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Major proteins synthesized in the hypopharyngeal gland of the worker honeybee change from bee-milk proteins to alpha-glucosidase in accordance with the age-dependent role change of the worker bee. Previously, we showed that the gene for alpha-glucosidase is expressed specifically in the forager-bee gland [Ohashi, K., Sawata, M., Takeuchi, H., Natori, S. & Kubo, T. (1996) Biochem. Biophys. Res. Commun. 221, 380-385]. Here, we describe the isolation and analysis of cDNAs for two bee-milk 56-kDa and 64-kDa proteins. The 56-kDa protein was a glycoprotein which shared 63.2% and 56.9% amino acid sequence identities with proteins encoded by cDNA for royal-jelly-related protein 57-1 (pRJP57-1) and pRJP57-2. The 64-kDa protein cDNA was identical to pRJP57-1. Thus, these bee-milk proteins seem to form a structurally related protein family. The gene for the 64-kDa protein/RJP57-1 was expressed specifically in the nurse-bee gland, whereas that for the 56-kDa protein was expressed in both the nurse-bee and forager-bee glands. mRNAs for the 56-kDa and 64-kDa proteins were detected by in situ hybridization in a whole acinus of the nurse-bee gland, whereas mRNAs for the 56-kDa protein and alpha-glucosidase were detected in that of the forager-bee gland. Therefore, the individual secretory cells of the acinus of the hypopharyngeal gland were shown to express these genes differently with the age-dependent role change of the worker bee.
The spermatheca of the honey bee queen is covered by a single-layered, uniform, polarised epithelium. The apical cell surface is greatly enlarged by protrusions and plasma membrane infoldings, the basal cell surface by numerous interdigitating, long, small processes. Cytoplasmic organelles are chiefly represented by mitochondria. Numerous microtubuli extend throughout the cytoplasm. Golgi and endoplasmic profiles are rare. The cells are subject to senile degeneration: with increasing age, a variety of cytoplasmic inclusions appear, among which are myelinated membranes, dense bodies and dense filamentous aggregates. The spermathecal epithelium does not seem to be involved in exocrine secretion related to nutrition of the long-term stored spermatozoa. The ultra-structure points, however, to ion transport functions and to an engagement in the maintenance of an adequate physicochemical environment ensuring the viability of the spermatozoa. Cellular junctions are represented by luminal zonulae adherentes, focal cell-cell adhering junctions and hemiadhering junctions along the basal plasmalemma. Desmosomal contacts and cytoskeletal intermediate filaments are missing. Along the lateral plasmalemma, gap junctions and septate junctions are found.
A synopsis about published methods and results on experiments to cultivate bee cells in vitro is given. Experimental investigations were performed with haemocytes of larvae of the L-5 stage using many different media and methods for the preparation of primary tissue culture. Monolayers could be prepared and a high rate of reproduction has been achieved, although subpassages could not be obtained. Haemocytes could be kept alive up to 27 days by using BML-TC/7A medium according to Gardiner and Stockdale, modified by Skatulla (pers. communic., 1987). Further experiments are necessary in order to study the suitability of bee cells to detect specific pathogens and toxic substances.
The plant flora has an important role in the ecology of Cryptococcus neoformans. It is estimated that the environmental spreading and contamination of human beings with this yeast occurs via contaminated particles of plants. Cultivation of canopy parts of plants in selective media is the most widely used isolation method of this yeast. Cryptococcus neoformans var. grubii was isolated from honeybee colonies in Eucalyptus forests but was not isolated from the places where this flora did not exist. Our results indicate that the occurrence of C. neoformans in honeybee colonies during the flowering season of Eucalyptus spp. trees can be an important bioindicator for environmental yeast presence. The screening of honeybee colonies is a practical and a rapid method for the monitoring of the C. neoformans presence in flowering plants.
The mushroom bodies (MBs) are central structures in the insect brain that have been associated with olfactory learning and memory. Here we used hydroxyurea (HU) to treat honeybee larvae and induce partial MB ablations at the adult stage. We studied olfactory learning in honeybees with unilateral loss of the median calyces of their MBs and compared their ability to solve different forms of olfactory discrimination. When odorants were delivered in a side-specific manner, ablated bees could not solve either discrimination of the unambiguous problem (Paradigm 1: A+, B- on one antenna, C+, D- on the other; A+B-/C+D-) whereas they could solve at least one of both discriminations of the ambiguous problem (Paradigm 2: A+B-/A-B+), namely that proposed to their intact brain side. Non-ablated bees could learn side-specific discriminations on both brain sides. When odorants were delivered simultaneously to both antennae (Paradigm 3: A+B-C+D-), HU-ablated bees learned slower than HU-normal bees. Thus, in all three paradigms, the unilateral loss of a median calyx affected olfactory learning. We propose that the MBs are required for solving elemental olfactory tasks whose complexity is increased by the number of stimuli involved and that MB ablations could have an effect on the inhibition of information exchange between brain hemispheres.
OBJECTIVE: To test a new system for the biotyping of Streptococcus mutans, based on the measurement of enzyme activity, and to investigate the relationship between biotype and in vitro susceptibility to seven clinically useful antibiotics. METHODS: In total, 160 oral isolates of S. mutans were classified into different biotypes with the APIZYM test for enzyme activity, excluding results that were positive or negative in >80% of the strains. The susceptibility of all 160 strains to amoxycillin, cefazolin, erythromycin, clindamycin, vancomycin, teicoplanin and imipenem was tested by dilution in a solid medium. Statistical analysis of susceptibility (mean minimum inhibitory concentrations (MICs)) was based on chi-squared tests. RESULTS: Eight different biotypes (1-8) were identified on the basis of three kinds of enzyme activity: valine aryl amidase, acid phosphatase and alpha-galactosidase. Biotype 5 was found to be the most common. The mean MIC values showed strains belonging to biotype 4 to be the most susceptible to amoxycillin, cefazolin and erythromycin, whereas biotype 1 was the least susceptible to teicoplanin. CONCLUSIONS: The proposed biotyping method, which is relatively fast and simple to perform, provided reproducible results, and may contribute to clinically effective treatment of S. mutans infections.
1 Peptide 401, a potent mast cell degranulating factor from bee venom, substantially inhibited the oedema provoked by subplantar injection of carrageenin or intra-articular injection of turpentine in the rat. The ED(50) of 401 was c. 0.1 mg/kg. The anti-inflammatory effect was assessed by measurement of the increased (125)I-albumin content of an injected site in comparison with an uninjected contralateral site.2 Peptide 401 also suppressed the increased vascular permeability due to intradermal injection of various smooth muscle spasmogens (histamine, bradykinin, 5-hydroxytryptamine (5-HT), and prostaglandins).3 Other comparable mast cell degranulating agents (48/80 and melittin) showed little evidence of anti-inflammatory activity when tested at comparable dosage on turpentine arthritis and carrageenin oedema.4 The anti-inflammatory effects were not abolished by pretreatment with mepyramine and methysergide, which abolished the increased vascular permeability produced by local injection of 401.5 The anti-inflammatory action of 401 was not affected by regional denervation or pretreatment with phenoxybenzamine, and was reduced but not abolished by adrenalectomy.6 Measurement of skin temperature, fractional extraction of (86)Rb and blood flow in perfused mesentery gave no evidence that the anti-inflammatory action of 401 was due to reduced tissue perfusion.7 It is concluded that 401 may exert its anti-inflammatory action directly by making the vascular endothelium anergic to phlogistic stimuli.
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1. Receptor potentials in honeybee drone retinula cells were recorded with intracellular micro-electrodes in the dorsal part of the superfused retina. The light stimuli were sufficiently weak that the response amplitude was proportional to the intensity. 2. Responses to stimuli of different wave-lengths, although of different amplitude, all had the same time course. 3. The maximal sensitivity in all the cells recorded from was to a wave-length between 450 and 460 nm. 4. Microspectrophotometry showed the presence of a pigment with two stable states, interconvertible by light, absorbing maximally at 445 nm (rhodopsin) and 505 nm (metarhodopsin). 5. There was a good match between the absorption spectrum of rhodopsin and the spectral sensitivity of retinula cells. 6. Transformation of a large fraction of rhodopsin to metarhodopsin by light reduced the sensitivity of the retinula cell but did not alter the shape of the relative spectral sensitivity curve or the time course of the responses. 7. It is concluded that for weak lights the receptor potential is determined only by the number of rhodopsin molecules that absorb photons: neither the presence of metarhodopsin nor its phototransformation to rhodopsin produces a detectable effect.
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A survey of six bee viruses on a large geographic scale was undertaken by using seemingly healthy bee colonies and the PCR technique. Samples of adult bees and pupae were collected from 36 apiaries in the spring, summer, and autumn during 2002. Varroa destructor samples were collected at the end of summer following acaricide treatment. In adult bees, during the year deformed wing virus (DWV) was found at least once in 97% of the apiaries, sacbrood virus (SBV) was found in 86% of the apiaries, chronic bee paralysis virus (CBPV) was found in 28% of the apiaries, acute bee paralysis virus (ABPV) was found in 58% of the apiaries, black queen cell virus (BQCV) was found in 86% of the apiaries, and Kashmir bee virus (KBV) was found in 17% of the apiaries. For pupae, the following frequencies were obtained: DWV, 94% of the apiaries; SBV, 80% of the apiaries; CBPV, none of the apiaries; ABPV, 23% of the apiaries; BQCV, 23% of the apiaries; and KBV, 6% of the apiaries. In Varroa samples, the following four viruses were identified: DWV (100% of the apiaries), SBV (45% of the apiaries), ABPV (36% of the apiaries), and KBV (5% of the apiaries). The latter findings support the putative role of mites in transmitting these viruses. Taken together, these data indicate that bee virus infections occur persistently in bee populations despite the lack of clinical signs, suggesting that colony disease outbreaks might result from environmental factors that lead to activation of viral replication in bees.