Nutritional, functional and microbiological properties of edible crickets enriched cereal-based complementary foods

doi:10.21203/rs.3.rs-4800906/v1

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Nutritional, functional and microbiological properties of edible crickets enriched cereal-based complementary foods

https://doi.org/10.21203/rs.3.rs-4800906/v1

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Edible insects are a compelling alternative source of animal protein that holds potential for diversifying and enriching complementary foods with minerals and proteins. This study aimed at formulating, developing and characterizing nutrient- dense complementary flour (CF) containing edible cricket containing maize and roasted finger millet flours (CECF). In addition, the effect of extrusion was evaluated on nutritional, functional and microbiological properties. Analysis of variance (ANOVA) with multiple comparisons using Tukey’s HSD (honestly significant difference) test (α = 0.05) and T- test were used to determine the significance of differences. Edible cricket had significantly high protein, fat, chitin/fibre, energy, iron, zinc and phosphorus. Contrary, cereal flour had high total carbohydrates, manganese and calcium. Phytates were undetectable in roasted finger millets but tannins remained highest at 494.1 mg/ 100g. Extrusion of CECF significantly (p > 0.05) improved the energy ration and densities of protein, carbohydrate but decreased the energy content and fibre content and density. Extrusion reportedly eliminated the phytates and reduced tannins content to acceptable levels for infant foods. Iron content were increased (4.64–6.22 mg/ 100g), whereas Ca: P remained low at 0.35–0.36. Extruded and blended CECFs had poor foaming and emulsion properties. Extrusion improved swelling power, bulking density and water related properties as well as protein digestibility. Microbial quality of the CECF was influenced by the initial microbial load, cross contamination and pre-processing conditions. The study suggests inclusion of edible cricket, pre-processing of ingredients, extrusion and a possible final cooking of CECF as well as observing hygienic and sanitary conditions for a safe and nutrient dense infant food. Further research is needed on evaluating the acceptability and the impact of feeding CECF on children’s growth and development.

Complementary food

extrusion

digestibility

safety

cricket

grasshopper

palm weevil larvae

Malnutrition is a global problem and almost every nation in Sub-Saharan Africa (SSA) with enormous challenges that impacts physical, general health, mental growth, immunity (Gillespie & van den Bold, 2017). According to world health statistics (WHS), malnutrition accounts to about 45% of deaths among children under 5 years of age annually particularly in the developing and underdeveloped countries (WHS, 2022). By 2020, sub–Saharan Africa had the highest prevalence of undernutrition of 264.2 million people (24.1% of population). A report by the Kenya Demographic and Health Survey (KDHS, 2022), showed that in Kenya the malnourished children-aged between 6 and 59 months are either stunted (12%), overweight (3%) and wasted children (5%). These undernourished children are in deficient of the additional nutrient requirements needed for their growth and development (Okoth et al., 2017) and thus affected by micronutrients such as zinc, iron and vitamins deficiencies as well as protein energy malnutrition (PEM) (WHO, 2018). This problem is worse among infants and children (Dewey, 2013) and begins when they are transitioned to high cereal and low animal complementary food products, around the age of six (Roohani et al., 2013).

Addressing the challenges of the growing global malnutrition particularly undernutrition, micronutrients deficiencies and protein energy malnutrition necessitate a transition towards nutrient-rich diets from sustainable alternative food sources (UNICEF &WHO, 2010). Edible insects are a nutritional rich animal source food with both macro and micro-nutrients enough to provide high- quality proteins to support growth and development, as well as alleviate deficiencies respectively (Pal & Roy, 2014; van Huis, 2016). They have less environmental impact compared to conventional livestock (Gahukar, 2016) and have potential to promote dietary diversification (Mishyna et al., 2021). Currently, edible crickets farming is wide-spread promoted for household consumption across East Africa especially in Kenya (Alemu et al., 2023). Studies show insects- enriched complementary flours (CFs), have alleviated many cases of child malnutrition such as protein energy malnutrition (PEM) (Govorushko, 2019) and micronutrient deficiencies (Konyole et al., 2019) especially in poor rural communities. Dehulling of cereal grains have been reported to reduce antinutrients in the resultant rain and their flours despite the possibility of losing specific micronutrients (Ikwebe et al., 2021). In addition, heat- treatment of ingredients such as roasting have been suggested to reduce antinutrients (Chinelo, et al., 2017). Also, processing of CFs through extrusion (Maseta et al., 2016) can render them nutrient rich and safe for infant consumption.

Maize and finger millet have been used in complementary feeding in Kenya over the years. This cereal based complementary foods are however energy dense hence offering low nutrient density to the children (Yohannes et al., 2020). In addition, maize has a relatively low protein content and is deficient in essential amino acids such as lysine and tryptophan (Abeshu, 2016). Although higher in protein than maize, finger millet also has limitations in essential amino acids, particularly lysine. Both maize and finger millet are deficient in certain micronutrients critical for infant and young children development, such as iron, zinc, and vitamin A (Abeshu, 2016; Liomba et al., 2018). This can lead to deficiencies unless the complementary foods are fortified or consumed with other nutrient-rich foods. Finger millet contains phytates and tannins which can inhibit the absorption of iron and other minerals, reducing the overall nutritional value of the complementary foods (Harvey et al., 2020; Pragya Singh, 2012).

However, maize and finger millet are popular in child feeding in Kenya and they also have promising properties of high stability during pasting and storage suitable which is suitable for low- viscosity products processing (Kiiru et al., 2024). Therefore, there is a need to improve their utilization in complementary feeding by blending them with foods that rich in high protein quality such as edible crickets and pre-processing the cereals using technologies that help reduce antinutrients such as roasting and extrusion cooking. The objective of this study was to formulate, develop and characterize a nutrient- dense complementary flour (CF) containing edible cricket containing maize and roasted finger millet flours (CECF) suitable for infants and young children. Subsequently, the study aimed to evaluate the effects of extruding CECF on nutritional, functional and microbiological properties.

Gluten- free cereals acquisition and processing

Finger millet (Eleusine coracana) grains (approximately 4000 grams) were purchased within Juja market, Kenya, and were dry- heat treated or roasted at 180°C for 1 minute (Mridula et al., 2016) on aluminium trays using a hot air oven (Memmert UF 110, Memmert, Schwabach. Germany). Roasted finger millet grains were milled using a two-speed Waring laboratory blender (Camlab, Over, Cambridge. UK) for 30 seconds with 10 seconds pause interval to avoid overheating. Dehulled maize (Zea mays) flour were locally sourced at Nafaka maize miller located in Kasarani, Kenya. Thereafter, both flours were sieved through a 0.45 mm sieve size to obtain uniform fine flours.

Edible crickets acquisition and processing

Field crickets (Gryllus bimaculatus) at 7th instar stage were harvested from Jomo Kenyatta university of agriculture and technology (JKUAT) insect farm, Kenya. They were reared on commercial chicken feeds and during harvesting (approximately 1500 grams) they were starved for 12 hours, washed and drowned before blanching at 60°C for 5 minutes (Cacchiarelli et al., 2022). Then, they were oven-dried on aluminium trays using a laboratory oven (Memmert UF 110, Memmert, Schwabach. Germany) at 45°C for 36 hours as recommended in Dobermann et al. (2019). The dried insects were milled individually into a powdery consistency using a two-speed Waring laboratory blender, (Camlab, Over, Cambridge. UK) at highest speed for 30 seconds with 10 seconds pause interval to avoid overheating. Then they were stored in air-tight containers at -20°C.

Formulation of edible cricket- enriched complementary flours

The data from proximate composition in Table 1 were used as input data in formulating the CFs. NutriSurvey program, (version 2007) was used to generate nutrient-dense formulations to meet the recommended daily allowance (RDA) for 6–11 months old. This included minimum of 400 Kcal, protein content of 8g/100g, fat content of 10-25g/100g, zinc content of 4.1mg/100g, iron content of 5.8mg /100g and calcium content of 500mg/100g (Codex, 2006; WFP, 2018; WHO, 2023). In addition, the formulations met an energy percentage (E%) limit for fat of 20–30%, a protein E% limit of 6–15% and a total carbohydrate E% limit of 55–75% (Codex, 2006). The resulting cricket- enriched complementary flour (CECF) blend comprised of 40% roasted finger millet, 30% maize, 0.2% vitamin/ mineral FBF-V-13 premix, 24% edible cricket and 5.8% sunflower oil. The blended CECF was then packaged in plastic containers and stored in a cool dry place awaiting analysis. A similarly formulated portion of blended CECF was extruded packaged in plastic containers and stored in a cool dry place awaiting analysis. Extrusion of CECF was then conducted and WFP approved corn soy blend plus (CSB+) and its requirements were used for comparison. CSB + is fortified cooked blend of milled, heat-treated corn (64.3%) and whole soya beans (24.0%), sugar (10.0%), soya bean oil (4.0%) and fortified with a vitamin and mineral premix (UNICEF, 2020). CSB + in form of a porridge or gruel is extensively used by USAID implementing partners as a food aid for food-insecure populations such as children to meet the nutritional needs (UNICEF, 2020; USAID, 2017).

Extrusion of edible cricket- enriched complementary flour

The CECF ingredients were blended using an automated mixer and preconditioned by mixing 2 liters of water and 50 kg of raw materials and then extruded in a twin-screw extruder which was locally fabricated. Feed moisture content and barrel temperature were adjusted from 15–16% and 128°C to 142°C for Zone I, and 105°C to 114°C for Zone II, respectively. The main motor and feeder speeds were 29.66 to 30.25 rpm and 09.00 to 11.82 rpm respectively. The resulting extrudates were milled using a two-speed Waring laboratory blender (Camlab, Over, Cambridge. UK) for 30 seconds with 10 seconds pause interval. Thereafter, the extruded flour was sieved through a 0.45 mm sieve size and flavoured using a caramel essence at the rate of 50g/100Kg flour blend. The flour was sealed in airtight plastic containers and preserved at 4°C for further analyses.

Nutritional and antinutritional composition analysis

Moisture content was determined gravimetrically using a forced-air oven (UF 110, Memmert, Schwabach, Germany) at 105°C till constant weight, according to AOAC (2008) method 967.08). Ash content was analyzed gravimetrically using incineration method according to AOAC (2008 method 942.05). Fat content was extracted and determined according to Bligh and Dyer (1959). Crude protein content was determined by digestion of samples, distillation and titration of the residue according to the Micro-Kjeldahl method no. 979.09 (AOAC, 2016). In addition, the nitrogen factor (kP) of specific insects were used; 5.00 for G. bimaculatus (Ritvanen et al., 2020). Crude fiber was determined by method of gravimetric, deproteinization using 1 M NaOH and subsequent demineralization with 1 M HCl as outlined in Liu et al. (2012). Total carbohydrates were calculated by difference as nitrogen- free extracts (NFEs) as described in WHO/FAO (2002). Total energy content and energy density were determined by calculation from carbohydrate and protein contents using conversion factors of 4 kcal/g, and fat content using conversion factor of 9 kcal/g (Guyot et al., 2007). Mineral content in exception for phosphorus analyses were conducted using atomic absorption spectrophotometer (AAS), (model AAS-7000; Shimadzu Corp. Kyoto) as outlined AOAC method 985.01 (AOAC, 2006). Phosphorus was analysed using UV spectrophotometer (PD- 3000 UV, Apel, Saitama) and ammonium molybdate method (Shyla & Nagendrappa, 2011). Phytates acid was determined using HPLC with an ODS C-18 column using a refractive index detector and 0.005M sodium Oxalate as a mobile phase at a flow rate of 0.5 mL per minute. A working standard of sodium phytate (inositol hexaphosphoric acid) solution (50-1000 ppm) was used for quantification at limit of quantification (LOQ) of 0.5ppm. Total condensed tannins (TCT) were determined by the vanillin assay according to Price et al. (1978) where a sample was mixed with 4% vanillin reagent and absorbance was measured after 20 minutes at 500 nm against a blank solution of 4% hydrochloric acid instead of vanillin. Catechin (CE) was used as a standard and results were expressed as mg CE/100 g on dry weight basis.

Determination of in-vitro crude protein digestibility (IVPD)

IVPD was determined following the pepsin enzyme method described in Mertz et al. (1984). The method involved determination of protein content before and after the digestion of samples using pepsin enzyme (1:3000, HOG Stomach, Loba Chemie). During pepsin digestion, 0.2g of sample in centrifuge tubes was mixed with 35 mL solution of pepsin solution (1.5 mg/ml) suspended in 0.1M phosphate buffer (pH 2.0). The mixture was incubated and shaken in a water bath for 2 hours at 37 ºC. The tubes were cooled in ice to 4 ºC for 30 minutes, followed by centrifugation (model 20000, Kokusan corporation, Tokyo) for 15 minutes at 1300 g and 4 ºC. Supernatant was discarded and 10mL of the phosphate buffer solution was added followed by shaking and centrifugation. Supernatant was discarded the residue filtered using a Whatman filter paper no. 3 and then washed with 5mLof the phosphate buffer. The residue was dried for 30 minutes and inserted into a Kjeldahl flask for digestion. A blank without sample flour was prepared in the similar procedure. Finally, the digested protein was calculated by subtracting residual protein from total protein of the sample as shown in Eq. 1.

\(\:\text{P}\text{r}\text{o}\text{t}\text{e}\text{i}\text{n}\:\text{d}\text{i}\text{g}\text{e}\text{s}\text{t}\text{i}\text{b}\text{i}\text{l}\text{i}\text{t}\text{y}\:\left(\text{%}\right)\:=(\text{A}\:-\:\text{B})/\text{A}\:\) …………………………..…..Eq. 1

where A and B represent % protein content of the sample before and after pepsin digestion, respectively.

Determination of flour bulk density

Bulk density (BD) of the flours was determined according to the method in Benitez et al. (2021). A sample of 25.00 g was weighed into a 50mL graduated plastic cylinder and the volume was recorded. Each sample was measured in triplicate. BD was calculated as the ratio of mass of the flour and the volume occupied in the cylinder (g/cm³).

Determination of flour tapped density

Tapped density (TD) was determined according to Kaushal et al. (2012) method. Flour samples were filled into tared 25 mL graduated plastic cylinders. The cylinders were gently tapped onto a wooden block until there was no further diminution of the sample level. Each sample was measured in triplicate and volume occupied was recorded. TD was calculated as mass of sample per unit volume of sample (g/cm³).

Determination of water absorption capacity (WAC) of the flour

Water absorption capacity was determined following the method in Villanueva et al. (2018). A sample of 2 g of flour was put in a graduated centrifuge tube and was filled with 30 ml of distilled water. The tube was vortexed for 30 seconds and left to hold for 10 minutes at room temperature and pressure (rtp). Samples were centrifuged for 25 minutes at 3000 g thereafter the supernatant was discarded. The tubes were carefully tilted at 45° to drain residual liquid for 5 minutes then dried in hot air oven at 45°C for 25 min. WAC (%) was calculated as shown in Eq. 2.

\(\:WAC\%=\frac{\left(Centrifuge\:tube\:weight+\:residue\right)-Centrifuge\:tube\:weight}{Sample\:weight}\:x\:100\) .. ..Eq. 2

Determination of oil absorption capacity (OAC) of the flour

Oil absorption capacity was analysed according to the method of Villanueva et al. (2018), where flour samples each 2 g were put in pre-weighed centrifuge tubes, then mixed with 30 mL of corn oil and vortexed for 30 seconds. Following a holding period of 10 minutes, the tubes were centrifuged at 3000g for 25 minutes. The separated oil on top was drained out by inverting the tubes at 45°C for 5 minutes prior to reweighing. The oil absorption capacity was expressed as gram of oil bound per gram of the sample on a dry basis as shown in Eq. 3.

\(\:OAC\:\%=\frac{\left(Centrifuge\:tube\:weight+residue\right)-Centrifuge\:tube\:weight}{Sample\:weight}\:x\:100\) ……….Eq. 3

Determination of water absorption index (WAI), water solubility index (WSI) and swelling power (SP) of the flour

WAI, WSI and SP were determined according to the method of Harasym et al. (2020). A 2 g of flour was dispersed in 30 mL of distilled water. The mixture was cooked at 90 ºC for 10 minutes in a water bath and finally cooled to room temperature before being transferred to tared centrifuge tubes. The sample was then centrifuged for 10 minutes at 3000 g and the supernatant was decanted into a tared evaporating dish. For WAI, the sediment was weighed, while as for WSI and SP the weight of solid contents following evaporating overnight at 110°C was recorded. WSI, WAI and SP were calculated by the equations 4, 5 and 6.

\(\:WAI\:\%=\frac{\left(Weight\:of\:Sediment\right)}{Sample\:weight}\:x\:100\) …………………………….….....Eq. 4

\(\:WSI\:\%=\frac{\left(Weight\:of\:dissolved\:solids\:in\:supernatant\right)}{Sample\:weight}\:x\:100\) ……………...Eq. 5

\(\:SP\:\%=\frac{\left(Weight\:of\:sediment\right)}{(Sample\:weight-dissolved\:solids\:weight)\:}\:x\:100\) ………………..Eq. 6

Determination of foaming capacity of the flour

The foaming capacity (FC) and foaming stability (FS) of the samples were determined as described in Harasym et al. (2020). The dispersions of sample of 1.5 g in 50 mL distilled water in graduated centrifuge tubes were homogenized, using a vortex mixer (model VM-2000-C, Taiwan) at 1200 g for 3 minutes. The volume was recorded before (V0) and following homogenizing (V1). FC and FS were calculated as presented in equations 7 and 8.

\(\:FC\:\%=\frac{\left(V1-V0\right)}{V0}\:x\:100\) ……………………………………………...….Eq. 7

\(\:FS\:\%=(Foaming\:changes\:at\:interval\:of\:20,\:40,\:60\:and\:120\:\text{m}\text{i}\text{n}\text{u}\text{t}\text{e}\text{s})\) ..Eq. 8

where V0 is the initial volume and V1 is the volume after homogenizing

Determination of dispersibility of the flour

Dispersibility (DS) of the flours was measured using the method of Kurkarni et al. (1991) where a sample weight of 15 g was placed in a 100mL measuring cylinder and the volume was made to 100 mL by adding distilled water followed by stirring for 1 and a half minutes. The sample was allowed to settle for 15 minutes and the difference was calculated by subtraction of volume of the settled particles from 100 mL and difference reported in percentage as shown in Eq. 9.

\(\:DS\:\%=\frac{\left(Volume\:after\:settling-Initial\:volume\right)}{Initial\:volume}\:x\:100\) ……………….…..Eq. 9

Determination of emulsifying activity (EA) and emulsifying stability (ES) of the flour

EA and ES were determined following the method in Harasym et al. (2020), about 1 g of sample was mixed with 30 mL of water and 30 mL of corn oil in centrifuge tubes. The mixture was homogenized for 60 seconds using a vortex mixer (model VM-2000-C, Taiwan). Emulsifying activity at room temperature (25°C) and at 80°C for 5 minutes was obtained by centrifuging the tubes at 3000 g for 5 minutes. Finally, EA was calculated by dividing the volume of the emulsified layer by the volume of the emulsion before centrifugation in percent as shown in equations 10 and 11.

\(\:EA\:\%=(Ve/V0)\:\times\:\:100\:\) ……………………………………………..Eq. 10

\(\:ES\%=(Vh/Ve)\:\times\:\:100\) ……………………………………………....Eq. 11

where V0 is total volume of tube contents, Ve is volume of the emulsified layer and Vh is volume of the emulsified layer after heating.

Determination of the microbiological content of the flour

A 5g sample was mixed with 45 mL sterile Peptone water and homogenized in a stomacher at normal speed for 1 minute. The homogenate was then diluted with sterilized peptone water (OXOID LP0034) to decimal dilutions of between 10^− 1 to 10^− 8 before surface plating (Dijk et al., 2007). Microbiological procedures on the samples included; total plate count using total plate count agar (PCA), Escherichia coli count using mac Conkey agar (MCA), presence of staphylococcus species count using Mannitol salt agar (MSA), yeast and molds count using Rose Bengal Agar (RBA). These specific microbial analyses were selected since they serve as indicators of food safety. The homogenates of sample dilutions (0.1mL) were aseptically aliquoted and inoculated onto 20 mL of the respective sterile and solidified media. Then it was spread over the agar plate using a sterile glass spreader. Enumeration of total viable count (TVC), the PCA plates were incubated at 37°C for 48 hours. Following the incubation period, the observed colonies were counted counted (Klunder et al., 2012). Enumeration of Escherichia coli, the MCA plates were placed in an inverted position to avoid water condensing into the medium. The plates were incubated at 37°C for 24 hours. Following the incubation period, the pink colonies were counted counted (Nyangena et al., 2020). Staphylococcus species was determined by placing the plates in an inverted position to avoid water condensing into the medium. The plates were incubated at 37°C for 48 hours. Following the incubation period, the yellow colonies were counted (Ramashia et al., 2020). Enumeration of yeasts and molds was conducted by inoculating the homogenates samples onto the surface of already solidified RBA that had already been prepared 24 hours in advance. This was then evenly spread on the surface by use of a glass rod. Samples were incubated at 25ºC for 72 hours and all the visible colonies were counted (Nyangena et al., 2020).

Statistical analysis

Data was analysed with R software version 4.0.3 (2020). Data from nutritional analysis, was analysed for analysis of variance (ANOVA) with multiple comparisons using Tukey’s HSD (honestly significant difference) test (α = 0.05) was used to determine the significance of differences. Data from functional properties and microbiological analysis was subjected to a Students t-test. All analyses were analysed in triplicates

Nutritional composition of raw materials

The nutritional content and energy content of the insect powder and cereal flours were significantly different (p < 0.05) as presented in Table 1. Edible cricket was significantly high in protein, fat, fibre, ash and energy whereas cereal flours had high amount of carbohydrates. Iron (Fe), zinc (Zn) and phosphorus (P) were also high in edible cricket, whereas calcium (Ca) and manganese (Mn) were high in finger millet. In terms of antinutrients, phytates were high in maize while tannins were higher in finger millet. The crude protein of the edible cricket insects was greater than some plant protein sources such as beans (23.5%), pea (20–25%) (Ulbek et al., 2017) and soy (41.1%) (Blásquez et al., 2012). In comparison to animal sources, protein content was lower than those of some animal protein sources such as fish (81%), beef (54%) but comparable to chicken (43–46%) (Blásquez et al., 2012; Teffo et al., 2007). The caloric value of edible cricket was 568 Kcal higher and it was higher than the average Orthoptera (nymphs and adults) of 20 species (~ 418 Kcal) as reported in Ramos-Elorduy & Piao (1990). The caloric between insects have been reported to vary with changes of proximate contents brought about by different diets, life stages or ages (Kipkoech et al., 2021). Overall, the insects had higher caloric values than the cereal flours and most of non-insect foods such as pork, soyabeans and beef as reported in Ramos-Elorduy & Piao (1990). This could be due to the high fat content in insects which also translates to formulated complementary foods (Agbemafle et al., 2020). Chitin was high in edible cricket at 7.06%, has been reported to be deacetylated to chitosan which can function as a prebiotic with potential to eliminate pathogenic microbes in human gut (Kipkoech et al., 2021). Edible cricket showed to be a good source of essential minerals such as Fe and Zn at 13.2 mg/100g and 20.8 mg/100g. The amounts however can be vary by age, diets and other proximate composition such as chitin where some are bound (Rumpold & Schluter, 2013). Generally, the findings suggest that edible crickets could be a good source essential macro- and micronutrients that can enrich maize and finger millet flours in formulation of complementary flours.

Phytates were below detectable limits of quantification (< 5.0 mg/ 100g) in the finger millet flour probably due to the dry-heat treatment also referred to as roasting. Studies have shown elimination of phytic acid at 180°C for 1 minute (Mridula et al., 2016), 115°C for 15 minutes (Oluwole et al., 2020) and reductions at 100°C for 120 minutes (Bhati et al., 2016). Phytic acid in food chelates with divalent metals ions especially key minerals such as iron and zinc and significantly reduces their absorption and utilization (Hurrell et al., 2002). Tannins were high in finger millet compared to maize flour. The reported tannin contents in finger millet was slightly high to that in Singh et al. (2018) at 314.2 mg/100g. Tannins and phenols are commonly high in dark brown or red pigmented grain crops such as millets, sorghums (Dykes & Rooney, 2006) and have been to confirmed to increase or reduce with heat treatment such as roasting (Dykes & Rooney, 2006; Oluwole et al., 2020). Hence, roasting can be recommended to reduce phytates yet, further processing is needed to reduced tannins in maize and finger millet flours.

Table 1

Nutrient and antinutrient composition of edible crickets and cereal flours on dry basis
Ingredient	Macro nutrients (g/100g) and energy (Kcal/ 100g)							Minerals (mg/100g)					Antinutrients (mg/ 100g)
Ingredient	Protein	Crude Fat	Carbohydrates	Chitin/ Fibre	Ash		Kcal	Fe	Zn	Ca	Mn	P	Phytates	Tannins
Edible cricket	44.88 ± 0.03^a	26.60 ± 0.21b^a	17.18 ± 0.26^c	7.06 ± 0.19^a	4.28 ± 0.20^a	568.98 ± 2.17^a		13.22 ± 1.29^a	20.85 ± 1.11^a	176.70 ± 14.01^b	5.07 ± 0.19^b	1094.60 ± 2.00^a	n.a	n.a
Maize	6.53 ± 0.49^b	4.09 ± 0.05d^b	86.28 ± 0.21^a	1.34 ± 0.34d^c	0.95 ± 0.03^c	416.44 ± 2.03^b		2.34 ± 0.80^c	2.06 ± 0.11^b	13.72 ± 1.10^c	0.73 ± 0.24^c	349.88 ± 0.99^b	18.60 ± 1.54^a	47.36 ± 12.86^b
Finger millet	5.67 ± 0.30^b	0.91 ± 0.08^c	87.07 ± 0.28^a	3.71 ± 0.61^b	2.28 ± 0.08^b	395.11 ± 2.24^b		5.18 ± 1.43^b	3.11 ± 0.11^b	302.00 ± 33.20^a	26.38 ± 5.79.^a	386.61 ± 50.24^b	n.d ^b	494.15 ± 83.23^a
Values are means ± standard deviation on dry weight basis, n = 6; Values with different superscripts (^{a, b, c}) on the same column are significantly different, - p ≤* 0.05, *** - p ≤ 0.001; n.a- not available, n.d- not detected

Nutritional composition of cricket- cereal complementary flours

The nutritional compositions and nutrient density of CECFs varied significantly (p < 0.05) different as presented in Table 2. The nutrient contents and nutrient density of CFs were compared to CSB + as well as against recommendations for complementary foods for older infant and young children. Nutritional content and density of CECF after extrusion significantly (p < 0.05) improved in terms of protein, carbohydrates, iron. Conversely, fibre content and density as well as the energy, phytates and tannins content significantly (p < 0.05) decreased after extrusion. The thermal mechanical shearing during extrusion cooking, can cause physical, structural and chemical changes of the flour components (Guy, 2001). In the study, lipid would melt and stick onto surfaces which might account for the slight losses (p > 0.05), and possible low total energy in extruded CECF. Extrusion has been reported to reorganize insoluble dietary fibre by depolymerization, and reducing the molecular weight of complex starches including pectins and hemicellulose molecules into soluble dietary fibre portions (Brennan et al., 2008) resulting to reduced fibre content and increase in available carbohydrates. Further, changes in physical form and structure of proteins along with possible release of nitrogen from depolymerised insoluble fibre including chitin could account for the increase in protein (Zhang et al., 2020).

The high reduction of tannins by 85% and elimination of phytates (below limit of quantification of < 5.0 mg/ 100g) could be attributed to breakdown of these compounds or interaction, for instance tannins can interact with proteins such as prolamins (Emmambux & Taylor, 2003). Similar effects were reported in extruded sorghum-soybean composite products (Ndibalema, 2011). The tannins contents in extruded CECF of 27.06 mg/ 100g was equivalent to less than 0.002 urease activity units (Ndibalema, 2011), and this levels are within the recommended CSB + urease units of below 0.20 units (UNICEF, 2020). On the other hand, extrusion has been reported to have little effect on mineral stability hence no significant change on extrusion, but rather slight influence their bioavailability through thermal degradation of chelating agents such as phytates, fibres and proteins among others (Pismag et al., 2024; Sow et al., 2019). Overall, these findings suggest extrusion can be an effective step in improving the nutritional quality of CFs suitable for children consumption.

Table 2

Nutrient content and nutrient density of cricket- enriched complementary flours and CSB+
Nutrient/ Nutrient density	Complementary food			Standard/ Recommended levels
	Blended CECF	Extruded CECF	p-value	CSB + ⁸	Nutrient density	Nutrient content
Energy (Kcal/100g)	444.25 ± 2.27^a	430.04 ± 1.82^b	***	409.89	-	400.00¹
Protein (g/100g)	12.75 ± 0.28^b	14.41 ± 0.58^a	*	16.00		8⁶,9.1⁵,10⁷
Protein (g/100Kcal)	2.89 ± 0.72^b	3.35 ± 1.03^a	*	3.90	2.25–3.00²
Fat (g/100g)	7.76 ± 0.13^a	7.68 ± 0.24^a	ns	9.00		10.00¹
Fat (g/100Kcal)	1.74 ± 0.32^a	1.78 ± 0.22^a	ns	2.19	,4.40-6.00²
Carbohydrates (g/100g)	73.09 ± 0.17^b	75.82 ± 0.28^a	**	60.90		45⁵
Carbohydrates (g/100Kcal)	16.45 ± 0.04^b	17.63 ± 0.26^a	*	14.85	9.00-14.00²
Fibre (g/100g)	3.50 ± 0.30^a	2.61 ± 0.65^b	*	2.90		< 5¹
Fibre (g/100Kcal)	0.79 ± 0.04^a	0.61 ± 0.05^b	**	0.70	-
Ash (g/100g)	2.14 ± 0.14^a	2.12 ± 0.13^a	ns	4.20		-
Ash (g/100Kcal)	0.48 ± 0.08^a	0.49 ± 0.06^a	ns	1.02	-
Fe (mg/100g)	4.64 ± 0.08^b	6.22 ± 0.21^a	**	6.50		5.80^1,3
Fe (mg/100Kcal)	1.05 ± 0.07^b	1.44 ± 0.09^a	***	1.58	0.45–2.00²
Zn (mg/100g)	4.30 ± 0.15^a	4.44 ± 0.17^a	ns	5.00		4.10^1,4
Zn (mg/100Kcal)	0.96 ± 0.09^a	1.03 ± 0.06^a	ns	1.21	0.50- 1.50²
Ca (mg/100g)	165.87 ± 0.95^a	166.90 ± 1.98^a	ns	362.00		400^,1,6
Ca (mg/100Kcal)	37.33 ± 0.44^a	38.81 ± 0.80^a	ns	88.31	50-140²
Mn (mg/100g)	19.92 ± 0.47^a	21.20 ± 1.38^a	ns	-	-	-
Mn (mg/100Kcal)	4.48 ± 0.40^a	4.92 ± 0.66^a	ns			-
P (mg/100g)	471.22 ± 6.65^a	462.86 ± 10.44^a	ns	280.00	-	275–460⁶
P (mg/100Kcal)	106.06 ± 1.06^a	107.16 ± 1.67^a	ns	68.31
Phytates (mg/100g)	4.24 ± 0.93^b	n.d^b	***	-
Tannins (mg/100g)	188 ± 9.54^a	27.06 ± 2.81^a	***	-

Values are means values ± standard deviation, n = 6; Values with different superscripts (^{a, b, c}) on the same row are significantly different, ns- p > 0.05 *- p ≤ 0.05, **- p ≤ 0.01, ***- p ≤ 0.001; NFEs- Non-fat extract, CECF- Edible cricket- enriched complementary flour, CSB+- Super cereal plus; n.d- not detected. ¹Codex, (2017); ²Koletzko et al., (2005); ³10% dietary bioavailability; ⁴medium dietary bioavailability; ⁵WHO (1998), ⁶WFP (2018), ⁷WHO/FAO/UNU (2007), ⁸UNICEF,(2020).

The protein content of the CFs met the required amounts for 6–8 months, 9–11 months and 12–23 months which is 9 g/ day, 4 g/ day and 6.2g/ day, respectively (WHO/UNICEF, 2009). Additionally, the CECFs surpassed the recommended 10 g/100 g required for edible parts of food, enough to be referred as a “high protein food” (FAO/WHO/UNU, 2007). The protein density of the CECFs was in the range of 2.89–3.35 g/100kcal, and this indicates they were below the Codex (2017) standard but within the limit of the recommended levels in Koletzko et al. (2005). The protein density was slightly lower than that of CSB+, and that of CECF formulations (3.50 to 4.79 g/100 kcal) reported in Tenagashaw et al. (2017). The protein energy ration (PER) of CECFs was 7.46–8.71%, this was comparable to the recommended (6–15%) (Codex, 2006). These amounts could be attributed to the edible cricket inclusion, which is rich in essential amino acids such as methionine (Oibiokpa et al., 2018), compared to plant proteins that are low in lysine and tryptophan (Ogbonnaya et al., 2012).

The lipid content and lipid density of the CFs ranged between 7.68–7.76 g/100g and 1.74–1.78 g/ 100 Kcal respectively and were below that of CSB+. CECF did not meet the minimum requirement by Codex (2017) and Koletzko et al. (2005) and unaffected by extrusion. Thus it is recommended to use fat or oil during preparation to improve on energy density without resulting in high viscosity porridge (WHO/UNICEF, 1998). Tenagashaw et al. (2017) also reported low lipid content and lipid density in plant based CFs, but whereas Konyole et al. (2012) reported improved lipid content and density in CFs enriched with termites and dagaa. The ratio of energy derived from fat (fat E%) was 6.22–6.36%, and it was lower compared to recommended 20–30% % (Codex, 2006). However, with adequate breastfeeding the RDA of fat E% could be met for 6–8 months which is 0%, for 9–11 months which is 5–8%, but not for 12–23 months which is 15–20% (WHO, 2001). Dietary fats especially poly unsaturated fatty acids (PUFA) in edible insects (Ghosh et al., 2017) can play a crucial role in promoting good health.

The carbohydrate content of the CECFs was higher than that of CSB + and recommended amounts in WHO (1998). The carbohydrate density also surpassed the recommended levels (9–14 g/kcal) (Koletzko et al,. 2005). This high carbohydrate density is important particularly in older growing children of 12–23 months who rely on carbohydrates as the main source of energy source (WHO, 2001). In the extruded CECF, the increased carbohydrates were attributed to possible degradation of starch to simple sugars can be suitable to infants as it can impart sweetness (Amagloh et al., 2013). Fibre content and nutrient density of the extruded CECF were higher than in CSB+. Both CECFs had fibre contents within the recommended (< 5 g/100g) (Codex, (2017), indicating that CECFs might promote bowel movement but not impede nutrient bioavailability (Caballero et al., 2005). Most of the dietary fibre in this study came from insect chitin and according to Kipkoech et al. (2017), insect chitin can be deacetylated to chitosan which functions as a prebiotic with potential to eliminate pathogenic microbes in human gut. Hence, these insect- cereal CFs can be used in making healthy foods for improved gut health in children.

Minerals particularly iron, zinc, and calcium, are relatively low in breast milk compared to the requirements of 7–11 months old children making complementary foods essential to provide approximately 30–97% of these nutrients (WHO, 2001). Calcium content and its nutrient density was considerably low in the CFs compared to CSB + and the recommended levels indicated in WFP (2018) and Codex (2017), respectively. Edible cricket has been reported to contain low calcium due to lack of mineralised skeleton (Köhler et al., 2019). Conversely, phosphorus was twice as higher in CECFs than in CSB + and met the threshold for total phosphorus in diets as recommended in WFP (2018). The Ca:P ratio of the CECFs were 0.35–0.36, this was below the recommended level of above 0.5 (Jacob et al., 2015). A food is considered good or poor when the Ca:P ratio is more than 1 or less than 0.50, respectively (Alinnor & Oze, 2011). Low Ca:P levels are not favourable for calcium absorption which is key for growing children for the formation and maintenance of bones and teeth (WHO, 2001). Yet, low Ca:P ratio can be alleviated by continued intake of breast milk which provides at least 40% of the calcium requirements particularly to 15–18 months old children (WHO, 1998).

The iron and zinc contents in CECFs were slightly lower than the amounts in CSB+. Conversely, the iron and zinc densities of CECFs, in exemption of blended CECF met the recommended levels at medium and 10% dietary bioavailability, respectively (Codex, 2017). The increase in iron and zinc after extrusion has been associated with reductions in chelating agents and inhibitors such as polyphenols and phytates (Sow et al., 2019). Both non-haem iron and zinc in edible are typically bound to proteins like ferritin, holoferritin, transferrin, and other transport and storage proteins (Latunde-Dada et al., 2016; Mwangi et al., 2018). Study reported that bioaccessibility of iron in cornfield grasshoppers (Sphenarium purpurascens Charp) and G. bimaculatus was lower than haem ferritin in beef (Latunde-Dada et al., 2016). Also, the addition of insects to wheat flour (1:1) was superior to beef suggesting potential of insect-enriched cereal foods in curbing the iron deficiencies (Latunde-Dada et al., 2016). However, there is need to consider factors such as formulations, preparation, processing and preservation of these insect-enriched cereal foods (Mwangi et al., 2018). Indeed, enriching CFs with edible insects can be recommended due to their potential in supplementing nutrients and promote dietary diversification (Mishyna et al., 2021). The research suggests roasting and extrusion processing to improve the quality of the CECFs, however further research on the acceptability of the CECFs as well impact on growth and development of the children can be explored.

Functional properties of cricket - enriched complementary flours in comparison to CSB+

The functional properties including in vitro protein digestibility of the CFs are presented in Table 3. Results indicated that functional properties differed significantly (p < 0.05) among the complementary flours. Dispersibility of the CFs ranged between 72%- 86% in blended CECF and extruded CECF, respectively. This was comparable to the reported dispersibility in Birungi et al. (2023) on CF containing maize, sorghum, beans, sesame and groundnuts (76–77%). This high dispersibility is associated potential to create a smoother paste which is a desirable quality in infant foods (GAIN & UNICEF, 2021). Dispersibility of flours suggests how easily it can be dispersed or mixed into water to form a smooth, cohesive mixture (Anosike et al., 2020).

Bulk density (BD) is the amount in mass per volume of flour, was reportedly high in extruded CECF compared to blended CECF. BD of the CFs was lower to that (0.60- 0.68g/cm³) reported in Aboge et al. (2022) and (0.7-0.8g/cm³) (Tenagashaw et al., (2016). However, BD of blended CECF was comparable to CSB + as reported by Ewketu et al. (2024). Extrusion increased BD by 7.69%, and the feed moisture has been reported to increase BD through particle agglomeration and reduction in void spaces as observed in corn-based (Ding et al., 2005) and rice-based extrudates (Ding et al., 2005).

Table 3

Functional properties of edible cricket- enriched complementary flours
CF	Foaming capacity (%)	Dispersibility (%)	Tapped density (g/cm³)	Bulk density (g/cm³)	OAC (%)	WAC (%)	WAI (g/g)	WSI %	Swelling power (g/g)	IVPD
Blended CECF	1.88 ± 0.01^a	75.00 ± 1.09^b	0.87 ± 0.01^a	0.52 ± 0.01^b	71.18 ± 2.07^a	106.20 ± 1.10^b	8.26 ± 0.08^b	4.13 ± 0.68^b	6.10 ± 0.57 ^b	35.56 ± 1.39^b
Extruded CECF	1.82 ± 0.02^b	86.00 ± 0.69^a	0.77 ± 0.04^b	0.56 ± 0.01^a	68.02 ± 1.40^a	112.80 ± 0.20^a	8.48 ± 0.09^a	9.74 ± 0.81^a	8.94 ± 1.05 ^a	86.74 ± 2.01^a
Values are means ± standard deviation, n = 4; Values with different superscripts (^{a, b, c}) on the same column are significantly different, -p ≤ 0.05; OAC- Oil absorption capacity, WAC- Water absorption capacity, WAI- Water absorption index, WSI- Water solubility index; IVPD- In vitro* protein digestibility, CF- Complementary flour, CECF- Edible cricket- enriched complementary flour;

A higher bulk density (BD) is beneficial for complementary feeding as it allows a smaller portion of flour to provide sufficient nutrients, increasing gruel's nutrient concentration and enhancing overall nutrient intake. Tapped density also referred to the density of flour following a specified compaction process, can provide information about the flowability of flour particles or the packing behaviour (Awuchi et al., 2020). Both CFs recorded a high TD of ~ 0.82g/cm3 indicating a tight packing structure and possible reduced flowability. Extrusion of CECF reduced TD by 12.9% suggesting an improved flowability and compressibility of powder.

Water absorption capacity (WAC) was higher than oil absorption capacity (OAC) among the CECFs. WAC ranged between 106.2%- 112.8%, while OAC ranged between 68.02%- 71.18%. Higher WAC could be brought about by compositional variations, for instance absence of non-polar side chain proteins and high total carbohydrates which are hydrophilic components (Suresh et al., 2015). Water absorption index (WAI) which indicates the volume occupied by the granule or starch polymer swelling, ranged between (8.26–8.48%). WAI results were high to those (2.0- 4.9%) in Birungi et al. (2023) and Tenagashaw et al. (2017) for CECF blends. Extrusion of CECF resulted in significant increase in WAC and WAI possibly due to reduced water content of extrudate from the evaporation at die exit. In addition, a higher proportion of gelatinized starch (Wang et al., 2020) as well as increased low molecular sugars in the extruded flour which promotes water affinity (Dalbhagat et al., 2019). Overall, the extruded and blended CECF suggested potential for thicker consistency, enhanced flavour and mouth feel of resulting porridges

Water solubility index (WSI) represents the amount of soluble components released from the starch and an indicator of starch degradation and digestibility (Ding et al., 2005). WSI ranged between 4.13–9.74% and was significantly (p < 0.05) different among the CFs. The results for blended CECF were comparable to those (4–5%) of orange flesh sweet potato, sorghum, and soybean- based CF (Alawode et al., 2017). Additionally, the high WSI in extruded CECF was comparable to that (8.38–16.63%) of extruded complementary foods in Tenagashaw et al. (2016). Higher WSI of up to 20% have been reported in several extruded and instant porridge flour formulations (Mahgoub et al., 2020; M. Tenagashaw et al., 2017). Extrusion cooking can promote formation of low molecular weight and soluble molecules through fusion, depolymerization, and dextrinization of starches (Dalbhagat et al., 2019). This suggests better digestibility of the extruded CECF flours which is desirable characteristic for complementary feeding.

Foaming capacity (FC) of the CECFs ranged between 1.82–1.88%, further there was no observable foaming stability (FS), emulsifying capacity (EC) and emulsifying stability (ES). Insect powders (Akpossan et al., 2015), and insect proteins from five edible insect (Yi et al., 2013) have been reported to have poor or zero foaming activities, respectively. Conversely, a study on CFs made of soybean-wheat, rice-wheat and maize-millet flour blends had higher FC of 10.83%- 15.40% and EC of 35.05%- 50.95% (Godswill, 2019), and attributed the high protein and low lipids contents to the improved FC and EC, respectively. Further, the amount of oil and non-polar amino acids residues on the surface of protein or water as well as processing conditions have shown to affect emulsion and foaming activities (Zielińska et al., 2015). Thus, it can be hypothesized that inclusion of insect powders, the low protein contents (12–14%) of CFs as well as denaturation of proteins by high-temperature processing could have contributed to the degraded functional properties. This suggests that the CECFs may have a less creamy and denser texture with limited stability.

Swelling power (SP) ranged between 6.10–8.94 g/100g for blended CECF and extruded CECF, respectively. The high SP in starch that has been gelatinized have been shown to have a considerably high cold viscosity (Crosbie & Ross, 2007), possibly due to expanded structure and increased water holding capacity. SP have been correlated with viscosities, however extruded flours have been shown to have different cold and hot viscosities from unextruded flours (Crosbie & Ross, 2007). Hence further research is needed to confirm the viscosity of the cooked extruded CECF.

In vitro protein digestibility (IVPD of CECFs ranged from 35.5–86.7% in blended and extruded CECF, respectively. The IVPD of the blended CECF was considerably low compared to (78–88%) CFs made from maize, wheat and defatted soybean flour enriched with 10–70% Acheta domesticus as reported by Aboge et al. (2022). The IVPD of extruded CECF and effect of extrusion (~ 51% increase) was comparable to the reported IVPD of 87.6% and increase (~ 47.9%) after extrusion of CF made from teff fortified with soybean and orange-fleshed sweet potato (Tenagashaw et al., 2016). According to Gulati et al. (2020), extrusion and other thermal treatments could decrease the presence of protease inhibitors, induce denaturation and the breakdown of protein aggregates thereby enhancing IVPD. Edible cricket which was the main source of proteins have been reported to have high protein digestibility of 84–93% (Oibiokpa et al., 2018; Poelaert et al., 2018), and its inclusion into plant source foods, improves their protein digestibility (Kiiru et al., 2020). Subsequently, protein digestibility is a key determinant of protein quality, thus the study suggests that the extruded CECF could be suitable for child feeding for enhanced protein absorption and assimilation.

Microbiological quality of the CECFs

Table 4 summarizes the microbial counts for blended CECF and extruded CECF, and results showed significant differences (p < 0.05) in microbial counts among the samples. The raw edible cricket had a higher number of microbial counts compared to cereal flours except for yeast in molds in maize flour. The high nutritional profile of edible insects such as high mono- and polyunsaturated fatty acids, amino acids and micronutrients (Kinyuru et al., 2015; Rumpold & Schlüter, 2013) support a wider spectrum of microbes (Klunder et al., 2012). Currently, there no specific microbial criteria for unprocessed insects, however edible cricket had higher TVC than the recommended limit of raw minced meat of 6.70 log CFU/g. The TVC levels in raw G. bimaculatus of 9.02 log cfu/g were higher than the results of Klunder et al. (2012), which reported TVC of 7.2 log cfu/g. Variations in microbial load could be explained by the rearing or harvesting conditions (Stoops et al., 2017), instar stage (Klunder et al., 2012). Conversely, the TVC for cereal flours were within the lower limit according to East African standards, EAC (2011).

Table 4

Microbial counts of ingredients, blended and extruded complementary flours
Sample	Microbial load (Log CFU/g)
	Total viable count	Escherichia coli	Staphylococcus. aureus	Yeasts and molds
Raw ingredients
Edible cricket	9.02 ± 0.3^a	7.98 ± 0.4^a	4.64 ± 0.2^a	3.64 ± 0.1^b
Maize flour	4.51 ± 0.1^b	4.24 ± 0.2^b	4.12 ± 0.1^b	4.15 ± 0.2^a
Finger millet flour	4.23 ± 0.1^c	4.16 ± 0.2^b	4.05 ± 0.1^b	3.30 ± 0.1^b
Raw minced meat¹, cereal flours²	< 6.70, 5.00	< 2.70, nd	< 3.70, nd	< 4.00, < 4.00
Raw and dried edible cricket
Edible cricket	9.02 ± 0.3^a	7.98 ± 0.4^a	4.64 ± 0.2^a	3.64 ± 0.2^a
Dried edible cricket	4.32 ± 0.2^b	nd^b	nd^b	nd^b
Dried insects³, edible foods standard⁴	< 5.00, < 7.00	nd,<2.00	nd,<4.00	< 3.00,<6.00
Complementary foods
Blended CECF	4.28 ± 0.2^a	4.36 ± 0.3^a	3.52 ± 0.1^a	3.51 ± 0.2^a
Extruded CECF	3.52 ± 0.2^b	nd^b	3.00 ± 0.1^b	2.85 ± 0.1^b
CSB + ³; infant foods⁴	< 4.00, < 4.00	< 1.00, nd	< 1.00, nd	< 3.00, < 2.00

Values are means ± standard deviation, n = 3; Values with different superscripts (^{a, b, c}) on the same column are significantly different, *- p ≤ 0.05; CFs- Complementary flours, CECF- Edible cricket- enriched complementary flour, CSB+- Super cereal plus, nd - Not detected, ¹SHC & FASFC, (2014), ²EAC (2011), ³EAC (2023), ³WFP (2014) and ⁴EU (2007).

Escherichia coli and S. aureus counts in raw edible cricket were significantly (p < 0.001) higher than that cereal flours. The presence of E. coli, and S. aureus which primarily a human pathogenic bacteria was attributed to cross-contamination through feed or direct contact during handling steps including feeding, harvesting (Ochieng et al., 2023). Both edible cricket and cereal flours exhibited high levels of E. coli and S. aureus compared to recommended set thresholds. Yeast and molds were high in unprocessed maize and low in roasted finger millet flours. Yeast and molds are common in cereal grains as they can grow only at low water activity compared to E. coli and S. aureus. The microbial loads of dried edible cricket and roasted finger millet flours were within the recommended limits. Overall, flour from finger millet had low microbial loads compared to maize which could be a consequence of heat treatment during roasting of finger millet grains (Sperber et al., 2007).

Different thermal processing techniques on insects have been reported to have varying efficacy on destroying active cells and lowering microbial load (Nyangena et al., 2020). In this study, the blanching and hot air drying of edible cricket lowered the TVCs, E. coli and S. aureus and yeast and molds to respective recommended limit of dried insects and edible foods except for yeast and molds. Blanching (Cacchiarelli et al., 2022), and dry heat treatment (Nyangena et al., 2020) has been shown to decrease microbial load. For instance, oven- drying of Hermetia. illucens, Acheta domesticus, Ruspolia differens and Spodoptera littoralis reduced the TVCs with up to 0.5–2.3 log cycles (Nyangena et al., 2020). This confirms significance of pre-processing of raw ingredients to reduce the microbial load. Blended CECFs reported a slight increase in TVC, S. aureus, E. coli, yeast and molds above acceptable levels. This was not in agreement with findings of Agbemafle et al. (2020) which reported its formulated OFSP-based cricket CFs was safe for consumption by children. The incorporation of the unprocessed (only milled) maize flour coupled with human handling, unsanitary equipment and environment could be main source of contamination (Ssepuuya et al., 2019). Further, extrusion of the cricket CF significantly lowered all microbial loads, except for S. aureus below the set limit for CSB + and infant foods. Studies have reported that extrusion reduces microbial contamination though the thermal inactivation of microbial cells and lowing water activity, which renders the product safe and shelf stable (Filli et al., 2014; Oluwole et al., 2013). Notably, the possible reintroduction of S. aureus after extrusion suggests need for hygienic and sanitary handling of ingredients as well as a final cooking to potentially render the CECF safe for infant consumption.

The study reported that G. bimaculatus can be a good source of essential macro and micronutrients. Phytates and tannins were present in maize and finger millet flours, and were shown to reduce by roasting and extrusion processing. Developed CECF met the recommended protein amounts for 7–23 months. The energy ration and densities of the CECF’s protein and carbohydrate were comparable to recommendation for 7–23 months old children, whereas fat energy ratio did not meet this requirement. However, the CECF were low in iron, fat and Ca: P contents compared to the recommendation for infants. Extrusion of CECF showed potential to improve the nutritional content and density in terms of protein and carbohydrates and iron. Additionally, extrusion was shown to decrease the fibre content and density, and energy contents. Additionally. extrusion was shown to eliminate phytates and reduce tannins content acceptable levels of infant foods. The functional properties of CECFs improved after extrusion including dispersibility, swelling power, bulking density, water absorption capacity, water absorption index and water solubility index as well as protein digestibility. Conversely, the extruded and blended CECFs showed poor foaming and emulsion activities which suggests limited stability and creaminess. The microbial quality of the CECF depended on the initial microbial load, cross contamination and pre-processing conditions. The study recommends inclusion of G. bimaculatus, pre-processing of ingredients, extrusion and a final cooking of CECF as well as observing hygienic and sanitary conditions in making a safe and nutrient dense and safe infant food. Further research is needed on the acceptability of the CFs and nutrient slight influence their bioavailability as well impact on growth and development of the children.

Acknowledgement

The authors would like to thank The Danish International Development Agency (DANIDA), Ministry of Foreign Affairs of Denmark through the HEALTHYNSECT project (Grant number: 19-08-KU) and the Rockefeller Foundation for the financial support.

Competing interests

The authors declare there is no known competing of interests.

Data availability

The data from this study is available on request upon reasonable request.

Ethical guidelines

Ethics approval was not required for this study.

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Editorial decision: Minor revisions
24 Sep, 2024
Reviewers agreed at journal
25 Aug, 2024
Reviewers invited by journal
20 Aug, 2024
Editor assigned by journal
29 Jul, 2024
First submitted to journal
26 Jul, 2024

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Nutritional, functional and microbiological properties of edible crickets enriched cereal-based complementary foods

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Abstract

Introduction

Material and methods

Gluten- free cereals acquisition and processing

Edible crickets acquisition and processing

Formulation of edible cricket- enriched complementary flours

Extrusion of edible cricket- enriched complementary flour

Nutritional and antinutritional composition analysis

Determination of flour bulk density

Determination of flour tapped density

Determination of water absorption capacity (WAC) of the flour

Determination of oil absorption capacity (OAC) of the flour

Determination of foaming capacity of the flour

Determination of dispersibility of the flour

Determination of emulsifying activity (EA) and emulsifying stability (ES) of the flour

Determination of the microbiological content of the flour

Statistical analysis

Results and discussion

Nutritional composition of raw materials

Nutritional composition of cricket- cereal complementary flours

Microbiological quality of the CECFs

Conclusion and recommendation

Declarations

References

Status:

Version 1