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The Beginning	Meet The Team	About The Project	Parts Submitted to the Registry	Modelling	Notebook	Safety	Attributions

Safety

Biosafety: what exactly is it?

Biosafety is defined as the prevention of 1) Persons being exposed to organism as well as toxic chemicals that can cause disease 2) The accidental release of the aforementioned organisms or chemicals

The major biosafety concern tackled herein is the prevention of unwanted/detrimental effects on the environment caused by the release of genetically modified organisms.

Bio-safety of genetically modified plants: Why should we be concerned?

The overall reason to test genetically modified organisms before their release is because humankind's total impact on ecosystems services (i.e. ecological processes that benefit human kind)from previous introduction of new technologies is substantial (Millennium Ecosystem Assessment (MEA) 2005). It has been learnt from the past 100 years of industrial –technological development that all environmentally related technological development came with a price – many of which outweigh the benefits(Harremoës 2002. Consequently, all new environmental stresses need to be carefully assessed (Lövei et al 2007).

Approximately 70% of the African population is engaged in agriculture. Consequently, introduction of technologies such as genetically modified plants must be tested as their effects will directly affect the lives of the majority of the populace. To add to the importance of testing the plants is the fact that in tropical countries , agricultural producers depend on ecosystem services (ecological process that human kind benefit from) more closely than farmers in developed countries (Lövei et al 2007).

Team Nairobi biosafety approach

Risks to the safety and health of team members or others in the lab

The materials used in this project do not pose any risks to team members and others in the lab. The experiment proteins used from the bacteria are not harmful to any animals . To the best of our knowledge Metarhizium has little to no impact on humans or other mammals in the field (Siegel et al, 1997). However,

• Cases of sinusitis have been reported in man suspected to have been caused by M. anisopliae (Revankar et al, 1999). Thus far, this is among the few potential harmful effects Metarhizium can have on vertebrates.
• In humans, there are six reported cases of a disease caused by M. anisopliae. M. anisopliae var. anisopliae was isolated for the first time from the eye of a Colombian male as the aetiological agent of keratomycosis (Cepero de Garcia et al. 1997). A topical natamycin treatment was successful. These harmful effects were however also described as being very rare. (Howard et al, 2002).

Risks to the safety and health of the general public if released by design or accident

It has no grave risks except for allergic reactions in already allergy susceptible individuals. Such individuals should be fore warned before use of the insecticide. As described above, Metarhizium has little to no impact on humans or other mammals in the field (Siegel JP. 1997). Safety data on naturally occurring M. anisopliae were summarised by Siegel and Shadduck (1990) and Zimmermann (1993). Previously, there were no reports of M. anisopliae infecting mammals or humans. However, to date some cases of human and mammalian infections have been described but none was associated with the use of M. anisopliae as a bio control agent.

Cases of allergy reports and studies include the following:

• In humans, a severe dermal hyper allergic response caused by an isolate of M. anisopliae var. acridum was reported by Goettel et al (2001). It was attributed to the widespread use of M. anisopliae for biocontrol of various sugar cane pests in Brazil. Several persons showing asthmatic symptoms due to this fungus were reported (see Barbieri et al. 2005).
• On the other hand, no allergic effects on the researchers, manufacturing staff, formulators or on field workers working with mycoinsecticides based on various strains of M. anisopliae var. anisopliae (BIO 1020, strain FI-1045, strains ICIPE 30 and 69) and M. anisopliae var. acridum (strains IMI 330189 and FI-985) were observed (Copping 2004).
• First inhalation experiments of M. anisopliae conidia were conducted in rats, guinea pigs and mice (Schaerffenberg, 1968; Shadduck et al, 1982; El-Kadi et al, 1983). No allergic reactions were observed.
• Further investigations, however, demonstrated that M. anisopliae has an allergenic potential. Ward et al (1998, 2000) carried out a series of studies injecting crude allergenic extract inoculations obtained from a M. anisopliae strain, into mice and demonstrated that this extract contains components that induce immunologically mediated inflammatory responses. The histo-pathological effects seen were characteristic of an allergic reaction.
• Fractions of specific crude extract of M. Anisoplia, suspected to induce an allergic reaction were obtained by Barbieri et al (2005). M. anisopliae was found to have the ability of potentiating the hypersensitivity reaction to an allergen in mice and thus may also worsen allergy in susceptible individuals (Instanes et al, 2006).
• The first well documented case of mammalian infection by M. anisopliae var. anisopliae was described by Muir et al. (1998). An invasive mycotic rhinitis was diagnosed in a cat with a 4-month history of nasal discharge and subcutaneous swelling of the nasal bridge. The infection was treated with orally administered itraconazole.

Similar products already in existence

There are similar products though none are transgenic. There are commercial biocontrol products in the market that use Metarhizium as their reactive agent. Below is a list of commercial biopesticides that use fungi as their active agents according to Gisbert Zimmerman, 2007; Table II. Mycopesticides of Metarhizium anisopliae registered or under commercial development (Butt et al, 2001; Wraight et al, 2001; Copping, 2004; Kabaluk, 2005; Zimmermann, 2005).

Product/Trade name	Company/Producer	Country/Origin
BioBlast	EcoScience	USA
Bio-Cane Granules	Becker-Underwood	Australia
Bio-Catch-M	Stanes	India
Bio-Green Granules	Becker-Underwood	Australia
Bio-Magic	Stanes	India
BioPath	EcoScience	USA
Cobican	Probioagro	Venezuela
Gran Met-P	Kwizda/Agrifutur	Austria/Italy
Green Guard	SC Becker-Underwood	Australia
Green Guard ULV	Becker-Underwood	Australia
Green Muscle	CABI Bioscience/NPP	UK/France
Metaquino	-	Brazil
Metarhizium Schweizer	Lbu (formerly Eric Schweizer Seeds)	Switzerland
Metathripol	ICIPE	Kenya
Muchwatox (proposed)	ICIPE	Kenya
Pacer SOM	Phytopharma	India
Taenure Granular Bioinsecticide	Novozymes Biologicals (formerly Earth BioSciences)	USA
TAE-001 Technical Bioinsecticide	Novozymes Biologicals (formerly Earth BioSciences)	USA
Tick-Ex EC	Novozymes Biologicals (formerly Earth BioSciences)	USA
Tick-Ex G	Novozymes Biologicals (formerly Earth BioSciences)	USA

Risks to environmental quality if released by design or accident

The risk to environmental quality is very low. There is however a slight risk to the following:

• Developing embryos of the inland silverside fish (Fred J. Genthner and Douglas P. Middaugh, 1995).
• Juvenile mosquito fish, Gambusia affinis, (Genthner et al, 1998). No mortalities or adverse effects were observed in adult Gambusia affinis
• Parasitic hymenoptera under laboratory condition an effect that was not confirmed under cage (Stolz I. 1999) or field conditions (Peveling R et al. 1999).
• Developing embryos of the grass shrimp Palaemonetes pugio (Genthner et al. 1997, 1998).
• Metarhizium anisopliae has antagonistic properties against some phytopathogenic fungi, but is also suppressed by several mycoparasites. The fungus was shown to be antagonistic to two strains of Ophistoma ulmi (=Ceratocystis ulmi), the cause of Dutch elm disease (Gemma et al. 1984)

Risks to security through malicious misuse by individuals, groups or states

There are no major concerns attributable to any of the parts used. The deployment of transformed strain of M. anisopliae did not depress the culturable indigenous fungal microflora (Hu et al, 2002) had no phytopathogenic or phytotoxic effects on leaves or plant roots, as a couple of studies sought to find out. (Zimmermann, 1981, Stenzel, 1992).

• There were no harmful effects on soil organisms like the collembolans, Folsomia candida). The collembollans consumed and inactivated the insect pathogen without any harmful effects. (Reinecke et al. 1990, Broza et al, 2001).
• There were no effects when fed to the leopard frog, Rana pipiens. (Donovan-Peluso et al, 1980) nor frog embryos were exposure to it. (Genthner et al, 1998)
• Avian safety studies were conducted with the Japanese quail by Wasti et al (1980). There was no mortality or abnormal behaviour in the experimental birds.
• Ring-necked pheasants were also demonstrated to not be toxically susceptible. (Smits et al. 1999, Johnson et al. 2002).
• Destruxin B is a major metabolite produced by M. anisopliae (Buchwaldt et al, 1992) causes necrotic and chlorotic symptoms both on host and nonhost plants. However, there are no observations on phytotoxic reactions in plants after application of M. anisopliae, which demonstrate that destruxin B is produced only after successful infection of the plant by A. brassicae.

How we address safety issues

To ensure safety of team members and others in the lab, basic lab safety measures are adhered to like use of gloves, face masks and goggles while working with Metarhizium spp. Later, a disclaimer will accompany the product upon completion as to the potential danger of the transgenic fungus to select organisms like fish. In the drafting of the methodology all risks were assessed and proper machinery used. The least antigenic strains were selected and also there are no animal or fish testing phases of the project.

Safety, security, health and/or environmental issues

Of importance are the potential allergic effects on humans who are immunologically susceptible to adverse hypersensitivity reactions. (Barbieri et al, 2005, Instanes et al, 2006).

Biosafety provisions we operate under

The project and all its undertaking are operated under the national biosafety guidelines(Kenya).

Our institution also has its own biosafety rules which are based off the national biosafety rules. Availability to them is however, at this point and time not possible. The institution has its own biosafety committee. However, we were unable to discuss the project with them.

Biosafety and lab training received

We received biosafety and lab training before this project. This came to use as part of our course work as we are had taken units that specifically deal with lab training. The units are called laboratory techniques and laboratory methods in microbiology. The relevant training we received pertaining to biosafety included: Laboratory safety ;containment and disinfection; physical containment levels- their laboratories equipment and respective work practices in each level; isolation and culture of microorganisms and sterilisation techniques

National biosafety guidelines

Below are the national biosafety guidelines.

National Guidelines for Release of Genetically Modified Organisms (GMOs) into the Environment:

Biosafety procedures for genetic modification and release are well established in most industrialized countries, thus reflecting the level of biotechnology activities undertaken. In a developing country like Kenya, research in genetic modification is being performed on a smaller scale in government funded research institutions and universities.

Nevertheless, release experiments of genetically modified organisms (GMOs) is envisaged in the future. Kenya also envisages that there will be release experiments performed by multi-national companies. Taking these into consideration, there is an urgent need for developing biosafety procedures at the national level and providing advice to the Government on biosafety policies.

The establishment of biosafety regulations is to ensure monitoring and regulation of international projects pertaining to genetic modification and release, commercial activities and technologies, as well as development, release and large scale use of GMOs. The proposed National Guidelines for release of GMOs into the environment have been developed from the existing principles derived from relevant regulations and guidelines at national, regional and international levels.

[http://www.biosafetykenya.co.ke/documents/ChecklistforInspectionRELEASEOFGMOsinKenya.pdf| Full document here]

Guidelines for Monitoring and Inspecting GMOs in Kenya.

Introduction

Inspection, Monitoring and Evaluation is a fairly new discipline all over the world. Monitoring is a regular, systematic and consistent assessment of the progress achieved in the implementation of an activity that is aimed at meeting set objectives, to ensure accountability, cost effectiveness, timeliness and quality and must include taking corrective measures.

Inspection is the official examination of a regulated article to determine if genetically modified organisms are present and or to determine compliance with the laws and regulations. The basic assumption in monitoring and evaluation is that, effective implementation of a project will have a positive impact on the economy, welfare of people, etc.

The main objective of monitoring and inspection is to ensure that the development, transport, use, transfer and release of any LMO are undertaken in a manner that prevents or reduces the risks to biological diversity taking also into account risks to human health. [http://www.biosafetykenya.co.ke/documents/ChecklistforInspectionRELEASEOFGMOsinKenya.pdf| Full document here]

[http://www.biosafetykenya.co.ke/documents/ChecklistforInspection1containeduselaboratory.pdf|Inspection Guidelines for Contained Use (Laboratory Activities) for GMOs in Kenya.]

[http://www.biosafetykenya.co.ke/documents/ChecklistforInspections3containeduseglasshouse.pdf|Inspection Guidelines for Contained Use (Glass-houses & Growth-rooms) for GMOs in Kenya (Biosafety Level 1-3).]

[http://www.biosafetykenya.co.ke/documents/ChecklistforInspections4Fieldtrials.pdf|Inspection Guidelines for Field Trials of GMOs in Kenya.]

Our ideas on how to deal with bio-safety

As the number of parts increase the registry needs to document any possible interactions between all available parts.

iGEM could also come up with standard biosafety rues that should be sent out to participating teams when they receive their parts. These can be used in addition to the national biosafety rules… Also a special part of the competition can be dedicated to the development of safe parts!

How could parts, devices and systems be made even safer through bio safety engineering?

Through the creation of switches, if the part leaves a predetermined environment, a self destruct (apoptotic-like) mechanism can be triggered which will ensure that the part eases to function.

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References

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Revankar, S. G., D. A. Sutton, S. E. Sanche, J. Rao, M. Zervos, F. Dashti, and M. G. Rinaldi. 1999. Metarrhizium anisopliae as a cause of sinusitis in immunocompetent hosts. J. Clin. Microbiol. 37: 195Ð198

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