Ergonomics in stingless bees: changes in intranidal behavior after partial removal of storage pots and honey inMelipona favosa (Hym. Apidae,Meliponinae)

Summary The ergonomics of task allocation in stingless bees was examined in two laboratory colonies ofMelipona favosa containing individually marked workers. Performance of intranidal behavior patterns, brood cell production and the processing and storage of nectar, were studied during a control and a wax-deprived period. Experimental wax deprivation consisted of regular controlled removal of storage pots and their contents. Stored carbohydrate concentration and storage pot mass declined during the wax-deprived period. Behavior was measured by sampling for a complete ethogram at seven intranidal areas.In comparison withApis mellifera, M. favosa appeared to be more sensitive to stress, with a distinct reallocation of effort between tasks. The significant increase under stress of brood production and brood-rearing behavior patterns observed was previously also found inApis. Self-oriented behavior patterns declined under stressed circumstances.Involucrum construction declined in wax-deprivedMelipona colonies. Relatively low rates of walking behavior in general inMelipona suggest a low mean free distance between tasks. These results are of particular interest in relation to the basically different nest structure of stingless bees, e.g. very distinct areas for brood production and the processing and storage of food. This architecture of stingless bee nests appeared to influence strongly their specific responses to wax deprivation.     

Occurrence and morphometry of the hypopharyngeal glands inScaptotrigona postica Lat. (Hymenoptera, Apidae, Meliponinae)

As compared toApis mellifera where only workers have hypopharyngeal glands, inScaptotrigona postica, these glands occur in workers, queens and males. They are composed of two long axial ducts with many unicellular secretory alveoli interconnected by secretory canaliculi. The axial ducts are longer in males than in workers, but the alveolar areas of queens and males are generally smaller. In workers the alveoli have their greatest size in the nurses or middle-aged individuals while in queens and males they are larger in newly emerged individuals. The results indicate that the glands in workers may produce food for the brood as inA. mellifera, since they are well developed in the nurse workers. However, the function of the glands in queens and males remains to be clarified since these individuals have no part in brood care.     

Influence of age and caging upon protein metabolism,hypopharyngeal glands and trophallactic behavior in the honey bee (Apis mellifera L.)

Summary Different-aged honey bees were either kept in a cage together with young sisters for eight days or lived in their colony. Following an injection of¹⁴C-phenylalanine (Phe) we measured incorporation of¹⁴C-Phe into head protein and total protein, as well as the size of the hypopharyngeal glands. While confined in a cage for four hours, injected bees (from colony or cage) dispensed the¹⁴C-labelled protein-rich products of their hypopharyngeal glands to recipients. Eight-day-old colony bees had well developed hypopharyngeal glands, whereas at the age of sixteen days the glands had already decreased in size. Young caged bees had smaller hypopharyngeal glands. Colony bees had higher incorporation rates into total protein and head protein than bees living in a cage. Bees of different age classes, irrespective of caging, fed the same number of recipients; but the amount of¹⁴C-labelled protein-rich jelly distributed by caged bees was significantly smaller than that distributed by colony bees. Our results indicate that trophallaxis between young donor workers and newly emerged recipient worker bees is not the key factor for regular development and activity of the hypopharyngeal glands.Dedicated to Achim Lass's daughter Katrin.     

The evolution of food-producing glands in eusocial bees (Apoidea,Hymenoptera)1

A food-producing role for cephalic exocrine glands has arisen independently in both taxa of highly eusocial bees, Apis and Meliponini. With several exceptions, there is little evidence that food is produced by glands of solitary bees or by most bees at lower levels of sociality. We suggest that this association with sociality is due to four adaptive features of these glands: (1) food from the glands allows feces from queens and larvae to have a small volume, (2) the queen's fecundity can be increased, (3) nutrient recovery via cannibalism can be facilitated, and (4) rearing of emergency replacement queens is accelerated. Acceleration of the rearing of other castes and of queens in the normal process of colony fission is not clearly an advantage ascribed to these glands. Trophic eggs produced by meliponine colony workers are analogous to the secretions from food-producing glands in Meliponini and Apis workers.     

Experimental studies on the larval development of the shrimpsCrangon crangon andC. allmanni

Larvae of the shrimpsCrangon crangon L. andC. allmanni Kinahan were reared in the laboratory from hatching through metamorphosis. Effects of rearing methods (larval density, application of streptomycin, food) and of salinity on larval development were tested only inC. crangon, influence of temperature was studied in both species. Best results were obtained when larvae were reared individually, with a mixture ofArtemia sp. and the rotiferBrachionus plicatilis as food. Streptomycin had partly negative effects and was thus not adopted for standard rearing techniques. All factors tested in this study influenced not only the rates of larval survival and moulting, but also morphogenesis. In both species, in particular inC. crangon, a high degree of variability in larval morphology and in developmental pathways was observed. Unsuitable conditions, e.g. crowding in mass culture, application of antibiotics, unsuitable food (rotifers, phytoplankton), extreme temperatures and salinities, tend to increase the number of larval instars and of morphological forms. The frequency of moulting is controlled mainly by temperature. Regression equations describing the relations between the durations of larval instars and temperature are given for bothCrangon species. The number of moults is a linear function of larval age and a power function of temperature. There is high variation in growth (measured as carapace length), moulting frequency, morphogenesis, and survival among hatches originating from different females. The interrelations between these different measures of larval development in shrimps and prawns are discussed.     

A mutant affecting the crystal cells inDrosophila melanogaster

Summary Black cells (Bc, 2-80.6±) mutant larvae ofDrosophila melanogaster have pigmented cells in the hemolymph and lymph glands. In this report we present evidence that these melanized cells are a mutant form of the crystal cells, a type of larval hemocyte with characteristic paracrystalline inclusions.Bc larvae lack crystal cells. Furthermore, the distribution pattern of black cells inBc larvae parallels that of experimentally-blackened crystal cells in normal larvae (phenocopy).InBc/Bc zygotes black cells appear during mid embryonic development but inBc ⁺/Bc zygotes pigmented cells are not found until late in the first larval instar.Crystal cells are present in the heterozygous larvae until this time, and paracrystalline inclusions can be seen in some of the cells undergoing melanization in these larvae.The rate of phenol oxidase activity inBc ⁺/Bc larval cell-free extracts is less than half that ofBc ⁺/Bc ⁺extracts whereas enzyme activity is undetectable inBc/Bc larvae. We propose that theBc ⁺gene product is required for maintaining the integrity of the paracrystalline inclusions; inBc/Bc larvae either the product is absent or nonfunctional so an effective contact between substrate and enzyme results in melanization of the cells.Phenol oxidase itself is either destroyed or consumed in the melanization process accounting for the absence of enzyme activity inBc/Bc larvae. These studies confirm that the crystal cells store phenolic substrates and are the source of the hemolymph phenol oxidase activity in the larva ofD. melanogaster.     

Interspecific competition and coexistence of the two chrysomelids,Gastrophysa atrocyanea motschulsky andGalerucella vittaticollis baly (Coleoptera: Chrysomelidae), under limited food resource conditions

Field experiments were conducted in order to investigate the mode of exploitation of food resources and the mechanism of coexistence of mixed larval populations of the two chrysomelids,Gastrophysa atrocyanea andGalerucella vittaticollis, under limited food resource conditions. The larval survival rates seemed high enough to assure coexistence when hatchlings of the two species were released in 1∶1 and 1∶3 ratios on a host plant. However, the survival rate became almost nil for both species when a 3∶1 ratio was employed, suggesting asymmetrical interspecific competition. Wasted food consumption was much higher inG. atrocyanea larvae. The population ofG. atrocyanea seemed to be regulated more by intraspecific competition, while on the other hand, the population ofG. vittaticollis was considered to be more likely affected by the interspecific competition withG. atrocyanea, depending on the initial ratio and density of the two species.     

Mechanisms of dance orientation in the Asian honey beeApis florea L.

Summary Early studies of dance communication inApis florea had shown that waggle dances are not performed on a vertical plane and oriented to gravity, as in the other species ofApis, but instead take place on the flattened top of the exposed comb and are oriented to celestial cues directly. More recent experiments showed thatA. florea can dance in the absence of a view of the sun or blue sky, but did not establish what mechanism permitted this orientation. I now report that dances can be oriented directly to landmarks visible from the nest, the first evidence of an environmental feature other than celestial cues or gravity being involved in dance orientation. Landmarks near the nest are probably used to refer to celestial cues, in a fashion analogous to the use of broad features of the landscape by honeybees in order to learn the sun's course, which permits them to determine their flight angle on overcast days or at night, and to compensate accurately for solar movement.Apis florea may therefore be able to learn the sun's course with respect to two sets of landmarks.In other experiments I have examined the influence of slope onA. florea's dance orientation to visual references. In the first extensive observations of its dances on a vertical plane, I have amply confirmed that this species cannot transpose light and gravity in setting its dance angle, as the other species ofApis can. Nor do dancers orient so as to match visual information seen during the dance with that remembered from the flight. Patterns in the data when the same patch of sky was presented from different angles suggest instead thatA. florea continues to orient to projections of celestial cues onto the horizontal plane even when dancing on a steep slope. This compensation for slope may involve an ability to detect gravity and factor it out in aligning the dance to celestial cues.These insights suggest thatA. florea's dance orientation system has been adapted to requirements imposed by its nesting behavior, and has diverged sharply from the system shared by the other species ofApis.     

Cell cycle events during the development of the silk glands in the mulberry silkworm<Emphasis Type="Italic"> Bombyx mori</Emphasis>

Silk glands of the mulberry silkworm Bombyx mori are long and paired structures originating from the labial region and are anatomically and physiologically divided into three major compartments, the anterior, middle and posterior silk glands. The silk gland morphogenesis is complete by 8 days post egg laying. Extensive growth of silk glands during the larval stages is due to increase in tissue mass and not cell number. The cells in a completely formed silk gland pursue an endoreplicative cell cycle, and the genome undergoes multiple rounds of replication without mitosis or nuclear division. The expression patterns of cyclin B (mitotic cyclin) and cyclin E (G1 cyclin, essential for G1/S transition in both mitotic and endoreplicative cell cycles) in the course of silk gland development revealed that mitotic cell divisions take place only in the apex of the growing silk gland. However, the persistence of another mitotic focus in the middle silk gland even when the growing apex has moved well past this zone suggested the continued operation of mitosis for a while in this restricted region. The lack of cyclin B expression and abundance of cyclin E in the rest of the areas confirmed an alternation of the G1 and S phases of the cell cycle without an intervening mitotic phase. No expression of cyclin B was noticed anywhere in the silk glands after stage 25 of embryogenesis, indicating a complete switch over to the endomitotic mode of the cell cycle. The onset of expression of various genes encoding different silk proteins correlated with the onset of endomitotic events.Edited by D. Tautz     

Copulation duration in the spiderPhilodromus vulgaris (Hentz) (Araneae: Philodromidae) and its influence on the evolution of host transfer behavior during cannibalism byMantispa uhleri banks (Neuroptera: Mantispidae)

Matings and acts of cannibalism were observed in the crab spiderPhilodromus vulgaris, some of which were carrying a first-instar larva of the mantispidMantispa uhleri. Larval mantispids did not transfer during the copulation of their spider hosts but did switch from spider to spider during cannibalism. Twenty-three of 27 total cannibalisms in which the cannibalized spider carried a larva resulted in larval transfers, for a transfer rate of 85%. Copulation times inP. vulgaris were found to be extremely short in comparison to lycosids such asSchizocosa ocreata, whose long copulation time supports the larval transfer ofClimaciella brunnea. The time required for larval transfer during cannibalism inP. vulgaris was measured. Cannibalisms were much longer than copulations but larvae transferred to the cannibalizing spider in a comparatively short period of time.     

Genetic and phenotypic variation for flight ability in the cactophilicDrosophila species,D. aldrichi andD. buzzatii

The flight ability ofDrosophila aldrichi (Patterson & Crow) andD. buzzatii (Patterson & Wheeler) using tethered flights, was measured with respect to age-related changes, genetic variation and adult body size variation induced by rearing at different larval densities.Drosophila buzzatii flew for much longer thanD. aldrichi, especially females, but age-related changes in flight duration were significant only forD. aldrichi. Effects of body size on flight ability were significant inD. buzzatii, but not inD. aldrichi. InD. buzzatii, there was a significant genotype-environment interaction (larval density × line) for flight duration, with short and average flight duration isofemale lines showing longer flights, but a long flight duration line shorter flights as body size decreased (i.e., as larval density increased). Heritability estimates for flight duration were similar in the two species, but flight duration showed no significant genetic correlations with developmental time, body size or wing dimensions (except for one wing dimension inD. buzzatii). Although not significantly different between the species, heritabilities for life-history traits (adult size and developmental time) showed contrasting patterns — with higher heritability for body size (body weight and thorax length) inD. buzzatii, and higher for developmental time inD. aldrichi. In agreement with limited previous field evidence,D. buzzatii is better adapted for colonization than isD. aldrichi.     

Analysis of nuclear proteins from silk glands ofBombyx mori

A gentle method for the isolation of nuclei from developing silk glands ofBombyx mori has been standardized. The nuclei, whether isolated or directly visualizedin situ within the silk glands, exhibit complex morphology. The nuclei occupy almost the entire volume of the gigantic silk gland cells. Although the isolated nuclei still retain their ramified morphology, being polyploid they are fragile and often become fragmented. The histone and low-salt-extractable proteins from nuclei isolated from the middle and posterior silk glands on different days of the fourth and fifth instars of larval development have been analysed. The histones did not show any stage- or tissue-specific variations whereas the low-salt-extractable proteins showed some developmental stage specific variation. Using the antibody raised against one such protein, its absence in the early stage of development has been confirmed by Western blotting techniques. This developmental stage specific protein may be functionally linked to some activities responsible for boosting up the production of silk or silk-related proteins during the fifth instar of larval development.     

Larval secretions and food odors affect orientation in femalePlodia interpunctella

Substrates contaminated by wandering fifth instar larvae ofPlodia interpunctella (Hübner) (Lepidoptera: Pyralidae) elicit oviposition by conspecific female moths, and larval rearing diet enhances oviposition and also induces upwind flight. Two-choice oviposition assays determined that four-day-old gravid femaleP. interpunctella preferred to lay eggs on dishes containing cornmeal-based rearing diet compared to empty dishes. Pieces of cheesecloth contaminated by fifth instar larvae elicited more oviposition than untreated cheesecloth or dishes with food. The combination of larval contamination and food was preferred over food only or larval contamination only in both two- and four-choice experiments. The factor(s) in larval contamination responsible for eliciting oviposition in female moths was extracted in hexane, confirming that organic semiochemicals are responsible for the effect. The oviposition-eliciting activity of larval contamination was retained on cheesecloth for up to 30 days following treatment with larvae, suggesting the active component(s) is stable and of low relative volatility. In two-choice windtunnel bioassays female moths initiated flight only when rearing food was present in one of the treatments, and they displayed the highest landing responses to a combination of larval contamination and food. Earlier work onP. interpunctella and related pyralid species found that larval contamination due to secretions from the mandibular glands acted as both a spacing pheromone for wandering larvae and as a kairomone for host-seeking parasitoid wasps. The present study suggests that the same or a similar secretion acts as an oviposition-eliciting pheromone for conspecific females.     

Yeast communities of the cactusPilosocereus arrabidae as resources for larval and adult stages ofDrosophila serido

The feeding behavior ofDrosophila serido on the yeast communities of necrotic stem tissue ofPilosocereus arrabidae were studied in a sand dune ecosystem of Rio de Janeiro, Brazil. The prevalence of cactophilic yeasts includingPichia barkeri, Candida sonorensis andGeotrichum sp. in the crops and external surfaces ofD. serido reflected its association with the cactus habitat. The effective number of yeasts vectored on the surface of flies was higher than that in the crops. Also overlap between the yeasts from stems and from crops was partial suggesting selective feeding by the flies in the substrates visited. The females had a higher effective number of yeast species and a lower similarity than males with the yeast community ofP. arrabidae. This was probably related to the search for oviposition sites by females. The presence ofPichia thermotolerans-like andPichia amethionina varpachycereana in the flies, but not inP. arrabidae stems, indicated thatD. serido was not limited to this cactus species. The larvae and adults lived in different patches with the adults feeding in patches with higher yeast species richness. The larvae had a narrower feeding niche and higher overlap withP. arrabidae, and preferredP. barkeri andPichia cactophila as food. Adult flies fed on patches with the most frequent yeasts except forP. cactophila. Pichia caribaea was found in higher frequency in the adult crops than in the stems. Our data suggested that there was food selection and diet partitioning between adult and larval stages ofD. serido.     

Ultrastructural analysis on the gland secretion in the extracellular ducts of the hypopharyngeal glands of the honeybee infected by Nosema apis

The hypopharyngeal glands of the honeybee secrete the larval food royal jelly, along with enzymes that are used in the production of honey from nectar. The composition of this secretion mixture varies with the physiological condition of the honeybee, and the final secretion mixture is conveyed through the extracellular ducts to the large excretory duct. The secretion products in the extracellular duct of both healthy and infected honeybees, was studied by transmission electron microscopy and image analysis. These products consisted of a densely packed, faintly visible granular substance. The granules were clearly defined by the computer enhanced image. The secretion products from the Nosema-infected honeybees exhibited the same granular structure, but were much less concentrated, thereby reducing the overall electron density of the secretion. These observations suggest that the composition of the secretion products from the hypopharyngeal glands in the Nosema-infected honeybees, was altered considerably.     

Sialic acids in the salivary glands of some insects with different feeding habits

Summary Eight insects (some adult and some larval forms) are studied for the presence of sialic acids in the cells of the salivary glands. This was sought by staining with alcian blue and Azure A at different pH accompanied by acid hydrolysis, sialidase digestion and methylation-saponification.On the basis of susceptibility to acid hydrolysis and sialidase digestion, different sialic acids are discernable. Though there is apparently no correlation between the secretion of sialic acid and the feeding habits of these insects, there is an interesting correlation between these two in the case of nectar and pollen eating habit of Apis.Presented at the 56th Session of Indian Science Congress. 2–9 January, 1969, Bombay, India.     

Chemotaxonomic studies onArachniodes (dryopteridaceae) I. Phloroglucinol derivatives of japanese species

Twelve species ofArachniodes from Japan were examined for phloroglucinol derivatives and glandularity of rhizome and petiole bases. For comparison 3 species ofDryopteris, 1 species ofCtenitis, 4 species ofPolystichum, and 1 species ofAcystopteris were also investigated. Phloroglucinol derivatives were detected for the first time by sensitive TLC in the followingArachniodes, Dryopteris andPolystichum species:A. amabilis, A. dimorphophylla, A. exilis, A. hasseltii, A. maximowiczii, A. miqueliana, A. mutica, A. nipponica, A. okinawensis, A. simplicior incl. var.major, D. hayatae, D. sparsa, andP. tsus-simense. On the other hand,A. pseudo-aristata, A. standishii, P. polyblepharum, P. pseudomakinoi, P. retroso-paleaceum, Ctenitis maximowicziana, andAcystopteris japonica were totally lacking phloroglucinols. The phloroglucinol containing species ofArachniodes andPolystichum contained external glands, whereas those ofDryopteris contained internal and/or external glands. External glands are common inDryopteris sect. Nephrocystis. In taxa lacking phloroglucinols no glands were present. The homologous iso-aspidins BB and AB were detected in ferns for the first time.