Fungi Uses in Pest Management

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-[[Image:98503.jpg|thumb|right|300px|This image shows different species]]+[[Image:Lecanicillium2.jpg|thumb|right|250px|''Lecanicillium lecanii'' on hemlock woolly adelgid cadaver<ref>Svetlana Y. Gouli, University of Vermont, Bugwood.org</ref>]]
-This is a background of fungi uses in pest management. Testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing, testing...+ 
 +[[Image:Beauveria bassiana2.jpg|thumb|right|250px|''Beauveria bassiana'' on pecan weevil (''Curculio caryae'')<ref>Louis Tedders, USDA Agricultural Research Service, Bugwood.org</ref>]]
 + 
 +[[Image:Trichoderma2.jpg|thumb|right|250px|''Trichoderma spp.'' on Norway spruce (''Picea abies'')<ref>Andrej Kunca, National Forest Centre - Slovakia, Bugwood.org</ref>]]
 + 
 +Fungi are useful biological controls for pests in crops and [[Fungi uses in Urban Environments|urban environments]] and have been commercially available since 1981.<ref name ="Rechcigl"> Rechcigl, E.J., & Rechcigl N.A. (2000). ''Biological and Biotechnological Controls of Insect Pests.'' (1st ed.). Boca Ranton (FL): Lewis.</ref> They can be used as [[Fungi used as a Herbicide|herbicides]], [[Fungi used as a Insecticide|insecticides]] and [[Fungi used as a Fungicide|fungicides]]. Mycopesticides are interesting organic alternatives to chemical pesticides due to their enhanced safety and their high specificity for the non-targeted species such as humans, pets and desired insects.<ref name="textbook fungi">Moore, D., Robson, G.D., & Trinci, A.P.J. (2011). ''21st Century Guidebook to Fungi''. (1st ed.) New York (NY): Cambridge</ref> They share [[#Advantages and Disadvantages|advantages and disadvantages]] with other biopesticides.<ref name="textbook fungi"/> [[Lecanicillium lecanii|''Lecanicillium lecanii'']], [[Species|''Beauveria bassiana'']] and [[Species|''Trichoderma'' spp.]] are amongst the most commonly used [[Species|species]] and have been widely studied in spite of the short [[#History|history]] of this field of study. With the growing enthusiasm for organic commercial farming and justified environmental concerns in the population, mycopesticide have a definitely a bright [[#Novel Uses of Mycopesticides|future]].<ref name ="CPL"> CPL Business Consultants. (2010). ''The 2010 Worldwide Biopesticides Market Summary.'' Wallingford: CAB International.</ref>
 + 
==History== ==History==
- <p> The use of Entomopathogenic fungi in pest management covers a wide variety of Eumycota subdivisions. These subdivisions of Eumycota include; Mastigomycotina, Zygomycotina, Ascomycotina, and Deuteromycotina <ref>BIOLOGICAL and BIOTECHNILOGICAL CONTROL of INSECT PESTS (2000). Rechcigl. E.J. and Rechcigl. N.A. (Pg 49) Pg</ref> In 1981, the first mycoinsectiside registered in the U.S. was ''Hirsutella thompsonii'' and was given the registered name; Mycar. This species has been noted to cause [[Definitions|epizootics]] as early as 1920 in some species of spider mites <ref>Biological and Biotechnological Controls of INSECT PESTS Rechcigl E.J., Rechcigl N.A. Pg 49-51.</ref> +For over 100 years, scientists have been studying many different species of fungi with [[Definitions|entomopathogenic]] characteristics but the use of these organisms in the field has been relatively elusive. Many of these fungal organisms are not only entomopathogens, but also aid in general plant growth. Since the late 1940's, chemical insecticides have been the main tool that growers have utilized, in order to control potentially damaging pests and diseases. Some of the earliest recorded studies of entomopathogenic fungi occurred in the early 1800 when the silkworm industry in France was devastated by [[Species|''Beauveria bassiana'']]. As a consequence of this insight, further research began looking into the use of fungal insect pathogens to manage a wide range of pests.<ref name="Vega">Vega, F.E., Goettel, M.S., Blackwell, M., Chandler, D., Jackson M.A., Keller, S., et al. (2009). Fungal entomopathogens: new insights on their ecology. ''Fungal Ecology'' 2(4), 149-59.</ref>
 + 
 +Most recently, DNA sequence analysis has lead to a much better understanding between the differences in entomopathogenic fungi and other types of fungi. This new research has revealed that some of these entomopathogenic fungi also play the role of beneficial rhizosphere associates, plant growth promoters, as well as plant endophytes. These findings have now lead to further research to determine if some of these entomopathogenic fungi can be used for more than just one purpose.
-<p>With the rising cost of synthetic chemical pesticides and increasing cases of pesticide resistance, the search for natural biologically based forms of pest management has been a key area of research for the last decade. With the world population rising over 7 billion and the drastic effects of climate change, the need for alternative forms of pest control will become essential. Although insects are key in the performance of many ecosystems, they play a large role in the 35 billion dollars worth of crop that is lost in the United States each year <ref>Entomopathogenic fungi as biological control agents. P. A. Shah, J. K. Pell. Applied Microbiology and Biotechnology June 2003, Volume 61, Issue 5-6, pp 413-423.</ref> 
-==Biopesticides in Agriculture==+The use of Entomopathogenic fungi in pest management covers a wide variety of Eumycota subdivisions. These subdivisions of Eumycota include; [[Definitions|Mastigomycotina]], [[Definitions|Zygomycotina]], [[Definitions|Ascomycotina]],and [[Definitions|Deuteromycotina]].<ref name="Rechcigl"/> In 1981, the first mycoinsectiside registered in the U.S. was [[Species|''Hirsutella thompsonii'']] and was given the registered name; Mycar.<ref>Kenneth, R., Muttath, T.I. & Gerson, U. (1979). ''Hirsutella thompsonii'', a fungal pathogen of mites. I. Biology of the fungus in vitro. ''Annals of Applied Biology'', 91(1), 21–28.</ref> This species has been noted to cause [[Definitions|epizootics]] as early as 1920 in some species of spider mites.<ref name="Rechcigl"/>
- <p> Insect and weed management in important crops... sustainable agriculture, as a replacement for +With the rising cost of synthetic chemical pesticides and increasing cases of pesticide resistance, the search for natural biologically based forms of pest management has been a key area of research for the last decade. With the world population rising over 7 billion and the drastic effects of climate change, the need for alternative forms of pest control will become essential. Although insects are key in the performance of many ecosystems, they play a large role in the $35 billion worth of crop that is lost in the United States each year.<ref> Shah, P.A., & Pell, J.K. (2003). Fungi as Biological Control Agents. ''Applied Microbiology and Biotechnology'', 61(5-6), 413-23.</ref>
-===Fungi used as a Herbicide=== 
-'''''Introduction''''' 
-The presence of weeds can result in the loss of up to half of a grower’s expected crop yield. The lost yield results in economic losses which is a problem in the agriculture industry. Fungi can be used as a herbicide to gain control of weeds and solve this issue. Fungal herbicides have been legal in Canada since 1973<ref> Misra, H.P. Weed Management Through Fungal Herbicides. November 2005. Orissa Review. http://orissa.gov.in/e-magazine/Orissareview/nov2005/engpdf/Weed_Management_Through_Fungal_Herbicides.pdf</ref>. In order to use a fungi for this purpose, a [http://en.wikipedia.org/wiki/Mycoherbicide mycoherbicide] is created with the fungi as its active ingredient. This mycoherbicide must be able to inhibit weed growth without harming the crop or the living things surrounding it. A benefit is that the fungi used are host specific and will reside on the soil or weed host, while a commercial chemical herbicide would need to be reapplied to the crop. Certain mycoherbicides work better on certain strains of weeds. There are currently 37 mycoherbicides and 8 techniques of applying them <ref> Misra, H.P. Weed Management Through Fungal Herbicides. November 2005. Orissa Review. http://orissa.gov.in/e-magazine/Orissareview/nov2005/engpdf/Weed_Management_Through_Fungal_Herbicides.pdf</ref>. The fungi species most commonly used as herbicides in North America are ''Collectrichum gloeosporioides'' (Collego®) and ''Phytophthora palmivoraa'' (De Vine®).+There is enormous potential for pest and vector control in pest management in the future due to an increasing amount of knowledge in the area of mycology.<ref>Seigfried, K. (1998). Use of Fungi for Pest Control in Sustainable Agriculture. ''Phytoprotection'', 79(4), 56-60.</ref>
-'''''Process'''''+==Types of Mycopesticides==
- Mycoherbicides act by releasing [[Definitions|phytopathogens]] to suppress weed growth. The phytopathogens release [http://en.wikipedia.org/wiki/Phytotoxin phytotoxins] that can kill weeds in up to five weeks time. The pathogens released have the ability to produce spores for asexual reproduction and are tested to be stable and temperature tolerant. Fungal herbicides are easily made once the anti-herb properties have been testing as well as the living conditions for that fungi. They are easily applied by spraying. Once applied, the spores will germinate and penetrate the herb tissue and the phytotoxins will start to work. However, there are reasons fungal herbicides may not be used in place of a harmful chemical. In North America, fungal herbicides have to be registered and approved by the Environmental Protection Agency [http://en.wikipedia.org/wiki/EPA (EPA)] which can take years. The anti-herb capacity of fungi is generally less than that of a chemical herbicide. Fungal herbicides are not commonly used because their success rate depends on the environmental conditions and because they are so host specific.  
-====Phytotoxins====+===Mycoherbicide===
-Phytotoxins are low molecular weight secondary metabolites that cause plant disease. In this case, the plant being infected is a weed plant. Phytotoxins may also be responsible for visual symptoms of disease such as wilting or [[Definitions|necrosis]]. A common phytotoxin found in the fungi species ''Penicillium'' is vulculic acid. Vulculic acid works against weeds by increasing leaf membrane permeability, inducing lipid peroxidation and damaging the cell membrane. Phytotoxins have a limited stable state because of their short half-life and sensitivity to the environment. Phytotoxins are difficult to produce for commercial sale because they are produced in very small quantities by fungi and difficult to isolate. +''See main article'': [[Fungi used as a Herbicide|Fungi used as a Herbicide]]
 +Fungi can be used as an herbicide to exterminate specific weeds and other plants that are detrimental to the effective growth of crops. In order to use a fungus for this purpose, a mycoherbicide is created with the fungi as its active ingredient. The use of mycoherbicides focus on the theory of the disease triangle. The disease triangle breaks down the conditions that need to be apparent for infection to occur, those being the environment, pathogen and host. The following will fall under one or more of these categories. Mycoherbicides act by releasing phytopathogens to suppress weed growth. The phytopathogens release phytotoxins that can kill weeds in up to five weeks time. A common phytotoxin found in the fungi species ''Penicillium'' is vulculic acid.<ref name="Misra"> Misra, H.P. (2005). ''Weed Management Through Fungal Herbicides''. Orissa Review. 53-56</ref> The fungi species most commonly used as herbicides in North America are [[Species|''Collectrichum gloesporioides'']] (Collego®) and [[Species|''Phytophthora palmivoraa'']] (De Vine®).<ref name="Misra"/> A specific weed strain commonly known as arrowhead (''Sagitaria trifolia'') is the cause of the largest rice plantation problem in Iran. Arrowhead is notably resistant to chemical herbicides. Collego® is used in the USA as the solution to arrowhead.<ref name="Javadzadeh"> Motlagh, M.R.S. & Javadzadeh, A. (2010). Study of Alternaria pellucida as a promising mycoherbicide for controlling Arrowhead (Sagitaria trifolia) in paddy fields. ''Proteomics Journal,'' 3(6), 172-6.</ref> Other types of fungi that can be used as mycoherbicides are rusts and mildews. These fungi use spore distribution to cause infection in their host plants.
-'''''Arrowhead'''''+===Mycoinsecticide===
 +''See main article'': [[Fungi used as a Insecticide|Fungi used as a Insecticide]]
-[[Image:Arrowhead.jpg|thumb|right|250px|Arrowhead Weed]]+Certain species of fungi can act as parasites of insect. When a fungus is used as insecticides, it is called mycoinsecticide. <ref name="Wraight and Carruthers"> Wraight, S.P. & Carruthers, R.I. (1998). Production, Delivery, and Use of Mycoinsecticides for Control of Insect Pests on Field Crops. ''Methods in Biotechnology,'' (5), 233-69.</ref> In recent years, crop protection has been trending towards integrated pest management (IPM) using bacteria and fungi as insecticides. Approximately 750 species of fungi are pathogenic to insects and only 12 are being utilized as insecticides.<ref name="Wraight and Carruthers"/> Two prominent species of fungi used as insecticides are [[Species|''Beauveria bassiana'']], and [[Species|''Metarhizium anisopliae'']]. Mycoinsecticides function by first being applied to the insects in spray form. The fungi then use their hyphae to burrow into the insects. The hyphae spread the insectotoxins throughout the insect to activate them, eventually leading to the insect’s death.
 +===Mycofungicide===
 +''See main article'': [[Fungi used as a Fungicide|Fungi used as a Fungicide]]
- A specific weed strain commonly known as arrowhead (''Sagitaria trifolia'') is the cause of the largest rice plantation problem in Iran. Arrowhead is notably resistant to chemical herbicides. Collego® is used in the USA as the solution to arrowhead. The fungi ''Alternaria pellucida'' was tested and proved that it could successfully be used as a mycoherbicide against arrowhead in Iran. This fungi works by preventing early growth of the weed. The fungi species was tested for its anti-herb properties on three indigenous cultivars of rice, two bred cultivars and lastly on an arrowhead infested cultivar, all cultivated separately in a greenhouse. ''Alternaria pellucida'' significantly altered the height of the arrowhead weed and was concluded to be a mycoherbicide able to control the arrowhead weed. +There are many cases of mycoparasitism in nature. In fact, a large amount of dormant spores like sclerotia or chlamydospore are found in the wild already parasitized by another fungi.<ref name="Jeffries">Jeffries, P. (1995). Biology and Ecology of Mycoparasitism. ''Canadian Journal of Botany''. 73(1), 1284-1290.</ref> This is made possible by the ability of most fungi to release lytic enzymes like chitinase that allows them to break down chitinuous cell walls. The parasitism of a fungi on another one occurs generally through four main stages: chemotaxi, recognition, attachment and penetration.<ref name ="Plant-Microbe">Boland, G. J., & Kuykendall, L. (1998). ''Plant-microbe interactions and biological control.'' (1st ed.). New York (NY): Marcel Dekker</ref>The type of relationship that will follow such an invasion will be either necrotrophic or biotrophic.<ref name="textbook fungi"/> The most commonly used species are in the [[Species|''Trichoderma'']] genus.<ref name="tricho on botry">Cheng, C., Yang, C., & Peng, K. (2012). Antagonism of ''Trichoderma harzianum'' ETS 323 on ''Botrytis cinerea'' Mycelium in Culture Conditions. ''Phytophatology'' 102(11), 1054-63.</ref>
-(Not done, don't worry -Emily)+===Mycopesticides in Urban Environments===
 +''See main article'': [[Fungi uses in Urban Environments]]
-===Fungi used as an Insecticide===+[[Definitions|Entomopathogens]] are key components for integrated pest management solutions.<ref name="Milner and Pereira">Milner, R.J. & Pereira R.M. (2007). Microbial control of urban pests - cockroaches, ants and termites. ''Field Manual of Techniques in Invertebrate Pathology'', 20(2), 695-711</ref> There is a large market for urban pest control that has been dominated by the use of pesticides and insecticides.<ref name="Milner and Pereira"/> These chemical pesticides pose a danger not only to the person applying them, but other people, or pets, which may come into contact with the chemicals in an urban environment.<ref name="Milner and Pereira"/> Previously used in agricultural settings as [[Definitions|deterrents]], fungi such as [[Species|''Beauveria bassiana'']], [[Species|''Metarhizium anisopliae'']] and [[Species|''Paecilomyces fumosoroseus'']] may have possible pest control applications in urban settings as well.<ref name="Lenz">Lenz, M. (2005). Biological control in termite management: the potential of nematodes and fungal pathogens. ''Proceedings of the Fifth International Conference on Urban Pests.'' 47-52</ref>
-===Fungi used as a Fungicide===+==Advantages and Disadvantages==
-===Biopesticides in Urban Environments===+''See main article'': [[Advantages and Disadvantages|Advantages and Disadvantages]]
-<p>[[Definitions|Entomopathogens]] are key components for integrated pest management solutions <ref name="Milner and Pereira"> Milner, RJ. and Pereira RM. 2007. Microbial control of urban pests - cockroaches, ants and termites. Field Manual of ''Techniques in Invertebrate Pathology''. 20(2):695-711</ref>. There is a large market for urban pest control that has been dominated by the use of pesticides and insecticides <ref name="Milner and Pereira"/>. These chemical pesticides pose a danger not only to the person applying them, but other people, or pets, which may come into contact with the chemicals in an urban environment <ref name="Milner and Pereira"/>. Previously used in agricultural settings as [[Definitions|deterrents]], fungi such as ''Beauveria bassiana'',''Metarhizium anisopliae'' and ''Paecilomyces fumosoroseus'' may have possible pest control applications in urban settings as well <ref name="Lenz">Lenz, M. 2005. Biological control in termite management: the potential of nematodes and fungal pathogens. ''Proceedings of the Fifth International Conference on Urban Pests''. 47-52</ref>. </p>+
-<p>Treatment of pest insects with fungi relies on the natural spread of the disease organism, whether through direct contact with nest mates, or ingestion <ref name="Milner and Pereira"/>. [[Definitions|Biopesticides]] have the largest potential for treatment of social insects such as cockroaches, ants and termites <ref name="Milner and Pereira"/>. Application methods include the use of [[Definitions|bait stations]] or direct application to a located nest <ref name="Milner and Pereira"/>. In order to be effective, pathogenic [[Definitions|conidia]] must come into contact with the insect cuticle, or [[Definitions|exoskeleton]] <ref name="Milner and Pereira"/>.</p>+The negative impacts of chemical pesticides is what has guided new attention towards increased emphasis on biological control agents with respect to using fungi through the use of naturally occurring pathogens of arthropods. Many of these naturally occurring pathogens produced by fungi are already being employed, mostly on a small scale controlling arthropod pests in greenhouse crops, orchards, ornamentals, turf and lawn grasses, stored products and forests, and also for moderation of vectors of animal and human diseases.<ref name="Lacey et al">Lacey, L.A., Frutos, R., Kaya, H.K., & Vail, P. (2001). Insect Pathogens as Biological Control Agents: Do They Have a Future? ''Biological control,'' 21(3), 230-48</ref>
-<p>Pathogenic fungi may have a greater [[Definitions|repellant]] effect, and can be applied to plants or mixed with soil, or even mixed with paint and painted onto timber <ref name="Milner and Pereira"/>. These fungus treatments may reduce the infestation by deterring pests rather than eliminating them <ref name="Milner and Pereira"/>. Studies indicated similar success with fungal spores to those effects achieved by bifenthrin and triflumeron insecticides <ref name="Milner and Pereira"/>.</p> 
-<p> Overcoming this repellency can be achieved by mixing the conidia with agar or cellulose, diluting the conidia and masking the factors inducing repellency <ref name="Milner and Pereira"/>. Additionally, adding the repellent spores to an attractant such as honey water may entice the insects to visit the biopesticide trap <ref name="Milner and Pereira"/>.</p> 
 +Although the utilization of the full potential of fungal biocontrol agents against arthropod pests has had limited commercial success, the major advantages of these biocontrol agents are:<ref name="textbook fungi"/>
 +* Effectiveness and potentially high specificity;
 +* Acceptable naturalness;
 +* Safety for humans and other non target organisms;
 +* Reduced chemical pesticide use and consequential reduction of residues in food and the environment;
 +* Protection of natural enemies of the pest;
 +* Protection of biodiversity in managed ecosystems.
-[[Image:German-cockroach-md.jpg|thumb|left|150px|The German Cockroach, ''Blatella germanica'', a common household pest]]'''''Cockroaches'''''+Despite these promising advantages of using fungi as biological control agents in pest management, there are some disadvantages as well such as:<ref name="textbook fungi"/>
-<p>Cockroaches are not just a nuisance pest; they are also capable of transmitting a variety of human pathogens and diseases, which makes them a public health concern <ref name="Milner and Pereira"/>. Cockroach infestations are notoriously hard to treat with baited pesticide traps, as they are able to detect and avoid both chemical and microbial pesticides <ref name="Milner and Pereira"/>. Research in the field of cockroach infestation control through the use of biopesticides, specifically fungi, is extremely limited. However, it has been determined that ''Cordyceps blattae'' is highly pathogenic for the German cockroach (''Blattella germanica'') <ref name="Milner and Pereira"/>. Unfortunately, dead cockroaches infected with fungi are not cannibalized, which suggests avoidance by the healthy cockroaches <ref name="Milner and Pereira"/>. </p>+* Can be very costly to produce for commercial use and in quantity;
 +* They can have a short shelf life;
 +* The pest must be present before the pathogen can be usefully applied thus making preventative treatment difficult.</P>
 +Furthermore, the disadvantages of the use of entomogenous fungi as biocontrol agents against arthropod pests has been lacking by the need for specific environmental conditions (humidity over 80% and above) during the prolonged period in which the fungi are required to have spores germinate and then penetrate the surface of the arthropods cuticle. This is different from bacterial pathogens in which they go through the host’s digestive tracts.<ref name="textbook fungi"/> Researchers have been trying to overcome this problem through the development of oil based and other formulations of fungal spores for use in biological control. Thus we can see that a intricate set of interacting processes, both environmental and biotic, are necessary for or inhibitory to development of epizootics caused by entomopathogenic fungi. These include microbial antagonists; host behavior, physiological condition, pathogen vigor and age; presence of pesticides; and appropriate temperature, humidity, and inoculum thresholds.<ref name="Lacey et al"/>
 +==Novel Uses of Mycopesticides==
 +Throughout history, there have been many instances of harmful pesticides being released into the environment without fully understanding the implications of such actions. One example of this was the use of DDT, a chemical that has an extremely long half-life and a high degree of bioaccumulation directly linked to a high lipophilicity, which causes eggshell thinning in several birds, including birds of prey <ref name ="DDT"> Tucker, R.K. & Haegele H.A. (1970). Eggshell Thinning as Influenced by Method of DDT Exposure. ''Bulletin of Environmental Contamination and Toxicology.'' 5(3), 191-4.</ref>. With global environmental awareness increasing, there has been a greater demand for less harmful pesticides. Alternatives to pesticides, biopesticides, generally have a lower toxicity, increased safety, as well as a high efficacy in pest control <ref name ="Market Publishers"> Market Publishers. (2010). ''Global Biopesticides Market Trends & Forecasts (2012 - 2017).'' Birmingham: Market Publishers Report Database.</ref>.
-[[Image:Fireant.jpg|thumb|left|150px|The Southern Fire Ant, ''Solenopsis xyloni'', an invasive species]]'''''Ants'''''+Consequently, the demand and commercial application of microbial biopesticide has increased proportionally to 0.98% of the total worldwide pesticide market, growing at a rate of just over 13% per annum, representing an increase of 47% between 2004/2005 and 2007/2008. Fungal biopesticides represent a large proportion of the rapidly growing sector of pest management <ref name="CPL"/>.
-<p>Black or brown ants are those which may invade houses, causing no structural or other types of damage, but annoy homeowners <ref name="Milner and Pereira"/>. Ants that live inside structures may also include Carpenter ants that can damage the structural integrity of buildings <ref name="Milner and Pereira"/>. Ants which are a problem in urban landscapes, such as fire ants (''Solenopsis xyloni'') may attack humans or domestic animals, damage landscape plants or other materials outside human structures <ref name="Milner and Pereira"/>. ''B. bassiana'' conidia have been effectively used to control the southern fire ant and reduce their populations in urban environments <ref name="Milner and Pereira"/>. [[Definitions|Mycelium]] stage of ''M. anisopliae'' (pre-sporulation) is readily taken back to the nest by black and brown ants, where it will later produce spores <ref name="Lenz"/>.</p>+
-[[Image:Termite.jpg|thumb|left|150px|Dry-wood termites can cause considerable damage to wooden structures]]'''''Termites'''''+Several commercial applications of mycopestocides have been investigated, as described below.
-<p>Termites are social insects living in colonies comprising of a king and queen together with numerous workers and soldiers. Their life cycle is [[Definitions|hemimetabolous]] with the queen laying eggs that hatch into larvae that can develop into workers, soldiers or new reproductives <ref name="Lenz"/>. It is necessary to deposit large quantities of ''M. anisopliae'' conidia within the central portion of the nest to infect and kill the entire termite colony <ref name="Grace">Grace, JK. 1997. Biological control strategies for suppression of Termites. ''J. Agric. Entomol''. 14(3):281-289</ref>. Control of colonies of pest species of termite can be achieved within 3 months with a single treatment between 1-10g of conidia applied directly to the nest, and spores will remain active in nests for at least 2 years <ref name="Lenz"/>.</p>+===''Culicoides nubeculosus'' Biting Midge and the Bluetongue virus===
 +[[Image:Bluetongue.png|thumb|right|250px|Sheep infected with Bluetongue Virus<ref>P. Mellor, Institute of Animal Health, bluetonguevirus.org</ref>]]
-<p>With application of fungi as a biopesticide, termite workers may be found in the nest for many weeks after most of the colony has been killed. There is a possibility that the colony can produce neotenic reproductives and can recover despite the loss of the royal pair <ref name="Milner and Pereira"/>. The use of fungi for elimination of a termite colony may not be effective, but perhaps the greatest potential for application of biological control to suppression of termite populations is the use of pathogenic microorganisms or conidia as deterrents to protect timber <ref name="Lenz"/> <ref name="Grace"/>.</p>+The Bluetongue Virus (BTV), transmitted through populations of cattle by ''Culicoides nubeculosus'' causing Bluetongue disease, is a potentially death-inducing virus<ref name="Mellor"> Mellor, P.S., & Wittmann, E.J. (2002). Bluetongue Virus in the Mediterranean Basin 1998±2001. ''The Veterinary Journal,'' (164), 20-37.</ref><ref name="Ansari"> Ansari. M.A., Pope, E.C., Carpenter, S., Scholte, E., & Butt T.M. (2011). Entomopathogenic Fungus as a Biological Control for an Important Vector of Livestock Disease: The Culicoides Biting Midge. ''PlosOne,'' 6(1), 1-8.</ref>. When an animal is infected, it can develop a fever, high salivation, and cyanosis of the tongue, hence the name 'Bluetongue disease'. Losses have been estimated of causing upwards of $3 billion per year<ref name="Mellor"/>.
-'''''Problems'''''+BTV awareness has increased in recent years due to expansion of its range into newly colonized areas, exacerbated by climate change. As a result of the expanding range of the virus, direct losses from sheep and cattle as well as indirect losses from quarantining have caused great economical losses. Although vaccination can eventually control an outbreak such as this, a vaccine for a recent outbreak only made it into the field 18 months after the initial epidemic<ref name="Ansari"/>. [[Species|''Metarhizium anisopliae'']] strain V275 and several other fungi were tested to see their efficacy of control on ''Culicoides nubeculosus'', the disseminator of BTV. ''M. anisopliae'' strain V275 ensured a 100% mortality rate within the colonies of the biting midge, with sporulation occurring on the midge cadavers<ref name="Ansari"/>. The high infectiousness and virulence of the fungi will limit the amount of blood meals taken, therefore reducing the probability of the disease vector being acquired and dispersed. ''Metarhizium anisopliae'' strain V275 is already commercially available in the USA, sold as F52 by Novozymes<ref name="Ansari"/>. The initial infrastructure requirement is low, so this method can be a cost-effective way of limiting arboviruses. The fungi poses no risk to other wildlife, including humans, birds, fish, and the like, so environmental damage is limited to the pest itself. Although the fungus shows great efficacy as a method of biting midge management, further large-scale trials are needed to better determine the possibility as well as method of using ''Metarhizium anisopliae'' strain V275 as a mycoinsecticide in a commercial environment<ref name="Ansari"/>.
-<p>Though the potential for control is greatest in the social insects, there are a number of behavioural traits of ants and termites that limit the effectiveness of fungi as pest control. These behaviours include nest hygiene, self-grooming and [[Definitions|allogrooming]], and isolation of affected individuals; all of which can either remove or spread entomopathogens <ref name="Milner and Pereira"/>. Similarly, due to the low genetic diversity between individuals in social insect nests, effectiveness of pathogens can be either compromised or enhanced <ref name="Milner and Pereira"/>. For these reasons, the objective of control tactics must be the elimination of the reproductive potential of the nest <ref name="Milner and Pereira"/>.</p>+
-'''''Development'''''+===Adult Housefly management in poultry farms using ''Beauveria bassiana''===
-<p>Entomopathogens formulated as biopesticides have to be mass produced and applied in large amounts <ref name="Milner and Pereira"/>. Strains selected have to be virulent, be able to tolerate temperatures above 30C, pose no health threats to humans and higher animals, and have long-lived and robust spores <ref name="Lenz"/>. Additionally, quality control, shelf life and field longevity of biological agents as pest control must be assessed in order for biopesticides to become marketable <ref name="Lenz"/>.</p>+[[Image:Bassiana.jpg|thumb|right|250px|Fly colonized by ''Beauveria bassiana''<ref>N. Meyling, Sevas Educational Society, sbioinformatics.com</ref>]]
-<p>Since pathogens normally have little or no mobility on their own, a mobile or readily distributed and possibly self-perpetuating control agent should mean more complete and less labor intensive pest control <ref name="Grace"/>. Entomopathogens are naturally slow-acting and therefore ideal for use in bait stations, where the forager is not immediately killed <ref name="Milner and Pereira"/>. This allows the active ingredient to be transferred to nest mates, juveniles and reproductives in the nest <ref name="Lenz"/>. ''B. bassiana'' shows the highest potential for control of several urban pests <ref name="Lenz"/>. Powders and granular baits are recommended for application directly onto the nest, if it can be located <ref name="Lenz"/>. If the nest cannot be located, it is recommended to use bait stations that contain pathogenic fungal conidia mixed with an attractant, such that the foragers return to the nest with the pathogen <ref name="Grace"/>.</p>+The common housefly is a constant pest to most people, having a short life cycle and being able to reproduce large amounts of offspring<ref name="Axtell"> Axtell, R.C., (1999). Poultry integrated pest management: Status and future. ''Integrated Pest Management Reviews,'' (4), 53–73.</ref>. Poultry manure, being slightly moist, is an optimal habitat for the development of large populations of the housefly. When flies occur in a large-scale, high rise, caged-layer poultry facilities, exponential population expansion can occur<ref name="Kaufman"> Kaufman, P.E., Reasor, C., Rutz, D.A., Ketzis J.K., & Arends, J.J. (2005). Evaluation of Beauveria bassiana applications against adult house Xy, Musca domestica, in commercial caged-layer poultry facilities in New York state. ''Biological Control,'' (33), 360-367.</ref>. The large numbers of flies are potentially released into the environment, causing distress for neighbours and local businesses<ref name="Kaufman"/>. Flies also carry several pathogenic organisms affecting humans, poultry and several other species, and can act as a vector for their dispersal<ref name="Axtell"/>. The defecation and regurgitation that flies perform can also cause blotching on the structure and equipment in the poultry farm, on the light fixtures, and also on the eggs<ref name="Axtell"/>, therefore reducing the potential marketability of the final product.
-<p>Integrated pest control measures are needed whereby the pathogen provides short-term cessation of damage while changes are made to the environment to make the building less attractive to pests. Alternatively pathogens could be used in baits over a long period of time to eliminate the colony <ref name="Milner and Pereira"/>. However, both the insect and the environments in which they live can significantly limit the success of fungal pathogens <ref name="Lenz"/>.</p>+As an attempt to reduce the amount of flies present in a commercial poultry facility, a commercially available product, balEnce, was used to determine the efficacy [[Species|''Beauveria bassiana'']] as a mycopesticide<ref name="Kaufman"/>. Since the fungus is non-pathogenic to humans, poultry, and many other organisms, ''B. bassiana'' was sprayed throughout a facility. This allowed efficacy of ''B. bassiana'' to be compared to another well-known insect neurotoxin, pyrethrin<ref name="Kaufman"/>. Adult fly populations were lower in the ''B. bassiana'' facility vs. the pyrethrin facility, as well as reducing the number of fly larvae by over one-half when compared to the pyrethrin<ref name="Kaufman"/>. When the fungi is applied in the manure pits, it colonizes the pit completely and is applied when flies are breeding, killing any flies that come in contact with the manure, as well as any emerging from it. The use of ''B. bassiana'' also does not negatively impact the levels of beneficial insects in the population, many of which are natural enemies of the fly<ref name="Kaufman"/>. With adulticides like pyrethrin, the chemical does not affect the larvae; therefore, repeated applications are required<ref name="Kaufman"/>. Therefore, it can be determined that fewer applications of ''B. bassiana'' will lead to a better pest management system, reducing costs. ''B. bassiana'' answers the questions on the minds of many large-scale poultry producers: how to kill large amounts of household flies without pesticide residue to damaging my livestock<ref name="Kaufman"/>.
-=== Pro et Contra===+==Species==
-===Novel Uses of Fungi as Biopesticides===+''See main article'': [[Species]]
-==[[Species|Species]]==+Effective pest management relies on the appropriate matching of a target species with the pathogenic fungal species<ref>Singh, A., Parmar, N., & Kuhad, R.C. (2011). ''Bioaugmentation, biostimulation and biocontrol.'' (28th ed.). Noida (India): Springer</ref>. Many species and target combinations have been included within the wiki and are summarized in the article above.
-==[[Definitions|Definitions]]==+==Definitions==
 +''See main article'': [[Definitions]]
==Notes and References== ==Notes and References==
<references/> <references/>

Current revision

Lecanicillium lecanii on hemlock woolly adelgid cadaver
Lecanicillium lecanii on hemlock woolly adelgid cadaver[1]
Beauveria bassiana on pecan weevil (Curculio caryae)
Beauveria bassiana on pecan weevil (Curculio caryae)[2]
Trichoderma spp. on Norway spruce (Picea abies)
Trichoderma spp. on Norway spruce (Picea abies)[3]

Fungi are useful biological controls for pests in crops and urban environments and have been commercially available since 1981.[4] They can be used as herbicides, insecticides and fungicides. Mycopesticides are interesting organic alternatives to chemical pesticides due to their enhanced safety and their high specificity for the non-targeted species such as humans, pets and desired insects.[5] They share advantages and disadvantages with other biopesticides.[5] Lecanicillium lecanii, Beauveria bassiana and Trichoderma spp. are amongst the most commonly used species and have been widely studied in spite of the short history of this field of study. With the growing enthusiasm for organic commercial farming and justified environmental concerns in the population, mycopesticide have a definitely a bright future.[6]

Contents

[edit] History

For over 100 years, scientists have been studying many different species of fungi with entomopathogenic characteristics but the use of these organisms in the field has been relatively elusive. Many of these fungal organisms are not only entomopathogens, but also aid in general plant growth. Since the late 1940's, chemical insecticides have been the main tool that growers have utilized, in order to control potentially damaging pests and diseases. Some of the earliest recorded studies of entomopathogenic fungi occurred in the early 1800 when the silkworm industry in France was devastated by Beauveria bassiana. As a consequence of this insight, further research began looking into the use of fungal insect pathogens to manage a wide range of pests.[7]


Most recently, DNA sequence analysis has lead to a much better understanding between the differences in entomopathogenic fungi and other types of fungi. This new research has revealed that some of these entomopathogenic fungi also play the role of beneficial rhizosphere associates, plant growth promoters, as well as plant endophytes. These findings have now lead to further research to determine if some of these entomopathogenic fungi can be used for more than just one purpose.


The use of Entomopathogenic fungi in pest management covers a wide variety of Eumycota subdivisions. These subdivisions of Eumycota include; Mastigomycotina, Zygomycotina, Ascomycotina,and Deuteromycotina.[4] In 1981, the first mycoinsectiside registered in the U.S. was Hirsutella thompsonii and was given the registered name; Mycar.[8] This species has been noted to cause epizootics as early as 1920 in some species of spider mites.[4]

With the rising cost of synthetic chemical pesticides and increasing cases of pesticide resistance, the search for natural biologically based forms of pest management has been a key area of research for the last decade. With the world population rising over 7 billion and the drastic effects of climate change, the need for alternative forms of pest control will become essential. Although insects are key in the performance of many ecosystems, they play a large role in the $35 billion worth of crop that is lost in the United States each year.[9]


There is enormous potential for pest and vector control in pest management in the future due to an increasing amount of knowledge in the area of mycology.[10]

[edit] Types of Mycopesticides

[edit] Mycoherbicide

See main article: Fungi used as a Herbicide

Fungi can be used as an herbicide to exterminate specific weeds and other plants that are detrimental to the effective growth of crops. In order to use a fungus for this purpose, a mycoherbicide is created with the fungi as its active ingredient. The use of mycoherbicides focus on the theory of the disease triangle. The disease triangle breaks down the conditions that need to be apparent for infection to occur, those being the environment, pathogen and host. The following will fall under one or more of these categories. Mycoherbicides act by releasing phytopathogens to suppress weed growth. The phytopathogens release phytotoxins that can kill weeds in up to five weeks time. A common phytotoxin found in the fungi species Penicillium is vulculic acid.[11] The fungi species most commonly used as herbicides in North America are Collectrichum gloesporioides (Collego®) and Phytophthora palmivoraa (De Vine®).[11] A specific weed strain commonly known as arrowhead (Sagitaria trifolia) is the cause of the largest rice plantation problem in Iran. Arrowhead is notably resistant to chemical herbicides. Collego® is used in the USA as the solution to arrowhead.[12] Other types of fungi that can be used as mycoherbicides are rusts and mildews. These fungi use spore distribution to cause infection in their host plants.

[edit] Mycoinsecticide

See main article: Fungi used as a Insecticide

Certain species of fungi can act as parasites of insect. When a fungus is used as insecticides, it is called mycoinsecticide. [13] In recent years, crop protection has been trending towards integrated pest management (IPM) using bacteria and fungi as insecticides. Approximately 750 species of fungi are pathogenic to insects and only 12 are being utilized as insecticides.[13] Two prominent species of fungi used as insecticides are Beauveria bassiana, and Metarhizium anisopliae. Mycoinsecticides function by first being applied to the insects in spray form. The fungi then use their hyphae to burrow into the insects. The hyphae spread the insectotoxins throughout the insect to activate them, eventually leading to the insect’s death.

[edit] Mycofungicide

See main article: Fungi used as a Fungicide

There are many cases of mycoparasitism in nature. In fact, a large amount of dormant spores like sclerotia or chlamydospore are found in the wild already parasitized by another fungi.[14] This is made possible by the ability of most fungi to release lytic enzymes like chitinase that allows them to break down chitinuous cell walls. The parasitism of a fungi on another one occurs generally through four main stages: chemotaxi, recognition, attachment and penetration.[15]The type of relationship that will follow such an invasion will be either necrotrophic or biotrophic.[5] The most commonly used species are in the Trichoderma genus.[16]

[edit] Mycopesticides in Urban Environments

See main article: Fungi uses in Urban Environments

Entomopathogens are key components for integrated pest management solutions.[17] There is a large market for urban pest control that has been dominated by the use of pesticides and insecticides.[17] These chemical pesticides pose a danger not only to the person applying them, but other people, or pets, which may come into contact with the chemicals in an urban environment.[17] Previously used in agricultural settings as deterrents, fungi such as Beauveria bassiana, Metarhizium anisopliae and Paecilomyces fumosoroseus may have possible pest control applications in urban settings as well.[18]

[edit] Advantages and Disadvantages

See main article: Advantages and Disadvantages

The negative impacts of chemical pesticides is what has guided new attention towards increased emphasis on biological control agents with respect to using fungi through the use of naturally occurring pathogens of arthropods. Many of these naturally occurring pathogens produced by fungi are already being employed, mostly on a small scale controlling arthropod pests in greenhouse crops, orchards, ornamentals, turf and lawn grasses, stored products and forests, and also for moderation of vectors of animal and human diseases.[19]


Although the utilization of the full potential of fungal biocontrol agents against arthropod pests has had limited commercial success, the major advantages of these biocontrol agents are:[5]

  • Effectiveness and potentially high specificity;
  • Acceptable naturalness;
  • Safety for humans and other non target organisms;
  • Reduced chemical pesticide use and consequential reduction of residues in food and the environment;
  • Protection of natural enemies of the pest;
  • Protection of biodiversity in managed ecosystems.

Despite these promising advantages of using fungi as biological control agents in pest management, there are some disadvantages as well such as:[5]

  • Can be very costly to produce for commercial use and in quantity;
  • They can have a short shelf life;
  • The pest must be present before the pathogen can be usefully applied thus making preventative treatment difficult.

Furthermore, the disadvantages of the use of entomogenous fungi as biocontrol agents against arthropod pests has been lacking by the need for specific environmental conditions (humidity over 80% and above) during the prolonged period in which the fungi are required to have spores germinate and then penetrate the surface of the arthropods cuticle. This is different from bacterial pathogens in which they go through the host’s digestive tracts.[5] Researchers have been trying to overcome this problem through the development of oil based and other formulations of fungal spores for use in biological control. Thus we can see that a intricate set of interacting processes, both environmental and biotic, are necessary for or inhibitory to development of epizootics caused by entomopathogenic fungi. These include microbial antagonists; host behavior, physiological condition, pathogen vigor and age; presence of pesticides; and appropriate temperature, humidity, and inoculum thresholds.[19]

[edit] Novel Uses of Mycopesticides

Throughout history, there have been many instances of harmful pesticides being released into the environment without fully understanding the implications of such actions. One example of this was the use of DDT, a chemical that has an extremely long half-life and a high degree of bioaccumulation directly linked to a high lipophilicity, which causes eggshell thinning in several birds, including birds of prey [20]. With global environmental awareness increasing, there has been a greater demand for less harmful pesticides. Alternatives to pesticides, biopesticides, generally have a lower toxicity, increased safety, as well as a high efficacy in pest control [21].

Consequently, the demand and commercial application of microbial biopesticide has increased proportionally to 0.98% of the total worldwide pesticide market, growing at a rate of just over 13% per annum, representing an increase of 47% between 2004/2005 and 2007/2008. Fungal biopesticides represent a large proportion of the rapidly growing sector of pest management [6].


Several commercial applications of mycopestocides have been investigated, as described below.

[edit] Culicoides nubeculosus Biting Midge and the Bluetongue virus

Sheep infected with Bluetongue Virus
Sheep infected with Bluetongue Virus[22]

The Bluetongue Virus (BTV), transmitted through populations of cattle by Culicoides nubeculosus causing Bluetongue disease, is a potentially death-inducing virus[23][24]. When an animal is infected, it can develop a fever, high salivation, and cyanosis of the tongue, hence the name 'Bluetongue disease'. Losses have been estimated of causing upwards of $3 billion per year[23].

BTV awareness has increased in recent years due to expansion of its range into newly colonized areas, exacerbated by climate change. As a result of the expanding range of the virus, direct losses from sheep and cattle as well as indirect losses from quarantining have caused great economical losses. Although vaccination can eventually control an outbreak such as this, a vaccine for a recent outbreak only made it into the field 18 months after the initial epidemic[24]. Metarhizium anisopliae strain V275 and several other fungi were tested to see their efficacy of control on Culicoides nubeculosus, the disseminator of BTV. M. anisopliae strain V275 ensured a 100% mortality rate within the colonies of the biting midge, with sporulation occurring on the midge cadavers[24]. The high infectiousness and virulence of the fungi will limit the amount of blood meals taken, therefore reducing the probability of the disease vector being acquired and dispersed. Metarhizium anisopliae strain V275 is already commercially available in the USA, sold as F52 by Novozymes[24]. The initial infrastructure requirement is low, so this method can be a cost-effective way of limiting arboviruses. The fungi poses no risk to other wildlife, including humans, birds, fish, and the like, so environmental damage is limited to the pest itself. Although the fungus shows great efficacy as a method of biting midge management, further large-scale trials are needed to better determine the possibility as well as method of using Metarhizium anisopliae strain V275 as a mycoinsecticide in a commercial environment[24].

[edit] Adult Housefly management in poultry farms using Beauveria bassiana

Fly colonized by Beauveria bassiana
Fly colonized by Beauveria bassiana[25]

The common housefly is a constant pest to most people, having a short life cycle and being able to reproduce large amounts of offspring[26]. Poultry manure, being slightly moist, is an optimal habitat for the development of large populations of the housefly. When flies occur in a large-scale, high rise, caged-layer poultry facilities, exponential population expansion can occur[27]. The large numbers of flies are potentially released into the environment, causing distress for neighbours and local businesses[27]. Flies also carry several pathogenic organisms affecting humans, poultry and several other species, and can act as a vector for their dispersal[26]. The defecation and regurgitation that flies perform can also cause blotching on the structure and equipment in the poultry farm, on the light fixtures, and also on the eggs[26], therefore reducing the potential marketability of the final product.

As an attempt to reduce the amount of flies present in a commercial poultry facility, a commercially available product, balEnce, was used to determine the efficacy Beauveria bassiana as a mycopesticide[27]. Since the fungus is non-pathogenic to humans, poultry, and many other organisms, B. bassiana was sprayed throughout a facility. This allowed efficacy of B. bassiana to be compared to another well-known insect neurotoxin, pyrethrin[27]. Adult fly populations were lower in the B. bassiana facility vs. the pyrethrin facility, as well as reducing the number of fly larvae by over one-half when compared to the pyrethrin[27]. When the fungi is applied in the manure pits, it colonizes the pit completely and is applied when flies are breeding, killing any flies that come in contact with the manure, as well as any emerging from it. The use of B. bassiana also does not negatively impact the levels of beneficial insects in the population, many of which are natural enemies of the fly[27]. With adulticides like pyrethrin, the chemical does not affect the larvae; therefore, repeated applications are required[27]. Therefore, it can be determined that fewer applications of B. bassiana will lead to a better pest management system, reducing costs. B. bassiana answers the questions on the minds of many large-scale poultry producers: how to kill large amounts of household flies without pesticide residue to damaging my livestock[27].

[edit] Species

See main article: Species

Effective pest management relies on the appropriate matching of a target species with the pathogenic fungal species[28]. Many species and target combinations have been included within the wiki and are summarized in the article above.

[edit] Definitions

See main article: Definitions

[edit] Notes and References

  1. Svetlana Y. Gouli, University of Vermont, Bugwood.org
  2. Louis Tedders, USDA Agricultural Research Service, Bugwood.org
  3. Andrej Kunca, National Forest Centre - Slovakia, Bugwood.org
  4. 4.0 4.1 4.2 Rechcigl, E.J., & Rechcigl N.A. (2000). Biological and Biotechnological Controls of Insect Pests. (1st ed.). Boca Ranton (FL): Lewis.
  5. 5.0 5.1 5.2 5.3 5.4 5.5 Moore, D., Robson, G.D., & Trinci, A.P.J. (2011). 21st Century Guidebook to Fungi. (1st ed.) New York (NY): Cambridge
  6. 6.0 6.1 CPL Business Consultants. (2010). The 2010 Worldwide Biopesticides Market Summary. Wallingford: CAB International.
  7. Vega, F.E., Goettel, M.S., Blackwell, M., Chandler, D., Jackson M.A., Keller, S., et al. (2009). Fungal entomopathogens: new insights on their ecology. Fungal Ecology 2(4), 149-59.
  8. Kenneth, R., Muttath, T.I. & Gerson, U. (1979). Hirsutella thompsonii, a fungal pathogen of mites. I. Biology of the fungus in vitro. Annals of Applied Biology, 91(1), 21–28.
  9. Shah, P.A., & Pell, J.K. (2003). Fungi as Biological Control Agents. Applied Microbiology and Biotechnology, 61(5-6), 413-23.
  10. Seigfried, K. (1998). Use of Fungi for Pest Control in Sustainable Agriculture. Phytoprotection, 79(4), 56-60.
  11. 11.0 11.1 Misra, H.P. (2005). Weed Management Through Fungal Herbicides. Orissa Review. 53-56
  12. Motlagh, M.R.S. & Javadzadeh, A. (2010). Study of Alternaria pellucida as a promising mycoherbicide for controlling Arrowhead (Sagitaria trifolia) in paddy fields. Proteomics Journal, 3(6), 172-6.
  13. 13.0 13.1 Wraight, S.P. & Carruthers, R.I. (1998). Production, Delivery, and Use of Mycoinsecticides for Control of Insect Pests on Field Crops. Methods in Biotechnology, (5), 233-69.
  14. Jeffries, P. (1995). Biology and Ecology of Mycoparasitism. Canadian Journal of Botany. 73(1), 1284-1290.
  15. Boland, G. J., & Kuykendall, L. (1998). Plant-microbe interactions and biological control. (1st ed.). New York (NY): Marcel Dekker
  16. Cheng, C., Yang, C., & Peng, K. (2012). Antagonism of Trichoderma harzianum ETS 323 on Botrytis cinerea Mycelium in Culture Conditions. Phytophatology 102(11), 1054-63.
  17. 17.0 17.1 17.2 Milner, R.J. & Pereira R.M. (2007). Microbial control of urban pests - cockroaches, ants and termites. Field Manual of Techniques in Invertebrate Pathology, 20(2), 695-711
  18. Lenz, M. (2005). Biological control in termite management: the potential of nematodes and fungal pathogens. Proceedings of the Fifth International Conference on Urban Pests. 47-52
  19. 19.0 19.1 Lacey, L.A., Frutos, R., Kaya, H.K., & Vail, P. (2001). Insect Pathogens as Biological Control Agents: Do They Have a Future? Biological control, 21(3), 230-48
  20. Tucker, R.K. & Haegele H.A. (1970). Eggshell Thinning as Influenced by Method of DDT Exposure. Bulletin of Environmental Contamination and Toxicology. 5(3), 191-4.
  21. Market Publishers. (2010). Global Biopesticides Market Trends & Forecasts (2012 - 2017). Birmingham: Market Publishers Report Database.
  22. P. Mellor, Institute of Animal Health, bluetonguevirus.org
  23. 23.0 23.1 Mellor, P.S., & Wittmann, E.J. (2002). Bluetongue Virus in the Mediterranean Basin 1998±2001. The Veterinary Journal, (164), 20-37.
  24. 24.0 24.1 24.2 24.3 24.4 Ansari. M.A., Pope, E.C., Carpenter, S., Scholte, E., & Butt T.M. (2011). Entomopathogenic Fungus as a Biological Control for an Important Vector of Livestock Disease: The Culicoides Biting Midge. PlosOne, 6(1), 1-8.
  25. N. Meyling, Sevas Educational Society, sbioinformatics.com
  26. 26.0 26.1 26.2 Axtell, R.C., (1999). Poultry integrated pest management: Status and future. Integrated Pest Management Reviews, (4), 53–73.
  27. 27.0 27.1 27.2 27.3 27.4 27.5 27.6 27.7 Kaufman, P.E., Reasor, C., Rutz, D.A., Ketzis J.K., & Arends, J.J. (2005). Evaluation of Beauveria bassiana applications against adult house Xy, Musca domestica, in commercial caged-layer poultry facilities in New York state. Biological Control, (33), 360-367.
  28. Singh, A., Parmar, N., & Kuhad, R.C. (2011). Bioaugmentation, biostimulation and biocontrol. (28th ed.). Noida (India): Springer
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