxide is the only commercially available chemical able to control the infection, but can only kill immature worms on the gills. Neoheterobothrium hirame infects the gills and wall of the buccal cavity of the Japanese flounder, Paralichthys olivaceus (Paralichthyidae). Since the first known occurrence of this monogenean in 1993, the species has been recorded from almost all areas where the host is distributed. Neoheterobothrium hirame has the potential to produce 781 eggs per day at 20 degreeC. In the western Sea of Japan, wild young-of-the-year flounder became infected in early summer, followed by a sharp increase in prevalence in late summer. By late summer, juvenile flounder have nearly disappeared from the area, strongly suggesting that N. hirame is responsible for mortality of young fish. This is in good agreement with the recent decline in the local flounder population. Neoheterobothrium hirame has also been considered the causative agent of anaemia among wild Japanese flounder since the late 1990s.     

The effect of desiccation and low temperature on the viability of eggs and emerging larvae of the tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodidae)

Process-based population models need sound and comprehensive data on an animal's response to climatic factors if they are to function reliably under a wide range of climatic conditions. To this end, different aged egg masses of the livestock tick, Rhipicephalus (Boophilus) microplus, were either desiccated in atmospheres with saturation deficits of 5, 10, 15 or 20 mmHg at 20 or 26 degrees C, or chilled at temperatures of 5, 10 or 14 degrees C with a saturation deficit of 1 mmHg for varying periods. The survival rate of the eggs through to hatching was related to the initial age of the eggs, the severity of the treatments and the duration of exposure. We established a relationship between desiccation and weight loss of eggs and, secondarily between weight loss and mortality. Mortality increased with weight loss until it reached 100% when the weight loss was about 35%. Low temperatures were increasingly detrimental to eggs as they reduced from 14 to 5 degrees C. Freshly laid eggs were more susceptible to both low temperatures and desiccation than were older eggs. Larvae emerging from eggs that were stressed by either cold or desiccation lived for a shorter time under optimal conditions than did larvae from eggs incubated under optimal conditions. Larvae from eggs with the same hatching rate had the same viability, whether the stress was induced by desiccation or low temperatures. Models were developed to describe the dynamics of weight loss of eggs with desiccation, the accumulation of cold stress of the eggs, and their effects on egg survival and larval viability. These data provide a sound basis for the development of predictive models for use under field conditions, although the response of different aged eggs to low temperatures was too variable to allow us to develop an accurate model to describe that relationship. Field models will also need to take diurnal temperature fluctuations into account.     

Bemerkungen zur Biologie sowie Haltung und Vermehrung des Sokotra-Riesengeckos, Haemodracon riebeckii (Peters, 1882)

The Socotra Giant Gecko, Haemodracon riebeckii, is the largest species of lizard on Socotra Island. The nocturnal, arboreal and rupiculous living geckos are omnivorous. Two pairs were kept in terrariums and were fed with various insects (crickets, locusts, cockroaches), sweet fruits and other feeding stuff (such as meat, fish). Temporarily H. riebeckii was kept together with other lizards (Eublepharis macularius, Trachylepis socotrana), without any signs of aggressive behaviour. Juveniles and adults of both sexes are able to produce a sound. These acoustic signals seem to be related to predators, because never any intraspecific function could be observed. Within seven years of captive breeding two females produced 253 eggs. Usually two white and sticky soft-shelled eggs were laid within one clutch, more rarely a single egg was laid. The two eggs of a clutch were always laid on the same day. H. riebeckii belongs to the geckos that bury their eggs and practice some brood care, but no special parental care. The female is able to proof with her hind legs the deep and shape of a hollow in the substrate to bury the eggs, which were buried in a sticky and soft-shelled condition. They are oval in shape (egg length 16.4-19.8 mm, egg width 12.4-17.8 mm, quotient EL:EW 1.22±0.05) and have in the beginning a weight ranging from 1.7100 to 2.5201 g. As typical for geckos with hard-shelled eggs the egg weight decreases during the incubation period. The loss can be between 5.59 to 30.29%. The development of eggs up to hatching of young depends upon temperature and the germinal stage in the laid egg. The time difference between the hatching of the young within one clutch of two eggs was usually 1 to 5 days. In some cases there were, however, longer differences (up to 61 days), which are probably caused by different developmental stages of the embryos during the time of egg laying. The shortest incubation period recorded during our investigations was 83 days for eggs incubated at constant temperature of 28 to 29.5 °C and the longest 359 days at 26 to 26.5 °C. Constant high incubation temperatures caused a premature hatching of young. In normal hatched young were the yolk sac retracted and the navel closed. In premature hatched young were the yolk not resorbed and the mortality within the first three month comparatively high. The snout-vent length (SVL) of newly hatched young is from 27 to 39 mm and the tail length (TL) from 25 to 38 mm (SVL:TL index 0.90-1.27), the weight is from 0.7688 to 1.5366 g. Young specimens are distinguished from adults by the brown/white striped lower jaw and the white-banded tail. Young which hatched in the terrarium were eaten by the adults. A loss of young can be avoided if they are raised individually.     

Effect of temperature on life history of the predatory mite Amblyseius (Neoseiulus) californicus (Acari: Phytoseiidae)

Amblyseius (Neoseiulus) californicus is an indigenous mite in Japan that feeds on many spider mite species. We evaluated the development, survivorship and life-history parameters of A. californicus on a diet of eggs of Tetranychus urticae (red form). More than 97.3% of A. californicus eggs hatched and more than 81.6% of newly hatched larvae attained maturity at temperatures between 15 and 35 degrees C. Females oviposited at 37.5 degrees C, but no eggs hatched. At 40 degrees C, no females laid eggs. The lower threshold temperature from egg to oviposition was 10.3 degrees C and the thermal constant was 86.2 degree-days. Based on these data, the maximum number of generations that could complete development in a year under field conditions in Ibaraki, central Japan, would be between 21 and 28. At 25 degrees C, females laid a mean of 41.6 eggs during a mean oviposition period of 19.4 days. The intrinsic rates of natural increase (rm) were 0.173 at 20 degrees C, 0.274 at 25 degrees C and 0.340 at 30 degrees C.     

Laboratory studies of factors affecting egg hatch of triops longicaudatus (LeConte) (Notostraca : Triopsidae)

Egg hatch was greatest (78.33%) for eggs not previously desiccated. A reduction in numbers hatched occurred as the relative humidity at which they were dried decreased. Some eggs hatched (0.67–79.33%) at pH levels of 3.10–10.01 with the highest hatch at pH 5.60. Water temperature greatly affected egg hatch. No hatch occurred until temperatures were above 14°C. A constant 29°C significantly inhibited hatching. Egg hatch increased 13.00 to 43.42% as salinity decreased from 2200 to 9.24 micromhos/cm. As little as 13 mm of flooded soil covering the eggs prevented them from hatching for 14 days. Eighteen percent hatch resulted when soil and eggs were redistributed to a 1 mm soil layer. Egg samples from the same parent, even though treated similarly, often hatched at greatly varying rates and only rarely was hatching 100% within a replication.     

Reproductive Strategies of the Insidious Fish Ectoparasite,Neobenedenia sp. (Capsalidae: Monogenea)

Fish monogeneans are lethal parasites in aquaculture. We provide the first experimental evidence that a notorious fish monogenean, Neobenedenia sp., can produce viable eggs in isolation for three consecutive generations. We infected individual, isolated, farmed barramundi, Lates calcarifer (Bloch) with a single oncomiracidium (larva) of the hermaphroditic monogenean Neobenedenia sp. Isolated parasites reached sexual maturity at day 10 post-hatch (24°C, 35‰) and laid ∼3,300 embryonated eggs over 17 days. Egg production rapidly increased following sexually maturity on day 10 (58±15 eggs) and peaked on day 15 (496±68 eggs) before gradually decreasing. Neobenedenia sp. exhibited egg laying and egg hatching rhythms. Parasites laid eggs continuously, but egg production increased in periods of darkness (64.3%), while the majority of oncomiracidia (81%) emerged from eggs in the first three hours of light. Eggs laid by isolated ‘parent’ parasites hatched and individual emerging oncomiracidia were used to infect more individual, isolated fish, with three consecutive, isolated, parasite generations (F1, F2 and F3) raised in the laboratory. Infection success and egg hatching success did not differ between generations. Our data show that one parasite, in the absence of a mate, presents a severe threat to captive fish populations.     

Relatively low upper threshold temperature in lizards from cool habitats

We used the slender forest skink (Scincella modesta) as a model animal to test for the hypothesis that the upper threshold of incubation temperature is relatively low in lizards using shaded (and thus, cool) habitats. Eight gravid females were collected in early May 2005 from a population in Hangzhou, Zhejiang (eastern China). All females laid a single clutch of 7–13 eggs between mid-May and early June. Eggs were incubated at 24, 28 and 30 (±0.2) °C. None of eggs incubated at 30 °C hatched. Eggs incubated at 24 and 28 °C differed in incubation length but not in hatching success. The incubation length at 24 and 28 °C averaged 22.3 and 20.3 days, respectively. Hatchlings from eggs incubated at 24 and 28 °C did not differ in all examined morphological traits, but hatchlings from eggs incubated at 28 °C performed apparently worse in the racetrack than did their counterparts from eggs incubated at 24 °C. The temperature of 28 °C is close to the upper thermal threshold for successful embryonic development in S. modesta. Compared to other oviparous lizards using open (and thus, warm) habitats, the upper thermal threshold and the range of optimal temperatures for embryonic development are both lower in S. modesta. Our study supports the previous conclusion that species living in thermally different habitats may differ in the upper thermal threshold and the range of optimal temperatures for embryonic development.     

Environmental constraints influencing survival of an African parasite in a north temperate habitat: effects of temperature on egg development

Factors affecting survival of parasites introduced to new geographical regions include changes in environmental temperature. Protopolystoma xenopodis is a monogenean introduced with the amphibian Xenopus laevis from South Africa to Wales (probably in the 1960s) where low water temperatures impose major constraints on life-cycle processes. Effects were quantified by maintenance of eggs from infections in Wales under controlled conditions at 10, 12, 15, 18, 20 and 25°C. The threshold for egg viability/ development was 15°C. Mean times to hatching were 22 days at 25°C, 32 days at 20°C, extending to 66 days at 15°C. Field temperature records provided calibration of transmission schedules. Although egg production continues year-round, all eggs produced during >8 months/ year die without hatching. Output contributing significantly to transmission is restricted to 10 weeks (May-mid-July). Host infection, beginning after a time lag of 8 weeks for egg development, is also restricted to 10 weeks (July-September). Habitat temperatures (mean 15·5°C in summer 2008) allow only a narrow margin for life-cycle progress: even small temperature increases, predicted with 'global warming', enhance infection. This system provides empirical data on the metrics of transmission permitting long-term persistence of isolated parasite populations in limiting environments.     

Adaptive difference in daily timing of hatch in two locust species,Schistocerca gregaria and Locusta migratoria: The effects of thermocycles and phase polyphenism

This study examined the effects of temperature and phase polyphenism on egg hatching time in the desert locust, Schistocerca gregaria, and the migratory locust, Locusta migratoria. The two species exhibited differences and similarities in hatching behavior when exposed to different temperature conditions. In 12-h thermocycles of various temperatures, the S. gregaria eggs hatched during the cryoperiod (low temperature period), whereas L. migratoria eggs hatched during the thermoperiod (high temperature period). The eggs of both species hatched during the species-specific period of the thermoperiod in response to a temperature difference as small as 1 °C. Furthermore, the locusts adjusted hatching time to a new thermal environment that occurred shortly before the expected hatching time. In both species, the hatching of the eggs was synchronized to a specific time of the day, and two hatching peaks separated by approximately 1 day were observed at a constant temperature after the eggs were transferred from thermocycles 3 days before hatching. Eggs laid by gregarious females hatched earlier than those laid by solitarious females in S. gregaria but this difference was not observed in L. migratoria.     

Abiotic factors influencing embryonic development, egg hatching, and larval orientation in the reindeer warble fly, Hypoderma tarandi

Wild-caught, tethered females of the reindeer warble fly, Hypoderma tarandi (L.) (= Oedemagena tarandi (L.)), (Diptera, Oestridae) were stimulated to oviposit on hairs of a reindeer hide. Newly laid eggs incubated at constant temperatures and relative humidities hatched within 3 days to 2 weeks, depending on the experimental conditions. Over a range of 7-40 degrees C, hatching only occurred between 20 and 37 degrees C. Eggs held at 100% relative humidity had lower hatchability and longer time to hatch relative to eggs held at 77% relative humidity. The average number of degree-days for hatching was 50.35. Between 20 and 33 degrees C there was a temperature-dependent linear trend in developmental rate, and the proportion of eggs hatching was highest, and least variable, at the mid-temperature ranges. The temperature range found in the natural host micro-habitat where H. tarandi commonly affix their eggs (close to the skin at the base of a host hair) was consistent with the experimental temperature treatments that produced the highest hatching rate. Newly emerged larvae displayed positive thermotaxis, while showing no phototaxic or geotaxic behaviour. Results indicate that constraints of the host environment, coupled with temperature-dependent hatching success, may impose a selective pressure on oviposition behaviour.     

Changes in the reproductive biology of Biomphalara glabrata infected with different doses of Echinostoma paraensei miracidia

The egg-laying rate, number of egg masses, number of eggs/mass, number of eggs hatched/snail and egg viability of Biomphalaria glabrata exposed to different doses (5 and 50) of Echinostoma paraensei miracidia were analyzed as indicators of reproductive activity. Polystyrene plates were placed in aquariums containing the snails and every other day for four weeks after infection the plates were removed to count the number of egg masses and eggs laid. After this, the plates were numbered individually and placed in new aquariums free of snails and the egg masses were observed daily to determine the hatching rate. On average there was an increase in the parameters evaluated in the infected snails in relation to the controls (uninfected snails), except for egg viability, which was significantly lower in the groups infected with 50 miracidia. These findings indicate that when infected, this host snail is able to increase its reproductive activity, suggesting an ecological strategy to maintain the species.     

Growth rate, size, and sex ratio of last-laid, last-hatched offspring in the tree swallow Tachycineta bicolor

In tree swallows Tachycineta bicolor, last‐laid eggs typically hatch one to two days after the other eggs in the clutch hatch, putting last‐hatched offspring at a disadvantage when competing for food delivered by parents. We studied the biology of last‐laid, last‐hatched tree swallow offspring over two years in a Wyoming, USA, population. Our first objective was to compare the growth of last‐hatched offspring to that of their earlier‐hatched nestmates. One previous study had suggested that last‐hatched, competitively disadvantaged offspring grow feathers faster than senior nestmates, even at the expense of other aspects of growth. This may allow last‐hatched offspring to fledge with senior nestmates and avoid abandonment by parents. A second objective was to determine the sex of nestlings from last‐laid eggs. If last‐laid eggs typically produce undersized, weak adults that are poor competitors for resources, and if the fitness costs of being undersized/weak are more severe for males than for females, then selection may favour having offspring from last‐laid eggs to be female. In this study, last‐laid eggs hatched in 63 of 66 (94%) nests and hatched last in 93% of cases. At hatching, offspring from last‐laid eggs weighed, on average, 63% as much as their three heaviest nestmates (range: 26–107%). Offspring from last‐laid eggs fledged from 71% of the nests that produced at least one fledgling and apparently starved to death in remaining nests. Last‐hatched offspring who were presumably at a substantial competitive disadvantage (those whose mass at hatching was no more than about 75% of the mean mass of their three heaviest nestmates), gained mass more slowly than their senior nestmates but they eventually attained the same peak mass before fledging. Last‐hatched offspring grew primary feathers more slowly than their senior nestmates although the difference in growth rate was slight (0.2 mm/d) and only marginally significant. As a group, offspring from last‐laid eggs did not differ from offspring from all other eggs in either maximum mass attained before fledging or tarsus length at fledging. This is atypical for species with asynchronous hatching and is possibly the result of another unusual trait: the tendency of parent tree swallows to distribute food equally among young within broods. The sex ratio of offspring from last‐laid eggs did not deviate from 1:1 (22 males, 21 females). Given that last‐hatched eggs do not routinely produce undersized/weak individuals in our study population, there should be little selection on parent females to bias the sex ratio of last‐laid offspring towards females.     

A possible role for FRM-1, a C. elegans FERM family protein, in embryonic development

FRM-1 is a member of the FERM protein superfamily containing a FERM domain, which is a highly conserved protein-protein interaction module found in most eukaryotes. Although FRM-1 is thought to be involved in linking intracellular proteins to membrane proteins, the specific role for FRM-1 remains to be elucidated. In an effort to explore the biological function of FRM-1, we examined the phenotype of frm-1(tm4168) mutant worms. We observed that frm-1(tm4168) worms have a delayed hatching phenotype. Twelve hours after being laid, when virtually all wild-type eggs had hatched, only 64% of frm-1(tm4168) eggs had hatched. About 3% of frm-1(tm4168) eggs failed to hatch, even 3 days after they had been laid. We also found that frm-1(tm4168) mutants displayed a temperature-sensitive sterility phenotype. About 13% of frm-1(tm4168) worms were unable to produce eggs or produced nonviable eggs at 25°C. In contrast, less than 1% of wild-type animals were sterile at this temperature. At 20°C, neither the mutant nor wild type appeared to be sterile. Western blot analysis indicates that FRM-1 is expressed throughout the developmental stages with the strongest expression at the egg stage. Immunostaining experiments revealed that FRM-1 is mainly localized to the plasma membrane of most if not all cells at an early embryonic stage and to the plasma membrane of P cells during the late embryonic stages. GFP fusion experiments showed that FRM-1 can be expressed in the pharynx and intestine at the larval and adult stages. Our data suggest that FRM-1 may participate in diverse biological processes, including embryonic development.     

The effects of temperature, diet, and other factors on development, survivorship, and oviposition of Aethina tumida (Coleoptera: Nitidulidae)

Developmental rate and survivorship of small hive beetle, Aethina tumida Murray (Coleoptera: Nitidulidae), life stages were measured across different temperatures (21, 25, 28, 32 and 35 degrees C) and diets, which included natural and artificial pollen, honey, and bee pupae. Temperature affected hatch success, time to hatching, and larval growth. Eggs hatched in 61 h at 21 degrees C but in < 22 h at 35 degrees C. Larvae achieved peak weight in < 8 d at 35 degrees C but needed 17 d at 21 degrees C. Diet had comparatively little effect on larval survivorship or maximum weight, although larvae fed only bee pupae had lower survivorship. Access to soil influenced pupation success. Duration of the life stage spent in the soil, during which pupation occurs, was also affected by temperature: adults emerged after 32.7 d at 21 degrees C but after only 14.8 d at 35 degrees C, albeit with high mortality. Minimum temperature for development was estimated at 13.5 degrees C for eggs, and 10.0 degrees C for larvae and pupae. Temperature influenced adult longevity and oviposition: on a honey and pollen diet average adult lifespan was 92.8 d at 24 degrees C but only 11.6 d at 35 degrees C. Beetles lived longer at 28 degrees C or lower but produced the most eggs per female, regardless of diet, at 32 degrees C. Beetle density influenced fecundity: beetles kept at three pairs per vial laid 6.7 times more eggs per female than those kept as single pairs. Overall, beetles fared best at 28-32 degrees C with mortality of all stages highest at 35 degrees C.