INFLUENCE OF ACTIVE IMMUNIZATION ON MELANIZATION OF THE BLOOD OF WAX MOTH LARVAE

1962 ◽  
Vol 8 (5) ◽  
pp. 597-602 ◽  
Author(s):  
June M. Stephens

Larval blood of the wax moth, Galleria mellonella (L.), immunized against Pseudomonas aeruginosa does not melanize on exposure to air as does normal blood. Larval blood samples taken 20 to 24 hours after vaccination against either of the pathogens P. aeruginosa or Proteus mirabilis did not melanize; blood of insects vaccinated against the nonpathogenic Shigella dysenteriae strain K 629 or a nonpathogenic strain of Pseudomonas did melanize. Injection of nonspecific agents, such as sodium chloride or egg albumen, did not inhibit melanization. The oxidation–reduction potential of immune blood was markedly lower than that of normal blood. Enzymes from both normal and immune blood were precipitated with cold acetone. The enzyme preparations were nontoxic to normal larvae. Both normal and immune blood enzyme preparations produced melanin from tyrosine, though the immune blood enzyme acted more slowly. Sodium thioglycollate, ascorbic acid, benzoin oxime, and phenylthiourea each inhibited the melanization of normal blood in vitro but did not confer bactericidal activity. These inhibitors did not produce any change in the rate of melanization, bactericidal activity, or immune properties of the blood when injected into larvae.

2006 ◽  
Vol 387 (5) ◽  
pp. 549-557 ◽  
Author(s):  
Gregor Langen ◽  
Jafargholi Imani ◽  
Boran Altincicek ◽  
Gernot Kieseritzky ◽  
Karl-Heinz Kogel ◽  
...  

Abstract A cDNA encoding gallerimycin, a novel antifungal peptide from the greater wax moth Galleria mellonella, was isolated from a cDNA library of genes expressed during innate immune response in the caterpillars. Upon ectopic expression of gallerimycin in tobacco, using Agrobacterium tumefaciens as a vector, gallerimycin conferred resistance to the fungal pathogens Erysiphe cichoracearum and Sclerotinia minor. Quantification of gallerimycin mRNA in transgenic tobacco by real-time PCR confirmed transgenic expression under control of the inducible mannopine synthase promoter. Leaf sap and intercellular washing fluid from transgenic tobacco inhibited in vitro germination and growth of the fungal pathogens, demonstrating that gallerimycin is secreted into intercellular spaces. The feasibility of the use of gallerimycin to counteract fungal diseases in crop plants is discussed.


1962 ◽  
Vol 8 (4) ◽  
pp. 491-499 ◽  
Author(s):  
June M. Stephens

The blood of normal wax moth larvae is not bactericidal for Pseudomonas aeruginosa. The blood becomes moderately bactericidal when larvae are actively immunized against P. aeruginosa. This activity was measured by a modification of Nagington's technique for the estimation of typhoid antibody. Bactericidal activity appears to be the only measurable type of antibody response against P. aeruginosa. Cell-free blood was as active as whole blood; 0.02 ml of immune serum kills about 1000 organisms. The blood of normal wax moth larvae is bactericidal for Shigella dysenteriae but the blood of insects immunized against either this organism or P. aeruginosa shows no increase in activity against S. dysenteriae. A number of non-specific agents, both protein and non-protein, did not stimulate bactericidal activity in serum after their injection into normal larvae. Immune sera prepared against some strains of P. aeruginosa were not active against other strains. Storage at 37 °C or absorption with zymosan both result in blackening of immune blood and loss of bactericidal activity. Bactericidal activity is evident only during the immune state of the insect, i.e. from about 18 hours until the third day after vaccination; it develops at the same time that inhibition of melanization was observed in the blood from vaccinated larvae.


Sociobiology ◽  
2020 ◽  
Vol 67 (1) ◽  
pp. 89
Author(s):  
Daniele Maria Telles ◽  
Gabriel Moreno Martineli ◽  
Maurice Fabian Scaloppi ◽  
Marina Pagliai Ferreira Luz ◽  
Samir Moura Kadri ◽  
...  

Honey bees (Apis mellifera L.) have great global socioeconomic and environmental importance. However, the greater wax moth (Galleria mellonella L.) is a pest that causes serious worldwide damage to honey bee colonies. Good beekeeping practices and physical, chemical, or natural methods can be used to control wax moths. The use of natural products is a more sustainable option because of their lower toxicity to the environment and the colony. Therefore, we evaluated the efficiency of four natural products for greater wax moth control: neem oil (Azadirachta indica), eucalyptus oil (Eucalyptus spp.), tobacco extract (Nicotiana tabacum), and malagueta pepper extract (Capsicum frutescens). We also evaluated their effects on adult bees and on the population growth of colonies. The 4th instar wax moths and adult bees were subjected to in vitro bioassays of different concentrations of the products. The results allowed usto establish a concentration for each product that was safe for the bees and effectively controlled the moth. Then, we sprayed them on bee colonies to evaluate their effects on population growth. The neem and eucalyptus oils caused wax moth mortality at low concentrations, but did not affect colony population growth. However, they did have a toxic effect on adult bees. The tobacco and pepper extracts efficiently controlled the moth, but did not cause adult bee mortality or interfered with the population growth of the colonies. Therefore, the tobacco and pepper extracts could efficiently control the greater wax moth, without damaging honey bees.


2019 ◽  
Vol 97 (Supplement_1) ◽  
pp. 83-84
Author(s):  
Emily Bechtold ◽  
Surendranath Suman ◽  
Smita Mohanty ◽  
Suman Mazumder ◽  
Sadagopan Krishnan ◽  
...  

Abstract Myoglobin is the primary sarcoplasmic protein responsible for meat color. Previous research has reported that myoglobin oxidation is species-specific. Metmyoglobin reducing activity is an inherent property to limit myoglobin oxidation. However, limited research has determined species specificity in metmyoglobin reducing properties. The objective of current study was to compare metmyoglobin reducing properties of eight different species such as beef, porcine, bison, deer, emu, equine, goat, and sheep in vitro. Myoglobin was isolated from eight different species via ammonium sulfate precipitation. The pH of the myoglobin was adjusted by passing through a column pre-calibrated with 50 mM phosphate buffer at pH 5.6. All species myoglobin were converted to metmyoglobin, and the metmyoglobin reduction was determined by two different approaches, non-enzymatic metmyoglobin reducing activity (NMRA) and oxidation-reduction potential (ORP). In the first method, NADH (electron donor), EDTA, and methylene blue (electron carrier), were added in a cuvette and increase in absorbance at 582 nm was monitored using a UV-Vis spectrophotometer. In the second method, the ability of the heme to get reduced was determined using an RedoxSys analyzer, in which electron was directly transferred to heme. The NMRA and ORP experiments were replicated five times. The data were analyzed using the Mixed Procedure of SAS, with species as the fixed effect. There were species-specific differences (P < 0.05) in NMRA and ORP. Bovine myoglobin had the greatest (P < 0.05) NMRA compared with sheep, equine, goat, deer, bison, pork, and emu. There were no differences (P > 0.05) in NMRA among equine, goat, deer, bison, pork, and emu. ORP studies indicated that beef and porcine myoglobins had the greatest ability to get reduced (P < 0.05) compared with other species. Hence, use of different techniques and approaches will help to elucidate the mechanistic basis of metmyoglobin reduction.


2019 ◽  
Vol 97 (Supplement_1) ◽  
pp. 28-29
Author(s):  
Emily Bechtold ◽  
Surendranath Suman ◽  
Smita Mohanty ◽  
Suman Mazumder ◽  
Sadagopan Krishnan ◽  
...  

Abstract Myoglobin is the primary sarcoplasmic protein responsible for meat color. Previous research has reported that myoglobin oxidation is species-specific. Metmyoglobin reducing activity is an inherent property to limit myoglobin oxidation. However, limited research has determined species specificity in metmyoglobin reducing properties. The objective of current study was to compare metmyoglobin reducing properties of eight different species such as beef, porcine, bison, deer, emu, equine, goat, and sheep in vitro. Myoglobin was isolated from eight different species via ammonium sulfate precipitation. The pH of the myoglobin was adjusted by passing through a column pre-calibrated with 50 mM phosphate buffer at pH 5.6. All species myoglobin were converted to metmyoglobin, and the metmyoglobin reduction was determined by two different approaches, non-enzymatic metmyoglobin reducing activity (NMRA) and oxidation-reduction potential (ORP). In the first method, NADH (electron donor), EDTA, and methylene blue (electron carrier), were added in a cuvette and increase in absorbance at 582 nm was monitored using a UV-Vis spectrophotometer. In the second method, the ability of the heme to get reduced was determined using an RedoxSys analyzer, in which electron was directly transferred to heme. The NMRA and ORP experiments were replicated five times. The data were analyzed using the Mixed Procedure of SAS, with species as the fixed effect. There were species-specific differences (P < 0.05) in NMRA and ORP. Bovine myoglobin had the greatest (P < 0.05) NMRA compared with sheep, equine, goat, deer, bison, pork, and emu. There were no differences (P > 0.05) in NMRA among equine, goat, deer, bison, pork, and emu. ORP studies indicated that beef and porcine myoglobins had the greatest ability to get reduced (P < 0.05) compared with other species. Hence, use of different techniques and approaches will help to elucidate the mechanistic basis of metmyoglobin reduction.


Author(s):  
М. G. Markova ◽  
Е. N. Somova

Work on the clonal micropropagation of strawberries comes down to the search for new growth regulators, which include a biologically active substance - the waste product of the wax moth Galleria mellonella L. The effect of the waste product of the wax moth on the efficiency of clonal micropropagation of strawberries (Fragaria х ananassa duch) in vitro and in vivo conditions in 2018-2020 is shown. The object of research is micro-cuttings, rooted micro-cuttings and adapted micro-plants of garden strawberries of the Korona variety and of the remontant strawberries of the Brighton variety. It was revealed that at the proliferation stage, the propagation coefficient of the Korona variety increased significantly with the introduction of the waste product of the wax moth in doses of 4.0 mg/L and 6.0 mg/L and amounted to 4.2 and 3.8 pcs./explant, respectively; for Brighton variety, the coefficient increased significantly when the dose of the waste product of the wax moth 2.0 mg/L and amounted to 4.6 pcs./explant. The introduction of the waste product of the wax moth in doses of 4.0 mg/L and 6.0 mg/L into the nutrient medium had a significant effect on the yield of Brighton micro-cuttings suitable for rooting: the yield was 95.5 and 94.1%, respectively 87.7% in the control. For the Korona variety, no significant positive effect of the waste product of the wax moth on this indicator was noted. The rooting of micro-cuttings of strawberries of both varieties significantly increased with the introduction of the waste product of the wax moth into the nutrient medium in all studied doses and amounted to 86.4-100% in the Korona variety, and 88.9-100% in the Brighton variety.  The survival rate of adaptable micro-cuttings of Corona variety strawberries when sprayed with an aqueous solution of the waste product of the wax moth at a dose of 4.0 mg/L was 100%; the maximum survival rate of micro-cuttings Brighton variety is 99.8% in the variant with spraying with an aqueous solution of the waste product of the wax moth at a dose of 6.0 mg/L.


2021 ◽  
Vol 8 (6) ◽  
pp. 304-309
Author(s):  
Abdul Majid ◽  
Frans Salesman

This study aimed to analyze the bacteriostatic and bactericidal activity of Acalypha indica L. extract against Shigella dysenteriae type 1 bacteria in vitro. This research is a laboratory experimental study consisting of six concentration treatments, namely: (K = 0/ml, P1 = 100/ml, P2 = 200/ml, P3 = 400/ml, P4 = 800/ml, P5 = 1600 mg/ml ), and five replications each. The results showed that the herbal extract of Acalypha indica L. was bacteriostatic (inhibiting) at a concentration of 400 mg/mm, and bactericidal (killing) the bacteria Shigella dysenteriae type 1, which was tested in vitro at a concentration of at least 800 mg/ml. It is necessary to study the effect of Acalypha indica L. herbal extract from Timor in the serum of Rattus norwegicus against bacteria that cause bloody diarrhea (EHEC O157:H7, and Shigella dysenteriae type 1), MRSA bacteria, and other pathogenic bacteria.


2021 ◽  
Author(s):  
Amanda Michelle Starr ◽  
Masoud Zabet-Moghaddam ◽  
Michael San Francisco

Abstract The fungus, Batrachochytrium dendrobatidis, is the causative agent of chytridiomycosis and a leading cause of global decline in amphibian populations . The first stages of chytridiomycosis include: inflammation, hyperkeratosis, lethargy, loss of righting reflex, and disruption of internal electrolyte levels leading to eventual death of the host. Previous work indicates that B. dendrobatidis can produce immunomodulatory compounds and other secreted molecules that regulate the growth of the fungus. In this study, filtrates of the fungus grown in media and water were subjected to ultra performance liquid chromatography-mass spectrometry and analyzed using Compound Discoverer 3.0. Identification of cyclo(phenylalanyl-prolyl), chitobiose, and S-adenosylmethionine were verified by their retention times and fragmentation patterns from B. dendrobatidis supernatants. Previous studies have analyzed the effects of B. dendrobatidis on amphibian models, in vitro, or in cell culture. We studied the effects of live B. dendrobatidis cells, spent culture filtrates containing secreted metabolites, and cyclo(pheylalanyl-prolyl) on wax moth larvae ( Galleria mellonella) . Concentrated filtrates caused melanization within 24 hours, while live B. dendrobatidis caused melanization within 48 hours. Our results indicate B. dendrobatidis produces secreted metabolites previously unreported. These findings provide another alternative for the use of a non-amphibian model system to study pathogenicity traits in this fungus.


Melanin is formed with in the coelomic amoebocytes of Diadema when exposed to air in vitro by a process of oxidation involving enzymes. It appears in association with spheroidal inclusions. An enzyme system with both mono- and polyphenolase activity has been demonstrated in the amoebocytes, which is affected by the same factors, and in the same way, as the production of pigment. Although the enzyme system and its substrate co-exist in the amoebocytes, pigment is not formed in them whilst they are in the perivisceral coelom. A heat-labile factor inhibiting the action of the phenolase system and the formation of melanin is present in the coelomic fluid but not in the amoebocytes. There is some evidence indicating that the enzyme system is localized in or around the spheroids, an area of relatively high oxidation-reduction potential, in which black pigment appears, especially after exposure to ultra-violet light. In the light, of these facts, a concept of melanogenesis in the sea urchin is advanced.


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