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.     

Effect of avocadofurans on larval survival, growth, and food preference of the generalist herbivore, Spodoptera exigua

We examined the effect of two avocadofurans, 2-(pentadecyl)furan and 2-(heptadecyl)furan, from avocado idioblast oil cells on maturation and larval feeding behavior of a generalist insect herbivore, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). Experiments were conducted using two larval sizes: early-stadium larvae refer to those larvae from experiments initiated with neonates while late-stadium larvae refer to those larvae from experiments initiated with third instars. In order to use selected sublethal doses for developmental and behavioral studies on early- and late-stadium larvae, log-dose probit lines were determined using diet incorporation bioassays. Both avocadofurans had similar toxicities to early-stadium larvae [LC50=2.2 and 1.9 moles/g of diet for 2-(pentadecyl)furan and 2-(heptadecyl)furan, respectively] and late-stadium larvae (LC50=3.0 and 3.4 moles/g of diet, respectively). In diet bioassays extending from egg hatch to adult emergence, the avocadofurans significantly prolonged larval developmental times and reduced S. exigua pupal weights. In 7 d no-choice bioassays initiated with cohorts of newly-molted third instars, the avocadofurans significantly reduced larval weights at various sublethal concentrations (below LC50 values). To test larval feeding deterrence effects of these avocadofurans, choice tests were conducted using early and older instar larvae. A significantly higher proportion of early-stadium larvae preferred control diet over diet treated with either avocadofuran at several sublethal concentrations. Similarly, choice tests with late-stadium larvae showed greater proportions of larvae on control diet than treated diet even at concentrations below the LC50. Moreover, late-stadium larvae consumed significantly more of the control diet than the treated diet. Thus, the avocadofurans may act as feeding deterrents as well as toxicants in plant protection against non-adapted insect herbivores.     

Artificial diets for rearing the coconut hispine beetle Brontispa longissima and its parasitoid Asecodes hispinarum

We tested artificial diets for rearing the coconut hispine beetle, Brontispa longissima, a serious invasive pest of coconut (Cocos nucifera) in Southeast Asia. We examined three artificial diets that were identical except for their agar content. The survival rate from hatching to adult emergence was 26.0% when beetles were reared on a ‘soft diet’ (20 g/l agar), 16.0% on a ‘hard diet’ (40 g/l agar), and 41.0% on a ‘mixed diet’, in which the hard diet was used for the first instar and the soft diet for later instars. Females raised as larvae on the mixed diet and then as adults on the soft diet laid few eggs, which did not hatch. However, females reared on the mixed diet as larvae and then on fresh leaves as adults consistently laid eggs that hatched. We then examined the suitability of B. longissima larvae reared on the mixed diet as hosts for the larval parasitoid Asecodes hispinarum, a specialist parasitoid of this beetle. Of the oviposited hosts, 75.0% became mummified and 41.7% produced adult wasps. These results indicate that the mixed diet may be useful for rearing B. longissima larvae as hosts for the rearing of A. hispinarum.     

The in vitro development from egg to prepupa of Campoletis sonorensis (Hymenoptera: Ichneumonidae) in an artificial medium: importance of physical factors

A further attempt to improve the in vitro rearing of the solitary larval endoparasitoid Campoletis sonorensis (Hymenoptera: Ichneumonidae) was made after third instars were obtained in previous studies. Respiration was an important factor for the parasitoid's continued development: both small diet volumes that exposed the larval cuticle to air and O(2)greatly improved growth and development of C. sonorensis. Fifth instars were obtained when the parasitoid larvae were reared in medium volumes of 0.25ml and 0.1ml per well. Increasing the O(2) tension of the air resulted in larger larvae and prepupae. The agar layer under the nutritional medium was designed to slowly remove water from the diet. The results indicated that a drier environment was important for the parasitoid at the terminal stage. One percent and 0.8% agar gels, which removed water from the diet more rapidly, provided a drier environment and resulted in improved larval growth compared to the 0.6% agar gel. The detrimental effect of higher concentration of chicken egg yolk mixed in the diet was eliminated by using a dry egg yolk layer covered by an agar layer. This sandwich technique resulted in more parasitoid larvae molting to the final larval instar (5th) and the formation of many prepupae. However, the development of prepupae was premature and none of the fifth instar larvae successfully formed a normal cocoon.     

Effect of larval density on development and diapause of Sesamia nonagrioides (Lef.) (Lep., Noctuidae) under laboratory conditions

The effect of larval density on development and diapause of Sesamia nonagrioides (Lef.) was examined. Larvae were grown on an artificial diet at densities of five, 10, 20, 30 and 40 larvae per box, under two environmental (diapause-including and diapause-averting) conditions. High larval densities resulted in increased mortality under both experimental conditions tested. Although crowded larvae showed a higher pupation rate than those reared under low densities, small differences occurred between mean larval developmental time at different densities under developmental condition. In spite of this, crowding proved effective on the intensity of diapause of this species and the duration of diapause was shorter for crowded larvae. Pupal weights of both sexes were significantly reduced at high densities while there were significant differences between pupal weights formed by larvae reared under diapause conditions and those formed by larvae reared under nondiapause conditions at all density treatments. The pupal period was significantly longer under diapause conditions than that under development conditions but density was not found to play a role on the time of the emergence of the adults produced by larvae reared under developmental or diapause conditions.     

粘虫幼虫密度对成虫能源物质含量的影响

通过粘虫Mythimna separata幼虫密度(1头/瓶、20头/瓶、40头/瓶)对成虫水分、甘油酯和糖原含量影响的研究结果。不同幼虫密度处理的初羽化成虫水分含量无明显差异,但1～5日龄40头/瓶的雄蛾及1～3日龄雌蛾高于单头处理的;20、40头/瓶的初羽化成虫甘油酯含量没有显著的差异,但均显著地高于单头饲养的;20头/瓶羽化的雌、雄成虫l～5日龄甘油酯含量随日龄的增加而增加,到5日龄达到最大值后才开始下降,40头/瓶的在3日龄达到最大值后即开始下降,而单头饲养羽化的在1日龄达到较高值,2日龄降至最低后再缓慢回升。幼虫密度对初羽化成虫及1～7日龄雄蛾的糖原含量没有显著的影响,但20、40头/瓶条件下羽化的1～7日龄雌蛾糖原含量随日龄的增加而增加,而单头饲养的则随日龄的 增加而下降。这些结果表明幼虫密度不仅影响到初羽化成虫能源物质的含量,而且也可能影响到成虫能源物质特别是甘油酯的代谢。     

Black soldier fly (Diptera: Stratiomyidae) larval heat generation and management

Mass production of black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae), larvae results in massive heat generation, which impacts facility management, waste conversion, and larval production. We tested daily substrate temperatures with different population densities (i.e., 0, 500, 1000, 5000, and 10 000 larvae/pan), different population sizes (i.e., 166, 1000, and 10 000 larvae at a fixed feed ratio) and air temperatures (i.e., 20 and 30 °C) on various production parameters. Impacts of shifting larvae from 30 to 20 °C on either day 9 or 11 were also determined. Larval activity increased substrate temperatures significantly (i.e., at least 10 °C above air temperatures). Low air temperature favored growth with the higher population sizes while high temperature favored growth with low population sizes. The greatest average individual larval weights (e.g., 0.126 and 0.124 g) and feed conversion ratios (e.g., 1.92 and 2.08 g/g) were recorded for either 10 000 larvae reared at 20 °C or 100 larvae reared at 30 °C. Shifting temperatures from high (30 °C) to low (20 °C) in between (∼10-d-old larvae) impacted larval production weights (16% increases) and feed conversion ratios (increased 14%). Facilities should consider the impact of larval density, population size, and air temperature during black soldier fly mass production as these factors impact overall larval production.     

Effect of larval diet on cat flea (Siphonaptera: Pulicidae) developmental times and adult emergence.

The natural diet of cat flea, Ctenocephalides felis (Bouche), larvae is primarily adult flea feces, but dried bovine blood may be substituted in the laboratory. Percentage adult emergence (79.4% on feces; 78.9% on blood) and developmental times (20.6 d on feces; 17.1 d on blood) did not significantly differ for the two diets. The drying temperature of blood determined its quality; blood dried at 120 degrees C was unsatisfactory for larval development. The dietary value of dried bovine blood was not enhanced when supplemented with brewer's yeast, rodent chow, or a combination of those constituents. Blood particle size ranging from less than 180 to greater than 500u did not affect the value of blood as a diet. Rodent chow, yeast, albumen, hemoglobin, and mixtures of these constituents were unsuitable as larval diets.     

In Vitro Rearing of the Parasitoid Exorista larvarum (L.) (Diptera: Tachinidae) on Meat Homogenate-Based Diets

The tachinid Exorista larvarum (L.), a polyphagous gregarious larval endoparasitoid of Lepidoptera, was reared from egg to fecund adult on media containing commercial meat homogenates for babies as the main ingredient. Four media, each containing a diverse homogenate supplemented with extract of Galleria mellonella L. pupae, were tested first. Despite the difference in nutrient content, the kind of homogenate did not significantly affect the adult yields (30.2 to 40.7%) or puparial weights. Two other diets free of host materials (I and II) were then tested. Both were based on Gerber veal homogenate combined with different amounts of yeast extract and chicken egg yolk and were supplemented with wheat germ (I) or saccharose (II). Adult yields (28.7 to 32.7%) and puparial weights did not differ significantly between the two diets. Fly longevity and fecundity of the females obtained on diet I were comparable to those of the females emerged from puparia formed in G. mellonella larvae. Male and female puparial weights were, however, higher and development times longer on the diet than in the host.     

Toxicity and synergism in the feeding deterrence of some coumarins on Spodoptera frugiperda Smith (Lepidoptera: Noctuidae)

The phagodepression activity of five coumarins (= 2H-1-benzopyran-2-ones), 6-hydroxy-7-isoprenyloxycoumarin (1), 6-methoxy-7-isoprenyloxycoumarin (2), 6,7-methylenedioxycoumarin (3), 5-methoxy-6,7-methylenedioxycoumarin (4), and 6-methoxy-7-(2-hydroxyethoxy)coumarin (5), from the Argentine native herb Pterocaulon polystachyum, was tested against Spodoptera frugiperda (Lepidoptera: Noctuidae) larvae. Two analogs, scopoletin (6) and 2-methoxy-2-methyl-3,4,5,6,7,8-hexahydro-3H-chromen-5-one (7), synthesized in our laboratory, were also evaluated for comparison. The compounds were added to an artificial diet at doses ranging from 50 to 200 microg per g of diet. Natural coumarins induced 100% of phagodepression when 200 microg were added per g of diet. Binary equimolar mixtures of the natural coumarins were phagodepressors against S. frugiperda surpassing the expected additive responses, indicating that these compounds can act synergistically against S. frugiperda larvae. Compounds 1 and 3 (non-methoxylated coumarins), and the equimolar mixture of both, displayed the strongest phagodepression. Additionally, 50 microg/g of 1 and 3 incorporated to the larval diet caused 80 and 50% of pupal mortality, respectively, while a 100 microg/g dose of compounds 2, 4, 6, and 7 produced 60, 50, 10, and 80% pupal mortality, respectively. Larval growing rate during the early larval instars was significantly reduced by treatments with the methylenedioxycoumarins 3 and 4. Coincidentally, the larval period duration was significantly increased by the latter compounds.     

Nutrient balance in medfly, Ceratitis capitata, larval diets affects the ability of the developing insect to incorporate lipid and protein reserves

The Mediterranean fruit fly [Ceratitis capitata Wiedemann (Diptera: Tephritidae)], or medfly, is mass produced in many facilities throughout the world to supply sterile flies for sterile insect technique programs. Production of sterile males requires large amounts of larval and adult diets. Larval diets comprise the largest economic burdens in the mass production of sterile flies, and are one of the main areas where production costs could be reduced without affecting quality and efficacy. The present study investigated the effect of manipulating diet constituents on larval development and performance. Medfly larvae were reared on diets differing in the proportions of brewer's yeast and sucrose. We studied the effect of such diets on the ability of pupating larvae to accumulate protein and lipids, and on other developmental indicators. Except for diets with a very low proportion of brewer's yeast (e.g., 4%), pupation and adult emergence rates were in general high and satisfactory. The ability of pupating larvae to accumulate lipid reserves and proteins was significantly affected by the sucrose and yeast in the diet, and by the proportion of protein to carbohydrates (P/C). In contrast to previous nutritional studies conducted with other insects, low P/C in medfly larval diets (with excess dietary carbohydrates) resulted in pupating medfly larvae having a relatively reduced load of lipids; medfly larvae protein contents in these diets were, as expected, relatively low. Similarly, high P/C ratios in the diet produced larvae with high protein and lipid contents. Differences with other insects may be due to differential post‐ingestion regulation where a high dietary carbohydrate diet reduces the lipogenic activity of the larvae, and induces a shift from lipid to glucose oxidation. Larvae reared on low P/C diets spent more time foraging in the diet than larvae maintained on a high P/C diet, suggesting a compensatory mechanism to complement nutrient intake. The results suggest that the content of brewer's yeast, the most expensive diet component, could be fine‐tuned without apparently affecting fly quality.     

Food preferences and the value of animal food for the carabid beetle Amara similata (Gyll.) (Col., Carabidae)

Abstract:  Several studies have shown that the mainly granivorous carabid beetles, e.g. Amara spp., include animal food in their diet to a considerable extent. We therefore hypothesized that the performance of these beetles would be enhanced by dietary mixing including both seeds and animal food. In order to test this, we conducted laboratory feeding experiments with adults and larvae of Amara similata . Both adults and larvae were subjected to different diet treatments including: seeds, houseflies, grasshoppers, earthworms, slugs and snails in pure and mixed diets. Larval survival, development time, pupal and teneral weights were used as indicators of food quality for the larvae. For the adult beetles, mass change was used as an indicator of food quality. We found seeds to be high-quality food, while all pure animal diets were of low quality for both adults and larvae. Animal foods added to the seed diet had both positive and negative effects. A mixed diet of all foods enhanced the mass gain of adults compared with the seed diet, but reduced larval performance dramatically. Earthworms and grasshoppers added to seeds increased the pupal and teneral weights, while reduced larval survival. Thus, A. similata is omnivorous with a mainly granivorous feeding habit. It may gain benefits on some fitness parameters and incur costs on others from preying or scavenging on animal food. Therefore, the overall fitness consequences of a mixed seed-animal diet are uncertain.     

In vitro rearing of the screwworm fly Wohlfahrtia magnifica

Wohlfahrtia magnifica (Schiner) (Diptera: Sarcophagidae) is the main agent of traumatic myiasis in many European, African and Asian countries. Although it can be reared in vivo without technical difficulty, such rearing presents ethical problems. Studies were therefore made of in vitro rearing to facilitate development of laboratory colonies that could be used in a wide range of biological, physiological and applied studies of W. magnifica, particularly in the long period of the year when natural populations of the fly are unavailable for study. Parental colonies of W. magnifica were established from larvae collected from natural infestations of sheep and cattle in central Hungary. First stage larvae were harvested from gravid females and were reared in groups of 5-20 on one of six artificial diets. The diets were based on various combinations of five to seven of eight ingredients: water, agar, blood (heparinized or dried), ground meat, egg yolk, low-fat milk powder, yeast and 10% formol. The larvae were incubated on the diets at 37 degrees C. There was no mortality of first stage larvae, which appeared to feed together in foci, in a natural manner. However, during the second stage, and especially after renewal of diet associated with disturbance of the larvae, many larvae began to disperse, crawling over the surface of the media and feeding less intensively. Mortality of larvae during all larval stadia was 64-98%, compared to 33% in batches of third stage larvae collected from natural infestations. The mean weights of puparia from artificial diets ranged from 38.7 to 59.3 mg, compared to 92.2 mg of puparia from larvae collected from natural infestations. There was a high mortality in the pupal stage, from 61 to 100%. Only a maximum of 6% of first stage larvae were successfully reared to the adult stage. Further studies are needed to identify factors present or absent in the diets that contributed to the present poor development of W. magnifica in vitro.     

幼虫密度对二点委夜蛾生长发育及繁殖的影响

【目的】在不同幼虫密度饲养条件下,研究二点委夜蛾Athetis lepigone生长发育及繁殖的情况,明确幼虫密度对该害虫的室内种群增长的影响。【方法】本实验设置5个幼虫饲养密度即1,5,10,20和30头/瓶(750 mL),分别观察5个饲养密度下该虫的各个龄期及整个幼虫发育历期及存活率、蛹重、蛹期以及成虫生殖情况。【结果】幼虫密度对该虫幼虫各龄期及整个幼虫发育历期及存活率、蛹重、蛹期以及成虫生殖情况均有显著性影响。整个幼虫发育历期随着密度的增加而缩短,10头/瓶达到最短(18.27 d),之后随着幼虫密度的增加而显著延长;幼虫至蛹的存活率随着密度增高而显著下降,30头/瓶最低(39.37%)。蛹期随着密度的增加而延长(10头/瓶除外)。蛹重和每雌产卵量均以1头/瓶最高,随着幼虫密度的增加而显著下降。雌雄蛾寿命均以10头/瓶最长,与1和5头/瓶没有显著性差异。生命表分析显示:二点委夜蛾的种群增长指数以5头/瓶最高,幼虫密度过低或者过高均不利于种群增长。【结论】幼虫密度是影响二点委夜蛾种群增长的重要因子之一。     

Effect of neem seed extract on feeding,growth, survival,and reproduction of Diaprepes abbreviatus (Coleoptera: Curculionidae)

A commercially available neem seed extract, Neemix 4.5, containing 4.5% azadirachtin (AZA), was assessed for biological activity against the root weevil Diaprepes abbreviatus (L.), an important exotic insect pest of Florida citrus. Laboratory bioassays against neonatal and 3-wk-old larvae fed sliced carrot treated with Neemix produced dose-dependent larval mortality and reduced fresh weights among survivors of treatments. The weight response was greater than the mortality response for both larval age groups. Neonates treated with 45 mg/liter AZA weighed 60% less than those in the control after 4 wk. Three-week-old larvae treated with 45 mg/ liter AZA weighed 30% less than those in the control after 5 wk. When neonates were exposed to insect diet incorporated with Neemix, reductions in larval survival and weight were observed at concentrations as low as 4.8 mg/liter AZA after 6 wk. Larval growth was inhibited by >97% with 42.9 mg/liter AZA in the diet. A soil drench containing 30 mg/liter AZA reduced the survival and weight gain of neonates added to potted citrus and provided protection to the roots in a greenhouse experiment. A concentration of 90 mg/liter AZA was required to provide protection of citrus roots against 4-wk-old larvae. Reproductive effects were observed when adult weevils were fed foliage treated with Neemix. The numbers of larvae hatching per egg mass were reduced by 27% and 68% at 30 and 90 mg/liter AZA, respectively. These results suggest that Neemix should be further evaluated for use in integrated pest management (IPM) programs of citrus.     

Variations in the life-history parameters of Hemipyrellia ligurriens (Diptera: Calliphoridae) in response to larval competition for food

Abstract. 1. Larval rearing densities of Hemipyrellia ligurriens (Wiedemann) (Diptera: Calliphoridae) in standardized carrion were manipulated in order to investigate changes in life-history parameters in response to larval competition for food.
2. Competition was of the typical scramble type. Survivorship remained high at densities up to 32 larvae g liver^-1 but decreased rapidly as larval density increased further.
3. Emergent adults were undersized with reduced fecundity and longevity. Variations in adult body size apparently reduced the effects of competition on larval mortality.
4. Females of dry weight corresponding to only 10.4% of the potential maximum emerged at the highest rearing densities of 128 larvae g liver-1. However, these females had a nearly four-fold increase in reproductive investment (per unit weight) when compared to the largest individuals.
5. The duration of larval development declined when competition was intense (i.e. at high larval densities).
6. The short adult life of H.ligurriens, combined with the unpredictability of larval habitat availability, may reduce the value of long-range dispersal so that females 'do better' by maintaining reproductive investment despite a concomitant decline in dispersal ability.     

幼虫密度对草地螟生长发育及繁殖的影响

为了明确幼虫密度对草地螟Loxostege sticticalis种群增长的影响, 对室内条件下（温度22±1℃, RH 70%±5%, 光周期16L∶ 8D）不同幼虫密度[1, 10, 20, 30和40头/瓶（650 mL）］饲养的草地螟生长发育及繁殖进行了研究。结果表明: 幼虫密度对草地螟幼虫体色、 发育历期和存活率, 以及蛹重和成虫生殖等有显著影响。随着幼虫密度的增加, 幼虫体色黑化程度呈增强趋势, 幼虫密度大于10头/瓶时的体色黑化值均显著大于幼虫密度为1头/瓶时的体色黑化值（P<0.05）。20头/瓶的幼虫和蛹历期最短, 且随幼虫密度的增加而显著延长（P<0.05）。幼虫存活率以10头/瓶最高, 其他幼虫密度的幼虫存活率显著较低（P<0.05）。蛹重以1头/瓶的最重, 并随幼虫密度增加而显著下降（P<0.05）。成虫产卵量和交配率分别以1和20头/瓶的幼虫密度最高, 幼虫密度升高则产卵量、 交配率逐渐降低。成虫产卵历期随着幼虫密度的增加逐渐缩短。雌、 雄蛾寿命分别以10和20头/瓶幼虫密度时最长, 幼虫密度过高时雌雄蛾寿命又显著缩短（P<0.05）。生命表分析表明, 幼虫密度对草地螟种群增长指数有显著影响, 以10头/瓶幼虫密度的种群增长指数最高, 幼虫密度过高或过低时种群增长指数下降。据此认为, 幼虫密度是影响草地螟种群增长的重要因子之一。     

Intraspecific competition effects on Aethina tumida (Coleoptera: Nitidulidae)

Two kinds of experiments were conducted with Aethina tumida Murray larvae over four temperatures: "consumption" experiments, in which larvae and diet were weighed to determine food consumption rates under conditions of unlimited food and few conspecifics; and "competition" experiments, in which varying numbers of larvae were presented with the same amount of honey and pollen diet, and larval weight at final instar was used to determine competition effects. In consumption experiments temperature, diet and their interaction all had significant effects on the ratio of larval weight to the weight of food consumed, which was higher at 24 degrees C than at any other temperature. In competition experiments, three relationships were examined and modeled: that between the number of larvae per experimental unit and the average weight of those larvae; that between average larval and adult weights; and that between average adult weight and survivorship to adult (emergence rate). An exponential decay function was fit to the relationship between number of larvae per experimental unit and their average weight. Average adult weight was linearly correlated with larval weight. Likewise, emergence rates for adults < 11.6 mg in weight were linearly correlated with adult weights, but no significant relationship was observed for heavier adults. Using these relationships, the reproductive potential for A. tumida were estimated for a frame of honey and pollen. Information on resource acquisition by A. tumida will be useful in evaluating the impact of different factors on beetle population dynamics, such as bee hygienic behavior or control strategies used by the beekeeper.     

Laboratory evaluation of the chemosterilant lufenuron against the fruit flies Ceratitis capitata, Bactrocera dorsalis, B. cucurbitae, and B. latifrons

Four species of tephritid fruit flies, Ceratitis capitata, Bactrocera dorsalis, B. cucurbitae, and B. latifrons were evaluated for toxic, developmental, and physiological responses to the chemosterilant lufenuron. No significant mortality of laboratory strains of the first three species was observed after their exposure up to 50 μg/mL of lufenuron in agar adult diet, whereas B. latifrons adults fed with 50 μg/mL of lufenuron in the diet caused significant mortality compared to the control. Fertility of C. capitata adults fed on 50 μg/mL lufenuron-fortified diet between 7 and 12 days of age was approximately 46% of the no lufenuron control. Fertility of B. dorsalis and B. latifrons adults fed on 50 μg/mL lufenuron-incorporated diet was about 45% and 62% of the control, respectively. Lufenuron did not significantly affect fertility of B. cucurbitae adults. Lufenuron did not affect fecundity of C. capitata and B. dorsalis. Fecundity of B. cucurbitae and B. latifrons was not evaluated due to difficulty to count the eggs laid deep in the agar diet. Larvae fed on a liquid larval diet with ≤ 0.1 μg/mL of lufenuron were also evaluated. Pupal recovery, adult emergence, adult fliers, mating, egg hatch, and egg production of C. capitata were significantly decreased, while for B. dorsalis, pupal recovery, larval duration and adult emergence were affected. No effect of lufenuron on B. cucurbitae larvae was observed. B. latifrons was not performed because shortage of eggs at the time of this research. Lufenuron is a potential agent for management and control of C. capitata and B. dorsalis.     

New larval agar-based diet for laboratory rearing of Mediterranean fruit fly <Emphasis Type="Italic">Ceratitis capitata</Emphasis> (Diptera,Tephritidae)

Five larval diets for laboratory rearing of Ceratitis capitata were tested. These diets were based on wheat bran, microcellulose, potato starch and agar. To evaluate the quality of diet, pupal rearing efficacy and pupal weight were checked. The best results were obtained with an agar based diet used for Manduca sexta laboratory rearing. To simplify the preparation and to reduce the cost of this diet, a new formulation was developed. Larvae reared on the new agar-based diet achieved higher pupal rearing efficacy than larvae reared on bran diet recently used in medfly mass rearing facilities. Heat treated medflies reared on the new agar-based diet achieved similar pupal rearing efficacy with heat treated medflies reared on bran diet. When testing population density, higher pupal rearing efficacy was again achieved on new diet. The highest pupal rearing efficacy was achieved with 100 eggs per 25 g of diet, lowest with 500 eggs per 25 g of diet. Concerning pupal weight, there was no difference in results achieved on Petri dishes with different larval population densities. Larvae reared on new agar-based diet reached better results than larvae reared on bran diet. The preparation of the new diet is simple and the cost is low, so it is good for laboratory tests and rearing.