Effect of fatty acid-enriched black soldier fly larvae meal combined with chitinase on the metabolic processes of Nile tilapia.

The aim of this study is to determine to what extent the addition of chitinase to black soldier fly (BSF) larval meal enriched or not with long-chain PUFA (LC-PUFA) could improve growth, protein digestion processes and gut microbial composition in Nile tilapia. Two different types of BSF meal were produced, in which larvae were reared on substrates formulated with vegetable culture substrate (VGS) or marine fish offal substrate (FOS). The BSF raised on VGS was enriched in α-linolenic acid (ALA), while that raised on FOS was enriched in ALA + EPA + DHA. Six BSF-based diets, enriched or not with chitinase, were formulated and compared with a control diet based on fishmeal and fish oil (FMFO). Two doses (D) of chitinase from Aspergillus niger (2 g and 5 g/kg feed) were added to the BSF larval diets (VGD0 and FOD0) to obtain four additional diets: VGD2, VGD5, FOD2 and FOD5. After 53 d of feeding, results showed that the BSF/FOS-based diets induced feed utilisation, protein efficiency and digestibility, as well as growth comparable to the FMFO control diet, but better than the BSF/VGS-based diets. The supplementation of chitinase to BSF/FOS increased in fish intestine the relative abundance of beneficial microbiota such as those of the Bacillaceae family. The results showed that LC-PUFA-enriched BSF meal associated with chitinase could be used as an effective alternative to fishmeal in order to improve protein digestion processes, beneficial microbiota and ultimately fish growth rate.

Does Consumption of Baker's Yeast (Saccharomyces cerevisiae) by Black Soldier Fly (Diptera: Stratiomyidae) Larvae Affect Their Fatty Acid Composition?

Fatty acids are important compounds for insects, but the requirements for essential fatty acids may differ between insect species. Most of the fatty acids are acquired through the insect's diet; therefore, supplementing the diet with baker's yeast (Saccharomyces cerevisiae Meyen ex E.C. Hansen), which produces unsaturated fatty acids, was predicted to affect the fatty acid composition of the insect. The tested insect was the black soldier fly (BSF) (Hermetia illucens L.), that is used as a source of protein and fat in feed. Therefore, there is importance for BSF larvae (BSFL) nutritional composition, especially the unsaturated fatty acids content, which is one of the nutritional limitations for mammalian diets. The dominant fatty acids of the tested BSFL were the saturated fatty acids: lauric, myristic, and palmitic acids, as found in other BSF studies. Oleic acid (c18:1) and linoleic acid (C18:2) were the abundant unsaturated fatty acids in the BSFL. The proportion of linoleic acid was higher in the substrate with the supplemental yeast; however, this did not affect its proportion in the larvae. The higher proportion of linoleic acid may have been exploited as a source for production of saturated lauric acid. Therefore, providing unsaturated fatty acids to the substrate through supplemental baker's yeast is not the most efficient way to increase the proportion of unsaturated fatty acids in the larvae.

Transfer of Lauric and Myristic Acid from Black Soldier Fly Larval Lipids to Egg Yolk Lipids of Hens Is Low.

Implementing insects, such as the black soldier fly larvae (BSFL), as animal feed commonly includes the previous removal of substantial amounts of fat. This fat may represent an as yet underutilized energy source for livestock. However, transfer of lauric and myristic acid, prevalent in BSFL fat and undesired in human nutrition, into animal-source foods like eggs may limit its implementation. To quantify this, a laying hen experiment was performed comprising five different diets (10 hens/diet). These were a control diet with soybean oil and meal and a second diet with soybean oil but with partially defatted BSFL meal as protein source. The other three diets were based on different combinations of partially defatted BSFL meal and fat obtained by two different production methods. Lauric acid made up half of the BSFL fat from both origins. Both BSFL fats also contained substantial amounts of myristic and palmitic acid. However, in the insect-based diets, the net transfer from diet to egg yolk was less than 1% for lauric acid, whereas the net transfer for myristic and palmitic acid was about 30% and 100%, respectively. The net transfer did not vary between BSFL originating from production on different larval feeding substrates. The results illustrate that hens are able to metabolize or elongate very large proportions of ingested lauric acid and myristic acid, which are predominant in the BSFL lipids (together accounting for as much as 37 mol%), such that they collectively account for less than 3.5 mol% of egg yolk fatty acids.

Black soldier fly (Hermetia illucens) larvae meal in diets of European seabass: Effects on antioxidative capacity, non-specific immunity, transcriptomic responses, and resistance to the challenge with Vibrio alginolyticus.

Black soldier fly larvae meal (BSFLM) has been successfully demonstrated as a promising fish meal (FM) replacer in diets of several fish species including European seabass (Dicentrarchus labrax). However, its impacts on antioxidant capacity, and immune responses of treated fish are still poorly understood. A 60-day feeding trial was carried out to evaluate the effects of partial substitution of FM with different levels of dry BSFLM on the antioxidative status, non-specific immunity, transcriptomic responses, and resistance of European seabass to the challenge with Vibrio alginolyticus. Four isoproteic (45%) and isolipidic diets were formulated by replacing 0.0%, 25%, 35%, and 50% of the dietary FM. Each diet was randomly assigned to four fish groups (in triplicates) (initial mean body weight, 12.1 ± 0.21 g) (20 fish per aquarium) (n = 240). Fish were fed three times daily to the apparent satiation. At the end of the feeding trial, serum antioxidant biomarkers such as malondialdehyde levels, and catalase, superoxide dismutase, and glutathione peroxidase enzyme activities were significantly increased in all BSFLM groups in a dose-dependent manner compared to the control group (P < 0.05). The non-specific immune indices, including phagocytic activity, phagocytic index, serum lysozyme and respiratory burst activities were significantly elevated in BSFLM groups compared to those in the control group (P < 0.05). Significant upregulation of the mRNA expression levels of hepatic heat shock protein 70, interleukin-1beta and interleukin-10 genes were observed in all BSFLM groups compared to the control group (P < 0.05). Additionally, after the challenge with V. alginolyticus, the relative percent of survival was significantly elevated in fish groups fed on diets containing graded levels of BSFLM over the control group (P < 0.05). Conclusively, the present study suggests the potential efficacy of partial replacement of dietary FM protein for up to 50% by BSFLM without negative effects on fish health with possible potentiation of the antioxidative status, and the immune responses of the European seabass.

Insect (black soldier fly, Hermetia illucens) meal supplementation prevents the soybean meal-induced intestinal enteritis in rainbow trout and health benefits of using insect oil.

Black solider fly larvae (BSFL) and their oils (BSFLO) are receiving increasing attention as sustainable ingredients in fish feeds, but mostly as replacements to marine sources. There were two aims to this study; in exp. 1, soybean meal (SBM)-based diets were formulated to contain BSFL as supplements at 0 (SBM), 8 (SBM + BSFLlow) or 16% (SBM + BSFLhigh) with a control diet being fishmeal-based (FM). In exp. 2, diets included only fish oil (FO), soybean oil (SBO), BSFLO or BSFLO + bile acid (BA), and all lipid sources were added at 16%. Both experiments were run at the same time and fed to rainbow trout (32 g) with each treatment being triplicated. After 10 weeks the fish were sampled for liver and distal intestine histology, expression of genes responsible for inflammation in the intestine and kidneys, and serum peroxidase and lysozyme activities. In exp. 1, supplementations of BSFL effectively prevented SBM-induced intestinal enteritis, down-regulated intestinal prostaglandin and interferon regulatory factor 1 (IRF-1), while the SBM + BSFLhigh diet significantly increased serum lysozyme activity. In exp. 2, BSFLO caused no histomorphological change to the liver or intestine, but kidney interluekin-8, tumor necrosis factor and IRF-1 were significantly upregulated along with significantly higher serum peroxidase activity. The inclusion of BA in the BSFLO diets significantly upregulated intestinal prostaglandin gene expression. Overall, BSFL supplementations of 8 or 16% prevented SBM-induced intestinal enteritis based on histological observations, which was supported by a down-regulation in pro-inflammatory genes and enhanced innate immunity. Meanwhile, the use of BSFLO showed some immunological benefits. Therefore, these sustainable resources are recommended in the diets of rainbow trout, especially when using elevated levels of plant-based proteins.

Lactobacillus plantarum in black soldier fly (Hermetica illucens) meal modulates gut health and immunity of freshwater crayfish (Cherax cainii).

Probiotic supplements are being used to improve the growth and immune performance of aquaculture species over the last couple of decades. In recent times, black soldier fly (BSF) is considered as one of the promising sources of alternative protein to fishmeal protein in aqua-diets. Since the freshwater crayfish, marron (Cherax cainii), a Western Australian's native and iconic freshwater crayfish species, grows fairly slow under commercial farming environment, this study was aimed to investigate the supplemental effect of BSF and BSF with probiotic bacteria Lactobacillus plantarum (BSFLP) on overall health and immune performance of marron after 56 days of feeding under laboratory conditions. The post-trial data revealed insignificant influences of any diets on growth performance, however, both BSF and BSFLP based diets significantly improved some haemolymph parameters and gut health of marron. High throughput sequence data revealed that both BSF and BSFLP diets significantly improved the diversity of microbial communities including some beneficial bacteria for crustaceans in the hindgut of marron. Further analysis showed that both BSF and BSFLP diets upregulated the expression of some genes in the gut tissue and haemocytes associated with the innate immune response of marron at 48 h post injection. The up-regulation of some immune genes in BSFLP diet group was found significantly linked to OTU abundance for Lactobacillus. The findings of this study could be helpful for improving overall health status of marron.

Black soldier fly larvae oil as an alternative fat source in broiler nutrition.

The present study was conducted to investigate growth performance, carcass characteristics, short-chain fatty acids, fatty acid composition in abdominal fat, and serum parameters in broiler chickens fed diets containing corn oil, coconut oil, or black soldier fly larvae (BSFL) oil at the level of 50 g per kg of diet during the 30-day-feeding period. A total 450 one-day-old male broiler chicks (Ross 308) were randomly allocated to one of 3 dietary groups. Each treatment had 10 replicates with 15 chicks per replicate. Feed conversion ratio was decreased in the coconut and BSFL oil group compared with the corn oil group. Dietary BSFL oil increased ileal weight-to-length ratio at day 30 after hatch. Dietary BSFL oil increased significantly ileal branched-chain fatty acid (P < 0.05) and moderately total short-chain fatty acid in 15-day-old broilers (P = 0.074). At day 30, ileal propionate was highest in the coconut oil group but cecal propionate was highest (P < 0.05) in the BSFL oil group. Fatty acid composition of abdominal fat was affected by dietary fat sources. Especially, chickens fed diets containing coconut oil or BSFL oil had higher contents (P < 0.05) of saturated fatty acid being dominant in lauric and myristic acids compared with those fed on corn oil. On the other hand, the reverse trend was noted (P < 0.05) as to polyunsaturated fatty acids being dominant in corn oil compared with coconut oil and BSFL oil. Coconut oil vs. corn oil significantly increased total and high-density lipoprotein cholesterol. Finally, BSFL oil vs. corn oil significantly increased total antioxidant capacity in chickens. It is concluded that dietary BSFL oil improves feed conversion ratio and increases the incorporation of medium-chain fatty acids into abdominal fat pad and serum antioxidant capacity in broiler chickens.

Development and Characterization of Nano-emulsions Based on Oil Extracted from Black Soldier Fly Larvae.

Insect-based biorefinery is seen as a potential alternative approach to manufacturing foods, feeds, and fuel because of the increasing demand for renewable and sustainable products. Insect oil and protein are the two major components that can be quantitatively obtained from insect farming. However, very few attempts have been conducted to utilize insect oil for the production of value-added products. In this study, the oil extracted from the black soldier fly (Hermetia illucens) larvae (BSFL) was used as a novel feedstock for preparing nano-emulsions. The nano-emulsions were prepared with BSFL oil, hydrogenated lecithin (HL), and d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) in water using pre-homogenization followed by ultrasonication. The morphology and the particle size of nano-emulsions were affected by ratios of HL to TPGS. Moreover, the nano-emulsions showed a nearly Newtonian liquid behavior and the presence of TPGS was able to improve the storage stability of HL nano-emulsions. The addition of TPGS could eliminate the phase transition region of HL nano-emulsions but did not provide a significant change for the molecular mobility in the HL nano-emulsions. In summary, the BSFL oil could be used as a renewable feedstock for formulating nano-emulsions from the aspect of high value-added applications and physicochemical characteristics of the nano-emulsions could be adjusted by the mixed surfactant ratio, surfactant to oil ratio, and oil content. Graphical Abstract The physicochemical characteristics and optimization of nano-emulsions based on black soldier fly larvae oil were investigated.

Determination of antibacterial activity and minimum inhibitory concentration of larval extract of fly via resazurin-based turbidometric assay.

BACKGROUND: Antimicrobial resistance is currently a major global issue. As the rate of emergence of antimicrobial resistance has superseded the rate of discovery and introduction of new effective drugs, the medical arsenal now is experiencing shortage of effective drugs to combat diseases, particularly against diseases caused by the dreadful multidrug-resistant strains, such as the methicillin-resistant Staphylococcus aureus (MRSA). The ability of fly larvae to thrive in septic habitats has prompted us to determine the antibacterial activity and minimum inhibitory concentrations (MICs) of larval extract of flies, namely Lucilia cuprina, Sarcophaga peregrina and Musca domestica against 4 pathogenic bacteria [Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), Pseudomonas aeruginosa and Escherichia coli] via a simple and sensitive antibacterial assay, resazurin-based turbidometric (TB) assay as well as to demonstrate the preliminary chemical profile of larval extracts using gas chromatography-mass spectrophotometry (GC-MS). RESULTS: The resazurin-based TB assay demonstrated that the L. cuprina larval extract was inhibitory against all tested bacteria, whilst the larval extract of S. peregrina and M. domestica were only inhibitory against the MRSA, with a MIC of 100 mg ml-1. Subsequent sub-culture of aliquots revealed that the larval extract of L. cuprina was bactericidal against MRSA whilst the larval extracts of S. peregrina and M. domestica were bacteriostatic against MRSA. The GC-MS analysis had quantitatively identified 20 organic compounds (fatty acids or their derivatives, aromatic acid esters, glycosides and phenol) from the larval extract of L. cuprina; and 5 fatty acid derivatives with known antimicrobial activities from S. peregrina and M. domestica. CONCLUSION: The resazurin-based turbidometric assay is a simple, reliable and feasible screening assay which evidently demonstrated the antibacterial activity of all fly larval extracts, primarily against the MRSA. The larval extract of L. cuprina exerted a broad spectrum antibacterial activity against all tested bacteria. The present study revealed probable development and use of novel and effective natural disinfectant(s) and antibacterial agent(s) from flies and efforts to screen more fly species for antibacterial activity using resazurin-based TB assay should be undertaken for initial screening for subsequent discovery and isolation of potential novel antimicrobial substances, particularly against the multi-drug resistant strains.

First Record of Larval Secretions of Cochliomyia macellaria (Fabricius, 1775) (Diptera: Calliphoridae) Inhibiting the Growth of Staphylococcus aureus and Pseudomonas aeruginosa.

Maggot debridement therapy (MDT) consists on the intentional and controlled application of sterilized larvae of the order Diptera on necrotic skin lesions with the purpose of cleaning necrotic tissue and removing pathogenic bacteria. During MDT, a marked antimicrobial activity has been reported in literature specially associated with antibacterial substances from Lucilia sericata (Meigen); however, regarding Cochliomyia macellaria (Fabricius), little is known. This study aimed to evaluate in vitro inhibition of bacterial growth of Pseudomonas aeruginosa and Staphylococcus aureus in contact with excretions and secretions (ES) from C. macellaria larvae. Larval ES were extracted in sterile distilled water and divided in three groups: ES, containing 400 µL of autoclaved ES; ES+BAC, containing 400 µL of autoclaved ES+0.5-µL bacterial inoculum; and CONT-BAC, containing 400 µL of sterile distilled water +0.5 µL of bacterial inoculum. Aliquots of each experimental group were plated by spreading onto Petri dishes. Seedings were made at 0, 1, 2, 4, and 12 h after the extraction of ES. In ES+BAC groups, inhibition of S. aureus was verified between times 1 and 2 h and P. aeruginosa was inhibited between 0 and 4 h. There was no growth observed in any ES group. In the CONT-BAC groups, the number of colonies from time 4 h became countless for S. aureus and decreased for P. aeruginosa. As reported in the literature, we note here that ES have excellent bactericidal activity for both gram-positive and gram-negative bacteria, and this study shows for the first time the action of the bactericidal activity of exosecretions of C. macellaria against S. aureus and P. aeruginosa.

Long-term monitoring of arsenic, copper, selenium, and other elements in Great Salt Lake (Utah, USA) surface water, brine shrimp, and brine flies.

This paper presents long-term monitoring data for 19 elements with a focus on arsenic (As), copper (Cu), and selenium (Se), in surface water (2002-2011), brine shrimp (2001-2011), and brine flies (1995-1996) collected from Great Salt Lake (GSL, Utah, USA). In open surface waters, mean (±standard deviation [SD]; range; n) As concentrations were 112 (±22.1; 54.0-169; 47) and 112 µg/L (±35.6; 5.1-175; 68) in filtered and unfiltered surface water samples, respectively, and 16.3 µg/g (±5.6; 5.1-35.2; 62) dry weight (dw) in brine shrimp. Mean (±SD; range; n) Cu concentrations were 4.2 (±2.1; 1.3-12.5; 47) and 6.9 µg/L (±6.6; 1.9-38.1; 68) in filtered and unfiltered surface water samples, respectively, and 20.6 µg/g (±18.4; 5.4-126; 62) dw in brine shrimp. Finally, mean (±SD; range; n) dissolved and total recoverable Se concentrations were 0.6 (±0.1; 0.4-1.2; 61) and 0.9 µg/L (±0.7; 0.5-3.6; 89), respectively, and 3.6 µg/g (±2.2; 1.1-14.9; 98) dw in brine shrimp. Thus, Se in open lake surface waters was most often in the range of 0.5-1 µg/L, and concentrations in both surface water and brine shrimp were comparable to concentrations measured in other monitoring programs for the GSL. Temporally, the statistical significance of differences in mean dissolved or total recoverable As, Cu, and Se concentrations between years was highly variable depending which test statistic was used, and there was no clear evidence of increasing or decreasing trends. In brine shrimp, significant differences in annual mean concentrations of As, Cu, and Se were observed using both parametric and nonparametric statistical approaches, but, as for water, there did not appear to be a consistent increase or decrease in concentrations of these elements over time.

Determination of effective miRNAs in wound healing in an experimental Rat Model.

The larvae of Lucilia sericata have been used for centuries as medicinal maggots in the healing of wounds. The present study aimed to screen potential microRNAs related to ES-induced wound healing in rat skin wounds and to investigate the potential mechanisms contributing to accelerated wound healing. Healthy, male, 12 weeks old Wistar albino rats weighing 250-300 g were supplied by the Animal Experimental Center. All animal studies were performed in accordance with the NIH Guide for the Care and Use of Laboratory Animals. Wistar albino rats were treated by ES after post wounding and the differentially expressed miRNAs in wound biopsies were screened by microarray analysis at the end of treatments for 4,7 and 10 days. In addition, bioinformatics approaches were used to identify the potential target genes of differentially expressed miRNAs and the functions of their target genes. We found a significant up-regulation of rno-miR-99a* and rno-mir-877 in response to ES treatment. Further investigation of rno-miR-99a* and rno-mir-877 and their target genes (TGFa, TNF, TAGLN, MAPK1, MMP-9) implicated in present study could provide new insight for an understanding lead to the development of new treatment strategies. The identified miRNAs can be new biomarkers for ES- induced wound healing.

[Extraction of fatty constituents from Tabanus bivittatus and their analysis by GC-MS].

OBJECTIVE: To optimize the extracting condition of fatty constituents from Tabanus bivittatus and analyse by GC-MS. METHODS: Taking fatty yield as index, the extraction technology parameters of fatty constituents of Tabanus bivittatus ere optimized by single factor test and orthogonal test. RESULTS: The optimum extraction parameters were as follows: the ratio of acetone to petroleum ether 1: 2, extraction time 2 h, solid-liquid ratio 1: 50. Twenty-one fatty acids were identified by GC-MS. CONCLUSION: The process is reasonable and with good reproducibility. The main components of fatty acids in Tabanus bivittatus are palmitoleic acid, palmitic acid, linoleic acid, oleic acid and stearic acid.

Inhibition of bacterial motility and spreading via release of cranberry derived materials from silicone substrates.

The motility of bacteria plays a key role in their colonization of surfaces during infection. Derivatives of cranberry fruit have been shown to interfere with bacterial motility. Herein, we report on the incorporation of cranberry derived materials (CDMs) into silicone substrates with the aim of impairing bacterial pathogen motility and spreading on the substrate surface. The release of CDMs from the silicone substrates when soaking in an aqueous medium was quantified for a period of 24h. Next, we showed that CDMs released from two silicone substrates remain bioactive as they downregulate the expression of the flagellin gene of two key uropathogens - Escherichia coli CFT073 and Proteus mirabilis HI4320. Furthermore, we demonstrate that CDM-modified silicone inhibits the swarming motility of P. mirabilis, an aggressive swarmer. The bioactive, CDM-modified substrates can find broad applications in the medical device and food industries where the impairment of bacterial colonization of surfaces is of paramount importance.

In vitro antibacterial activity and physicochemical properties of a crude methanol extract of the larvae of the blow fly Lucilia cuprina.

The emergence of multidrug-resistant bacterial strains has prompted the reintroduction of maggot therapy in the treatment of chronic, infected wounds. Many previous studies have demonstrated the potent antibacterial activity of larval excretions/secretions of the blowfly Lucilia sericata (Meigen) (Diptera:Calliphoridae) against bacteria. However, the antibacterial activity of its sibling species, Lucilia cuprina (Wiedemann) (Diptera:Calliphoridae) against a wide range of pathogenic bacteria has never been determined. The aim of this study was to develop a new procedure to produce whole body extract of larvae of L. cuprina via methanol extraction as well as to demonstrate the in vitro antibacterial activity of this extract against seven selected wound pathogens (Staphylococcus aureus, methicillin-resistant S. aureus, S. epidermidis, Streptococcus pyogenes, Klebsiella pneumoniae, Pseudomonas aeruginosa and Escherichia coli). The turbidimetric assay demonstrated that L. cuprina larval extract was significantly potent against all bacteria tested (P < 0.001). Additionally, colony-forming unit (CFU), agar well diffusion and minimum inhibitory concentration assays have confirmed the apparent potency of larval extract against P. aeruginosa. The reconstituted larval extract was highly robust and thermally stable. These observations substantiated the feasibility of the methanol extraction method in the production of larval extract.

[Functional identification and application of horsefly as a representative form of traditional Chinese medicine].

As important traditional Chinese medicine materials, medicinal animals have been highly appreciated due to their strong bioactivities. Among these, medicinal insects have been thought to be significant, especially in preventing and treating modern diseases and tumors. Some of the most famous medicinal insects, such as horseflies, blister beetles and American cockroaches (Periplaneta americana) have been well known due to their reported effects in anti-thrombosis and fighting cancer. In general, identifying the medicinal functions and active components of medicinal insects has been a gradual processes. Originally, these medicinal insects were collected from open fields and usually their whole bodies were adopted. But, currently, most medicinal insects are under large-scale artificial propagations and only their purified active components are in use. In this article, we reviewed the historical process of the application of the horsefly in traditional Chinese medicine and tried to provide useful references for the modernization of traditional Chinese medicines via discussing the interrelationship between traditional Chinese medicine and western medicine.

Maggot metabolites and their combinatory effects with antibiotic on Staphylococcus aureus.

BACKGROUND: Maggot therapy has been in practice for effective debridement, disinfection and healing of chronic wounds. Due to their antiseptic action during wound healing, their metabolites have been investigated in the past for antibacterial activity. They have been particularly useful for treatment of wounds infected with multi-drug resistant Staphylococcus aureus. Antibiotics, on the other hand, can predispose bacteria to develop resistance. Substances that are able to modulate or delay the occurrence of resistance in bacteria are under investigation by many researchers around the world. In the present study, antibacterial activity in excretions/secretions (ES) from maggots of Lucilia cuprina blowfly was demonstrated. The extracts were also screened in combination with antibiotic, ciprofloxacin. METHODS: L. cuprina blowfly maggots were reared for extraction of its metabolites. The ES extracted was screened against S. aureus, alone and in combination with ciprofloxacin, both for short term and long term exposure analysis. A microchannel-based device and system was used for experiments instead of conventional techniques. RESULTS: The original ES had shown partial bacterial growth inhibition. However, in combination with ciprofloxacin, at sub-inhibitory concentrations, certain combinations revealed anti-staphylococcal activity, with bacterial reduction of up to 50%, after 24 hours. The six day study on S. aureus exposed to ES-ciprofloxacin combination suggested a potential delay in development of adaptive resistance as opposed to when ciprofloxacin was used as single agent. CONCLUSIONS: The combination effect of ES and ciprofloxacin at sub-MIC levels showed enhanced antibacterial activity compared to the effect of ES and ciprofloxacin as single agents. Based on the results of ES-ciprofloxacin combinations, a more effective means of treatment for S. aureus can be proposed.

Selective antimicrobial activity of maggots against pathogenic bacteria.

Maggot therapy, also known as biosurgery, is an ancient method for the healing of chronic infected wounds. Although clinicians have reported on the beneficial activities of the Lucilia sericata larvae that have been used for healing chronic wounds, the selectivity of this therapy against the different pathogenic micro-organisms that are found in chronic wounds has never been analysed. In the present study, we have investigated the in vitro activities of larval excreta/secreta both against selected bacterial strains that frequently occur in chronically infected wounds, and against bacteria isolated directly from the larvae and their excreta/secreta. Additionally, the antibacterial activities were investigated in in vivo studies, by comparing bacterial diversity in wounds before and after the application of L. sericata larvae. In conclusion, larval therapy is highly recommended, particularly for the treatment of wounds infected with Gram-positive bacteria, like Staphylococcus aureus, but less so for wounds infected with Gram-negative bacteria, especially Proteus spp. and Pseudomonas spp. strains. Bacteria from the genus Vagococcus were resistant to the maggot excreta/secreta.

[Chemical constituents from Faeces bombycis].

OBJECTIVE: To study the chemical constituents from Faeces bombycis. METHOD: Isolation and purification were carried out on silic gel, Sephadex LH-20 and RP-18 column chromatography. The chemical structures of the constituents were elucidated on the basis of physicochemical properties and spectral data. RESULT: Fifteen compounds were identified as 3S, 5R-dihydroxy-6R, 7-megstigmadien-9-one (1), 3S, 5R-dihydroxy-6S, 7-megstigmadien-9-one (2), (6R, 9R)-3-oxo-alpha-ionol-beta-D-glucopyranoside (3), (6R, 9S)-3-oxo-alpha-ionol-beta-D-glucopyranoside (4), blumenol C glucoside (5), byzantionoside B (6), alangionoside L (7), lutein (8), pipecolic acid (9), betaine (10), alanine (11), glutamic acid (12), phenylalanine (13), leucine (14), isoleucine (15). CONCLUSION: All the compounds were separated from Faeces bombycis for the first time.

Antibacterial properties of whole body extracts and haemolymph of Lucilia sericata maggots.

OBJECTIVE: To partially characterise maggot-secreted antibacterial substances and determine their range of activity against different bacteria. METHOD: Sterile and non-sterile maggots maintained in the laboratory and taken from chronic wounds of treated patients were used. Whole body extracts and haemolymph were fractionated and their range of activity against bacteria was tested using the zone of inhibition assay. The mode of action of bacterial destruction was examined by viable counts, influx of K+ and changes in the membrane potential by scanning electron microscope (SEM). RESULTS: Extracts of sterile and non-sterile maggots showed an activity of 200 arbitrary units (AU)/ml and 400AU/ml respectively. Maggots removed from chronic wounds had an activity of 1200AU/ml. Injuring sterile maggots with a sterile needle doubled the antibacterial activity within 24 hours, while the antibacterial activity of haemolymph increased fourfold after injury with a sterile needle and sixteenfold with an infected needle. The fractions with a molecular weight of < 1kDa and 3-10kDa showed antibacterial activity against Gram-positive and Gram-negative bacteria including Pseudomonas aeruginosa, Klebsiella pneumoniae and methicillin-resistant Staphylococcus aureus (MRSA) isolated from wounds. The fraction with a molecular weight of < 1kDa lysed over 90% of the bacteria within 15 minutes by causing an influx of K+ and changing the membrane potential of bacteria. CONCLUSION: The nature of the antibacterial materials extracted from maggots not only indicates their ability to ingest the necrotic tissue on the wound, but also their potential significance in wound healing,