Cryptic changes in the genetic structure of a highly clonal coral population and the relationship with ecological performance

Elkhorn coral, Acropora palmata, relies heavily on clonal propagation and often displays low genotypic (clonal) diversity. Populations in the Florida Keys experienced rapid declines in tissue cover between 2004 and 2006, largely due to hurricanes and disease, but remained stable from 2006 to 2010. All elkhorn colonies in 150 m² permanent study plots were genotyped in 2006 (n = 15 plots) and 2010 (n = 24 plots), and plots sampled in both years were examined for changes in allelic and genotypic diversity during this period of stable ecological abundance. Overall, genetic diversity of Florida plots was low and declined further over the 4-yr period; seven of the 36 original genets and two of 67 alleles (among five microsatellite loci) were lost completely from the sampled population, and an additional 15 alleles were lost from individual reefs. In 2010, Florida plots (~19 colonies) contained an average of 2.2 ± 1.38 (mean ± SD) genets with a significant negative correlation between colony abundance and genotypic diversity. When scaled to total tissue abundance, genotypic diversity is even lower, with 43 % of genets below the size of sexual maturity. We examined the hypothesized positive relationship of local genotypic diversity with ecological performance measures. In Florida plots (n = 15), genotypic diversity was not significantly correlated with tissue loss associated with chronic predation, nor with acute disease and storm-fragmentation events, though this relationship may be obscured by the low range of observed diversity and potential confounding with abundance. When more diverse plots in Curaçao (n = 9) were examined, genotypic diversity was not significantly correlated with resistance during an acute storm disturbance or rate of recovery following disturbance. Cryptic loss of genetic diversity occurred in the apparently stable Florida population and confirms that stable or even increasing abundance does not necessarily indicate genetic stability.     

Assessing Common Bean Cultivars for Resistance to the Soybean Looper <Emphasis Type="Italic">Chrysodeixis includens</Emphasis> (Lepidoptera: Noctuidae)

The soybean looper Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae) is known as an important pest of leguminous plants worldwide. In Brazil, this pest species is gaining importance to producers of the common bean Phaseolus vulgaris L. (Fabaceae) because it limits field production of the crop. Chemical control is still the primary method of insect control. However, due to the possible harmful effects of pesticides to humans and the environment, alternative and less aggressive practices are being investigated. For this reason, the use of resistant plant genotypes represents a valuable tool in insect control. This study evaluated the biological aspects of larvae of C. includens confined to 14 bean genotypes under laboratory conditions (26 ± 2°C; 65 ± 10% RH; photoperiod of 14 h L:10 h D). The duration of the instars, total duration of the larval phase, consumption while in the larval phase, weight of the fifth instar larvae, larval viability, duration of the pre-pupal and pupal phases, pupal weight, pupal viability, pupal deformity, caterpillar-to-adult cycle, duration of the pre-oviposition and oviposition periods, and total number of viable eggs per female were evaluated. The genotypes “IAC Boreal,” “IAC Harmonia,” and “IAC Formoso” expressed antibiosis, prolonging the caterpillar-to-adult cycle and reducing the larval viability; however, each of these genotypes also experienced high leaf consumption. “IAC Jabola” expressed moderate levels of antibiosis and/or antixenosis (feeding), while the genotype “BRS Horizonte” expressed antixenosis (feeding). The data obtained with IAC Boreal, IAC Harmonia, IAC Formoso, IAC Jabola, and BRS Horizonte are promising and may help with the improvement of programs aimed at managing C. includens damage to this leguminous agricultural crop.     

Effect of tropical storms on sexual and asexual reproduction in coral Pocillopora verrucosa subpopulations in the Gulf of California

Pocillopora verrucosa is a branching, reef-building coral, and a simultaneous hermaphrodite that reproduces sexually and asexually by fragmentation. In the Gulf of California, local P. verrucosa populations have mixed modes of reproduction which vary in frequency by site. Sexual and asexual reproductions were assessed using multi-locus genotypes deriving from six microsatellite loci at every location. Clone frequencies varied from 0.30 at Loreto to 0.96 in the San Lorenzo Channel. Isla Espíritu Santo and the San Lorenzo Channel were mostly asexual subpopulations, presented the lowest genotypic richness (N _g /N = 0.1–0.12) and genotypic diversity (G _o /G _e  = 0.04), and were dominated by one or two multi-loci genotypes (G _o /N _g  = 0.35–0.45). Loreto, El Portugués, and Cabo Pulmo were mostly sexual with high Ng/N (0.80–0.74) and G _o /G _e (0.52–0.58) and did not show domination by a single multi-locus genotype (G _o /N _g  = 0.70–0.74). There was a significant relationship (P < 0.05) between tropical storm frequency and the genotypic indexes of richness and diversity modeling an inverted U-shape, which indicates that the sites where storm frequencies were the highest had mostly clonal populations; sites exposed to intermediate or low storm frequencies had mostly sexual populations. The study included a restored area (San Lorenzo Channel) where genotypic analyses showed a high level of clonality similar to natural conditions occurring in a nearby subpopulation (Isla Espíritu Santo), which demonstrates the low natural genetic diversity of the area. This study showed that a species with mixed reproduction modes has different maintenance strategies at a regional and even local level among populations indicating the crucial role that storms play in population structure.     

Larval settlement preferences and post-settlement survival of the threatened Caribbean corals Acropora palmata and A. cervicornis

The settlement specificity of two threatened Caribbean corals, Acropora palmata and A. cervicornis, was tested by measuring their rates of larval metamorphosis in response to crustose coralline algae (CCA) and other substrata. In the no-choice experiments, the coral larvae were placed in six treatments: filtered seawater (FSW), a fragment of biofilmed dead skeleton of A. palmata, or a fragment of one of four species of CCA (Hydrolithon boergesenii, Porolithon pachydermum, Paragoniolithon solubile, and Titanoderma prototypum). Within each CCA treatment, there were three different substrata on which to settle and metamorphose: (1) the CCA surface, (2) the rock under the CCA, or (3) the plastic dish. The 5-day-old larvae of both A. palmata and A. cervicornis had similar rates of total metamorphosis (all substrata combined) in every treatment (excluding FSW) even in the absence of CCA. However, there were differences in larval behavior among the CCA species since the larvae settled and metamorphosed on different substrata in the presence of different CCA species. In the no-choice experiments the larvae of both corals had higher rates of metamorphosis on the top surfaces of H. boergesenii and/or T. prototypum than on P. pachydermum. In the choice experiments, the coral larvae were offered two species of CCA in the same dish. When given a choice, both species of coral larvae had more settlement and metamorphosis on the surface of H. boergesenii or T. prototypum or clean rock than onto the surface of P. solubile. After 6 weeks in the field, transplanted A. palmata recruits had approximately 15% survival on both T. prototypum and H. boergesenii, but A. cervicornis recruits only survived on T. prototypum (13%). Some, but not all, CCA species facilitated the larval settlement and post-settlement survival of these two threatened corals, highlighting the importance of benthic community composition for successful coral recruitment.     

Growth and morphological development of laboratory-reared larvae and juveniles of the Laotioan indigenous cyprinid Hypsibarbus malcolmi

The morphological development, including the pigmentation, body proportions, fins, and survival rate for 30 days after hatching, of laboratory-reared larval and juvenile Hypsibarbus malcolmi is described. Body lengths (BL) of larvae and juveniles were 2.0 ± 0.2 (mean ± SD) mm at 1 h after hatching (day 0) and 9.2 ± 0.6 mm on day 16, reaching 12.1 ± 0.9 mm on day 30. Yolk volume decreased linearly, with the yolk being completely absorbed by day 3 in all preflexion larvae (all specimens >3.2 mm BL). Feeding was observed on day 2 in fish which had rapidly undergone complete yolk absorption following mouth and anus opening on day 1, and on day 3 in all remaining fish. Myomere numbers were 20–21 + 11–12 = 31–33, although they were not clearly visible in juveniles. Melanophores were few on the body during days 0–2, but increased with growth and covered the entire upper dorsal body surface during the juvenile stage. Body proportions tended to become constant in juveniles. Notochord flexion began in larvae >5.2 mm BL on day 8, and was completed in larvae >8.4 mm BL on day 14. Specimens with full fin ray complements were initially observed on day 22 (10.4 mm BL in juveniles). All specimens >11.5 mm BL had attained the juvenile stage. A high survival rate of 92.7% was estimated on day 30.     

Involvement of reactive oxygen species in the electrochemical inhibition of barnacle (Amphibalanus amphitrite) settlement

The role of reactive oxygen species (ROS) in electrochemical biofouling inhibition was investigated using a series of abiotic tests and settlement experiments with larvae of the barnacle Amphibalanus amphitrite, a cosmopolitan fouler. Larval settlement, a measure of biofouling potential, was reduced from 43% ± 14% to 5% ± 6% upon the application of pulsed electric signals. The application of ROS scavengers such as glutathione and catalase counteracted the inhibitory effects of the electric signals, allowing settlement, and thus indicating that ROS are antifouling agents. Based on the experimental evidence, the proposed mechanism for ROS-based fouling prevention with interdigitated electrodes involved the electrochemical generation of hydrogen peroxide by oxygen reduction, and its likely reduction to hydroxyl radicals. Either hydroxyl radicals or products of hydroxyl radical reactions appeared to be the main deterrents of larval settlement.     

Growth and morphological development of laboratory-reared larval and juvenile bighead catfish Clarias macrocephalus (Siluriformes: Clariidae)

Morphological development in laboratory-reared larval and juvenile bighead catfish Clarias macrocephalus is described. Body length (BL) of larvae and juveniles was 3.4 ± 0.3 (mean ± SD) mm just after hatching, reaching 11.3 ± 1.0 mm by day 16, and 24.2 ± 2.8 mm by day 40. Overall aggregate fin ray numbers (except for caudal fin procurrent rays) attained full complements by 15.2 mm BL. Gill buds appeared on day 0, those of barbels (four pairs) and primordial nostrils on day 1, and pectoral fins on day 3. All larvae began feeding by day 3. Conical teeth were observed on day 7. Notochord flexion began on day 2, the yolk being completely absorbed during days 7–9. Melanophores were scarce at hatching, increasing with growth to cover almost the entire body except the ventral surface of the abdominal cavity. Proportions of head length, pre-anal length, body depth, eye diameter, and maxillary barbel length became relatively constant after yolk absorption, those of snout length and upper jaw length increasing until ca. 12–13 mm BL and decreasing thereafter. Suprabranchial organ started developing in postflexion larval stage larger than ca. 11.0 mm BL (day 16), and air-breathing was suggested to be functional at that time.     

The effect of Achyranthes japonica extract on larval survival and development and oviposition behavior of Plutella xylostella L. (Lepidoptera: Plutellidae)

The extract of Achyranthes japonica was tested for effects on larval survival and development and the oviposition behavior of the diamondback moth, Plutella xylostella L. Chinese cabbage dipped in A. japonica extract solution showed 51–80% antifeedant activity for 5 days against P. xylostella larvae, and more larvae were also on untreated cabbage leaves 24 h after release. The mortality of P. xylostella larvae increased proportionally to the duration of dipping time in the extract, and both pupation and emergence rates of larvae feeding only on treated cabbage were lower than those for larvae raised on untreated or with a choice of cabbage. The 20-hydroxyecdysone (20E) concentration in leaves was approximately 549, 1232, 1275, and 1426 μg/g at 6, 12, 24, and 48 h after dipping treatment, respectively. Notably, naive females laid more eggs on untreated cabbage than on treated cabbage, and females from larvae raised on treated Chinese cabbage also preferred the non-treated leaves. Our results are in contrast to those from earlier studies using various insect models that confirmed most females prefer to lay eggs on the host type that was eaten in the larval stage (Hopkins host selection principle). Cabbage dipped in the A. japonica solution for 24 h caused 59% larval mortality and inhibited both pupation and emergence rates of the larvae when exposed to plants 15 and 22 days after planting in the field, with the 20E concentration in the treated cabbage leaves at 1600.9 ± 122.36 and 1386.8 ± 24.69 μg/g, respectively. Therefore, the biological effectiveness could be attributed to the 20E in the treated cabbage leaves.     

Larval settlement preferences of Acropora palmata and Montastraea faveolata in response to diverse red algae

Settlement specificity can regulate recruitment but remains poorly understood for coral larvae. We studied larvae of the corals, Acropora palmata and Montastraea faveolata, to determine their rates of settlement and metamorphosis in the presence of ten species of red algae, including eight species of crustose coralline algae, one geniculated coralline and one encrusting peyssonnelid. Twenty to forty percent of larvae of A. palmata settled on coralline surfaces of Hydrolithon boergesenii, Lithoporella atlantica, Neogoniolithon affine, and Titanoderma prototypum, whereas none settled and metamorphosed on Neogoniolithon mamillare. Larvae of M. faveolata had 13–25 % settlement onto the surface of Amphiroa tribulus, H. boergesenii, N. affine, N. munitum, and T. prototypum, but had no settlement on the surface of N. mamillare, Porolithon pachydermum, and a noncoralline crust Peyssonnelia sp. Some of these algal species were common on Belizean reefs, but the species that induced the highest rates of larval settlement and metamorphosis tended to be rare and primarily found in low-light environments. The shallow coral, A. palmata, and the deeper coral, M. faveolata, both had increased larval settlement rates in the presence of only a few species of red algae found at deeper depths suggesting that patterns of coral distribution can only sometimes be related to the distribution of red algae species.     

Assessing genetic structure in a rare clonal eucalypt as a basis for augmentation and introduction translocations

Genetics can provide information that is critical for planning translocations for conservation, such as levels of diversity and divergence of target and source populations. For clonal plants, assessing population characteristics (size, diversity, mortality, gene flow) that influence conservation values also requires identification of different genetic individuals. We used 12 microsatellite markers to guide germplasm source recommendations for augmentation and introduction translocations to conserve the critically endangered Eucalyptus cuprea that occurs in fragmented populations in the semi-arid shrublands of Western Australia. Ramet clumps with identical multilocus genotypes were identified in all populations but clonal richness (R = 0 ? 0.86) and heterogeneity (D = 0 ? 0.98) varied among populations. Genetic diversity was low to moderate in all populations (mean H _o = 0.61, mean A = 3.78) and did not differ significantly between localities. There was evidence of inbreeding in some populations but outcrossing (t _m = 0.495) in the small number of families available for study (N = 4) and genotypic diversity of the larger extant populations suggest the generation of novel genotypes is a component of the reproductive strategy. Most diversity was within populations and differentiation among populations was moderate (F _ST = 0.100) suggesting mixing of source population for translocation is unlikely to lead to outbreeding depression. Principal Co-ordinate and Bayesian analyses indicated the Northern population is distinct from Central/Southern populations. We recommend use of mixed germplasm to conserve the moderate diversity characterising larger remnant populations and to enable the production of recombinants through sexual reproduction. But given seed availability and the distinction of the Northern population, an initial precautionary approach to a translocation proposed for south of the geographical range may be to source germplasm from the Central/Southern locality.     

Functional Response of Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae) to Sugarcane Whitefly Aleurolobus barodensis (Maskell) in Laboratory Conditions

A functional response study of Chrysoperla carnea (Stephens) larvae to different densities of sugar cane whitefly Aleurolobus barodensis (Maskell) was conducted in test tubes at 26?±?2 °C, 65?±?5 % RH. Chrysoperla carnea showed two different types of functional response in larval instars. First instar exhibits type II. However, second and third larval instars revealed type III functional response. Based on modified Holling’s disk equation, the highest searching rates (a) of 0.82?±?0.0247 h^?1 was found for first instar larva. For second and third larval instars, the attack coefficient (b) were 0.002?±?0.030 and 0.0025?±?0.0424 respectively. The shortest handling time (T_h) per prey was observed at third instar stage (1.574?±?0.0568 h) followed by second and first instar with 1.72?±?0.0411 h and 1.919?±?0.0568 h respectively.     

Ascaris suum: Influence of embryonation temperature on the viability of the infective larva

• 1.1.|The infective larvae of Ascaris suum develop in the egg between the temperatures 16 ± 1°C and 34 ± 1°C. Within this temperature range, increases in temperature increase the rate of development. The maximum rate of egg development is attained at 31 ± 1°C.
• 2.2.|Eggs embryonated at 28 ± 1°C and above give rise to infective larvae which have less ability to hatch in vitro, shorter longevity when aged in phosphate buffered saline (pH 7.2) at 37°C, and more limited ability to penetrate tissue membranes in vitro, when compared with those larvae from eggs embryonated at lower temperatures.
• 3.3.|Maximum larval viability and ability to penetrate tissues in vitro was achieved when eggs were embryonated at 22 ± 1°C. These results suggest that the optimal temperature for rate of development and larval viability, or survivability are not the same, a point which has not previously been emphasised.
• 4.4.|The practical significance of these results are discussed in relation to the epidemiology of Ascarias and the known biology of the parasite.

     

The thermal tolerance of crown-of-thorns (Acanthaster planci) embryos and bipinnaria larvae: implications for spatial and temporal variation in adult populations

To understand the role of sea temperature on the population biology of the crown-of-thorns sea star Acanthaster planci, the thermal window for embryonic and larval development was investigated. In two experiments, the response of embryos and larvae across 12 temperatures from 19.4 to 36.5 °C was quantified as the percentage of individuals reaching cleavage stage embryos, blastula, gastrula, early-bipinnaria, late-bipinnaria larvae or abnormal. Measurements were made at 7 times up to 72 h post-fertilisation, with the morphometrics of larvae measured in the 72-h sample. Acanthaster planci developed at temperatures between 19.4 and 33.2 °C, with a thermal window for development to the late-bipinnaria stage between 25.6 and 31.6 °C. Development rate, normal development and larval size were optimal at 28.7 °C, with development rates remaining relatively constant up to 31.6 °C. Rates of abnormality increased steadily (early embryonic stages) above 28.7 °C and was 100 % at temperatures approaching 33 °C. These experiments provide a more detailed insight into the response of A. planci developmental stages to temperature. The present day distribution of the species in eastern Australia overlap with the optimal thermal window for development to the late-bipinnaria stage (≈25–32 °C), implying a role of temperature in controlling population distributions and abundances. Despite this, short- or long-term temperature increases may not be a major modulator of the crown-of-thorns recruitment success, population dynamics and distribution in the future as no significant change in development rates, larval survival and growth occurred within this thermal window. Therefore, moderate (1–2 °C) increases in sea temperatures caused by El Niño or near-future ocean warming may not drive an increase in developmental and settlement success. Indeed, without any acclimation to warmer temperatures expected under near-future warming (+2 to 4 °C), climate change could ultimately reduce larval survival due to elevated mortality above the optimal development temperature.     

Influence of temperature and light–dark cycle on hatching of <Emphasis Type="Italic">Eylais extendens</Emphasis>

Little research has been done on egg diapause and the embryonic development of water mites. The aim of this study was to check the impact of temperature and periods of light on hatching of larvae of Eylais extendens. Three batches of eggs which were spawned on 30 July were placed at one of three temperatures (4, 10 and 20 °C) and two periods of light (7 and 14 h per day). Egg hatching (both, percentage of hatched larvae and rate of hatching) was found to differ between 4 versus 10 °C and between 4 versus 20 °C, but not between 10 versus 20 °C. The periods of light had no influence on hatching. This synchronization of hatching, enabling the eggs to emerge from diapause in the spring, could be considered an evolutionary adaptation aimed at postponing hatching of late-spawned eggs until a time allowing for completion of the full development cycle, including the parasitic larval stage.     

Environmental induction of larval diapause and life-history consequences of post-diapause development in the Large Copper butterfly,Lycaena dispar (Lepidoptera: Lycaenidae)

Many insects in temperate zones withstand the adverse conditions of winter through entering diapause and the two most important environmental stimuli that induce diapause are photoperiod and ambient temperature. The Large Copper butterfly, Lycaena dispar Haworth (Lepidoptera: Lycaenidae), is a Palearctic butterfly that hibernates as larvae. Since this butterfly is a near threatened species in some regions, there has been a growing need for a standardized protocol for mass rearing of this butterfly based on the adequate knowledge of its ecology. In the present study, we first identified that L. dispar larvae were sensitive to the photoperiodic induction of diapause during their first larval instar. We then investigated to what extent the diapause-inducing effects of photoperiod could be modified by ambient temperatures in L. dispar larvae by exposing them to the range of day-lengths (L:D 14:10, 12:12, 10:14 and 8:16) at three different temperatures (15, 20 and 25 °C). All larvae were induced to enter diapause at low ambient temperature (15 °C) regardless of photoperiod, whereas most of them (86 %) exhibited direct development when temperature was high (25 °C). The photoperiodic induction of diapause was evident when day-length was shorter than 14 h at intermediate temperature (20 °C). Pre-diapause development was prolonged at low temperatures. Finally, we found that post-diapause development of L. dispar larvae was determined by both the chilling temperature experienced by diapausing larvae and the duration of larval diapause. Adult emergence was enhanced when larvae were chilled at 8 °C and when they had been under the state of diapause for 20 days before they were treated to terminate diapause.     

Patterns of genotypic diversity suggest a long history of clonality and population isolation in the Australian arid zone shrub Acacia carneorum

For plants capable of both sexual and clonal reproduction, the relative frequency of these reproductive modes is influenced by genetic and ecological factors. Acacia carneorum is a threatened shrub from the Australian arid zone that occurs as a set of small, spatially isolated populations. Sexual reproduction appears to be very rare: despite regular flowering, only two populations set seed. It is not known whether this reflects an ancient pattern, or results from rapid land use changes following arrival of Europeans in the region 150 years ago. We assessed genotypic variation throughout the range of A. carneorum using AFLP markers, to elucidate the relative importance of clonal and sexual reproduction in this species’ history. Clonal diversity (CD) within populations ranged from 0 to 0.820 (mean CD = 0.270, SE = 0.094), but the relative abundances of genets were typically highly skewed. On average, the two fruiting populations had higher CD (mean CD = 0.590, SE = 0.265) than non-fruiting populations (mean CD = 0.179, SE = 0.077) (t = 2.315, p = 0.049), but most populations contained multiple genets. All genets were population-specific, and there was substantial divergence among populations (Φ _ST = 0.690), implying a long history of isolation. We conclude that clonality has predominated in A. carneorum populations, with occasional sexual recruitment, and that current failure of most populations to set seed likely reflects both a long history of asexual reproduction and effects of habitat disturbance. Conservation of this species may benefit from translocations to increase genotypic diversity within populations.     

Acute toxicity of ammonia and nitrite to different ages of Pacific cod (Gadus macrocephalus) larvae

Abstract

This study was conducted to evaluate the acute toxicity of ammonia and nitrite to three developmental stages of Pacific cod (Gadus macrocephalus) larvae (11, 22, and 35 days after hatching, with mean total lengths of 4.63 ± 0.14, 5.83 ± 0.17, and 7.46 ± 0.23 mm, respectively). The results showed for the first time that the acute toxicity of ammonia and nitrite is closely related to the age of Pacific cod larvae, and the acute toxicity of ammonia or nitrite increased with increased Pacific cod larval growth. Lethal concentrations (LC₅₀) of un-ionized ammonia nitrogen (UIAN) for a 48-h exposure in 11-day post-hatch, 22-day post-hatch, and 35-day post-hatch Pacific cod larvae were 1.72, 0.69, and 0.32 mg L^?1, respectively. The 48-h LC₅₀ of nitrite nitrogen to Pacific cod larvae 11-day post-hatch, 22-day post-hatch, and 35-day post-hatch were 831.76, 269.15, and 223.87 mg L^?1, respectively. The present findings demonstrate that the acute toxicity of ammonia for Pacific cod larvae is much higher than that of nitrite.     

Phenology of sexual reproduction in the common coral reef sponge,Carteriospongia foliascens

Understanding processes that contribute to population maintenance is critical to the management and conservation of species. Despite this, very little is currently known about the reproductive biology of Great Barrier Reef (GBR) sponge species. Here, we established reproductive parameters including mode of sexuality and development, seasonality, sex ratios, gametogenesis, reproductive output, and size at sexual maturity for the common phototrophic intertidal sponge, Carteriospongia foliascens, in the central GBR over two reproductive cycles. A population sexual productivity index (PoSPi) integrating key reproductive parameters was formulated to compare population larval supply over time. This study shows that C. foliascens is reproductive all year round, gonochoric and viviparous, with larvae developing asynchronously throughout the mesohyl. The influence of environmental parameters relevant to C. foliascens reproduction [i.e., sea surface temperature (SST), photoperiod, and rainfall] was also examined, and SST was found to have the most significant effect on phenology. C. foliascens reproduction exhibited annual mono-cyclic patterns closely resembling SST fluctuations. Reproductive output was depressed at low SST (<23 °C) and increased at temperatures above 23 °C. Peak sperm release occurred at temperatures above 25 °C, while peak larval release occurred during the annual temperature maxima (>28 °C). A twofold increase in maximum larval production (PoSPi) in C. foliascens was observed in the second reproductive cycle, following a depressed PoSPi in the first cycle. This reduction in PoSPi in the first reproductive cycle was associated with elevated SST and rainfall, coinciding with one of the strongest La Niña events on record.     

Rearing bacteria and maggots concurrently: a protocol using Lucilia sericata (Diptera: Calliphoridae) as a model species

Maggot debridement therapy using live Lucilia sericata (Meigen) larvae is an efficient and cost-effective way to treat chronic wounds. The recent increase in studies to assess the antibacterial properties of L. sericata has created a need for a simple, low-cost, and comprehensible rearing and investigative method for researchers with little or no entomological experience. This paper describes and evaluates a reproducible protocol for sterilising and rearing blowfly larvae utilising two sterile artificial diets (blood–yeast agar and pre-prepared blood agar plates) that is suitable for directly investigating the effect of larvae on microbial growth. Using Lucilia sericata as a model, the results show that larval growth on the pre-prepared blood agar diet is detrimental to larval growth and survival, whereas larval growth and survival on the blood–yeast agar diet are comparable to those of larvae raised on porcine liver. This diet is proposed as a standard for blowfly and bacteria interaction studies investigating clinical microbial strains. Developmental data are provided for L. sericata larvae raised on both sterile and nonsterile diets so that researchers can determine the effect of treatment based on the length of time for larvae to reach the required life stage at 25 ± 2 °C. Information on larval ageing (instars at an average of 1, 2, 3 and 4 days), oviposition times (4–5 days after adult emergence) and adult longevity on the diets (102–116 days) is also given.     

Endogenous arabitol and mannitol improve shelf life of encapsulated <Emphasis Type="Italic">Metarhizium brunneum</Emphasis>

Successful commercialization of microbial biocontrol agents, such as Metarhizium spp., is often constrained by poor drying survival and shelf life. Here, we hypothesized that culture age would influence endogenous arabitol, erythritol, mannitol and trehalose contents in M. brunneum mycelium and that elevated levels of these compounds would improve drying survival and shelf life of encapsulated mycelium coupled with enhanced fungal virulence against T. molitor larvae. We found that culture age significantly influenced endogenous arabitol and mannitol contents in mycelium with highest concentrations of 0.6?±?0.2 and 2.1?±?0.2 µg/mg after 72 h, respectively. Drying survival of encapsulated mycelium was independent of culture age and polyol content with 41.1?±?4.4 to 55.0?±?6.2%. Best shelf life was determined for biomass harvested after 72 h at all investigated storage temperatures with maximum values of 59.5?±?3.3% at 5 °C followed by 54.5?±?1.6% at 18 °C and 19.4?±?1.3% at 25 °C after 6 months. Finally, high fungal virulence against T. molitor larvae of 83.3?±?7.6 to 98.0?±?1.8% was maintained during storage of encapsulated mycelium for 12 months with larval mortalities being independent of culture age and polyol content. In conclusion, our findings indicate beneficial effects of endogenous polyols in improving shelf life of encapsulated mycelium and this may spur the successful development of microbial biocontrol agents in the future.