Edible Insects in Africa in Terms of Food, Wildlife Resource, and Pest Management Legislation.

: Entomophagy is an ancient and actually African tradition that has been receiving renewed attention since edible insects have been identified as one of the solutions to improve global nutrition. As any other foodstuff, insects should be regulated by the government to ensure product quality and consumer safety. The goal of the present paper was to assess the current legal status of edible insects in Africa. For that, corresponding authorities were contacted along with an extensive online search, relying mostly on the FAOLEX database. Except for Botswana, insects are not mentioned in national regulations, although the definitions for "foodstuff" allow their inclusion, i.e., general food law can also apply to insects. Contacted authorities tolerated entomophagy, even though no legal base existed. However, insects typically appear in laws pertaining the use of natural resources, making a permit necessary (in most cases). Pest management regulation can also refer to edible species, e.g., locusts or weevils. Farming is an option that should be assessed carefully. All this creates a complex, nation-specific situation regarding which insect may be used legally to what purpose. Recommendations for elements in future insect-related regulations from the food hygiene point of view are provided.

Comparative Study of the Use of Insect Meal from Spodoptera littoralis and Bactrocera zonata for Feeding Japanese Quail Chicks.

A transformation of current livestock production towards a more sustainable operation is crucial to face nutritional and environmental challenges. There is an urgent demand for more sustainable high-quality feed sources to reduce environmental costs. Insects pose a potential alternative since they can be reared sustainably on food and feed residues. Know-how in mass rearing already exists for insect species used in biological pest control, such as the African cotton leafworm Spodoptera littoralis and the peach fruit fly Bactrocera zonata. The impact of a replacement of 50% of soybean meal by S. littoralis and B. zonata meal, respectively, on seven-days-old Japanese quail chicks was investigated in feeding trials. Concomitantly, the chemical compositions of the two insect meals and soybean meal were determined and compared. It was observed that the insect meals had higher protein and fat contents, lower carbohydrate contents and contained more saturated fatty acids than soybean meal. They also had higher methionine, and S. littoralis had a higher lysine content. Feeding trials resulted in improved growth, feed performance parameters, carcass characteristics, and biochemical indices for both insect meals. Consequently, both insect meals represent a promising alternative to soy in the feed of Japanese quail chicks.

Metallothionein in Hermetia illucens (Linnaeus, 1758) larvae (Diptera: Stratiomyidae), a potential biomarker for organic waste system.

Black soldier fly, Hermetia illucens (Linnaeus, 1758), is an important economic fly as its larvae can be used for recycling organic waste, such as food waste and manure. H. illucens larvae (BSFL) could uptake Cd from substrates and accumulate it inside bodies, which need to be monitored during waste treatment. Metallothionein (MT) usually serve as biomarker because of its role in metal homeostasis, detoxification, and dose response of heavy metals. Therefore, a MT gene was cloned from H. illucens (HIMT) that encoded 40 amino acids with typical cysteine rich features, which had a high sequence identity with other insect MTs. The expression of HIMT and total MT protein was measured in BSFL fed by meals spiked with gradient dose of Cd (0, 5, 50, 500 mg/kg) for 24, 48, 72, and 96 h, respectively. Dose-associated response of HIMT and total MT were found and the possible correlative range of Cd was from 5 to 50 mg/kg. The expression of HIMT might be a potential biomarker for monitoring Cd contamination by H. illucens in terrestrial organic matters, which might further apply in waste transformation system.

Lupus vasculopathy: Diagnostic, pathogenetic and therapeutic considerations.

A rare form of vascular disease in systemic lupus erythematosus (SLE), lupus vasculopathy is characterized by necrosis and accumulation of immunoglobulins (IGs) and complements in the wall of arterioles and small arteries resulting in luminal narrowing. Lupus vasculopathy often accompanies lupus nephritis and portends a poor prognosis. Although there is general agreement on the treatment of lupus nephritis, effective treatment strategies for lupus vasculopathy remain to be defined. We report a 20-year-old woman with SLE who presented with generalized tonic-clonic seizure. Her immunosuppressive regimen consisted of mycophenolate mofetil, prednisone and hydroxychloroquine. On physical examination, she was Cushingoid in appearance and hypertensive. Laboratory tests indicated renal disease. Coagulation studies disclosed de novo lupus anticoagulant. Magnetic resonance imaging of the brain demonstrated acute focal cerebral hemorrhage. Echocardiography revealed reduced ejection fraction and severe mitral regurgitation. Despite high-dose glucocorticoids and mycophenolate mofetil, renal function remained poor. Kidney biopsy demonstrated lupus vasculopathy and glomerulonephritis. Plasma exchange therapy and intravenous cyclophosphamide were administered. Over the ensuing four weeks, renal function improved, complement levels increased, autoantibody titers decreased and lupus anticoagulant disappeared. In conclusion, lupus vasculopathy can occur in SLE despite a heavy immunosuppressive regimen. Antiphospholipid antibodies might be involved in the pathogenesis of lupus vasculopathy. Plasma exchange therapy in conjunction with intravenous cyclophosphamide may represent an effective treatment strategy for lupus vasculopathy.

Mitotic and polytene chromosomes analysis of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae).

The Oriental fruit fly, Batrocera dorsalis s.s. (Hendel) is one of the most destructive agricultural pests, belonging to a large group of difficult to distinguish morphologically species, referred as the B. dorsalis complex. We report here a cytogenetic analysis of two laboratory strains of the species and provide a photographic polytene chromosome map from larval salivary glands. The mitotic complement consists of six chromosome pairs including a heteromorphic sex (XX/XY) chromosome pair. Analysis of the polytene complement has shown a total of five polytene chromosomes (10 polytene arms) that correspond to the five autosomes. The most important landmarks of each polytene chromosome and characteristic asynapsis at a specific chromosomal region are presented and discussed. Chromosomal homology between B. dorsalis and Ceratitis capitata has been determined by comparing chromosome banding patterns. The detection of chromosome inversions in both B. dorsalis strains is shown and discussed. Our results show that the polytene maps presented here are suitable for cytogenetic analysis of this species and can be used for comparative studies among species of the Tephritidae family. They also provide a diagnostic tool that could accelerate species identification within the B. dorsalis complex and could shed light on the ongoing speciation in this complex. Polytene chromosome maps can facilitate the development of biological control methods and support the genome mapping project of the species that is currently in progress.

Concentration of rafts in platelet filopodia correlates with recruitment of c-Src and CD63 to these domains.

The molecular mechanism that causes non-adhesive, discoid platelets to transform into sticky dendritic bodies that form blood clumps is a complex series of events. Recently it has become clear that lipid microdomains--also known as rafts--play a crucial role in this process. We have used a non-cytolytic derivative of perfringolysin-O, a cholesterol binding cytolysin, that binds selectively to cholesterol-rich membrane domains, combined with confocal- and immunoelectron microscopy to visualize cholesterol-raft dynamics during platelet adhesion. In resting platelets cholesterol was uniformly distributed on the cell surface and confined to distinct intracellular compartments (i.e. multivesicular bodies, dense granules, and the internal membranes of alpha-granules). Upon interaction with fibrinogen, cholesterol accumulated at the tips of filopodia and at the leading edge of spreading cells. Stimulation with thrombin receptor activating peptide (TRAP) resulted in a similar redistribution of cholesterol towards filopodia. The adhesion-dependent raft aggregation was accompanied by concentration of the tyrosine kinase c-Src and the tetraspanin CD63 in these domains, whereas glycoprotein Ib (GPIb) was not selectively targeted to the raft clusters. c-Src, the tetraspanin CD63, and GPIb were recovered in biochemically isolated low-density membrane fractions. Disruption of rafts by depleting membrane cholesterol had no effect on platelet shape change but inhibited platelet spreading on fibrinogen and TRAP-induced aggregation. Our results demonstrate that cholesterol rafts in platelets are dynamic entities in the membrane that co-cluster with the tyrosine kinase c-Src and the costimulatory molecule CD63 in specialized domains at the cell surface, thereby providing a possible mechanism in functioning as signaling centres.

Selective binding of perfringolysin O derivative to cholesterol-rich membrane microdomains (rafts).

There is increasing evidence that sphingolipid- and cholesterol-rich microdomains (rafts) exist in the plasma membrane. Specific proteins assemble in these membrane domains and play a role in signal transduction and many other cellular events. Cholesterol depletion causes disassembly of the raft-associated proteins, suggesting an essential role of cholesterol in the structural maintenance and function of rafts. However, no tool has been available for the detection and monitoring of raft cholesterol in living cells. Here we show that a protease-nicked and biotinylated derivative (BCtheta) of perfringolysin O (theta-toxin) binds selectively to cholesterol-rich microdomains of intact cells, the domains that fulfill the criteria of rafts. We fractionated the homogenates of nontreated and Triton X-100-treated platelets after incubation with BCtheta on a sucrose gradient. BCtheta was predominantly localized in the floating low-density fractions (FLDF) where cholesterol, sphingomyelin, and Src family kinases are enriched. Immunoelectron microscopy demonstrated that BCtheta binds to a subpopulation of vesicles in FLDF. Depletion of 35% cholesterol from platelets with cyclodextrin, which accompanied 76% reduction in cholesterol from FLDF, almost completely abolished BCtheta binding to FLDF. The staining patterns of BCtheta and filipin in human epidermoid carcinoma A431 cells with and without cholesterol depletion suggest that BCtheta binds to specific membrane domains on the cell surface, whereas filipin binding is indiscriminate to cell cholesterol. Furthermore, BCtheta binding does not cause any damage to cell membranes, indicating that BCtheta is a useful probe for the detection of membrane rafts in living cells.

Scientists turn to journals in English.

Biomedical publications listed in Medline were analyzed based on publisher's location and first author's country of origin. In the present analysis I wished to determine the languages of biomedical publications and the publishers' locations.

Mechanism of dye binding in the protein assay using eosin dyes.

Eosin B and eosin Y have been used to estimate micro- and submicrogram quantities of proteins respectively as shown in our previous reports. In the present study we describe the mechanism of eosin binding to proteins. At pH lower than 3.0 the absorbance of unbound dye is greatly reduced. After the dye binds to protein, the absorption maximum of the dye changes from 514 to 530 +/- 5 nm. The absorbance and bathochromatic shift in absorption maximum of the protein-dye complex are proportional to the concentration of protein. The pH of the assay solution does not change due to protein. Arginine, histidine, and lysine (at both acidic and neutral pH) and tryptophan (at acidic pH) residues of a protein bind electrostatically to carboxylic and phenolic groups of the dye to produce a stable water-soluble protein-dye complex. The binding constants of eosin B with poly-L-arginine, poly-L-histidine, poly-L-lysine, and poly-L-tryptophan at pH 1.96 are 0.37, 0.32, 0.33 and 0.33 nmol/nmol of amino acid, respectively. The binding constants of eosin B and eosin Y with bovine serum albumin (BSA) at pH 1.96 are essentially the same, i.e., 0.82 nmol/nmol of reactive amino acid of BSA. The binding constant varies with solution pH so that a wide range of protein concentrations can be estimated. The reason for the higher absorbance of protein-eosin Y complex compared to that of protein-eosin B complex is discussed.

Estimation of submicrogram quantities of protein using the dye eosin Y.

Eosin B is used to estimate proteins above 1 microg/ml concentration [Waheed AA, Gupta, PD. Anal. Biochem. 1996:233:249-256; Waheed AA, Gupta PD. J. Biochem. Biophys. Meth. 1996;33:187-196]. In the present report we describe a method for estimating submicrogram quantities of proteins using the dye eosin Y. The increase in sensitivity of this assay is approximately two fold under optimal assay condition. The optimum concentration of eosin Y and citric acid for submicrogram assay is 0.01 and 0.05%; (final concentration) respectively. The protein-dye complex formation is completed within 2 min and its absorbance is stable up to 60 main with a variation of +/-4.0%. The interference due to sugars, reducing agents, glycerol and some neutral detergents like Triton X-100, NP-40 and Tween-20 is less than 12% whereas Brij-35, ethanol, acetone and chelators like EGTA and EDTA suppress the absorbance by about 12-18%. However, basic buffers like Tris, urea, CHAPS and NaN, interfere with the formation of the protein-dye complex. The increase in absorbance of protein-eosin Y complex compared to that of protein-eosin B complex is due to the higher extinction co-efficient of eosin Y compared to eosin B.

Lipid and fatty acid composition of brush border membrane of rat intestine during starvation.

Alterations in the lipid and fatty acid composition of brush border membrane (BBM) of small intestine were studied in well-fed, starved, and refed rats. The ratios of cholesterol/phospholipid (mol/mol), sphingomyelin/phosphatidylcholine (mol/mol), protein/lipid (w/w), and free fatty acids (w/w) decreased whereas the total phospholipid (w/w) ratio and the double-bond index increased in BBM of the intestine of the starved rat compared to that of the well-fed rat. Analyses of fatty acids showed higher percentage of stearic and arachidonic acids whereas oleic and linoleic acids decreased under starvation. The acyl chain of starved rat BBM was less ordered compared with that of well-fed rat BBM. On refeeding, these changes were restored to well-fed levels. The change in membrane state under starvation is associated with alterations in the lipid and fatty acid composition of BBM and may be responsible for functional changes that occur under nutritional stress.

Decreased densities of intramembranous particles and cytochemically detectable cholesterol in microvilli of starved rat enterocytes.

The densities of intramembranous particles (IMPs) and of sterol complexes induced by treatment of filipin were studied by freeze-fracture replication of intact intestine and/or isolated brush border membranes (BBM) of well-fed and starved rats. The density of IMPs and filipin-sterol complexes (FSCs) decrease considerably during starvation. Biochemical estimations show a decrease in the levels of cholesterol and proteins with respect to phospholipids during starvation which is in agreement with morphological findings. It is suggested that these changes may play a role in regulating membrane fluidity which in turn affects absorption of nutrients through BBM.

Changes in structural and functional properties of rat intestinal brush border membrane during starvation.

Changes in surface area of microvilli, fluidity of brush border membrane and transport of L-amino acids through intestinal epithelial cells were studied in wellfed and starved (2,4 and 6 days) rats. The surface area of microvilli per unit area of intestinal epithelial cells increased during starvation. Studies with fluoroprobes - pyrene, 1-anilinonaphthalene-8-sulphonate and 1,6-diphenyl-1,3,5-hexatriene, showed increased fluidity of brush border membrane on progressive starvation. Transport of five amino acids representing five different transport systems was studied during starvation in everted intestinal sleeves. Transport of L-proline, glycine and L-glutamic acid which represent imino, glycine and acidic systems respectively increased significantly in Na+-dependent pathway whereas transport of L-lysine representing basic system increased significantly in Na+-independent pathway during starvation.

Application of an eosin B dye method for estimating a wide range of proteins.

We describe a detailed procedure for estimating a wide range of proteins by the eosin B dye method. At acidic pH, eosin B binds to proteins and absorption of the protein-dye complex at 536-544 nm is proportional to the concentration of the proteins. Inorganic and organic acids are used for the assay, the optimum concentration of acid for each assay differs from acid to acid. This method provides minimal interference with commonly used compounds in protein purification and biomolecules like DNA, RNA, lipids, carbohydrates and minerals which are generally present in biological fluids. This method is applicable for estimating proteins in tissue homogenates, isolated proteins and various biological fluids.

Effect of starvation on glucose transport and membrane fluidity in rat intestinal epithelial cells.

Studies on the surface area of microvilli (MV), fluidity of brush border membranes (BBM) and D-glucose uptake were carried out in rat intestinal epithelial cells (IEC) during progressive starvation and under re-feed conditions. The surface area of MV, fluidity of BBM and D-glucose transport through IEC membranes showed an increase during starvation when compared to well-fed controls. Re-feeding experiments restored the control values of all the three parameters within a short time. The results showed that the increase in D-glucose transport through IEC membranes during starvation is due to increased surface area of MV and increased fluidity of BBM.