Alterations of the serotonin level in the central nervous system of Locusta migratoria during larval-adult transformation

Serotonin level was measured in the cerebral, suboesophageal, thoracic and abdominal ganglia of Locusta migratoria during larval-adult transformation. It has been established that the serotonin content shows a progressive increase in the period between the last larval stage and the first 7 days of adult life, being of the highest value in the abdominal ganglia (146%), followed by the thoracic (110%), and then by the cerebral and suboesophageal (75%) ganglia. No significant changes are manifested in the serotonin level in the course of moulting. Considerable increase is also detectable in the protein content of the ganglia during the studied development phase. No correlation is present between the protein and serotonin contents of the ganglia, the changes of the protein content was found to be faster as compared to that of serotonin.     

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

Abstract 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.     

Histochemical and histophysical aspects of silk secretion in Melipona quadrifasciata (Hym., Apoidea)

Summary Acid and neutral polysaccharides exist in the secretion of the silk glands of Melipona quadrifasciata during the entire functional stage, except in the late 5th instar. However, only from the 3rd instar on is a molecularly oriented structural protein present. The following evidence indicates that this protein is not collagen: (1) the optical sign of its birefringence does not become reversed after von Ebner's reaction, and (2) its secretion can be detected when the cell nucleus attains a ploidy degree optimal for secretory activity. This is at first observed in the distal part of the glands and then throughout the organ.Stipendiat der Alexander von Humboldt-Stiftung.     

Imidacloprid-induced impairment of mushroom bodies and behavior of the native stingless bee Melipona quadrifasciata anthidioides

Declines in pollinator colonies represent a worldwide concern. The widespread use of agricultural pesticides is recognized as a potential cause of these declines. Previous studies have examined the effects of neonicotinoid insecticides such as imidacloprid on pollinator colonies, but these investigations have mainly focused on adult honey bees. Native stingless bees (Hymenoptera: Apidae: Meliponinae) are key pollinators in neotropical areas and are threatened with extinction due to deforestation and pesticide use. Few studies have directly investigated the effects of pesticides on these pollinators. Furthermore, the existing impact studies did not address the issue of larval ingestion of contaminated pollen and nectar, which could potentially have dire consequences for the colony. Here, we assessed the effects of imidacloprid ingestion by stingless bee larvae on their survival, development, neuromorphology and adult walking behavior. Increasing doses of imidacloprid were added to the diet provided to individual worker larvae of the stingless bee Melipona quadrifasciata anthidioides throughout their development. Survival rates above 50% were only observed at insecticide doses lower than 0.0056 μg active ingredient (a.i.)/bee. No sublethal effect on body mass or developmental time was observed in the surviving insects, but the pesticide treatment negatively affected the development of mushroom bodies in the brain and impaired the walking behavior of newly emerged adult workers. Therefore, stingless bee larvae are particularly susceptible to imidacloprid, as it caused both high mortality and sublethal effects that impaired brain development and compromised mobility at the young adult stage. These findings demonstrate the lethal effects of imidacloprid on native stingless bees and provide evidence of novel serious sublethal effects that may compromise colony survival. The ecological and economic importance of neotropical stingless bees as pollinators, their susceptibility to insecticides and the vulnerability of their larvae to insecticide exposure emphasize the importance of studying these species.     

Modifications in the oviducts of workers and queens of <Emphasis Type="Italic">Melipona quadrifasciata anthidioides</Emphasis> (Hymenoptera: Apidae) with different ages

The study of morphological features of highly eusocial bees has helped to answer a series of questions concerning their biology. The labor division places the females into two castes, queen with reproductive function and worker with a wide variety of tasks. However, in different species and under different conditions, workers can develop ovaries and lay trophic eggs that are eaten by the queen or used to originate males. In this work, the development of the oviducts was monitored in workers and virgin queens of Melipona quadrifasciata anthidioides specimens of different ages to verify whether there is permanent sterility of these queens and workers due to aging. Lateral and common oviducts of virgin queens aged <7, 10, 15, 20, and 25 days old, physogastric queens, nurse, and forager workers were analyzed for histology and ultrastructure. Although the structural organization of the lateral and common oviducts were very similar, differences in width of the cuticle and the degree of chromatin condensation were observed, indicating differences in the development of this organ of the reproductive system between the castes. It was also demonstrated that electron-lucent vesicles appear to be related to the formation of the cuticle that lines the oviducts. Because no cellular death characteristics were found, it can be inferred that the absence of mating of the queens as old as of 25 days of age does not cause permanent sterility.     

The post-embryonic changes in Melipona quadrifasciata anthidioides Lep. (Hym. Apoidea). II. Development of the salivary glands system

The paper deals with the development of the salivary gland system in Melipona quadrifasciata anthidioides, which begins in the prepupal stage. The silk glands degenerate by autolysis at the end of the larval stage. Degeneration is characterized by cytoplasmic vacuolization and pycnosis of the nuclei of the secretory cells. The glandular secretory portion of degenerated silk glands separates from the excretory ducts. The salivary glands develop from the duct of the larval silk glands. The thoracic salivary glands develop from the ducts of the secretory tubules and the head salivary glands from the terminal excretory duct. The mandibular glands appear in the prepupa as invaginations of mandibular segments, and their differentiation to attain the adult configuration occurs during pupation. The hypopharyngeal glands have their origin from evaginations of the ventral anterior portion of the pharynx. A long tubule first appears with walls formed by more than one cellular layer. Then some cells separate from the lumen of the duct, staying attached to it by a cuticular channel in part intracellular. The initial duct constitutes the axial duct, in which the channel of the secretory cells opens. During the development of salivary and mandibular glands, they recapitulate primitive stages of the phylogeny of the bees. During the development of salivary glands system, mitosis accounts for only part of the growth. Most of the growth occurs by increase in size of cells rather than by cell division. In brown-eyed and pigmented pupae six days before emergence, the salivary gland system is completely developed, although not yet functioning.     

Characterization of cuticular hydrocarbons of diploid and haploid males, workers and queens of the stingless bee Melipona quadrifasciata

Males, queens and workers of stingless bees show differences in external morphology, behaviour and roles within a colony. In addition, each individual has a cuticular chemical signature responsible for mutual communication that is essential for maintaining the integrity of the colony. In this paper we characterize the cuticular hydrocarbon composition of newly emerged diploid and haploid males, workers and virgin queens of Melipona quadrifasciata by gas chromatography-mass spectrometry (GC/MS) analysis. This is the first time that the cuticular profile of diploid males in a species of stingless bee has been characterized. We found differences in the cuticular hydrocarbon composition among males, workers and virgin queens, recording both qualitative and quantitative differences among individuals of different phenotypes. However, no compound was found exclusively in diploid males. The cuticular chemical profiles of haploid and diploid males were very similar to those of workers. Moreover, the cuticular lipids of males and workers were significantly different from those of queens. Tricosane, pentacosene-2 and 7-methyl-heptacosane were the compounds responsible for this significant separation. This result correlates with the behavioural and morphological differences among these phenotypes.     

Evaluation of the neotropical stingless bee Melipona quadrifasciata (Hymenoptera: Apidae) as pollinator of greenhouse tomatoes

The Neotropical stingless bee Melipona quadrifasciata Lepeletier was evaluated for pollinating tomatoes (variety Rodas; long-life hybrid) in greenhouses under plastic and with a hydroponic system and organic concepts in Minas Gerais State, Brazil. Flowers not pollinated did not set any fruit. Pollination by bees plus manual pollination did not differ from either bee or manual pollination. Maximum fruit diameter, fruit height, and roundness (quotient between maximum fruit diameter and fruit height) were not significantly different between treatments, but fruit visited by M. quadrifasciata had 10.8% less seeds (dry mass) than manual pollination. This apparently low efficiency of M. quadrifasciata pollination was attributed to the overlap of only 30 min between highest bee foraging activity and highest flower stigma receptivity. Thus, it was concluded that M. quadrifasciata is a feasible pollinator of greenhouse tomatoes because of 1) the observed increase in fruit quality with lower mechanical injury than traditional manual pollination, 2) no significant decrease in fruit size, and 3) high price of such product in the market. Some considerations for sustainable use of M. quadrifasciata as greenhouse pollinator are presented. Although techniques for keeping captive colonies of M. quadrifasciata are currently available, the sole current method for acquiring new colonies is removing them from the forest, and if demand was created for large numbers of colonies for commercial use, techniques for captive rearing must be developed to prevent serious declines in wild populations.     

Histochemistry of the cuticle from proventriculus in stingless bee, Melipona quadrifasciata anthidioides (Hymenoptera, Apidae)

Histochemical study of the proventriculus from stingless bee Melipona quadrifasciata anthidioides revealed for the first time the presence of cysteine-rich proteins in the cuticle that covers the bulb of the proventriculus and its associated hair-like process. The presence of tyrosine-rich proteins was observed in the same structure, but not in hair-like projections. The cuticle of the crop and a small portion of the proventriculus contained no acid carbohydrates or fats and small amounts of neutral carbohydrates. These results are discussed in relation to different composition of insect cuticle depending on its different functions.     

Variation and genetic structure of Melipona quadrifasciata Lepeletier (Hymenoptera, Apidae) populations based on ISSR pattern

For a study of diversity and genetic structuring in Melipona quadrifasciata, 61 colonies were collected in eight locations in the state of Minas Gerais, Brazil. By means of PCR analysis, 119 ISSR bands were obtained, 80 (68%) being polymorphic. H_e and H _B were 0.20 and 0.16, respectively. Two large groups were obtained by the UPGMA method, one formed by individuals from Januária, Urucuia, Rio Vermelho and Caeté and the other by individuals from São João Del Rei, Barbacena, Ressaquinha and Cristiano Otoni. The Φst and θ^B values were 0.65 and 0.58, respectively, thereby indicating high population structuring. UPGMA grouping did not reveal genetic structuring of M. quadrifasciata in function of the tergite stripe pattern. The significant correlation between dissimilarity values and geographic distances (r = 0.3998; p < 0.05) implies possible geographic isolation. The genetic differentiation in population grouping was probably the result of an interruption in gene flow, brought about by geographic barriers between mutually close geographical locations. Our results also demonstrate the potential of ISSR markers in the study of Melipona quadrifasciata population structuring, possibly applicable to the studies of other bee species.     

Morphology of the oviducts and endometria of cynomolgus macaques during the menstrual cycle

We sampled the reproductive tracts of 27 cynomolgus macaques during the menstrual cycle and correlated the cytologic changes in the oviductal epithelium with changes in the serum levels of estradiol (E2) and progesterone (P) and with the histology of the ovaries and the endometria. We identified an orderly sequence of changes in the oviductal epithelium from the early follicular to the late luteal phase, and we classified this sequence into eight stages, named as follows: preciliogenic, ciliogenic, ciliogenic-ciliated, ciliated-ciliogenic, ciliated-secretory, early regression, late regression and full regression. The preciliogenic and ciliogenic phases were coincident with menses and the early follicular phase. The ciliogenic-ciliated, ciliated-ciliogenic and ciliated-secretory phases during which the oviductal epithelium became progressively more differentiated were coincident, respectively, with the midfollicular, late follicular and periovulatory phases of the cycle. The early, late and full regression stages during which the epithelium became progressively more atrophied, deciliated and nonsecretory were coincident, respectively, with the early, mid and late luteal phases of the cycle. The cyclic changes in the endometrium of cynomolgus macaques were similar to those reported for the rhesus macaque.     

新牧Ⅰ号杂花苜蓿异交机制与自花粉的生殖干扰

通过控制试验和荧光观测等手段对新牧Ⅰ号杂花苜蓿Medicago varia Martyn'Xinmu No.1'柱头可授期与最佳授粉时间、自花粉的堵塞效应、花粉萌发与花粉管生长状况等进行研究,结果显示:(1)苜蓿柱头可授期与单花寿命具高度的同步性,在单花开放的第1天授粉,传粉效率最高,结果率达65.170%±2.01%(P<0.01),为柱头的最佳授粉时期;在单花开放后第4天授粉,其结果率仍达34.25%±6.73%.(2)自花授粉比异花授粉有着较低的花粉萌发率和花粉管生长速度;清除柱头自花粉后再授异花粉,其结荚率/花序和英内种子粒数由原来的51.760%±5.37%和2.11±0.18粒,提高到72.31%±6.24%(P<0.01)和3.46±0.25粒(P<0.01),自花粉对柱头的堵塞效应非常明显.(3)在繁育系统中,苜蓿采取了雌雄异熟和自交不亲和两套机制来避免自交,但是由于其独特的花结构,即使雄蕊先熟,在保证苜蓿异交机制中也没有起到有效的作用,而其典型的自交不亲和系统,虽然保证了苜蓿较低的自交率,却无法避免自花粉的生殖干扰.