Techniques for the In Vitro Production of Queens in Stingless Bees (Apidae, Meliponini)

Considering the ecological importance of stingless bees as caretakers and pollinators of a variety of native plants makes it necessary to improve techniques which increase of colonies' number in order to preserve these species and the biodiversity associated with them. Thus, our aim was to develop a methodology of in vitro production of stingless bee queens by offering a large quantity of food to the larvae. Our methodology consisted of determining the amount of larval food needed for the development of the queens, collecting and storing the larval food, and feeding the food to the larvae in acrylic plates. We found that the total average amount of larval food in a worker bee cell of E varia is approximately 26.70 +/- 3.55 mu L. We observed that after the consumption of extra amounts of food (25, 30, 35 and 40 mu L) the larvae differentiate into queens (n = 98). Therefore, the average total volume of food needed for the differentiation of a young larva of F. varia queen is approximately 61.70 +/- 5.00 mu L. In other words; the larvae destined to become queens eat 2.31 times more food than the ones destined to become workers. We used the species Frieseomelitta varia as a model, however the methodology can be reproduced for all species of stingless bees whose mechanism of caste differentiation depends on the amount of food ingested by the larvae. Our results demonstrate the effectiveness of the in vitro technique developed herein, pointing to the possibility of its use as a tool to assist the production of queens on a large scale. This would allow for the artificial splitting of colonies and contribute to conservation efforts in native bees.

Development and evolution of caste dimorphism in honeybees - a modeling approach

The difference in phenotypes of queens and workers is a hallmark of the highly eusocial insects. The caste dimorphism is often described as a switch-controlled polyphenism, in which environmental conditions decide an individual's caste. Using theoretical modeling and empirical data from honeybees, we show that there is no discrete larval developmental switch. Instead, a combination of larval developmental plasticity and nurse worker feeding behavior make up a colony-level social and physiological system that regulates development and produces the caste dimorphism. Discrete queen and worker phenotypes are the result of discrete feeding regimes imposed by nurses, whereas a range of experimental feeding regimes produces a continuous range of phenotypes. Worker ovariole numbers are reduced through feeding-regime-mediated reduction in juvenile hormone titers, involving reduced sugar in the larval food. Based on the mechanisms identified in our analysis, we propose a scenario of the evolutionary history of honeybee development and feeding regimes.

Collection of Pollen Grains by Centris (Hemisiella) tarsata Smith (Apidae: Centridini): Is C. tarsata an Oligolectic or Polylectic Species?

Lia Goncalves, Claudia Ines da Silva, and Maria Luisa Tunes Buschini (2012) Collection of pollen grains by Centris (Hemisiella) tarsata Smith (Apidae: Centridini): Is C. tarsata an oligolectic or polylectic species? Zoological Studies 51(2): 195-203. Among pollinator species, bees play a prominent role in maintaining biodiversity because they are responsible, on average, for 80% of angiosperm pollination in tropical regions. The species richness of the bee genus Centris is high in South America. In Brazil, these bees occur in many types of ecosystems. Centris tarsata is an endemic species occurring only in Brazil. No previous studies considered interactions between plants and this bee species in southern Brazil, where it is the most abundant trap-nesting bee. Accordingly, the goals of this study were to investigate plants used by this species for its larval food supply and determine if this bee is polylectic or oligolectic in this region. This work was conducted in the Parque Municipal das Araucarias, Guarapuava (PR), southern Brazil, from Mar. 2002 to Dec. 2003. Samples of pollen were collected from nests of these bees and from flowering plants in grassland and swamp areas where the nests were built. All of the samples were treated with acetolysis to obtain permanent slides. The family Solanaceae was visited most often (71%). Solanum americanum Mill. (28.6%) and Sol. variabile Mart. (42.4%) were the primary pollen sources for C. tarsata in the study area. We found that although C. tarsata visited 20 species of plants, it preferred Solanum species with poricidal anthers and pollen grains with high protein levels. This selective behavior by females of C. tarsata indicates that these bees are oligolectic in their larval provisioning in this region of southern Brazil. http://zoolstud.sinica.edu.tw/Journals/51.2/195.pdf

Vitellogenin content in fat body and ovary homogenates of workers and queens of Melipona quadrifasciata anthidioides during vitellogenesis

Vitellogenin (Vg) is an egg yolk protein that is produced primarily in the fat body of most female insects. In the advanced social structure of eusocial honeybees, the presence of the queen inhibits egg maturation in the workers ovaries. However in the stingless bee Melipona quadrifasciata, the workers always develop ovaries and lay a certain amount of eggs while provisioning the brood cells with larval food during what is known as the worker nurse phase. The present work is a comparative study of the presence of Vg in homogenates of the fat bodies and ovaries of the nurse workers, and the virgin and physogastric queens of M. quadrifasciata. The presence of Vg was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting using Apis mellifera anti-egg antibody. Vg was not detected in the fat bodies or ovaries of the workers, but it was found in the ovaries of virgin and physogastric queens and in the fat body of physogastric queens. The results are discussed, taking into account the reproductive state of the individuals and the other possible roles of Vg, such as a storage protein for metoabolism of other organs.

Factors supporting the non-persistence of fruit fly populations in South Australia

For purposes of interstate and international fruit trade, it is necessary to demonstrate that in areas in which fruit fly species have not previously established permanent populations, but which are subject to introductions of fruit flies from outside the area, the introduced population once detected, has not become established. In this paper, we apply methodology suggested mainly by Carey (1991, 1995) to introductions of Mediterranean fruit fly (Medfly), Ceratitis capitata Weid., and Queensland fruit fly (QFF) Bactrocera tryoni Froggatt (Diptera: Tephritidae) to South Australia, a state in which these species do not occur naturally and in which introductions, once detected, are actively treated. By analysing historical data associated with fruit fly outbreaks in South Australia, we demonstrate that: (i) fruit flies occur seasonally, as would occur in established populations, except there is no evidence of the critical spring generation of either species; (ii) there is no evidence of increasing frequency of outbreaks, trapped flies or larval occurrences over 29 years; (iii) there is no evidence of decreasing time between catches of adult flies as the years progress; (iv) there is no decrease in the mean number of years between outbreaks in the same locations; (v) there is no statistically significant recurrence of outbreaks in the same locations in successive years; (vi) there is no evidence of spread of outbreaks outwards from a central location; (vii) the likelihood of outbreaks in a city or town is related to the size of the human population; (viii) introduction pathways by road from Western Australia (for Medfly) and eastern Australia (for QFF) are shown to exist and to illegally or accidentally carry considerable amounts of fruit into South Australia; and (ix) there was no association between the numbers of either Queensland fruit fly or Medfly and the spatial pattern of either loquat or cumquat trees as sources of larval food in spring. This analysis supports the hypothesis that most fruit fly outbreaks in South Australia have been the result of separate introductions of infested fruit by vehicular traffic and that most of the resultant fly outbreaks were detected and died out within a few weeks of the application of eradication procedures. An alternative hypothesis, that populations of fruit flies are established in South Australia at below detectable levels, is impossible to disprove with conventional technology, but the likelihood of it being true is minimised by our analysis. Both hypotheses could be tested soon with newly developed genetic techniques.

The roles of natural and sexual selection during adaptation to a novel environment

The net effect of sexual selection on nonsexual fitness is controversial. On one side, elaborate display traits and preferences for them can be costly, reducing the nonsexual fitness of individuals possessing them, as well as their offspring, In contrast, sexual selection may reinforce nonsexual fitness if an individual's attractiveness and quality are genetically correlated. According to recent models, such good-genes mate choice should increase both the extent and rate of adaptation. We evolved 12 replicate populations of Drosophila serrata in a powerful two-way factorial experimental design to test the separate and combined contributions of natural and sexual selection to adaptation to a novel larval food resource. Populations evolving in the presence of natural selection had significantly higher mean nonsexual fitness when measured over three generations (13-15) during the course of experimental evolution (16-23% increase). The effect of natural selection was even more substantial when measured in a standardized, monogamous mating environment at the end of the experiment (generation 16; 52% increase). In contrast, and despite strong sexual selection on display traits, there was no evidence from any of the four replicate fitness measures that sexual selection promoted adaptation. In addition, a comparison of fitness measures conducted under different mating environments demonstrated a significant direct cost of sexual selection to females, likely arising from some form of male-induced harm. Indirect benefits of sexual selection in promoting adaptation to this novel resource environment therefore appear to be absent in this species, despite prior evidence suggesting the operation of good-genes mate choice in their ancestral environment. How novel environments affect the operation of good-genes mate choice is a fundamental question for future sexual selection research.

On the methods of determining size/weight characteristics of food organisms of fish. (Determination of the length of Copepoda in larval stages by egg-laying females of the free living freshwater Copepoda). [Translation from: Voprosy Ikhtiologii 14, 182-194, 1960. ]

As is known, copepods play an important role in the nutrition of fish. Therefore with a view to facilitating research on the study of the quantitative side of feeding, there have recently appeared a considerable number of papers devoted to the development of methods for determining the wet. weight of these crustaceans. For the further facilitating of research in the nutrition of fish it would be of great interest to clarify the problem, is there not some kind of rule in the growth of the crustaceans during metamorphosis, and if there is such a rule is it not possible, to determine the length of the larvae at each stage, not by measuring them, but by using the formulae derived on the basis of these rules. This article examines the growth curves of different species of freshwater Copepoda, obtained on the basis of experimental observations in cultures or by way of measurement of mass material at all stages of development in samples from water-bodies. The authors study in particular the ratio of the mean diameter of the eggs to the mean length of the egg-bearing females.

Post-larval production of Macrobrachium rosenbergii using mass-cultured live food organisms

Macrobrachium rosenbergii post-larvae were produced in 1992 and 1993 using Artemia nauplii and cultured zooplankton Brachionus plicatilis (rotifier), Apocyclops dengizicus (copepod) and Moina sp. (cladoceran) supplemented with chopped Tubifex worms. In 1992 (first trial) two experiments were carried out under water temperature range of 24.5 to 28°C and 26.0 to 28.5 °C respectively and corresponding post-larval production was 5.6% and 86.3%. The duration of experiments was 58 and 40 days. During second trial in 1993 water temperature varied between 25.0 to 27.0°C. At the end of 59 days the post-larvae were found to be 44% of the total number of larvae stocked on the first day.

Viability of frozen algae used as food for larval penaeids

Freezing with added chemicals as flocculants and protectants was assessed as a means of preserving stock cultures of 4 algal species used for larval penaeid food Chaetoceros calcitrans, Skeletonema costatum, Tetraselmis chuii and Isochrysis galbana . The maximum storage effectability of the preservation techniques for each species was also determined.

Viability of frozen algae used as food for larval penaeids

Freezing with added chemicals as flocculants and protectants was assessed as a means of preserving stock cultures of 4 algal species used for larval penaeid food Chaetoceros calcitrans, Skeletonema costatum, Tetraselmis chuii and Isochrysis galbana. The maximum storage effectability of the preservation techniques for each species was also determined.

Survey of food supplementary (Pseudomonas fluorescence) and live food (Algae and brine shrimp) on Penaeus indicus larval stages

In this study ,the effects of Pseudomonas fluorescence obtained from generator pond water of Kolahi as supplementary and four algae consisting of : Chaetoceros sp, Chlorella and Skeletonema sp and Tetraselmis sp, three types of artemia as live food larval states from zoa to postlarvae (PL4 ) Penaeus indicus were investigated. The results indicate that Pseudomonas fluorescence has positive effect on Penaeus indicits larvae growth and their living food. Effective ranges at minimum and maximum were estimated. In most cases optimum dosage was approximately determined. Optimum dosage is between 50 -150 milligrams per liter for living food and Penaeus larval More than 200 milligram per liter resulted in a negative effect on the growth and survival. Also the results indicate Uromiana artemia. Requires a higher concentration of the bacteria the imported artemia. As a conclusion it is recommended to introduce Pseudonionas fluorescence as a new medium for the growth of some mentioned algae .

Digestion of natural food by larval and post-larval turbot Scophthalmus maximus

The digestion of natural, mainly crustacean zooplankton, by different age groups of turbot Scophthalmus maximus larvae was evaluated by comparisons of visual appearance, dry weight and carbon and nitrogen content of fresh food organisms with material recovered from faeces. Visually, the degree of digestion of food particles ranged from no discernible change of lamellibranch larvae, copepod eggs, intact copepod faecal pellets and some phytoplankton species, to varying degrees of removal of body constituents in copepods, cladocerans and decapod zoea. For crustaceans, the proportion of body constituents removed was related to the size and construction of their apparently indigestible exoskeleton. Uppon defaecation larger organisms showed the greatest percentage loss in dry weight and carbon. A high percentage of nitrogen was extracted from all organisms.

The influence of larval density, food availability and habitat longevity on the life history and population growth rate of the midge Chironomus riparius

Despite long-standing interest in the forms and mechanisms of density dependence, these are still imperfectly understood. However, in a constant environment an increase in density must reduce per capita resource availability, which in turn leads to reduced survival, fecundity and somatic growth rate. Here we report two population experiments examining the density dependent responses under controlled conditions of an important indicator species, Chironomus riparius. The first experiment was run for 35 weeks and was started at low density with replicate populations being fed three different rations. Increased ration reduced generation time and increased population growth rate (pgr) but had no effect on survival, fecundity and female body weight in the first generation. In the second generation there was a six-fold increase in generation time, presumably due to the greatly reduced per capita resource availability as the estimated initial densities of the second generation were 300 times greater than the first. Juvenile survival to emergence, fecundity, adult body weight and pgr declined by 90%, 75%, 35% and 99%, respectively. These large between-generation effects may have obscured the effects of the threefold variation in ration, as only survival to emergence significantly increased with ration in the second generation. These results suggest that some chironomid larvae survive a reduction in resource availability by growing more slowly. In the ephemeral habitats sometimes occupied by C. riparius, the effects of population density may depend crucially on the longevity of the environment. A second experiment was therefore performed to measure pgr from six different starting densities over an eight-week period. The relationship between pgr and density was concave, viewed from above. At densities above 16 larvae per cm(2), less than 1% of the population emerged and no offspring were produced. Under the conditions of experiment 2 - an 8-week habitat lifespan carrying capacity was estimated as 8 larvae per cm(2).