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Beta Bugs News

Leveraging Sustainable Genetic Technologies to Improve the Black Soldier Fly

Leveraging Sustainable Genetic Technologies 

to Expand and Improve the Black Soldier Fly Market and Products

 

Dr. Anna Melkov¹ – co-founder and CTO, Dr. Yehonatan Alcalay¹ – Head of R&D and Dr Thomas Farrugia² – CEO
1. BugEra Biotechnology
2. Beta Bugs Ltd

 

About BugEra BugEra (Bugheera) – Insect Biotechnology for a sustainable future – is a USA / Israeli start-up based in Beer Sheva, originating from Ben-Gurion University of the Negev. BugEra leverages state-of-the-art genetic engineering technologies to develop Modefied BSF (MBSF) platform to promotes the development of multiple strains for diverse verticals  BugEra’s first  novel genetically-edited BSF strain named BSFx2, addresses the biofuel need by doubling the oil volume that can be extracted from BSF larvae.

About Beta Bugs – Beta Bugs Limited, headquartered in Edinburgh, UK, is an insect genetics company that develops and distributes Black Soldier Fly breeds, using selective breeding, to the insect farming sector. The company’s mission is to increase the output of insect farmers through the use of its improved Black Soldier Fly genetics, HiPer-Fly®, which it supplies from its egg production facility, The Multiplier®. This enables the insect farming industry to produce a protein that is competitive on price point against existing feed ingredients.

Introduction

The global climate crisis and the growing human population present an existential threat never encountered before. Harnessing nature’s solutions like the BSF unique traits to mimic nature and create a circular bioeconomy with proper waste management systems is essential in addressing these challenges. Insect farming and BSF as the leading species in it, is evolving a new industry that should be named InsecTech. InsecTech refers to the utilization of insects in the industrial production for food and feed, novel applications (like biofuel), and waste valorization. All of the above is resulting in novel biomanufacturing processes and more resilient feed and food supply chains,  sustainable biomass to biofuel, and more.

 

“Harnessing nature’s solutions like the BSF unique traits to mimic nature and create a circular bioeconomy with proper waste management systems is essential.”

 

The BSF industry is currently focused on improving technologies for waste handling, cheaper and more efficient rearing processes, optimization of diets, product extraction methods and more. Implementing genetic programs in BSF facilities, including selective breeding and genetic engineering, offer a repertoire of solutions to optimize the bioreactor itself – the BSF. The importance of managing genetic diversity for the long-term viability and productivity of BSF colonies is becoming more and more prevalent in the scientific community. Here, we will review how selective breeding and genetic engineering can play a key role in the BSF InsectTech. Specifically, we will present the potential, challenges, and recent breakthroughs in employing these two solutions in the BSF industry.

Selective Breeding:

This solution acts on heritable genotypic variation of a certain trait, in favor of individuals – carrying a desired phenotype. Only the selected animals  are allow to mate during consecutive generations, resulting in strains with improvements in the desired phenotypes that are under selection. A strain refers to a wild or captive population displaying a stable characteristic over multiple generations that can be distinguished from other populations.

Similar to other insect species, BSF holds promising reproduction characteristics for selective breeding, namely high fecundity and short generation time. This enables development of a strain with phenotypic improvements using artificial breeding selection in a relatively short time when compared to other livestock or plants. The traits being selected depend on the breeding program’s goal and method of selection.

However, given the industry’s current status, the most important trait for BSF breeders are those which can improve bioconversion ratio to increase the yield of production while ensuring larval and fly fitness, across fertility, size and survival, is retained. This is important because improving larval growth rate along with feed conversion ratio will enable insect farms to increase their overall outputs whilst requiring less feedstock.

However, in some cases, artificial selection results in a negative correlation between life history traits and phenotypes – what is called trade-off effects. For example, a strain solely selected for increased larval mass could grow heavier but have a slower development time, which can lead to reduced farm output at a large scale. This means that careful and holistic phenotypic monitoring over multiple generations within selective breeding is required to avoid undesired outcomes.

In addition, BSF colonies undergoing any selective program should be consistently monitored for their genetic diversity, as a precautionary action to avoid genetic bottlenecks and eventually colony collapse. Such colonies might be also prone to inbreeding depression, characterized by prolonged mating among genetically close relatives especially when the population is founded by a small effective population size.

Beta Bugs has developed it’s HIPER-FLY® selective breeding programme by taking into account the principles outlined above:

  • Leveraging the reproductive characteristics of BSF, it is possible to have a large population from which to select the top performers at both individual and family levels. This is important because having insufficient top performers can lead to loss of genetic performance or the strain failing.
  • Selecting for multiple traits at each generation. Specifically, we focus on improving larval mass, reducing larval development time and improving fly fecundity. The combined improvements in these traits yield a higher-performing larvae for use in the insect farming industry. Trade-off in genetic performance is avoided by monitoring performance across these traits and associated life history parameters (Figure 1).
  • By structuring the breeding program appropriately and collecting both environmental and phenotypic data, we can determine a number of parameters that are key to the operation of selective breeding:
    • The inbreeding coefficient, which gives an indication of the level inbreeding within a given strain, ensuring that inbreeding is kept in check.
    • Correlations between specific traits and life history parameters – this information allows us to determine how selecting for one trait can impinge on the other, or conversely, how selecting for one trait can improve another.
    • The heritability of each trait, i.e., the ease with which the phenotype can be passed from generation to generation. By knowing heritability values it is possible to set the appropriate selection pressure per trait and predict the genetic gain that can be achieved under different breeding program configurations.
    • The overall genetic component for each trait – this is key to improving genetic performance because the measured phenotypes are a combination of both environment (which is held constant and also recorded within the breeding program) and genetics. By decomposing the phenotypes into genetic and environmental effects it is possible to validate that HIPER-FLY performance is increasing not because the larvae have been fed more food or reared in a higher temperature, but because we are improving underlying genetic performance.

 

Leveraging Sustainable Genetic Technologies 
Figure 1: Onwards and upwards – productivity gains achieved to date within the HiPer-Fly® breeding programme.

 

To run the HIPER-FLY® breeding programme, our technical team collects data on all key traits and life history parameters, and then cleans, processes and analyzes it using a suite of algorithms, data processing pipelines, and software tools. Selections are then made using selection criteria that fulfill the breeding goal. By running our selective breeding program on a generational basis within our genetic nucleus, we are able to improve Black Soldier Fly performance – this is then transferred to our in-house egg production facility, The Multiplier, from which we then supply our customers with HIPER-FLY® genetics.

 

Figure 2 – Beta Bugs – From Nucleus to Multiplication to Customer. HiPer-Fly® genetics are developed within a nucleus population, and then scaled up within it’s Multiplier unit, after which they are dispatched to customers for rearing within their own operations.

 

Whilst Beta Bugs’ work has been largely undertaken in-house, the current and previous version of our breeding programs have been enabled through working with our long-term academic collaborator, The Roslin Institute, center of excellence in animal genetics, along with UK InsecTech companies that form part of the £9M Insectrial Revolution project funded by InnovateUK, the UK’s Innovation Funding Service. By doing this, we have been able to ensure that we focus on developing a commercially-relevant product that provides benefit to insect farmers.

 

Genetic Engineering:

Recent establishment of genetic engineering techniques provides the capacity for manipulations via gene knock-out or knock-in of new genes to a certain genomic location. While selective breeding tackles highly complex innate population genetic interactions through several generations. Modern genetic engineering focuses on precise manipulation of a certain gene or a single trait associated with animal performance. This means that desired genetic modification can be achieved more rapidly with greater efficiency, compared to selective breeding.

However, in the case of animal feed production, selective breeding is favored in most countries as there should be strict regulation policies on products potentially entering the feed and food chains. Possible traits that can be manipulated using genetic engineering tools in BSF may include for instance delay of metamorphosis resulting in bigger larvae with improved feeding capacity or increasing the concentration of particular nutrients of the larvae (e.g., protein and fat).

In the scientific community, CRISPR/Cas9 system is the most popular technique to introduce mutations into the genome of an organism, due to its’ high specificity rates.  The most common technique to deliver CRISPR/Cas9 to BSF genome is via microinjection of freshly laid embryos (eggs). A major obstacle in the race for genetic engineering of BSF is the ability to rapidly screen and isolate suspected transgene individuals from the whole population.

It is usually conducted using selection markers, such as fluorescent protein genes or body phenotype (Figure. 4a-b). Moreover, cell cultures provide an efficient and fast system for early molecular screening of desired genetic elements prior to microinjection.  BugEra has developed various types of cell cultures and successfully used them to test different genetic elements in-vitro (Figure 3).

 

Figure 3
BSF cell culture developed by BugEra.
A – confocal micrograph showing fluorescent signal expressed by cells
B – visible light channel, showing cellular morphology
C – merged image showing florescent signal localized within the cells

 

BugEra’s first product is a novel strain named BSFx2, addresses the biofuel need by doubling the larvae oil volume. While the lipid profile of the maggot is a perfect fit for biofuels (biodiesel, renewable diesel, and sustainable aviation fuel (SAF) production, it is not commercialized due to relatively low volume and market bias to feed production. BSFx2 will enable the economic feasibility of using maggots as the most sustainable lipids for biofuel while upcycling none feed grade organic waste achieving the best carbon-credit score.

Our working pipeline begins with bioinformatic screening of target genes in BSF and model organisms. We than proceed to molecular work required for the construction and validation of genetic elements which bring the inherited genetic change in the offspring. Target genes also include selection markers (molecular or phenotypic) to allow identification of suspects after microinjection. These suspects are then allowed to mate and upscale and after molecular validation a stable novel strain can be announced.

Our genetic toolbox focuses on the implementation of CRISPR/Cas9 for introducing precisely targeted mutations. We recently developed two phenotypic lines of BSF, with a stable inheritance of the introduced mutations across multiple generations. While mutations introduced in the gene involved in BSF proper wing development cannot be used as a phenotypic marker because of reduced mating abilities (mating occurs during flight; Figure 4b).

Flies in the second genetically-edited strain exhibited a clear phenotypic change in eye coloration which can be used as a phenotypic marker (Figure 4). BugEra is also developing alternative solutions based on non-inherited gene downregulation techniques, such as RNA interference (RNAi). Such non-transgenic products might be of advantage in terms of regulation and simplicity of the delivery system, compared to laborious embryo microinjection needed for genetic engineering.

BugEra is also participating in the Israeli BSF consortium (financed by the Israeli Innovation Authority) aiming at the acceleration of circular economy solutions based on thoroughly crafted BSF farming. This consortium is based on joint research of academic and industrial companies. More specifically, we are operating in a multidisciplinary scientific approach combining nutritional, entomological, biochemical, agricultural, genomic, ecological, and microbiological research fields to generate a body of data across the value chain. AI-powered software will then process and reiterate this data to maximize the value of the knowledge.

 

A promising combination between selective breeding and genetic engineering:

The great synergetic effect of the combination of the two genetic approaches introduces almost endless opportunities for improved efficiency of InsecTech. For instance, the “adaptation” of the larvae to the waste stream could be achieved through selective breeding more efficiently than even advanced genetic engineering techniques. As no prior data is needed.  Academic research papers have already reported differences in the performance of different artificially bred BSF wild-type strains on the same substrate.

Further enhancements to selective breeding could be achieved through genetic engineering. Specifically, substrate-“adapted” strains could be introduced with “super”-abilities as for example increased food conversion ratio, enhanced fatty acid oil traits , or the ability to produce novel biomaterials. The opportunities for such biological solutions are endless and entirely novel biomanufacturing supply chains could be created shaping the transition to the bioeconomy of the future.

Leveraging Sustainable Genetic Technologies 
Figure 4
Genetically edited BSF phenotypic markers strains developed by BugEra. A -A’ Wild type BSF fly eyes micrographs (lateral view and ventral view) showing characteristic light green coloration with dark violet stripes. B-B’ a genetically engineered strain exhibiting a phenotype change in eye coloration to dark green. C – wild-type BSF showing normal wings morphology; C’ – genetically engineered BSF strain with defected wing morphology being curved, highlighted by red arrow.

 

Summary

Genetics-based solutions should play a major role in expanding the value proposition of InsecTech and the BSF market.  The above innovative opportunities armed with high-throughput sequencing technologies and powerful tools for genome editing are primed to yield promising results that will give substantial contributions to fighting climate crisis and food shortage, and position the BSF-based InsecTech at the forefront of bioeconomy and biomanufacturing.

 

To read the full Beta Buzz Science Edition click here 

 

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Beta Buzz #4
.

 

Thomas Farrugia introduces the long-awaited Beta Buzz #4,  it reflects two things: i) the rapid development of our sector and ii) holding true to our commitment to enabling new market entrants to join the insect farming sector.

Over the past half-year, our industry has continued to make strides towards scaling up, with a significant number of large-scale insect farms being completed recently and further involvement from agri food stakeholders who are either providing their own offerings or partnering with existing Black Soldier Fly companies.

Read the full Beta Buzz #4 issue here

The team at Beta Bugs are talented, committed, and excited to be working at the next frontier of animal breeding. In line with the company values, Beta Bugs have established a culture which encourages development, performance, accountability, teamwork and trust.

If you have any questions about Beta Bugs or require any further information on our products, please do get in touch at info@betabugs.uk, outlining your query and one of the Beta Bugs team members will get back to you.

Best regards,

Beta Bugs Limited