Larval Stadium Research Articles

Spirulina sebagai Pengganti Artemia untuk Meningkatkan Produksi Larva Ikan Patin Pangasius hypophthalmus

The provision of quality and sufficient shrimp fry for production is largely determined by the management of maintenance during larval stadia. This study aims to obtain the best treatment of natural feed in the form of Spirulina sp. and Artemia sp. as feed supplements to improve the growth performance and survival rate of patin fish larvae. The experiment used a Complete Randomized Design consisting of 4 feed treatments and 3 replicates: A (Artemia sp. 100%), B (Artemia sp. 100% and Spirulina sp. 100%), C (Artemia sp. 50% and Spirulina sp. 50%), and D (Spirulina sp. 100%). The larvae with a density of 50 heads/L were kept in the aquarium and fed according to the treatment. The density of Spirulina sp. administered was 104–105 cells/mL maintained until day 7 according to the treatment. The parameters measured included growth performance, which included specific growth rate (SGR), survival (SR), absolute length growth (PPM), and analysis of larval digestive enzymes (amylase, lipase, and protease). The data were analyzed using one way-analysis of variance and continued with Duncan's Multiple Range Test with a 95% confidence interval using SPSS version 22.0. The results showed that the administration of Spirulina sp. produced better SGR, SR, PPM, amylase enzyme, lipase, and protease activities compared to the control (100% administration of Artemia sp.). The best treatment was given by 100% Spirulina sp. treatment. Keywords: Artemia sp., digestive enzymes, patin fish larvae, Spirulina sp.

The larvae of Cora inca Selys, 1873 and Polythore gigantea (Selys, 1853) from Colombia (Odonata: Polythoridae), with a larval diagnoses of some genera in the family

The larvae of Cora inca Selys, 1873 and Polythore gigantea (Selys, 1853) were found in first and second order forested streams at the Tatamá Natural National Park in the Colombian Western Andes. Their final larval stadia are here described and figured. Cora inca differs from the larvae of C. cyane Selys, 1853 and C. marina Selys, 1868 by a combination of features including presence of an irregular patch of minute, gray scales to each side of the ventral midline on the ventral pad of hypopharynx, paraprocts with five projections, and the position of epiproct and paraproct projections. On the other hand, the larva of P. gigantea differs from the larva of P. spaeteri Burmeister & Börzsöny, 2003 by antennal scape slightly shorter than 3rd antennomere, prementum 0.10x longer than its widest part, and the position of epiproct and paraproct projections. Although most of the larvae in the family Polythoridae are still unknown, here we discuss the main morphological characteristics that help to diagnose some of the genera in this family, presenting a regional taxonomic key that includes the genera Cora Selys, 1853, Euthore Selys, 1859, Miocora Calvert, 1917, and Polythore Calvert, 1917.

Description of the final larval stadium of Miocora aurea (Ris, 1918) (Odonata: Polythoridae)

The larva of Miocora aurea (Ris, 1918) was found in first and second order forested streams at the Tatam National Park in the Colombian Western Andes. Here it is described and figured the final larval stadium. It differs from the larva of M. chirripa (Calvert, 1917) by a combination of features such as antennal pedicel (0.25x longer than 3rd antennomere), prementum (0.20x longer than its widest part), and posterior margin of tergite 10 with a moderate incision.

Life History of Talicada nyseus (Guerin) (Lepidoptera:Lycaenidae) on Bryophyllum pinnatum from Western Uttar Pradesh, India

Talicada nyseus, a small, stunning butterfly, is distributed throughout the Indian subcontinent and Southeast Asia. The life history and seasonal occurrence of T. nyseus were studied on the succulent host plant Bryophyllum pinnatum grown in the homestead garden in district Saharanpur, Uttar Pradesh, India. It passes through five larval stadia.

Locomotion and searching behaviour in the honey bee larva depend on nursing interaction

Although honey bee brood does not need to seek shelter or food and restricts its movements to small wax cells, larvae have some degree of motility. Previously, other studies described how honey bee larvae showed analogous behaviours to the wandering period in holometabolous insects. The current research aimed to measure locomotion of the honey bee brood at different conditions of food supply and larval stadia. Besides, we developed an actometry assay to describe the larval behaviour under laboratory conditions. Our results suggested that the satiety and developmental program of larvae modulated their locomotion. Before they pupated, larval speed increased sharply and then it dropped until quiescence. However, starvation also induced an increase in angular velocity of brood. Starved larvae were between three and five times faster than the satiated ones. Moreover, fifth instars left their wax cells after 2 h of starvation without nurse bees. In the actometry assay, larvae showed behaviours of dispersion and changes in their kinematic parameters after detecting a tactile stimulus like the edge of arenas.

Omnivory in predatory lady beetles is widespread and driven by an appetite for sterols

Abstract Animals maintain physiological and behavioural systems that allow them to detect and consume specific macro‐ and micronutrients to maximize their fitness. One common physiological system is the nutrient‐state‐dependent or demand‐driven appetite. These systems are well described for macronutrient regulation, but not for micronutrients. Sterols are essential micronutrients that all animals need to survive. They are the backbone of many hormones, important in cell signalling and an integral component of cell membranes. Lady beetles are globally distributed predators of insect herbivores. Adult sevenspotted lady beetles maintain a state‐dependent sterol appetite and consume plant tissues to obtain sterols, which improves their fitness. Additionally, sevenspotted lady beetles can detect sterols pre‐ingestion. We used lady beetle species distributed across the three clades of the Coccinellini to determine (a) whether other beetle species maintain a state‐dependent sterol appetite, (b) if sterol structure affects beetles' state‐dependent sterol appetite and (c) whether lady beetles consume foliage in a sterol‐state dependent manner. Additionally, we determined (a) what sensory organ beetles use to detect sterols, (b) their limit of detection and (c) when during development their appetite manifests. All six beetle species we tested maintained a state‐dependent appetite for sterols. Sterol structure affected beetles' propensity to feed on sterol‐treated disks, indicating that the number and position of double bonds in sterol molecules affects beetles' ability to detect or desire to feed on them. Only beetles in clade III fed on plant foliage in response to sterol limitation. Few beetles in any clade that were supplemented with sterols consumed plant tissue. Beetles' appetite for sterols first appeared during the second larval stadium, and the rate of sterol consumption increased with age. Ablations of sensory organs revealed that beetles use their labial palps to detect sterols, and that they detect them at concentrations as low as 1ppm. These data demonstrate that lady beetles across the Coccinellini maintain a state‐dependent appetite for an essential class of micronutrients. They also provide very strong evidence that lady beetles can taste sterols, raising the possibility that they maintain novel as‐yet undescribed gustatory receptors for an essential class of lipids. A free Plain Language Summary can be found within the Supporting Information of this article.

The life cycles of Boyeria irene and Onychogomphus uncatus (Odonata: Aeshnidae, Gomphidae) in western Spain: A biometric study

Co-occurrence of species with similar trophic requirements, such as odonates, seems to depend both on them occupying different microhabitats and differing in their life-cycles. The life cycles of the dragonflies Boyeria irene and Onychogomphus uncatus were studied in two consecutive years, mainly by systematic sampling of larvae in seven permanent head courses that constitute the upper basin of the River Agueda, western Spain, in the central part of the ranges of these two The size ranges of the last five larval stadia of both species were established based on biometric data. The eggs of the egg-overwintering aeshnid hatched in late and early and for the gomphid hatching peaked in middle-late summer. Both species showed mixed voltinism with cohort splitting. B. irene had a dominant three-year development (partivoltinism), with some developing in two years (semivoltinism). O. uncatus requires four, sometimes three years to complete development (all partivoltine). B. irene larvae spent the winter before emergence in the last three, maybe four stadia, as a summer species. O. uncatus mainly behaved as a spring species, most larvae spending the last winter in the final larval stadium.

Description of the larva of Orthemis ferruginea (Fabricius, 1775) (Odonata: Libellulidae).

The last larval stadium of Orthemis ferruginea (Fabricius, 1775) is described and illustrated in detail based on reared material collected in the municipality of Paso de Ovejas, Veracruz, Mexico. It is compared with larvae of other species of the genus previously described. The main structural features are the number of setae on palp, prementum and palp crenations, the size proportion of cerci respect to epi- and paraprocts, and the number of sclerites on abdominal sternum 5.

Amblyospora khaliulini (Microsporidia: Amblyosporidae): Investigations on its life cycle and ecology in Aedes communis (Diptera: Culicidae) and Acanthocyclops vernalis (Copepoda: Cyclopidae) with redescription of the species.

A multi-year study was conducted to examine the natural ecology of the microsporidium Amblyospora khaliulini and more fully characterize parasite development and histopathology in all stages of its primary mosquito host, Aedes communis and intermediate copepod host, Acanthocyclops vernalis with redescription of the species. A. khaliulini exhibits polymorphic development, produces three morphologically and functionally distinct spores, and is both horizontally and vertically transmitted. Development in A. vernalis is restricted to females, occurs within the ovaries and results in death of the host. Development is haplophasic with division by binary and multiple fission producing rosette-shaped sporogonial plasmodia and conical uninucleate spores that are orally infectious to Ae. communis larvae. Both sexes are equally susceptible and infections are confined to testes in males and ovaries in females. Initial stages of development include uninucleate schizonts that undergo karyokinesis forming diplokaryotic meronts that divide repeatedly by binary fission. Sporogony occurs in both host sexes, but sporogenesis does not progress normally in adult males and elliptical, thin walled binucleate spores that function in vertical transmission of the microsporidium via infection of the ovaries and eggs are formed in adult females only. Development of vertically acquired infections in larval Ae. communis hosts occurs within fat body tissue, leads to the production of meiospores in male hosts only and results in death during the 4th larval stadium. Initial development is characterized by merogonial multiplication of diplokarya by synchronous binary division producing additional diplokarya. The cessation of merogony and the onset of sporogony are characterized by the simultaneous secretion of a sporophorous vesicle and meiotic division of diplokarya resulting in the formation of octonucleate sporonts that undergo cytokinesis and sporogenesis to form eight uninucleate, broadly ovoid meiospores enclosed within a sporophorous vesicle. The natural prevalence of patent vertically acquired fat body infections in field populations of Ae. communis ranged from 1.6% to 3.6%. Yearly infection rates in A. vernalis copepods ranged from 57.1% to 15.0%. Prevalence rates of horizontally acquired infections in emerging adult Ae. communis ranged from 69.0% to 11.9% in males and 50.0% to 16.4% in females.

Effect of SSP – 11 on Weight and Tissue Composition of 4th Instar Larvae of Silkworm, Bombyx mori

The anti-juvenoid, SSP-11 ((E) 4-chloro-a, a, a-trifluro-N [ 1 (1 H-imidazole- 1 –y1)- 2-propoxy ethylyledene] O, toludine) was used to induce trimoulters from tetramoulter larvae of the silkworm, Bombyx mori (race NB7). The compound was fed for two days through an artificial diet at the concentration of 200 ppm to newly ecdysed 4th stadium larvae. Body and silkgland fresh weight were recorded daily in the 4th larval stadium and protein profiles of haemolymph, fat body and silkgland were determined at the end of the stadium. In larvae treated with SSP-11, feeding period in stadium 4 was prolonged by 3-4 days and 80-85% of the larvae started to spin as trimoulters. Eight and thirty fold increases in the larval body and silkgland weights respectively were the result of SSP-11. Further, total proteins, total carbohydrates, glycogen and nucleic acid (DNA & RNA) also increased. The activity levels of alanine aminotransferase and aspartate aminotransferasc increased significantly than in the control indicating increased mobilization of aminoacids into transamination activities.

THE EFFECT OF THE TRAMADOL ACCUMULATED IN RAT LIVER ON THE DEVELOPMENT OF THE IMMATURE STAGES OF THE FLESH FLY SARCOPHAGA ARGYROSTOMA (ROBINEAU-DESVOIDY, 1830) (DIPTERA: SARCOPHAGIDAE).

The present study investigated the effects of the accumulation of tramadol on the development rates of the larvae and pupae. Larvae of the flesh fly Sarcophaga argyrostoma (Robineau-Desvoidy, 1830) were reared on three groups of Wister albino rat livers. One group was administered the recommended or normal dose (Dl) and the second group was injected with higher over or double dose (D2) of tramadol. Tramadol was administered by stomach tube once a day, for 3 months. The third group was injected with normal saline solution as the control. Using GC-Ms analysis, Rat livers contained 0.72 and 1.62mg/g of tramadol, in case of Dl and D2, respectively. Larvae of S. argyrostoma fed on DI and D2 rat livers contained (0.11 & 0.1 8mg/g), respectively. The corresponding tramadol concentrations persisted in the produced pupae of S. argyrostoma were (0.07 & 0.09mg/g), respectively. The accumulation of tramadol in tissues of S. argyrostoma larvae reduced the durations of larval stadia from (7.0408 1.0198 days) in case of the control to (6.6383±0.4857 days) when fed on DI liver and (6.3438±0.4826 days) when fed on D2 livers. The pupal durations was altered to (14.3750±0.4919 days) for D2 and (14.9574±0.6580 days) for DI, compared to (13.9167±1.0071 days), in case of the control. The average weight of the 3rd day larva of S. argyrostoma decreased from 18.1807±1.4949mg, in case of the control to 14.7279±1.3366mg and 14.9560±3.8210mg, when fed on Dl and D2 tramadol treated livers. In contrast, corresponding weight produced pupae increased from 14.1750±0.4667mg (control). to 15.2449±0.302mg & 15.9062±0.2888mg, in case of D1 and D2, respectively.

Differences in pupal cold hardiness and larval food consumption between overwintering and non‐overwintering generations of the common yellow swallowtail,Papilio machaon(Lepidoptera: Papilionidae), from the Osaka population

AbstractTo clarify differences in pupal cold hardiness and larval food consumption between overwintering and non‐overwintering generations of the common yellow swallowtail,Papilio machaon, we reared larvae from the Osaka population under photoperiods of 16 h light : 8 h dark (LD 16:8) (long day) or LD 12:12 (short day) at 20°C. We examined the relationship between food consumption and weight during the final larval stadium and pupae, and measured the pupal supercooling point (SCP). Although the ratio of assimilation to consumption did not differ significantly between photoperiods, the ratio of assimilation to pupal weight differed significantly between individuals reared under long and short days. All diapausing pupae were brown, whereas 56% of non‐diapausing pupae were green with the remainder brown. The mean pupal body length (L), dorsal width (W1) and lateral width (W2) were larger in non‐diapausing than in diapausing pupae, and theW1/LandW1/W2ratios differed significantly between non‐diapausing and diapausing pupae. SCP was approximately –20°C and did not differ among pupae 5, 15 and 30 days after pupation under long‐day conditions. However, under short‐day conditions, mean SCP gradually decreased, stabilizing at approximately –24 to –25°C by 30 days after pupation. After freezing, some diapausing pupae emerged as adults, whereas all non‐diapausing pupae died. Both egestion and assimilation were greater under long‐day conditions. The results revealed that pupae of this papilionid exhibit seasonal polyphenism in physiological and morphological traits. Energy from food appears to be expended on increasing cold hardiness in the overwintering generation and on reproduction in the non‐overwintering generation.

STATISTIK DEMOGRAFI DIAPHANIA INDICA SAUNDERS (LEPIDOPTERA: CRAMBIDAE)

Demographic statistic of Diaphania indica Saunders (Lepidoptera: Crambidae). Diaphania indica is a pest of Cucumis sativus L. The information about the biology of D. indica is needed to determine appropriate pest management strategies. The aim of this study was to study the biology and life cycle of D. indica. The study was conducted by observing life stage development of D. indica, such as life cycle, head width and body length. The life table was determined by calculating mortality of D. indica individuals and fecundity at each age. Mortality and fecundity data of D. indica were then used to calculate the parameters of population growth including gross reproduction rate (GRR), net reproductive rate (Ro), mean generation time (T) and and intrinsic rate of increase (r ). The results show that the life cycle of D. indica is about 25.7 days, while egg stadium = 3.2 days, larval stadium = 13.5 days, pupal stadium = 7.9 days, imago stadium = 8.6 days, and preoviposition period = 3.9 days. GRR of D. indica is 263 individuals per generation. Ro = 51 individuals per female per generation. T is 31.3 days and the innate capacity for increase (r) = 0.1 individual per female per day.

EVIDENCE THAT ECDYSIS IN THE LARVAL COCKROACH, Periplaneta americana L. IS TRIGGERED BY AN INCREASE IN THE CONCENTRATION OF HEMOLYMPH SUGAR.

Ecdysis in insects can be defined as shedding of the cuticle at the end of a larval stadium. This event can only occur after the peak titer of ecdysteroid in the hemolymph has returned to a low level. In the cockroach Periplaneta americana, ecdysis is strongly correlated with a rise in the concentration of trehalose and glucose in the hemolymph, leading to the idea that a causal relationship may exist between both events. The objective in this study was to determine if an increase in hemolymph sugar level would shorten the time to ecdysis in cockroach larvae with experimentally delayed ecdysis. The last larval stadium of P. americana averages 33.5 days but this increases significantly if the larva is injected with a small volume of saline. Injection of 10 μl of saline on day 20 and on four successive days lengthened the stadium by as much as 2 weeks. If, however, trehalose or glucose is incorporated into the saline, approximately 40% of the treated larvae undergo ecdysis at the same time as uninjected larvae. Injection of Peram-AKH, the hypertrehalosemic hormone, also decreases the time for ecdysis to occur. This suggests that peak levels of ecdysteroid trigger the release of Peram-AKH, which then leads to activation of trehalose synthesis. The results support the hypothesis that elevated hemolymph sugar is a contributing factor in the removal of ecdysteroid from the hemolymph.

Culex quinquefasciatus larval microbiomes vary with instar and exposure to common wastewater contaminants.

Like many insects, mosquitoes, rely on endosymbionts to grow and develop. These can be acquired from the environment. We used next generation 454 pyrosequencing to discern the whole-body microbiome of the mosquito species Culex quinquefasciatus in various larval stadia and following exposure to common pharmaceutical and personal care products (PPCPs) found in wastewater. PPCP treatments included environmentally-relevant concentrations; 1) a combination of common antibiotics, 2) a combination of mammalian hormones, 3) a mixture of the antibiotic and hormone treatments plus acetaminophen and caffeine and, 4) an untreated control. Within control groups, the predominant families of bacterial symbionts change with each larval instar despite consistent diets and rearing conditions. This trend was also seen in hormone treatments but not in the antibiotic or the mixture treatments. Richness and evenness were reduced in both antibiotic and mixture treatments, suggesting that antibiotics remove certain bacteria or inhibit them from increasing to proportions seen in the control treatment. Interestingly, the mixture treatments had greater richness and evenness compared to antibiotic alone treatments, possibly due to the other contaminants facilitating growth of different bacteria. These findings illuminate the complexity of the microbiome of C. quinquefasciatus and may have implications for more effective control strategies.

Aspects of life history ofPlatycnemis subdilatata(Zygoptera: Platycnemididae) in Northeast Algeria

The determination of seasonal regulation is important to understand how species have adapted to their local environmental conditions. In this study, we investigate the life history of a North African endemic damselfly, Platycnemis subdilatata, in a northeast Algerian population. We combined field and laboratory investigations to assess the embryonic development, larval growth, emergence pattern and adult flight season. The embryonic development was direct and asynchronous, with 50% of all eggs hatching after three weeks of egg laying and a hatching period ranging from 13 to 51 days. Hatching success was 48.8%, and the causes of hatching failure were infertility and unhatchability. Larval population structure was quite asynchronous during the winter and less so before emergence. The occurrence of larval diapause is improbable due to the increase of the proportion of the last larval stadium in late winter. Emergence was asynchronous with half of the larval population (EM50) emerging after 44 days within an ...

Life history strategy and adult and larval behavior of Macrodiplosis selenis (Diptera: Cecidomyiidae), a species that induces leaf‐margin fold galls on deciduous Quercus (Fagaceae)

AbstractLife historical, behavioral and ecological traits of Macrodiplosis selenis, which induces leaf‐margin fold galls on Quercus serrata, Q. mongolica and Q. dentata (Fagaceae) in Japan and South Korea, were studied. Daily activity and larval development indicate that M. selenis is a diurnal and univoltine gall midge. In April, females lay their eggs both on upper and under surfaces of fresh leaves. The duration of the egg stage varies from 5 to 9 days, depending on daily temperatures. Hatched larvae crawl to the upper surface of the leaf margin, where they start to induce galls. Larvae become full‐grown in October, drop to the ground in November and overwinter in cocoons on the ground, while larvae of congeners mature in May and drop to the ground in June. A relatively long period of the second larval stadium from July to October on the host trees seems to be effective for M. selenis in avoiding summer mortalities caused by predation and aridity on the ground and by ectoparasitoids that attack mature larvae or pupae on the host leaves. The spatial distribution pattern of M. selenis leaf galls is contagious and the mean gall density per leaf is significantly correlated with the mean crowding. This study adds new insights of life history strategy and adult and larval behavioral pattern to the ecological knowledge of gall midges, and these kinds of information are essential for further studies of M. selenis population dynamics and interactions with other Quercus‐associated herbivores.

Temporal and Spatial Foraging Behavior of the Larvae of the Fall WebwormHyphantria cunea

During their first three larval stadia, caterpillars ofHyphantria cunea(Lepidoptera: Arctiidae) are patch-restricted foragers, confining their activity to a web-nest they construct in the branches of the host tree. Activity recordings of eight field colonies made over 46 colony-days showed that the later instars become central place foragers, leaving their nests at dusk to feed at distant sites and then returning to their nests in the morning. Colonies maintained in the laboratory showed that same pattern of foraging. In Y-choice laboratory experiments, caterpillars were slow to abandon old, exhausted feeding sites in favor of new food finds. An average of approximately 40% of the caterpillars in five colonies still selected pathways leading to exhausted sites at the onset of foraging bouts over those leading to new sites after feeding exclusively at the new sites on each of the previous four days. On returning to their nests in the morning, approximately 23% of the caterpillars erred by selecting pathways that led them away from the nest rather than toward it and showed no improvement over the course of the study. The results of these Y-choice studies indicate that, compared to other previously studied species of social caterpillars, the webworm employs a relatively simple system of collective foraging.

Unexpected Use of Pupal Stage of a Lepidopteran Host by the Ectoparasitoid Wasp Agrothereutes lanceolatus (Hymenoptera: Ichneumonidae)

In general, the majority of ectoparasitoids of Lepidoptera parasitize larval and/or prepupal stages of the host and do not use the pupal stage, suggesting the presence of ecological and physiological constraints in the evolution of host range. Here, the evidence is given that Agrothereutes lanceolatus, a solitary ectoparasitoid wasp, can exceptionally parasitize host pupae, succeeding to extend its host range to pupal hosts that have a hardened cuticle. Laboratory experiments confirmed that A. lanceolatus developed successfully to adult both on host prepupae and on young pupae in equal proportions. Development time and size of the resulting wasps did not differ between the host types. However, the percentages of hosts producing wasp offspring differed among age classes within the pupal stage. The offspring survival was high except the oldest class of host pupae. With increasing host pupal ages, the offspring size decreased whereas the development time increased. Older host pupae hence were less suitable for the development of offspring. The offspring mortality mostly occurred during the 1st larval stadium without signs of growth, suggesting a difficulty in feeding on host pupae from an external position. Thus, A. lanceolatus can use a wider range of hosts than expected from its ectoparasitic development. The fact may enable A. lanceolatus to be a common and widespread polyphagous species among ectoparasitic Cryptinae.

Spatial and temporal synthesis of Mamestra configurata peritrophic matrix through a larval stadium

The structure and synthesis of the Mamestra configurata peritrophic matrix (PM) was examined at various time points during a larval stadium. Bright field and confocal fluorescence microscopy revealed major differences between the PM of feeding and molting larvae. The PM from feeding larvae was thinner and composed of approximately 5–10 layers. In contrast, mid-molt larvae had a chitinaceaous PM composed of multiple thick layers which filled most of the midgut lumen. PM synthesis initiates in the anterior midgut, based on the expression of genes encoding chitin synthase-2 (CHS-2), coincident with the incorporation of the major structural PM proteins (McIIM1, McIIM2 and McPM1). This is followed by reinforcement with other PM proteins (McIIM3 and McIIM4) as it moves toward the posterior of the midgut.Chitin deacetylase (McCDA1) was associated only with the anterior PM. Collectively, these findings indicate that the structural properties of the PM differ along the length of the midgut. Genes encoding chitinolytic enzymes (McCHI and McNAG) were expressed and exochitinase activity was present when the PM had degraded (pre-molt) and when the new PM was forming (mid-molt), indicating that they are involved in either PM turnover and/or maintenance dependent upon the stage.