RESUMEN
Con motivo de conocer el estado de salud de poblaciones silvestres de la concha prieta Anadara tuberculosa se estudiaron los parámetros hemocitarios (viabilidad, conteo total y diferencial, estabilidad lisosomal, fragilidad osmótica) y la química sanguínea (hemoglobina-Hb, proteínas, triglicéridos, glucosa, lactato deshidrogenasa-LDH y catalasa-CAT). Se extrajo hemolinfa en reproductores salvajes aparentemente sanos colectados en la isla Corazón, río Chone, Ecuador. La viabilidad celular fue elevada, con presencia de 5 morfotipos: eritrocitos (74%), granulocitos traslucidos (6%), amebocitos (3%), hialinocitos (12%), blastocitos (5%). Los hemocitos presentaron membranas lisosomales estables al rojo neutro durante 240 min y una fragilidad osmótica media (FO50) de 4.8‰. Las concentraciones de Hb, proteínas, lípidos y glucosa denotan la función respiratoria y reservas energéticas durante los cambios de marea. La actividad de LDH está vinculada al metabolismo anaeróbico y CAT a la capacidad de mantener el equilibrio redox del sistema inmunitario. Los parámetros hemocitarios y química de la hemolinfa pueden servir como índices fisiológicos normales de referencia en reproductores de A. tuberculosa.
In this work we studied the health status of wild populations of the black shell Anadara tuberculosa, haemocyte parameters (viability, total and differential count, lysosomal stability, osmotic fragility) and blood chemistry (haemoglobin-Hb, proteins, triglycerides, glucose, lactate dehydrogenase-LDH and catalase-CAT). Haemolymph was extracted from apparently healthy wild broodstock collected from Corazón Island, Chone River, Ecuador. Cell viability was high, with 5 morphotypes present: erythrocytes (74%), colorless granulocytes (6%), amebocytes (3%), hyalinocytes (12%), blastocytes (5%). Haemocytes showed stable lysosomal membranes to neutral red for 240 min and a mean osmotic fragility (OF50) was 4.8‰. Hb, protein, lipid and glucose concentrations denote respiratory function and energy reserves during tidal changes. LDH activity is linked to anaerobic metabolism and CAT denotes the ability to maintain the redox balance of the immune system. The haemocitary parameters and haemolymph chemistry can serve as normal physiological reference indices in A. tuberculosa broodstock.
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The observation statistics suggested that the haemolymph melanization speed of larvae became fast and the growth inhibition of Escherichia coli was strong as the quantities of feeding on mulberry leaves increased. The RT-PCR result showed that the mRNA expressions of melanin biosynthesis enzyme BmTan, BmPo-1, BmYellow-f and BmDdc were high in the haemolyph of 5 L 3 d larvae. The qPCR analysis showed Bmtan, Bmddc, Bmyellow, Bmebony and Bmblack, especially Bmddc expression were significantly higher in black disease larvae than in normal larvae. Compared with control, Ddc inhibitors drastically inhibited the lipopolysaccharide-induced haemolymph melanization. In addition, the content of Dopa and Dopamine markedly rose after E. coli injection. These indicated that haemolymph melanization was linked to immune defenses and Bmddc may play a role in melanization response of haemolymph immune in silkworm.
Asunto(s)
Animales , Bombyx , Genética , Microbiología , Escherichia coli , Genes de Insecto , Hemolinfa , Química , Larva , MelaninasRESUMEN
Background: The increasing incidence of high blood pressure, its complications and associated fatalities has led to an upsurge in the use of alternate forms of medicaments in its management. The oral ingestion of the hemolymph of Archachatina marginata is commonly used as an antihypertensive by the Yoruba people of South West Nigeria. This study investigated the effect of oral administration of Archachatina hemolymph on normotensive and adrenaline induced hypertensive wistar rats. Methods: The hemolymph of Archachatina marginata was orally administered at doses of 22.8 and 45.6 mg/kg body weight to normotensive and adrenaline induced hypertensive rats for 7 days. Blood pressure parameters were measured via a polygraph. Histopathological assessment of the heart tissue was conducted. Data gathering and analysis were done in 2014 (February – August). Results: In this study, the orally administered hemolymph had no significant (p˂0.05) lowering effect on the systolic/ diastolic pressure, pulse pressure, mean arterial pressure or heart rate of either the normotensive or adrenaline induced hypertensive rats. Histopathological assessment of the cross section of the heart tissues shows the hemolymph had no adverse effect on the examined cross section of the heart tissue. Conclusion: Based on the data from this study, there is no justifiable reason for the use of the hemolymph of Archachatina marginata as a antihypertensive.
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Examined the day to day changes of amino acid profiles in the haemolymph of 5th instar silkworm Bombyx mori L. during the development of fungal pathogen Beauveria bassiana (Bals.) Vuill. For the qualitative determination of amino acids Thin Layer Chromatography (TLC) was performed. The qualitative changes were observed in amino acid at different lengths on 1st and 6th day after inoculation of Beauveria bassiana. More number of amino acid spots were noticed on 1st and 6th day of inoculated larvae compared to control.
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Background: The use of wild animals and their products are an integral and essential ingredients in the preparation of drugs/medication for various ailments and diseases in West African traditional medicine. Methods: This study investigated the effect of administering the haemolymph of Achachatina marginata on the biochemical parameters of the normal animals as well as the histopathological effect of the continuous administration of the haemolymph on the vital organs of the animals.. Results: The preliminary study revealed that the haemolymph of A. marginata exhibited a weight loss effect in the studied animals as well as significantly lowered the cholesterol and triglyceride levels in normal rats used in this study. Conclusion: Though this result appear promising, it will be necessary to evaluate the long term effect of the haemolymph of A. marginata effect on the biochemical parameters.
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The process of uptake of hexamerins during metamorphosis from insect haemolymph by fat body cells is reminiscent of receptor-mediated endocytosis. Previously, we had identified a hexamerin-binding protein (HBP) and reported for the first time that uptake of hexamerins is dependent on the phosphorylation of HBP partly by a tyrosine kinase, which is, in turn, activated by 20-hydroxyecdysone (20E). However, the exact nature of HBP and the mechanism of interaction are still unknown. Here we report the possibility of HBP being a GPIanchored protein in the fat body of Achaea janata and its role in the tyrosine-kinase-mediated phosphorylation signalling. Digestion of fat body membrane preparation with bacterial phosphatidylinositol-specific phospholipase C (PI-PLC), and the subsequent recognition by antibodies specific for the cross-reacting determinant (CRD), revealed that HBP is glycosylphosphatidylinositol (GPI)-anchored protein and, further, that the hexamerin binding to HBP was inhibited after digestion. Hexamerin overlay assay (HOA) of co-immunoprecipitated in vitro phosphorylated HBP showed exclusive binding to ~120 kDa protein. Lectin-binding analysis of hexamerins revealed the presence of N-acetylgalactosamine (GalNAc) and N-acetylglucosamine (GluNAc), whereas HBP showed the presence of GalNac alone. Mild chemical deglycosylation studies and binding interaction in the presence of sugars revealed that glycan moieties are possibly not involved in the interaction between HBP and hexamerins. Taken together, these results suggest that HBP may be a GPI-anchored protein, and interaction and activation of HBP is through lipid-linked non-receptor src tyrosine kinases. However, additional studies are needed to prove that HBP is a GPI-anchored protein.
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The present study investigated the effect of increasing temperature stress on the thermotolerance of B. mori crossbreed PM x CSR2 and tissue specific differential expression of heat shock proteins at IVth and Vth instars. The larvae reared at 25 ± 1oC and 70 ± 5% relative humidity were treated as control. Larvae were subjected to heat shock temperatures of 34, 38 and 42oC for 3 hr followed by 3 hr recovery. Expression of Heat shock protein 72 were analyzed by SDS-PAGE and confirmed by western blotting analysis. The impact of heat shock on commercial traits of cocoons was analyzed by following different strategies in terms of acquired thermotolerance over control. Resistance to heat shock was increased as larval development proceeds and increased thermotolerance is achieved with the induction of Heat shock protein 72 in the Vth instar larval haemolymph. Relative influence of heat shock temperatures on commercial traits corresponding to the generation of heat shock protein 72 was significantly improved over control. In PM x CSR2, cocoon and shell weight significantly increased to 9.90 and 11.90% over control respectively.
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Barylelphusa gureini, a fresh water male crab was exposed to sub-lethal concentration (1ppm) of copper sulfate, and the oxygen consumption and haemolymph sugar level of the animal were estimated to study the stress caused by this heavy metal toxicant. These animals are abundantly found in the outskirts of Pune district of Maharashtra State, mainly in and around paddy fields. They were brought to the laboratory from their natural habitat, and first subjected to acclimatization to the laboratory conditions. They were subsequently exposed for 0, 24, 48, 72 and 96 hours to the toxicant, and the total oxygen consumption was studied by Winkler's method, and the haemolymph sugar level was estimated by Enthrone method. In the present study, total oxygen consumption showed a gradual declining trend from 0 to 96 hours in the experimental animals exposed to copper sulfate, while the haemolymph sugar level recorded an elevation, with maximum increase at 48 hours.