Protein requirements in larvae and adults of Scaptotrigona postica (Hymenoptera: Apidia, Meliponinae): Midgut proteolytic activity and pollen digestion

The number and degree of digestion of pollen grains in the midgut and rectum, the midgut proteolytic activity and the time of pollen grain passage through the digestive tract in the stingless bee Scaptotrigona postica (Latreille) have been analyzed. The results show similar protein requirements among larvae, nurse bees and queens, as well as between forager bees and old males, but these requirements are higher in individuals from the former groups than in those from the latter. Although protein requirements have been demonstrated to vary according to a bee's activity in the colony, they are similar among bees from different castes or sexes. These changes in feeding behavior are related to the bee's function and to less competition for nourishment among individuals of the colony. It is also noted that pollen grains took between 6 and 28 h to pass through the digestive tract. Pollen grains are irregularly accumulated in the various regions of the midgut, which may reflect functional differentiation throughout the midgut. © 2001 Elsevier B.V.

Targeting Plasmodium ligands on mosquito salivary glands and midgut with a phage display peptide library

Despite vast efforts and expenditures in the past few decades, malaria continues to kill millions of persons every year, and new approaches for disease control are urgently needed. To complete its life cycle in the mosquito, Plasmodium, the causative agent of malaria, has to traverse the epithelia of the midgut and salivary glands. Although strong circumstantial evidence indicates that parasite interactions with the two organs are specific, hardly any information is available about the interacting molecules. By use of a phage display library, we identified a 12-aa peptide-salivary gland and midgut peptide 1 (SM1)-that binds to the distal lobes of the salivary gland and to the luminal side of the midgut epithelium, but not to the midgut surface facing the hemolymph or to ovaries. The coincidence of the tissues with which parasites and the SM1 peptide interact suggested that the parasite and peptide recognize the same surface ligand. In support of this hypothesis, the SM1 peptide strongly inhibited Plasmodium invasion of salivary gland and midgut epithelia. These experiments suggest a new strategy for the genetic manipulation of mosquito vectorial capacity.

Oviposition behavior of Ceratitis capitata wiedemann (Diptera: Tephritidae): Association between oviposition preference and larval performance in individual females

The oviposition preference and larval performance of females of Ceratitis capitata (Wiedemann) were examined. Wild or laboratory-reared females were tested for oviposition preference in relation to papaya fruit in different stages of ripeness, as well as to two different fruits: papaya and apple. The results demonstrated that both laboratory-reared and wild females preferred ripe papaya fruit compared to unripe fruit for oviposition, and that ripe papaya was the best for larval performance. When oviposition preference was tested with different fruits (papaya and apple), both wild and laboratory-reared larvae developed better in papaya. However, only wild females showed a preference for oviposition on papaya as opposed to apple. Laboratory-reared females showed no oviposition preference on fruits that resulted in better larval performance.

Number of larval instars of the ant Pachycondyla (=Neoponera) villosa (Formicidae: Ponerinae) determined by the rule of Dyar

In this work we present the number of larval instars in the Ponerinae ant Pachycondyla (=Neoponera) villosa. The analysis of maximal head capsule width measurement of 147 larvae was made. Four larval instars were measured: 1st instar the cephalic capsule varied from 0.18mm to 0.22mm; 2nd instar from 0.23mm to 0.27mm; 3rd instar from 0.30mm to 0.33mm and the 4th instar varied from 0.35mm to 0.38mm. The mean growth rate was 1.2375 according to the rule of Dyar. We also reviewed the number of larval instars for 35 ant species.

Chemical analysis of different larval instars and tissues of Pachycondyla (=Neoponera) villosa (Hymenoptera: Formicidae, Ponerinae)

Chemical analyses of complete larvae of the first to third instar and cuticle, fat body and salivary glands extracts of fourth instar larvae using gas chromatography and gas chromatography-mass spectrometry, were performed upon Pachycondyla villosa. The results revealed that P. villosa larvae do not produce a pheromone, as only fatty acids and n-alkanes were detected. After quantifying the identified compounds, it was determined that the fat body is the main place of storage and/or production of the cuticular hydrocarbons. It was also observed that the absolute quantity of cuticular hydrocarbons increases progressively during larval development. Inferences about the transport behavior of matured larvae to the pupation place and the colony odor are discussed.

Ultrastructure of the salivary glands of Pachycondyla (=Neoponera) villosa (Fabricius) (Formicidae: Ponerinae): Functional changes during the last larval instar

The cells of secretory region of the salivary glands of Pachycondyla (=Neoponera) villosa at the time of enzyme production presents the basal cellular membranes profusely folded and the intercellular junctional membranes present a few enlarged spaces. The rough endoplasmic reticulum and the Golgi bodies shift from being flat and small vesicular cisternae to enlarged vesicular cisternae according to the cell physiological state and characterize an asynchronic cell cycle. Enzymes are released into the lumen by microapocrine secretion. The stage of silk production is detected after a behavioral act, when the nurse worker separates the mature larva. At this time, the salivary gland cells present only one physiological state (synchronized secretory cycle): this state was characterized by basal cellular membrane poorly folded, intercellular junctions presenting some small spaces, rough endoplasmic reticulum compounded by flat cistenae, enlarged Golgi bodies with fibrous material inside and a few secretory vesicles containing silk, which undergo exocytosis. The silk in the lumen shows 2 forms: tactoid and flocculent material.

Ovarian growth during larval development of queen and worker of Apis mellifera (Hymenoptera: Apidae): a morphometric and histological study.

The present work reports the differences between the ovarian grow in queen and worker larvae of A. mellifera, from the start of differential feeding. The observations made of the growth rates in larvae of both castes showed that the queen and worker larvae have the same rates of cephalic capsule growth from one instar to another but the weight gain is greater in queens. In the same way, the draw areas of ovaries of queens increase more and continuously, while from the 5th instar on the ovaries of workers decrease in size. The decrease is due to a loss of ovariole numbers that starts early in the worker larvae and increases in the 4th-5th instar. The ovarian shape in queens and workers became different in the last larval instars.

Effects of nitrate concentration on larval development of the giant river prawn, Macrobrachium rosenbergii

The effect of nitrate concentration on giant river prawn, Macrobrachium rosenbergii, larvae was investigated. Survival rate, weight gain, and larval development were evaluated for different concentrations of nitrate in three experiments. The experiments were divided i n to two phases. In the first phase, larvae from stages I through VIII were analysed, while in the second phase larvae from stage VIII through post-larvae metamorphosis were analysed. Oxygen consumption was also determined for zoea I, II, and VIII exposed to 0, 700, and 1,000 mg/L of nitrate-N. No effect was observed for concentrations up to 180 mg/L NO3-N (experiments I and II), and nitrate levels as 1,000 mg/L NO3-N did not affect survival in the first phase of the third experiment. On the other hand, larval stage index (LSI) and weight gain decreased as nitrate-N concentration increased from 0 to 1,000 mg/L. In the second phase, survival and metamorphosis rate decreased as nitrate concentration increased, according to a linear model. The effect of nitrate levels on weight gain followed a curvilinear pattern. Larval respiration decreased in the water where nitrate was added, but only during stage II. The results demonstrated that nitrate presents extremely low toxicity for giant river prawn larvae, and data were related to the levels of nitrate that usually occur in larviculture systems also discussed. Therefore, nitrate is not a limiting factor for giant river prawn larviculture. © 2003 by The Haworth Press, Inc. All rights reserved.

Cloning and characterization of the cDNA for the Brazilian Cratomorphus distinctus larval firefly luciferase: Similarities with European Lampyris noctiluca and Asiatic Pyrocoelia luciferases

Studies on firefly (Lampyridae) luciferases have focused on nearctic species of Photinus and Photuris and Euroasiatic species of Lampyris, Luciola, Hotaria, and Pyrocoelia. Despite accounting for the greatest diversity of fireflies in the world, no molecular studies have been carried out on the highly diverse genera from the neotropical region. Here we report the luciferase cDNA cloning for the larva of the Brazilian firefly Cratomorphus distinctus. The cDNA has 1978 bp and codes for a 547-residue-long polypeptide. Noteworthy, sequence comparison as well as functional properties show the highest degree of similarity with Lampyris noctiluca (93%) and Pyrocoelia spp. (91%) luciferases, suggesting a close phylogenetic relationship despite the geographical distance separating these species. The bioluminescence emission spectrum peaks at 550 nm and, as expected, is sensitive to pH, shifting to 605 nm at pH 6. The kinetic properties of the recombinant luciferase were similar to those of other firefly luciferases. © 2004 Elsevier Inc. All rights reserved.

Larval development of the giant river prawn Macrobrachium rosenbergii at different ammonia concentrations and pH values

The effect of ammonia and pH levels on giant river prawn Macrobrachium rosenbergii larvae were evaluated to provide science-based information on safe levels of ammonia and pH for larviculture. Survival rate, developmental stage, and larval weight gain were determined for larvae kept in water with total ammonia (NH4-N) concentrations of 0, 1, 2, 4, and 8 mg\L and pH 7, 8, and 9. The trials were conducted in two phases: phase 1, larvae from stages I through VIII and phase 2, larvae from stage VIII until metamorphose. Oxygen consumption was determined for larvae in stages I and VIII at total ammonia concentrations of 0, 4, and 8 mg/L and pH 8. Survival rate up to stage VIII varied from 86 to 98% and did not differ for total ammonia concentrations in pH 7 and 8 and for 0 mg/L NH4-N in pH 9. Survival rate was significantly lower (0-20%) for total ammonia concentrations from 1 to 8 mg/L (0.43-3.41 mg/L of unionized ammonia) in pH 9. Larval stage indexes (7.9-8.0 range) and weight gain (1.572-2.931 mg range) of larvae at the end of phase 1 of the experiment did not differ for the different ammonia concentration solutions, but were significantly lower in pH 9. In phase 2, no parameter differed among treatments for pH 7 and 8; however there was total mortality at pH 9 until 96h. Respiration rates diminished when larvae were exposed to total ammonia concentrations of 4 and 8 mg/L (0.28 and 0.55 mg/L of unionized ammonia), but development remained unaltered. Therefore, M. rosenbergii larvae tolerate high levels of total ammonia, while toxicity depends primarily on unionized ammonia concentrations. In addition, alkaline pH (9) acted directly on the larvae, curbing development and causing severe mortality. Larval tolerance to high ammonia and pH levels decreases for the last zoeal stages. © Copyright by the World Aquaculture Society 2005.

Evolutionary modeling of larval dispersal in blowflies using non-uniform cellular automata

When the food supply flnishes, or when the larvae of blowflies complete their development and migrate prior to the total removal of the larval substrate, they disperse to find adequate places for pupation, a process known as post-feeding larval dispersal. Based on experimental data of the Initial and final configuration of the dispersion, the reproduction of such spatio-temporal behavior is achieved here by means of the evolutionary search for cellular automata with a distinct transition rule associated with each cell, also known as a nonuniform cellular automata, and with two states per cell in the lattice. Two-dimensional regular lattices and multivalued states will be considered and a practical question is the necessity of discovering a proper set of transition rules. Given that the number of rules is related to the number of cells in the lattice, the search space is very large and an evolution strategy is then considered to optimize the parameters of the transition rules, with two transition rules per cell. As the parameters to be optimized admit a physical interpretation, the obtained computational model can be analyzed to raise some hypothetical explanation of the observed spatiotemporal behavior. © 2006 IEEE.

Comparative morphology of the midgut of three species of Cephalotes (Hymenoptera, Formicidae, Myrmicinae)

Studies of histological nature, histochemistry and ultra-morphology of the midgut of three species of ants belonging to the genus Cephalotes (atratus, clypeatus and pusillus) were performed. In histology H-E was employed, and in histochemistry techniques for detection of carbohydrates, proteins in general and collagen and calcium fibers were used. The results obtained from this second technique made possible some discussion about the pH of the midgut of these insects. The species are identical in the characters under study. The digestive cells exhibited a secretion that is typical of ants, the apocrine type.

Interaction of Bacillus thuringiensis Cry1 and Vip3A proteins with Spodoptera frugiperda midgut binding sites

Vip3Aa, Vip3Af, Cry1Ab, and Cry1Fa were tested for their toxicities and binding interactions. Vip3A proteins were more toxic than Cry1 proteins. Binding assays showed independent specific binding sites for Cry1 and Vip3A proteins. Cry1Ab and Cry1Fa competed for the same binding sites, whereas Vip3Aa competed for those of Vip3Af. Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Pyrearinus termitilluminans larval click beetle luciferase: Active site properties, structure and function relationships and comparison with other beetle luciferases

Several beetle luciferases have been cloned and sequenced. However, most studies on structure and function relationships and bioanalytical applications were done with firefly luciferases, which are pH sensitive. Several years ago we cloned Pyrearinus termitilluminans larval click beetle luciferase, which displays the most blue-shifted bioluminescence among beetle luciferases and is pH insensitive. This enzyme was expressed in E. coli, purified, and its properties investigated. This luciferase shows slower luminescence kinetics, KM values comparable to other beetle luciferases and high catalytic constant. Fluorescence studies with 8-anilino-1-naphtalene-sulfonic acid (1,8-ANS) and modeling studies suggest that the luciferin binding site of this luciferase is very hydrophobic, supporting the solvent and orientation polarizability effects as determining mechanisms for bioluminescence colors. Although pH insensitive in the range between pH 6-8, at pH 10 this luciferase displays a remarkable red-shift and broadening of the bioluminescence spectrum. Modeling studies suggest that the residue C312 may play an important role in bioluminescence color modulation. Compared to other beetle luciferases, Pyrearinus termitilluminans luciferase also displays higher thermostability and sustained luminescence in a bacterial cell environment, which makes this luciferase particularly suitable for in vivo cell analysis and bioimaging. © The Royal Society of Chemistry and Owner Societies 2009.

Fluazuron-induced morphophysiological changes in the cuticle formation and midgut of Rhipicephalus sanguineus Latreille, 1806 (Acari: Ixodidae) nymphs

The present study demonstrated the effects of the arthropod growth regulator, fluazuron (Acatak®), in the formation of the integument and digestive processes of Rhipicephalus sanguineus nymphs fed on rabbits treated with different doses of this chemical acaricide. For this, three different doses of fluazuron (20, 40, or 80 mg/kg) were applied pour on to the hosts (groups II, III, and IV), as well as distilled water to the control group. On the first day after treatment (24 h), the hosts were artificially infested with R. sanguineus nymphs. After full engorgement (7 days), the nymphs were removed, placed on labeled Petri dishes, and kept in biochemical oxygen demand incubator for 7 days. The engorged nymphs were then taken for morphological, histochemical, and histological analyses. The results showed the occurrence of cytological, morphohistological, and histochemical alterations in the integument and midgut of nymphs from all the different treated groups. These alterations occurred at cuticular level in the subdivisions of the cuticle, related to the size of the digestive cells, amount of accumulated blood elements, and digestive residues, as well as the presence of vacuoles in the cytoplasm of the digestive cells. Thus, this study demonstrated that fluazuron acts on the integument and midgut cells of R. sanguineus nymphs fed on treated rabbits and pointed out the possibility of the use of this chemical - which is more specific, less toxic, and less harmful to the environment and nontarget organisms - in the control of R. sanguineus, at least in the nymphal stage of its biological cycle. © 2012 Springer-Verlag.