Theoretical approaches to forensic entomology: II. Mathematical model of larval development

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Larval predation by Chrysomya albiceps on Cochliomyia macellaria, Chrysomya megacephala and Chrysomya putoria

Chrysomya albiceps, the larvae of which are facultative predators of larvae of other dipteran species, has been introduced to the Americas over recent years along with other Old World species of blowflies, including Chrysomya megacephala, Chrysomya putoria and Chlysomya rufifacies. An apparent correlate of this biological invasion has been a sudden decline in the population numbers of Cochliomyia macellaria, a native species of the Americas. In this study, we investigated predation rates on third instar larvae of C. macellaria, C. putoria and C. megacephala by third instar larvae of C. albiceps in no-choice, two-choice and three-choice situations. Most attacks by C. albiceps larvae occurred within the first hour of observation and the highest predation rate occurred on C. macellaria larvae, suggesting that C. albiceps was more dangerous to C. macellaria than to C. megacephala and C. putoria under these experimental conditions. The rates of larvae killed as a result of the predation, as well as its implications to population dynamics of introduced and native species are discussed.

Larval aggregation and competition for food in experimental populations of Chrysomya putoria (Wied.) and Cochliomyia macellaria (F.) (Dipt., Calliphoridae)

In blowflies, larval aggregation in patches of food can be both intra- and interspecific, depending upon the degree to which competitors are clumped among the patches. In the present study, the implications of spatial aggregation for larval competition was investigated in experimental populations of the introduced blowfly Chrysomya putoria and the native Cochliomyia macellaria, using data from survival to adulthood in a range of single- and double-species larval cultures. The reduction in C. macellaria survival rate in the presence of C. putoria suggests that the former species is the inferior competitor. The results on survival to adulthood for both species in single- and double-species cultures can be explained in the light of the relationship between the level of intra- and interspecific aggregation and the efficiency of the larval feeding process. The possible implications of these results for the population biology of both species in natural environments are discussed.

Regression of the lateral oviducts during the larval-adult transformation of the reproductive system of Melipona quadrifasciata and Frieseomelitta varia

Toward the end of the larval phase (pre-pupa), the reproductive systems of Melipona quadrifasciata and Frieseomelitta varia workers are anatomically similar. Scanning electron microscopy showed that during this developmental phase the right and left ovaries are fused and form a heart-shaped structure located above the midgut. Each ovary is connected to the genital chamber by a long and slender lateral oviduct. During pupal development, the lateral oviducts of workers from both species become extremely reduced due to a drastic process of cell death, as shown by transmission electron microscopy. During the lateral oviduct shortening, their simple columnar epithelial cells show some signs of apoptosis in addition to necrosis. Cell death was characterized by cytoplasmic vesiculation, peculiar accumulation of glycogen, and dilation of cytoplasmic organelles such as mitochondria and rough endoplasmic reticulum. The nuclei, at first irregularly contoured, became swollen, with chromatin flocculation and various areas of condensed chromatin next to the nuclear envelope. At the end of the pupal phase, deep recesses marked the nuclei. At emergence, worker and queen reproductive systems showed marked differences, although reduction in the lateral oviducts was an event occurring in both castes. However, in queens the ovarioles increased in length and the spermatheca was larger than that of workers. At the external anatomical level, the reproductive system of workers and queens could be distinguished in the white- and pink-eyed pupal phase. The metamorphic function of the death of lateral oviduct cells, with consequent oviduct shortening, is discussed in terms of the anatomical reorganization of the reproductive system and of the ventrolateral positioning of adult worker bee ovaries. (C) 2000 Wiley-Liss, Inc.

Application of on-line C30 RP-HPLC-NMR for the analysis of flavonoids from leaf extract of Maytenus aquifolium

The application of on-line C30-reversed-phase high-pressure liquid chromatography-nuclear magnetic resonance spectroscopy is described for the analysis of tetraglycosylated flavonoids in aqueous and hydroalcoholic extracts of the leaves of Maytenus aquifolium (Celastraceae). Triacontyl stationary phases showed adequate separation for on-line 1H-NMR measurements at 600 MHz and allowed the characterisation of these flavonoids by detection of both aromatic and anomeric proton signals. Copyright (C) 2000 John Wiley and Sons, Ltd.

Larval development of Epialtus bituperculatus H. Milne Edwards, 1834 (Crustacea: Decapoda: Brachyura: Majidae) with comments on majid larvae from the southwestern Atlantic

The larval development of the spider crab Epialtus bituberculatus H. Milne Edwards which lives on rocky shores with algae such as Sargassum and Hypeneia, is described. Larvae were obtained from ovigerous females collected in Ubatuba, State of São Paulo, Brazil. Rearing was carried out at 24 ± 1°C, with an average salinity of 35%. Larval development consists of two zoeal stages and one megalopa. Zoeal development was completed in 9.5 days. Analysis indicated that zoeae of E. bituberculatus are very similar to those of E. brasiliensis Dana and Acanthonyx scutiformis (Dana). Differences noted between these species pertain to the setation of the carapace, maxillule and second maxilliped. The main morphological features useful for identification are presented together with a summary of features that characterize larvae of majid subfamilies in Brazil. A key for the identification of southwestern Atlantic majid zoeae to the family level is provided.

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.

Elevation of boiling point of coffee extract

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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.

Enzymatic activity of the hypopharyngeal gland extract from workers and males of Scaptotrigona postica Lat. (Hymenoptera, Apidae, Meliponini)

The objective of this research was to contribute to elucidation of the function of the hypopharyngeal glands of S. postica in enzyme production, using the Api Zym kit (Bio Mérieux). Dealing with a comparative analysis between the enzymatic content of the hypopharyngeal gland extracts from newly emerged, nurse and forager workers, as well as, newly emerged males and males mature for mating of S. postica. The hypopharyngeal glands from nurse workers of Apis mellifera, that produce part of the royal jelly, were used for comparison. While in A. mellifera, the hypopharyngeal glands are present only in workers, in S. postica, the hypopharyngeal glands are present and functioning in all adult individuals of the colony. The higher enzymatic activity was observed in the hypopharyngeal gland extracts from nurse workers and may be related to a larger demand for energy, compared to other individuals. The occurrence of large quantities of leucine arylamidase in all individuals may mean that protein processing is happening.

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.