L. Van Campenhout

Microbial community assessment of mealworm larvae (Tenebrio molitor) and grasshoppers (Locusta migratoria migratorioides) sold for human consumption.

In Western countries, the popularity of edible insects as an alternative animal protein source is increasing. Nevertheless, there is a lack of profound insight into the microbial safety and shelf life of living insects sold for human consumption. The purpose of this study was to characterise the microflora of fresh edible mealworm larvae and grasshoppers in a quantitative and qualitative way. Therefore, culture-dependent analyses (the total viable aerobic count, Enterobacteriaceae, lactic acid bacteria, yeasts and moulds, and bacterial endospores) and next-generation sequencing (454amplicon pyrosequencing) were performed. High microbial counts were obtained for both insect species. Different insect batches resulted in quite similar microbial numbers, except for bacterial endospores. However, the bacterial community composition differed between both insect species. The most abundant operational taxonomic unit in mealworm larvae was Propionibacterium. Also members of the genera Haemophilus, Staphylococcus and Clostridium were found. Grasshoppers were mainly dominated by Weissella, Lactococcus and Yersinia/Rahnella. Overall, a variety of potential spoilage bacteria and food pathogens were characterised. The results of this study suggest that a processing step with a microbiocidal effect is required to avoid or minimize risks involved with the consumption of edible insects.

Microbial counts of mealworm larvae (Tenebrio molitor) and crickets (Acheta domesticus and Gryllodes sigillatus) from different rearing companies and different production batches

The rising interest in insects for human consumption and the changing regulations in Europe require a profound insight into the food safety of insects reared and sold in Western society. The microbial quality of edible insects has only been studied occasionally. This study aimed at generating an overview of intrinsic parameters (pH, water activity and moisture content) and microbial quality of fresh mealworm larvae and crickets for several rearing companies and for several batches per rearer. In total, 21 batches obtained from 7 rearing companies were subjected to analysis of intrinsic parameters, a range of plate counts and presence-absence tests for Salmonella spp. and Listeria monocytogenes. The microbial counts of the fresh insects were generally high. Different rearing batches from a single rearing company showed differences in microbial counts which could not be explained by variations in intrinsic properties. The largest variations were found in numbers of bacterial endospores, psychrotrophs and fungi. Salmonella spp. and L. monocytogenes were not detected in any of the samples. Altogether, our study shows that large variations were found between batches from individual rearers. As a consequence, no overall differences between rearers could be observed.

Metagenetic analysis of the bacterial communities of edible insects from diverse production cycles at industrial rearing companies

Despite the continuing development of new insect-derived food products, microbial research on edible insects and insect-based foods is still very limited. The goal of this study was to increase the knowledge on the microbial quality of edible insects by comparing the bacterial community composition of mealworms (Tenebrio molitor) and crickets (Acheta domesticus and Gryllodes sigillatus) from several production cycles and rearing companies. Remarkable differences in the bacterial community composition were found between different mealworm rearing companies and mealworm production cycles from the same company. In comparison with mealworms, the bacterial community composition of the investigated crickets was more similar among different companies, and was highly similar between both cricket species investigated. Mealworm communities were dominated by Spiroplasma and Erwinia species, while crickets were abundantly colonised by (Para)bacteroides species. With respect to food safety, only a few operational taxonomic units could be associated with potential human pathogens such as Cronobacter or spoilage bacteria such as Pseudomonas. In summary, our results implicate that at least for cricket rearing, production cycles of constant and good quality in terms of bacterial composition can be obtained by different rearing companies. For mealworms however, more variation in terms of microbial quality occurs between companies.

Interaction between fat type and lysolecithin supplementation in broiler feeds

Lysolecithins are added to poultry diets to promote the intestinal absorption of nutrients, in particular of dietary fats. Lysolecithins contain a mixture of phospho- and lysophospholipids and differ in composition depending on the conditions and source of the lecithin used for its production. The importance of the lysolecithin composition and its interaction with the fat type was investigated in vitro in a fat digestion model and in vivo in a digestibility trial with broilers (24 to 28 d age). The in vitro digestion of soybean oil and pig lard was investigated without and with the inclusion of soybean or rapeseed lysolecithin. Correspondingly, for the digestibility trial, 108 Ross 308 male broilers were assigned to 6 dietary treatments: a basal diet with either soybean oil (5.3%) or pig lard (5.8%), each basal diet supplemented with 250 ppm soybean lysolecithin, and each basal diet supplemented with 250 ppm rapeseed lysolecithin. In vitro pig lard digestibility was significantly lower compared to soybean oil digestibility. Although in vivo no significant difference was observed for crude fat digestibility, broilers fed the basal diet with pig lard had a lower (P < 0.05) DM digestibility, nitrogen retention, and AMEn compared to those fed the basal diet with soybean oil. Lysolecithin supplementation showed a significant interaction with the fat type, both in vitro and in vivo. The in vitro hydrolysis of pig lard, but not of soybean oil, increased (P < 0.001) with supplementation of soybean and rapeseed lysolecithin. Moreover, soybean and rapeseed lysolecithin supplementation improved (P < 0.05) DM digestibility of the basal diet with pig lard by 5.1 and 5.7%, respectively; nitrogen retention by 2.8 and 3.1 g/kg, respectively; and AMEn by 182 and 199 kcal/kg, respectively. Despite the major difference in molecular composition, there was, however, no impact of the lysolecithin composition on in vitro and in vivo fat digestibility. This study demonstrates that the improvements that can be made with lysolecithin supplementation are highly dependent on the fat incorporated in broiler feeds.

Relationship between wheat characteristics and nutrient digestibility in broilers: comparison between total collection and marker (titanium dioxide) technique

Three wheat cultivars (Orpheus, Rustic, and Viscount) were used to formulate 3 test feeds (62.4% wheat) in a broiler digestibility trial. The diets were fed to male Ross 308 broiler chickens. The wheat cultivars mainly differed in their amount of non-starch polysaccharides ( NSP: ). The cultivar Orpheus was chosen to represent a high amount of NSP (102 g/kg DM), whereas the cultivars Rustic and Viscount represented low amounts of NSP (83.4 g/kg DM and 73.9 g/kg DM, respectively). Furthermore, the cultivars Orpheus and Viscount were feed quality wheat, whereas Rustic was a milling quality wheat. Nutrient digestibilities and AMEn contents of the diets were measured from 18 to 22-days-old by total excreta collection, or with the use of the indigestible marker titanium dioxide. In addition, the ileal viscosity was measured when the broilers were 25-days-old. Wheat cultivar affected N retention, DM digestibility, and AMEn. In general, the feed formulated with the high NSP wheat cultivar Orpheus resulted in the least favorable nutrient digestibilities and AMEn, whereas the results were better when the feed was formulated with the low NSP cultivars Viscount and Rustic. Feeding the Rustic cultivar caused the highest intestinal viscosity, although this was not reflected in the animal responses. Nutrient digestibilities and AMEn content of the diets were lower when calculated with the titanium dioxide marker than with the total excreta collection procedure. Moreover, the P-values of the effect of wheat cultivar on DM digestibility, N retention, crude fat digestibility and AMEn were lower with the use of the titanium dioxide marker. It can be concluded that wheat cultivar affected nutrient digestibility and AMEn, and that the observed differences were related to the amount of NSP. Furthermore, both the titanium dioxide marker and the total excreta collection methods showed the same trends despite the different values obtained. The titanium dioxide marker method was the simplest to use and produced valid results.

Microbial dynamics during production of lesser mealworms (Alphitobius diaperinus) for human consumption at industrial scale

In this study, the microbial dynamics during an industrial production cyle of lesser mealworms (Alphitobius diaperinus), sold for human consumption, were characterised. The microbial numbers as well as the microbial diversity were generally higher for the substrate, existing of remaining feed, faeces and exuviae, than for the larvae. Most of the species-level operational taxonomic units, identified using Illumina MiSeq sequencing, that were present in the feed were also detected in the larvae and vice versa. However, bacterial diversity decreased in the larvae during rearing. These results suggested that the feed is an important determinant of the insect bacterial community, but that some bacterial species show a competitive advantage inside the insect gut and become dominant. A blanching treatment of the larvae after harvest reduced most microbial counts, but the number of aerobic endospores remained at 4.0 log cfu/g. Whereas food pathogens Salmonella spp., Listeria monocytogenes, Bacillus cereus or coagulase-positive staphylococci were not detected in our study, fungal isolates corresponding to the genera Aspergillus and Fusarium were recovered. Therefore, it cannot be excluded that mycotoxins were present. The results of this study contribute to a better understanding of the microbial dynamics and food safety aspects during the production of edible insects.

Growth of Pseudomonas fluorescens in modified atmosphere packaged tofu

Aims:  The objective was to study the growth of Pseudomonas in a food product (tofu) where it typically occurs as a spoilage organism, and when this product is stored under modified atmosphere.

Fermentation of enset ( Ensete ventricosum ) in the Gamo highlands of Ethiopia: Physicochemical and microbial community dynamics

Enset (Ensete ventricosum) provides staple food for 15 million people in Ethiopia after fermentation into kocho. The fermentation process has hardly been investigated and is prone to optimization. The aim of this study was to investigate the physicochemical and microbial dynamics of fermentation practices in the Gamo highlands. These practices show local variation, but two steps were omnipresent: scraping of the pseudostem and fermenting it in a pit or a bamboo basket. Enset plants were fragmented and fermented for two months in order to investigate the physicochemical (temperature, moisture content, pH and titratable acidity) and microbial dynamics (total viable aerobic counts, counts of Enterobacteriaceae, lactic acid bacteria, yeasts and moulds and Clostridium spores counts, and Illumina Miseq sequencing). Samples were taken on days 1, 7, 15, 17, 31 and 60. The pH decreased, whereas the titratable acidity increased during fermentation. Of all counts those of lactic acid bacteria and Clostridium spores increased during fermentation. Leuconostoc mesenteroides initiated the fermentation. Later on, Prevotella paludivivens, Lactobacillus sp. and Bifidobacterium minimum dominated. These three species are potential candidates for the development of a starter culture.

Interactions between the concentration of non-starch polysaccharides in wheat and the addition of an enzyme mixture in a broiler digestibility and performance trial

ABSTRACT Two broiler trials were designed to investigate the relationship between the concentration of non‐starch polysaccharides (NSP) in wheat and 1) its nutritional value for broilers and 2) the efficacy of exogenous enzymes. In a balance trial, diets were formulated with 3 wheat cultivars (Rustic and Viscount—medium NSP, Centenaire—high NSP) and were tested with or without the addition of an exogenous enzyme mixture. The diets were fed to 144 male Ross 308 broiler chickens housed in digestibility cages. Total tract nutrient digestibilities and AMEn were measured from 18 to 22 d of age. In a performance trial, diets were formulated with wheat (medium NSP diet) or with wheat mixed with rye and barley (high NSP diet) and were tested with or without the addition of an exogenous enzyme mixture. The diets were fed to 960 male Ross 308 broilers housed in pens and broiler performance during starter, grower and finisher periods was measured. In the balance trial, wheat cultivar did not affect nutrient digestibility or AMEn. Enzyme addition caused a significant increase in nutrient digestibilities and AMEn for the diet formulated with the high NSP wheat Centenaire only. In the performance trial, feeding the high NSP diet resulted in a higher feed conversion ratio and lower final body weight compared to the medium NSP diet. The largest improvements by enzyme addition were observed in the high NSP diet. In conclusion, the study was not able to show a consistent relationship between the NSP concentration of wheat and its nutritional value, but did demonstrate that the effect of an enzyme mixture on nutrient digestibility or broiler performance depends upon the NSP concentration in the diet.

High-moisture extrusion with insect and soy protein concentrates: cutting properties of meat analogues under insect content and barrel temperature variations

Insect biomass production is recognised as one of the potential solutions for the problem of a lack of traditional protein sources (most feed protein sources are imported in Europe). It is perceive...