Agricultural compositions and methods

Abstract

The present invention relates to compositions and kits for treating or preventing bacterial diseases in terrestrial plants or animals, said compositions and said kits comprising a bacteriophage and / or an immunogenic agent and a flying insect attractant. The invention also relates to a method for attracting flying insects to bacteriophages and / or immunogenic agents. The invention also relates to the use of flying insects in methods of transferring bacteriophage and / or immunogenic agents to animals or plants, and methods of treating or preventing diseases in animals and plants using flying insects. The invention also relates to formulations of bacteriophages and / or immunogenic agents for the treatment and / or prevention of diseases in animals or plants.

Description

Technical Field

[0001] The present invention relates to compositions and kits for treating or preventing bacterial diseases in terrestrial plants or animals, the compositions and kits comprising bacteriophages and / or immunogenic agents and flying insect attractants. The present invention also relates to methods for attracting flying insects to bacteriophages and / or immunogenic agents. The present invention also relates to the use of flying insects in methods for transferring bacteriophages and / or immunogenic agents to animals or plants, and methods for treating or preventing diseases in animals and plants using flying insects. The present invention also relates to formulations of bacteriophages and / or immunogenic agents for treating and / or preventing diseases in animals or plants. Background Art

[0002] Flying insects such as flies are known to transmit bacterial, parasitic and viral diseases to animal and plant species. Flies ingest and carry microorganisms from surfaces they come into contact with, including, inter alia, human and animal excreta and decaying organic material. When flies subsequently come into physical contact with animals or plants, they transmit the microorganisms they carry to their animal and plant hosts, thereby acting as mechanical vectors of microorganisms.

[0003] An example of a fly that is a common carrier of bacterial diseases is the autumn housefly (Musca autumnalis), which is commonly referred to as the face fly and is a pest species of bovine and equine animals, native to Europe and Central Asia, and found in much of Canada and the United States. Other members of the Muscidae family include the Australian bush fly (Muscavetustissima) and the buffalo fly (Haematobia exigus), flies that can act as carriers of bacterial diseases in southern hemisphere regions such as Australia. Face flies emerge from their wintering grounds in the spring, and female flies lay eggs in fresh cow dung. Depending on the temperature, the life span of adult flies is 20 to 50 days, and from early autumn, as the surviving flies prepare to winter, the insect load on bovine and equine animals decreases. Although fruit flies can migrate several kilometers in search of hosts, once they find a suitable habitat, they tend to gather around cattle, especially near watering places.

[0004] Face flies, especially females, feed on the blood, sweat, saliva, nasal mucosa, feces and tears of cattle and horses. Feeding occurs only during daylight hours, meaning that face flies leave the herd each evening and redistribute each morning. During the summer daytime, the number of flies typically recorded on the face of each cattle is about 5 to 30.

[0005] One disease for which face flies often act as vectors is infectious bovine keratoconjunctivitis (IBK), the most common eye disease in cattle, also known as New Forest Eye and Pinkeye. Its primary etiology is the Gram-negative rod-shaped bacterium, Moraxella bovis, although other microorganisms including Moraxella catarrhalis, Moraxella bovoculi, and Mycoplasma bovoculi infections have also been implicated as potential causative agents in the pathogenesis of IBK.

[0006] Face flies are common carriers of M. bovis and, therefore, IBK. Specifically, the surfaces of face flies, including their mouthparts, body, and limbs, may be contaminated with M. bovis and remain viable for hours, and M. bovis ingested by face flies can survive in the fly digestive tract for up to 3 days, meaning that face flies carrying M. bovis can cause eye infections in cattle for days via droplets regurgitated from the foregut and midgut.

[0007] The economic impact of this disease is considerable, and IBK is the most common disease affecting beef and dairy cattle and the second most common disease in lactating calves older than three weeks of age.

[0008] Currently, the most commonly used treatment for IBK is antibiotic drugs, which are delivered by systemic injection, subconjunctival injection, intra-eyelid injection or topical application. Alternatively, preventive measures are relied upon when necessary, such as the use of ear tags containing insecticides. However, all of these treatment options are affected by the pharmacokinetics of drug delivery to the ocular surface and the need to handle and restrain the animal in order to administer the treatment or preventive measures. Therefore, there remains a need for an effective method to reduce the incidence of bacterial infections in animals, and specifically to reduce the incidence of face fly-induced IBK in cattle, which method does not require the handling and restraint of the animal.

[0009] Cattle like to rub against fence posts, trees or bushes at certain times of the day, during which they seem to enjoy and be satisfied. Observations of this behavior eventually led to the development of commercially produced brushes and body scrubbers that can be deployed in fields and fences to facilitate this activity. Specifically, rubbing the head causes face flies to temporarily leave the face of the cattle, causing the flies to find another place to rest or remain in the air until they can return to the face. It would be advantageous if this natural tendency of bovines could be used in strategies to mitigate fly-induced IBK or other bacterial infections in bovines.

[0010] Other fly species, including horn flies (Haematobia irritans) and stable flies (Stomoxys calcitrans) are also major pests of bovines. Like the common housefly (Musca domestica) and blowfly (Calliphora vomitoria), these fly species are vectors of bacterial transmission, and horn flies in particular have been implicated in the transmission of bacteria, including Staphylococci, which can lead to mastitis in bovines. Therefore, removing these nuisance flies from bovines and eliminating their bacterial load is beneficial in terms of cattle health and welfare.

[0011] Another host species susceptible to bacterial diseases carried by flying insects is the banana plant (part of the Musaceae family). Banana plants are susceptible to several bacterial diseases, including 'banana bacterial wilt' caused by the bacterium Xanthomonas campestris and 'Moko disease' caused by the bacterium Ralstonia solancearum. Parasitic flies, such as the Oriental fruit fly (Bactrocera dorsalis) and wasps are active in banana plantations and can therefore act as vectors that carry bacterial diseases to banana plants.

[0012] Likewise, olive trees (Olea europaea) are susceptible to bacterial diseases caused by bacteria carried by the olive fruit fly (Bactrocera oleae), including Xylella fastidiosa and Pseudomonas sawantii.

[0013] Honey bees, such as bumble bees (Bombus terrestris audax), can also carry bacterial infections to plant species, such as flowers. Specifically, many plants are susceptible to bacterial blight caused by one of several strains of Pseudomonas syringae, which has a very wide range of host species. Honey bees and other flying insects can carry and spread these pathogens. For example, as a means of increasing fruit yields, commercial beehives are often used to enhance the pollination of fruiting plants such as kiwifruit (Actinidia deliciosa). However, kiwifruit plants are highly susceptible to infection caused by Pseudomonas syringae pv. kiwifruit, which may be carried by honey bees. Therefore, it would be very advantageous to limit the bacterial load of honey bees in commercial beehives and allow them to distribute phages with activity against this pathogen into plants as a means of disease prevention. Currently, agricultural strategies for mitigating bacterial infections in plants rely heavily on introducing artificial chemicals into the soil or applying them to plant species, which destroys the surrounding ecosystem. Therefore, there remains a need for effective methods of reducing the incidence of bacterial infections in plants. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Shown is a top view of Muscidae flies feeding on an attractant consisting of a 50:50 mix of glucose syrup and water.

[0015] Figure 2 a shows a top view of Muscidae flies feeding on an attractant consisting of powdered sugar and a separate water source.

[0016] Figure 2 b shows a side view of a Muscidae fly feeding on an attractant consisting of powdered sugar and a separate water source.

[0017] Figure 3 Shown is the growth of M. bovis on agar plates after the plates were placed in enclosures containing flies, bacteriophage with lytic activity against M. bovis in sucrose solution, or both over the course of a day.

[0018] Figure 4 Shown is the growth of Salmonella enterica on agar plates after the plates were placed in enclosures containing flies, bacteriophage with lytic activity against Salmonella enterica in sucrose solution, or both over the course of a day.

[0019] Figure 5 Shown is the growth of the Salmonella enterica pathogen on agar plates placed in enclosures containing face flies, bacteriophage with lytic activity against Salmonella enterica in a sucrose solution, or both over the course of a day after transfer of the solution used to rinse isolated bovine eyes.

[0020] Figure 6 Shown is the growth of the Salmonella enterica pathogen on agar plates that were placed in enclosures containing honey bees, bacteriophages with lytic activity against Salmonella enterica in a sucrose solution, or both over the course of a day, following transfer of a solution used to rinse freshly cut flowers.

[0021] Figure 7 Shown are transmission electron microscopy images of bacteriophages active against M. bovis.

[0022] Figure 8 Shown are transmission electron microscopy images of bacteriophages active against M. bovis.

[0023] Fig. 9 Shown are transmission electron microscopy images of bacteriophages active against M. bovis.

[0024] Fig.10 Shown are transmission electron microscopy images of bacteriophages active against M. bovis.

[0025] Fig.11 a and 11b show the generation of Fig.10 Aggregates produced after serial passaging of a phage isolate. Summary of the invention

[0026] In a first aspect of the present invention, a method for attracting flying insects to a bacteriophage and / or an immunogenic agent is provided, the method comprising (i) combining the bacteriophage and / or the immunogenic agent with a flying insect attractant, or (ii) placing the bacteriophage or the immunogenic agent within the attraction radius of the flying insect attractant.

[0027] A second aspect of the invention relates to the use of flying insects in a method of transferring a bacteriophage and / or an immunogenic agent to a terrestrial animal or plant, wherein the flying insect acts as a carrier of the bacteriophage and / or immunogenic agent.

[0028] A third aspect of the invention relates to a method for treating or preventing a disease in a terrestrial animal or plant, the method comprising transferring a bacteriophage and / or an immunogenic agent to the animal or the plant using a flying insect, wherein the flying insect acts as a carrier of the bacteriophage and / or immunogenic agent.

[0029] In a fourth aspect of the present invention, there is provided a composition for treating or preventing bacterial diseases in terrestrial plants or animals, the composition comprising: (i) a bacteriophage and / or an immunogenic agent; and (ii) a flying insect attractant, the flying insect attractant being selected from one or more of the following: fecal matter, pheromones, volatile agents, odorous materials, materials that can reflect or emit light within the visible electromagnetic spectrum, materials that reflect or emit electromagnetic radiation outside the visible spectrum, flying insect nutrition sources, and combinations thereof. Also provided is an article comprising the composition of the present invention.

[0030] In a fifth aspect of the invention, a kit for treating or preventing bacterial diseases in terrestrial plants or animals is provided, the kit comprising: (i) a bacteriophage and / or an immunogenic agent; and (ii) a flying insect attractant, the flying insect attractant being selected from one or more of the following: fecal matter, pheromones, volatile agents, odorous materials, materials that can reflect or emit light within the visible electromagnetic spectrum, materials that reflect or emit electromagnetic radiation outside the visible spectrum, a flying insect nutrient source or a resting place to which flying insects are naturally attracted (such as a beehive in the case of bees), and combinations thereof.

[0031] In some embodiments, the present invention relates to the use of bacteriophages. The use of bacteriophages solves the above problems because bacteriophages can fight against bacteria naturally carried by flying insects so as to reduce the possibility of flying insects infecting terrestrial animals and plants. In addition, bacteriophages can treat or control bacteria that have already infected target animals and plants, and then the bacteriophages can be transferred to uninfected organisms by flying insects. In other words, flying insects that usually act as carriers of bacteria that cause diseases in terrestrial animals or plants become carriers of bacteriophages that can fight against bacteria that cause diseases. This provides a targeted method to reduce the incidence of bacterial infections in terrestrial animals and plants without any direct human intervention, including processing, manual manipulation or restriction.

[0032] In some embodiments, the present invention relates to the use of immunogenic agents. The present invention further provides a method for delivering immunogenic agents for stimulating immunogenic responses without the need for direct human intervention. This allows the transfer of immunogenic agents to flying insects by combining the immunogenic agent with a flying insect attractant, or placing the immunogenic agent within the attraction radius of the flying insect attractant. The subsequent transfer of the immunogenic agent to the target animal or plant provides a targeted means to induce a protective response in terrestrial animals and plants without the need for any direct human intervention, including processing, manual manipulation or restriction. It will be understood that immune responses through humoral or cell-mediated mechanisms do not occur in plants as in animals. Nevertheless, it is generally believed that plants have an immune system and are able to induce a protective response in response to pathogen molecules.

[0033] In a preferred embodiment, the bacteriophage and / or immunogenic agent is combined with a flying insect attractant that contains a nutrient source that the flying insect wants to feed on, thereby promoting uptake of the bacteriophage or immunogenic agent. The bacteriophage or immunogenic agent is then transferred to the target organism during regurgitation of the digestive tract contents that normally occurs during the flying insect's feeding activity.

[0034] definition

[0035] As used herein, the term "flying insect" refers to any insect having the ability to fly. Therefore, the term "flying insect" includes all insects having one or more pairs of wings.

[0036] As used herein, the term "fly" refers to any flying insect characterized by the use of only one pair of wings for flight. This includes, but is not limited to, Diptera insects.

[0037] As used herein, the term "Diptera" refers to insects of the order Diptera, and the term "Diptera" refers to a single insect of the order Diptera.

[0038] As used herein, the term "face fly" refers to the Fall Fly.

[0039] As used herein, the term "Australian bush fly" refers to the Australian bush fly.

[0040] As used herein, the term "buffalo fly" refers to the Oriental horn fly.

[0041] As used herein, the term "Oriental fruit fly" refers to Bactrocera dorsalis.

[0042] As used herein, the term "olive fruit fly" refers to the olive fruit fly.

[0043] As used herein, the term "blow fly" refers to any insect from the group Blowfly vomiting or Lucilia sericata.

[0044] As used herein, the term "bumblebee" refers to any insect of the species Bombus terrestris or Bombus terrestris.

[0045] As used herein, the term "syrphid fly" refers to any insect in the family Syrphidae.

[0046] As used herein, the term "horn fly" refers to the western horn fly.

[0047] As used herein, the term "stable fly" refers to stable flies.

[0048] As used herein, the term "immunogenic agent" refers to any agent capable of providing an immune response in a target terrestrial organism.

[0049] As used herein, the term "attractant" refers to any substance that attracts a target animal. Thus, for example, a "flying insect attractant" refers to any substance that attracts flying insects. It will be understood that the attractant can be a substance other than a nutrient source, it can be a substance that contains or contains a nutrient source, or it can be a location to which insects are naturally attracted, such as a beehive.

[0050] As referred to herein, a "terrestrial animal" is an animal that lives primarily or entirely on land. As referred to herein, a "terrestrial plant" is a plant that grows on, in or from land.

[0051] As used herein, the term "comprising" means that at least all the listed elements must be present, but other elements not mentioned may also be present.

[0052] In the context of the present invention, "attraction radius" means the area over which the target animal is attracted to the attractant. For example, for an insect attractant involving the use of odor to attract insects, the attraction radius is the area over which the insect can smell the attractant. It will be understood that the attraction radius for a particular attractant and target animal can be determined by routine experimentation. For example, the attraction radius can be determined by placing the attractant in a test chamber under normal atmospheric conditions and placing the target animal at increasing distances from the attractant to determine the maximum distance at which the target animal is attracted by the attractant.

[0053] As used herein, the term "combination" refers to the combination of X and Y, encompasses mixing of X and Y, and contacting X with Y. In some embodiments, the term "combination" means mixing. In some embodiments, the term "combination" means contacting a first component with a second component.

[0054] As used herein, the terms "IBK," "New Forest Eye," and "pink eye" all refer to infectious bovine keratoconjunctivitis.

[0055] According to the present invention, the term "virus" includes double-stranded or single-stranded RNA or DNA viruses that infect cells of bacteria, plants and / or animals.

[0056] The terms "phage" and "bacteriophage" are used interchangeably herein to refer to bacteriophages, which are viruses that infect specific bacteria.

[0057] According to the present invention, the term "substrate" is understood to mean any solid phase material to which viruses, such as bacteriophages, can be immobilized.

[0058] As used herein, the term "about", when applied to one or more values ​​of interest, refers to a value similar to a specified reference value. In some embodiments, the term "about" refers to a value range that falls within 25% of a reference value (greater than or less than a reference value) in either direction. In some embodiments, the term "about" refers to a value that falls within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less of the reference value (unless such numerical value exceeds 100% of a possible value). DETAILED DESCRIPTION

[0059] The present invention relates to the use of flying insects in a method for transferring bacteriophages and / or immunogenic agents to terrestrial animals or plants, wherein the flying insects act as carriers of the bacteriophages and / or immunogenic agents.

[0060] In addition, the present invention relates to a method for treating or preventing a disease in a terrestrial animal or plant, the method comprising transferring a bacteriophage and / or an immunogenic agent to the animal or plant using a flying insect, wherein the flying insect acts as a carrier of the bacteriophage and / or immunogenic agent. In a preferred embodiment, the method for treating or preventing a disease in a terrestrial animal or plant comprises transferring the bacteriophage and / or immunogenic agent directly to a site of infection (e.g., bacterial infection) on the terrestrial animal or plant.

[0061] The present invention also relates to a method for transferring bacteriophages and / or immunogenic agents to flying insects, the method comprising (i) combining the bacteriophages and / or immunogenic agents with a flying insect attractant; or (ii) placing the bacteriophages and / or immunogenic agents within the attraction radius of a flying insect attractant.

[0062] In the methods and use embodiments of the present invention, the flying insect attractant is preferably a flying insect attractant selected from the following: fecal matter, pheromones, volatile agents, odorous materials, materials that can reflect or emit light within the visible electromagnetic spectrum, materials that reflect or emit electromagnetic radiation outside the visible spectrum, resting places to which flying insects are naturally attracted, nutrient sources or combinations thereof.

[0063] The present invention further relates to a composition for treating or preventing bacterial diseases in terrestrial plants or animals, the composition comprising: (i) a bacteriophage and / or an immunogenic agent; and (ii) a flying insect attractant selected from fecal matter, pheromones, volatile agents, odorous materials, materials that can reflect or emit light within the visible electromagnetic spectrum, materials that reflect or emit electromagnetic radiation outside the visible spectrum, a flying insect nutrient source, or a combination thereof.

[0064] In addition, the present invention relates to a kit for treating or preventing bacterial diseases in terrestrial plants or animals, the kit comprising: (i) a bacteriophage and / or an immunogenic agent; and (ii) a flying insect attractant, the flying insect attractant being selected from fecal matter, pheromones, volatile agents, odorous materials, materials that can reflect or emit light within the visible electromagnetic spectrum, materials that reflect or emit electromagnetic radiation outside the visible spectrum, resting places to which flying insects are naturally attracted, flying insect nutrition sources, or combinations thereof.

[0065] In the context of the present invention, "prevention" or "preventing" bacterial diseases encompasses any means by which a bacteriophage or immunogenic agent transferred to a flying insect can combat bacteria, whether the bacteria are carried by the insect itself or are located on a terrestrial animal or plant. For example, prevention of bacterial diseases can be performed by combating bacteria present on a flying insect with a bacteriophage or immunogenic agent, thereby reducing the bacterial load of the flying insect and preventing the spread of the bacterial disease by the insect. Alternatively, bacterial diseases can be prevented when a flying insect contacts a terrestrial animal or plant, thereby depositing the bacteriophage or immunogenic agent on the terrestrial animal or plant and preventing bacterial infection on the animal or plant.

[0066] In a preferred embodiment, the bacteriophage and / or immunogenic agent is an antimicrobial agent.

[0067] In other preferred embodiments, the bacteriophage and / or immunogenic agent comprises a bacteriophage.More preferably, the bacteriophage is an engineered bacteriophage, ie a bacteriophage that has been prepared for the methods, compositions, kits and uses of the invention.

[0068] Phages are viruses that target specific bacteria, and after infecting their host bacteria, they can reproduce in either a lytic or lysogenic manner. Lytic phages work by infecting bacteria and using it to replicate and produce more viruses. Ultimately, after sufficient viral replication, the lysis of the bacterial cell releases a large number of new phages, which themselves can infect neighboring bacteria. Lysogenic phages work by integrating their DNA into bacterial chromosomes and replicating along the host chromosome to produce new copies of the integrated viral DNA. Advantageously, phages are specific to the bacteria they target, which means they can be used as antibacterial agents in the host without causing damage to other bacteria in the host, unlike less specific antibiotics.

[0069] It will be understood that the technician can isolate phages with activity against specific bacteria. For example, phages can be isolated by first culturing the relevant bacteria and plating them on agar to produce a growth lawn of bacteria. Then, liquid can be collected from a suitable phage source, and the source includes but is not limited to animal tissues and secretions, fecal matter, plant matter, soil or water samples. For example, bovine rumen fluid, nasal secretions and lacrimal gland secretions can be collected to separate phages with activity against bovine Moraxella (a bacterium found in bovine animals), and liquid can be collected from pig fecal matter to separate phages with activity against Salmonella enterica (a bacterium found in porcine animals). The bacterial lawn on agar is then exposed to aliquots of the collected liquid, and for samples where bacterial lysis is observed, phage lysates can be concentrated therefrom.

[0070] In a preferred embodiment, the phage is stabilized. For example, as described in EP 1496919 B1, the phage is stabilized by being fixed to a matrix. This is advantageous because it ensures that the phage retains its antimicrobial activity while also increasing its stability relative to free phage.

[0071] In some embodiments, as described in EP 1496919 B1, the phage is fixed to the substrate via a covalent bond formed between the substrate and the phage. In a preferred embodiment, the phage is fixed via its head, leaving the tail free. This ensures that the phage retains its activity, as the tail of the phage is associated with the ability of the phage to recognize and infect specific bacteria.

[0072] In a preferred embodiment, the flying insect is a Diptera insect. In a particularly preferred embodiment, the Diptera insect is a face fly, Australian bush fly, buffalo fly, horn fly or stable fly. This is advantageous because these flies are pest species to bovines and equines. Therefore, using these flies to carry phages and / or immunogenic agents provides an effective method for targeting bovines and equines with these agents because these flies will transfer the phages or immunogenic agents to bovines and equines. In a particularly preferred embodiment, the Diptera insect is a face fly.

[0073] In some embodiments, the bacteriophage and / or immunogenic agent has activity against bacteria associated with diseases in terrestrial animals or plants. In a preferred embodiment, the bacteriophage and / or immunogenic agent has activity against bacteria associated with diseases in terrestrial animals. In a preferred embodiment, the terrestrial animal or plant is a terrestrial animal. In a particularly preferred embodiment, the terrestrial animal or plant is a bovine or equine animal, preferably a bovine animal, such as a cow or a calf. In a particularly preferred embodiment, the bacteriophage and / or immunogenic agent has activity against pathogens associated with the cause of infectious bovine keratoconjunctivitis. In these embodiments, the bacteriophage and / or immunogenic agent may have activity against one or more of Moraxella bovis, Moraxella catarrhalis, Moraxella bovis and mycoplasma (preferably Moraxella bovis). It will be understood that bacteriophages can have direct activity against bacteria, while immunogenic agents indirectly exert their effects by stimulating an immune response against bacteria. However, it can be said that both have activity against bacterial species, and this will be understood by those skilled in the art. Thus, the methods, kits and uses of the present invention provide an effective means for delivering bacteriophages or immunogenic agents to animals to combat disease-causing pathogens through natural processes without the need for direct human intervention.

[0074] In some embodiments, the phage and / or immunogenic agent has activity against bacteria associated with diseases in terrestrial plants. In particularly preferred embodiments, the phage and / or immunogenic agent has activity against pathogens associated with the cause of bacterial wilt of banana. In these embodiments, the phage and / or immunogenic agent may have activity against Xanthomonas campestris. In other preferred embodiments, the phage and / or immunogenic agent has activity against pathogens associated with the cause of horse disease. In these embodiments, the phage and / or immunogenic agent may have activity against Ralstonia solanacearum. In some embodiments, the flying insect is a wasp. In some embodiments, the flying insect is an oriental fruit fly. In other preferred embodiments, the phage and / or immunogenic agent has activity against Xylella fastidiosa, Pseudomonas syringae, or Pseudomonas savarroa. In some embodiments, the flying insect is an olive fruit fly. In other embodiments, the insect is a honey bee, preferably a bumblebee. Thus, the methods, kits and uses of the present invention provide an effective way to deliver phages and / or immunogenic agents to plants to combat disease-causing pathogens through natural processes, by exploiting the natural movement of insect species across plants, without the need for direct human intervention.

[0075] Additionally, the present invention relates to a method of transferring a bacteriophage and / or an immunogenic agent to a flying insect, the method comprising combining the immunogenic agent with a flying insect attractant, or placing the bacteriophage and / or immunogenic agent within the attraction radius of a flying insect attractant.

[0076] In some embodiments, the attractant comprises one or more of: fecal matter, pheromones, volatile agents, odorous materials, materials that can reflect or emit light within the visible electromagnetic spectrum, materials that reflect or emit electromagnetic radiation outside the visible spectrum, or resting places to which flying insects are naturally attracted. Resting places to which flying insects are naturally attracted include, but are not limited to, brushes, beehives, and surfaces of terrestrial plants and animals. For example, flies are naturally attracted to the faces of cows, and bees are naturally attracted to flowers. It will be understood that, where appropriate, a nutrient source may also act as an attractant. Therefore, the attractant may preferably comprise a combination of two substances: one that is a nutrient source, and one that is not a nutrient source.

[0077] In a preferred embodiment, the composition or kit comprises a flying insect attractant, the flying insect attractant comprises a flying insect nutrient source, or the method comprises combining a bacteriophage and / or an immunogenic agent with a flying insect nutrient source. The nutrient source may be a sugar, preferably glucose or dry ice sugar. In some embodiments, the combination of the bacteriophage and / or an immunogenic agent and a nutrient source is presented in the form of a liquid. In some embodiments, the combination of the bacteriophage and / or an immunogenic agent and a nutrient source is presented in the form of a solid.

[0078] The use of a flying insect attractant comprising a nutrient source is advantageous because it increases the likelihood that the insect will ingest the phage and / or immunogenic agent when the insect is feeding on the nutrient source. This is advantageous because it will ensure that the phage and / or immunogenic agent is transferred to the digestive tract of the flying insect. This is particularly advantageous when the flying insect is a fly because the regurgitation of droplets of intestinal contents is a normal feature of fly feeding activity. Thus, the transfer of the phage and / or immunogenic agent to the animal or plant host is particularly high when the fly is shifting from feeding on a nutrient source to feeding on an animal or plant host.

[0079] Preferably, the bacteriophage and / or immunogenic agent is combined with a flying insect attractant comprising a nutrient source that the flying insects tend to feed on, thereby promoting uptake of the agent by the flying insects.

[0080] Figure 1 An exemplary liquid nutrient source for flies is shown, consisting of a 50:50 mixture of glucose syrup and water. Figure 2 2a and 2b show exemplary solid nutrient sources for flies, consisting of dry ice candy. These figures show flies feeding on each nutrient source, demonstrating that when a bacteriophage and / or immunogenic agent used to treat or control bacteria that cause disease in terrestrial organisms is combined with a nutrient source or within the attractant radius of a nutrient source, it will also be ingested by the fly by ingestion or inhalation or limb contamination.

[0081] Preferred attractants include fecal matter. Using fecal matter as an attractant is advantageous because insects are frequently exposed to fecal matter throughout their life cycle. Specifically, for insect species that lay eggs in feces, including face flies, they are exposed to fecal matter during each egg-laying cycle. Therefore, when fecal matter is used as an attractant, this provides an effective way to ensure that the immunogenic agent is picked up by dipteran insects and is therefore subsequently transferred to a plant or animal host when the dipteran insects move. In a preferred embodiment, the fecal matter includes equine fecal matter.

[0082] In a particularly preferred embodiment, the fecal material comprises bovine fecal material. This is advantageous for use in combination with bacteriophages and / or immunogenic agents that are active against bacteria that cause infections in bovine animals, such as dairy cows. In addition, this is particularly advantageous for insects that lay eggs in bovine fecal material. Taking face flies as an example, assuming that female face flies lay eggs in cow dung, if they are located in or on the cow dung, they are very likely to come into contact with bacteriophages and / or immunogenic agents in the cow dung. In addition, female face flies have a particularly high feeding rate on the blood, sweat, saliva, nasal mucosa, feces and tears of bovine and equine hosts. Therefore, methods involving bovine fecal material are particularly likely to effectively transfer immunogenic agents to cattle hosts via face flies.

[0083] In some embodiments, the bacteriophage and / or immunogenic agent is combined with a flying insect attractant. This is advantageous, for example, where the attractant comprises a flying insect nutrient source, because it ensures that when the flying insect contacts or consumes some of the nutrient source, it will also consume some of the bacteriophage and / or immunogenic agent.

[0084] In other embodiments, the bacteriophage and / or immunogenic agent is placed within the attractant radius of the attractant. This is advantageous in situations where the bacteriophage and / or immunogenic agent cannot be combined with the attractant, because its location within the attractant radius ensures that the bacteriophage and / or immunogenic agent will be transferred to the flying insect as it moves toward the attractant.

[0085] It will be understood that the attraction radius will vary depending on the type, concentration and efficacy of the attractant and the characteristics of the target animal or plant. The attraction radius of a particular combination of attractant and target animal or plant is determined by routine experimentation. For example, the attraction radius can be determined by placing the attractant in a test chamber under normal atmospheric conditions and placing the target animal at increasing distances from the attractant to determine the maximum distance at which the target animal is attracted by the attractant. An example of a study using poultry farms as an attractant to determine the attraction radius of houseflies is found in Nazni WA et al., Trop Biomed. 2005 June; 22(1):53-61.

[0086] In some embodiments, the luring radius can be within about one thousand meters of the attractant. In some embodiments, the luring radius can be within about five hundred meters of the attractant. In some embodiments, the luring radius can be within about one hundred meters of the attractant. In some embodiments, the luring radius can be within about ten meters of the attractant. In some embodiments, the luring radius can be within about five meters of the attractant. In some embodiments, the luring radius can be within about one meter of the attractant.

[0087] It will be appreciated that the optimal concentration of phage or immunogenic agent in combination with the attractant can be determined using challenge studies in which different concentrations are investigated. 2 pfu / ml, preferably >10 3 pfu / ml,>10 4 pfu / ml or most preferably >10 6 In a particularly preferred embodiment, the concentration of phage is about 10 6 pfu / ml.

[0088] In some embodiments, the composition or kit comprises two or more phages and / or immunogenic agents, or the use or method involves the use of two or more phages and / or immunogenic agents. In a particularly preferred embodiment, the composition comprises a combination of two or more phages. It is advantageous to use a combination of phages and / or immunogenic agents because bacteria have evolved many defense mechanisms against viruses and are known to develop resistance. Therefore, it is desirable that the formulation comprises a mixture of two or more active agents to minimize the effects of such resistance.

[0089] In some embodiments, composition of the present invention can be applied to plastic substrate, and described plastic substrate can then be attached to the friction post for animal such as bovine or equine, or be placed in the luring radius of described friction post.Alternately, plastic substrate can be in the form of liquid or liquid glass.When it is liquid preparation, it can be coated, absorbed or dispersed in other materials such as fecal matter.In some embodiments, composition of the present invention can be placed in the housing that protection composition is protected from external conditions such as meteorological conditions.

[0090] For example, the plastic substrate may be attached to a post, tree or other natural or man-made structure that an animal such as a bovine or equine may come into contact with, or placed within their entrapment radius, such as rubbing against or in their environment. The structure may preferably be a brush or body scrubber.

[0091] This is particularly advantageous in the case of bovines, since face flies act as vectors of disease. Specifically, given that face flies are dislodged from the face of a bovine when the bovine rubs its head against a post, tree, or other structure (such as a brush or body scrubber), these flies either remain in the air or seek another roosting site until they can return to the bovine's face. Therefore, combining a bacteriophage or immunogenic agent active against M. bovis with a fly attractant and deploying it near or in conjunction with the rubbing station will facilitate their ingestion by the flies. For example, a face rubbing brush with an integrated fly roosting site will encourage the flies to rest on a reservoir of bacteriophage / immunogenic agent, which contains a reservoir of bacteriophage / immunogenic agent, with or without a fly attractant, when flies are dislodged from the face of a bovine. The flies will then ingest the phage / immunogenic agent and transfer it to the bovine animal's face upon their return, thereby treating or preventing bacterial infections such as IBK. This strategy is beneficial because it exploits the natural behavior of both the cattle and the face flies and does not require direct human intervention.

[0092] Alternatively, applying the phage or immunogenic agent with or without a flying insect attractant directly to a facial rubbing brush will allow the phage / immunogenic agent to be transferred to the face of the cattle without the need for direct human interaction. Additional methods of transferring the phage / immunogenic agent to the face of a bovine without the need for direct human interaction include adding the phage / immunogenic agent with or without a flying insect attractant to the rim of a suitably sized container containing a food source for the bovine. When the muzzle of the bovine comes into contact with the rim of the container, foraging and grasping for food from such a container will ultimately allow the phage / immunogenic agent to be transferred to the face of the bovine. In both cases, the flying insects can then transfer the phage / immunogenic agent from the face of the animal to the area of ​​the animal to be treated, such as the eyes. This is particularly advantageous when a flying insect attractant is present, as this encourages the flying insects to ingest the phage from the face of the cattle and transfer them to the site of infection.

[0093] In some embodiments, the composition of the present invention can be incorporated into a wearable article for a terrestrial animal, such as an ear tag for a bovine or equine animal. Alternatively, the composition of the present invention can be adhered directly or indirectly to the skin of the animal. In these embodiments, if the terrestrial animal is nomadic, the composition will be carried by the terrestrial animal, rather than being fixed in one place. This is advantageous because the composition will move with the animal and continue to attract flying insects to the animal even if the animal moves. Ear tags are particularly advantageous because the composition is located near the animal's face, thereby providing easy access to flying insects that are attracted to the animal's face, such as face flies.

[0094] In all of the above embodiments, the final form of the composition will be in a form that enables flying insects to ingest and uptake the bacteriophage and / or immunogenic agent.

[0095] Example

[0096] Experiments were conducted to investigate the ability of flying insects to carry phages from a source to a distantly located infection site. In these proof-of-principle experiments, the reproducibility and practicality of the concept were demonstrated using three different flying insect vectors and two different phage / pathogen systems. More specifically, experiments were conducted with face flies (Fall Housefly), blowflies (Blowfly vomiting), and bumblebees (Bombus fulvidraco) in combination with phages specific for either Moraxella bovis or phages specific for Salmonella enterica. In the case of experiments involving Salmonella enterica, phages previously isolated against Salmonella enterica serotype S. Typhimurium SL1344 were used. These Salmonella enterica phages were isolated by exposing Salmonella enterica growth lawns on agar to aliquots of liquid collected from pig fecal material that had been suspended in a sterile water buffer, and then observing plaques in the bacterial lawn after a period of incubation at 37°C.

[0097] In the case of Moraxella bovis, the isolation of phages is as follows. Moraxella bovis was purchased from the National Collection of Type Cultures (NCTC 9426) in the United Kingdom and cultured in brain heart infusion (BHI) liquid medium supplemented with 5% horse blood. The bacteria were plated onto BHI agar supplemented with 5% horse blood and the plates were incubated at 37°C for 48 hours. Phages with lytic activity against Moraxella bovis were isolated from samples of bovine rumen fluid and bovine nasal and lacrimal secretions collected from slaughterhouses, from which phage lysates were concentrated. This was achieved by introducing the lysate into Moraxella bovis plates in blood agar and observing bacterial lysis after a further period of incubation. The addition of triphenyltetrazolium chloride to the blood agar plates facilitated the observation of lysis of bacteria by phages. Plaques were collected for enrichment in Moraxella bovis / BHI liquid medium supplemented with 5% horse blood. Four separate cultures were serially passaged to obtain phage clonal populations, which were used for transmission electron microscopy (TEM). Figure 7 , 8 , 9 and 10 show TEM images of four phages isolated with activity against M. bovis. Fig.11 a and 11b show the generation of Fig.10 Aggregates of phage isolates produced after serial passage. It can be seen that all phages in the isolate have the same morphology, consistent with the clonal population after serial passage. Further, it can be seen that each isolated phage has a different morphology. Although Figure 7 The phages in the Fig.10 The phages in the genome have significantly different morphologies. Figure 8 and 9 The phages in have very similar morphologies to each other.

[0098] Example 1

[0099] A model system was established for experiments investigating the ability of flies to carry phages from a source to a distantly located site of infection. In this model, the site of infection is represented by the growth of M. bovis on blood agar or the growth of S. enterica on agar, and the flying insects used are either face flies (Fall flies) or blow flies (Callus vomitus). The face flies used in the experiments were samples captured extensively from the vicinity of cattle. In the case of the blow flies, these were 3-day-old adult flies that had recently hatched from pupae and were incubated at 23°C under a 12:12 light:dark cycle. Bacteriophages with lytic activity against M. bovis or S. enterica were added to a 20% sucrose solution that acted as a fly insect attractant. In the case of the M. bovis experiments, a sucrose solution was used. Figure 7-10 A mixture of the four phages shown. The concentration of phages with lytic activity against M. bovis was about 10 6 pfu / ml, and the concentration of phages with lytic activity against Salmonella enterica was about 10 11 pfu / ml.

[0100] An enclosure was set up containing the phage in a sucrose solution, five adult flies, and an agar plate on which the pathogen was grown, placed away from the sucrose solution. Thus, within the enclosure, the flies were able to feed on the sucrose solution, consuming the phage while doing so, and travel to the infection site on the agar plate located away, where they could feed additionally.

[0101] In addition, other closures were set up as controls containing:

[0102] 1. Control 1: 20% sucrose solution without phage but containing flies and a distant agar plate grown with either M. bovis or S. enterica.

[0103] 2. Control 2: phage in 20% sucrose solution and agar plates located at a distance growing M. bovis or S. enterica but without flies.

[0104] Table 1 summarizes this experimental setup for each of the M. bovis and S. enterica pathogen experiments.

[0105] Table 1: Overview of the experimental setup for Example 1

[0106]

[0107] Each enclosure was left at room temperature for one day. Agar plates were then removed from each enclosure and analyzed to determine if there was any effect on pathogen growth.

[0108] Figure 3 Each agar plate at the end of the Moraxella bovis experiment is shown. Figure 3 A) and control 2 ( Figure 3 In the iterations of the experiment with flies and phages in the sucrose solution ( Figure 3 C), Zones of depletion of M. bovis growth are observed due to phage-mediated cell lysis. Visualization of this lytic activity was enhanced by the addition of triphenyltetrazolium chloride.

[0109] Figure 4 Each agar plate at the end of the Salmonella enterica experiment is shown. Figure 4 A) and control 2 ( Figure 4 In the iterations of the experiment with flies and phages in the sucrose solution ( Figure 4 C), Zones of depleted growth of S. enterica are observed due to phage-mediated cell lysis. Visualization of this lytic activity was enhanced by the addition of triphenyltetrazolium chloride.

[0110] The results of Example 1 demonstrate that flying insects can act as vectors to transfer phage from one location to another and therefore, in the presence of a bacterial infection, they can be a means of ameliorging the infection at that second location.

[0111] Example 2

[0112] A model system was established for experiments to study the ability of flying insects to carry bacteriophages from a source to a remotely located cow's eye. In this model, the flying insect used was the face fly (Fall Housefly). The face flies used in the experiment were caught in the wild and were maintained in a 20% sucrose solution under ambient conditions with a 12:12 light:dark lighting pattern prior to the experiment. The cow's eye was an ex vivo cow's eye from a slaughterhouse. Bacteriophages with lytic activity against Salmonella enterica were added to the 20% sucrose solution that served as a flying insect attractant. The concentration of phages with lytic activity against Salmonella enterica was about 10 11 pfu / ml.

[0113] An enclosure was set up containing phage in a sucrose solution, 5 adult flies, and a bull's eye placed away from the sucrose solution. Thus, within the enclosure, the flies were able to feed on the sucrose solution, consuming the phage while doing so, and travel to the bull's eye.

[0114] In addition, other closures were set up as controls containing:

[0115] 1. Control 1: 20% sucrose solution without phages but containing flies and isolated bull's eyes located at a distance;

[0116] 2. Control 2: phage in 20% sucrose solution, isolated bull's eye at a distance, but no flies.

[0117] Table 2 summarizes this experimental setup.

[0118] Table 2. Overview of the experimental setup for Example 2

[0119]

[0120] Each block was left at room temperature for one day. The bovine eye was then removed from each block and the surface of the eye was rinsed with 1 ml of sterile PBS solution, 100 μl of which was transferred to a lawn of Salmonella enterica growth on agar to be incubated overnight at 37° C. The agar plate from each eye was then analyzed to determine if there was any effect on pathogen growth.

[0121] Figure 5 Each agar plate at the end of the experiment is shown. Figure 5 A) and control 2 ( Figure 5 B) Addition of PBS rinses from isolated bovine eyes on agar plates caused disturbance of the pathogen lawn, but no areas of bacterial lysis were detected. Figure 5 C), Addition of PBS rinses from isolated eyes caused disorganization of the pathogen lawn, and additional lytic activity was observed where fluid pooled.

[0122] The results of this experiment suggest that flies are able to transfer phages from one location to the bull's eye, where they have the potential to restrict or eradicate pathogens of sensitive bacterial species.

[0123] Example 3

[0124] A model system was established for experiments to study the ability of flying insects to carry phages from one source to a distantly located plant species. In this model, the flying insect used was a bumblebee (Bombus luteus). The plant species was fresh cut flowers. Phages with lytic activity against Salmonella enterica were added to a 20% sucrose solution that served as an attractant for the flying insects. The concentration of phages with lytic activity against Salmonella enterica was about 10 11 pfu / ml.

[0125] An enclosure was set up containing the phage in a sucrose solution, five bumble bees, and fresh cut flowers placed away from the sucrose solution. Thus, within the enclosure, the bees were able to feed on the sucrose solution, consuming the phage while doing so, and travel to the flowers.

[0126] In addition, other closures were set up as controls containing:

[0127] 1. Control 1: bees, flowers located far away, but no phage in 20% sucrose solution;

[0128] 2. Control 2: phage in 20% sucrose solution, flowers located far away, but no bees.

[0129] Table 3 summarizes this experimental setup.

[0130] Table 3. Overview of the setup for Example 3

[0131]

[0132] Each enclosure was left at room temperature for one day. The flowers were then removed from each enclosure and rinsed with 1 ml of sterile PBS solution, 100 μl of which was transferred to a lawn of Salmonella enterica growth on agar to be incubated overnight at 37° C. The agar plates from each flower were then analyzed to determine if there was any effect on pathogen growth.

[0133] Figure 6 Each agar plate at the end of the experiment is shown. Figure 6 A) and control 2 ( Figure 6 B) Iteration in which the addition of PBS solution rinses from flowers did not produce any visible areas of bacterial lysis. In contrast, for the replicates where all variables were present ( Figure 6 C), Addition of PBS flower rinse to a growing lawn of Salmonella enterica resulted in significant lytic activity.

[0134] The results of this experiment demonstrate that insect vectors are able to transfer phages from a location to plant species where they have the potential to limit or eradicate pathogens of susceptible bacterial species. Furthermore, the results of this experiment reveal that honey bees are good vectors for the transmission of phages to flowering plants, which has favorable implications for crop protection during plant pollination. In fact, it is reasonable to conclude that adding phages with activity against specific pathogens to commercial beehives will reduce the presence of pathogens in the bee colonies of the hives and on the plants where the bees spend time.

[0135] Example 4

[0136] A model system was established for experiments investigating the ability of flying insects to carry bacteria from a source to a remote location.

[0137] The bacteria was Moraxella bovis, which was cultured in BHI medium supplemented with 5% horse blood. A portion of this liquid medium was added to sucrose to produce a 20% solution containing the bacteria. The flying insect used was the blowfly (Blowfly vomiting).

[0138] An enclosure was set up containing the bacteria in a sucrose solution, 5 blowflies, and a sterile blood agar plate placed away from the sucrose solution. Thus, within the enclosure, the flies were able to feed on the sucrose solution, consuming the bacteria while doing so, and travel to the sterile agar plate.

[0139] Additionally, another enclosure was set up as a control, which contained bacteria in a sucrose solution and a sterile blood agar plate placed away from the sucrose solution, but without blowflies.

[0140] Table 4: Overview of the setup for Example 4

[0141]

[0142] Each enclosure was left at room temperature for one day. Then, the blood agar plates were removed and incubated at 37°C. Blood agar plates removed from enclosures without flies failed to show bacterial growth. However, blood agar plates removed from enclosures containing flies produced visible bacterial colonies.

[0143] This experiment demonstrates that flies are able to transfer bacteria from a source to a distant location. It will generally be understood that bacteria are immunogenic, and indeed vaccines can be developed from inactivated or killed bacteria or bacterial components such as cell membrane proteins. This therefore means that flying insects are able to carry immunogenic substances from one location to another, and it will generally be understood that providing such substances in combination with fly attractants together with flying insect carriers can be used to spread these immunogenic substances, including to animals and plants.

[0144] Numbered embodiments of the present disclosure

[0145] 1. A composition for treating or preventing bacterial diseases in terrestrial plants or animals, the composition comprising:

[0146] (i) bacteriophage and / or immunogenic agent; and

[0147] (ii) Flying insect attractants.

[0148] 2. A method of attracting flying insects to bacteriophage and / or immunogenic agents, the method comprising:

[0149] (i) combining the bacteriophage and / or immunogenic agent with a flying insect attractant; or

[0150] (ii) placing the bacteriophage and / or immunogenic agent within the attraction radius of a flying insect attractant.

[0151] 3. The method of embodiment 2, wherein the bacteriophage and / or immunogenic agent is transferred to the flying insect when the flying insect moves toward or comes into contact with the attractant.

[0152] 4. A kit for treating or preventing bacterial diseases in terrestrial plants or animals, the kit comprising:

[0153] (i) bacteriophage and / or immunogenic agent; and

[0154] (ii) Flying insect attractants.

[0155] 5. Use of a flying insect in a method for transferring a bacteriophage and / or an immunogenic agent to a terrestrial animal or plant, wherein the flying insect acts as a carrier of the bacteriophage and / or immunogenic agent.

[0156] 6. The use according to embodiment 5, wherein the bacteriophage and / or immunogenic agent has been transferred to the flying insect by the method according to embodiment 2 or 3.

[0157] 7. A method of treating or preventing a disease in a terrestrial animal or plant, the method comprising transferring a bacteriophage and / or an immunogenic agent to the animal or plant using a flying insect, wherein the flying insect acts as a carrier of the bacteriophage and / or immunogenic agent.

[0158] 8. The method of embodiment 7, wherein the bacteriophage and / or immunogenic agent has been transferred to the flying insect by the method of embodiment 2 or 3.

[0159] 9. The composition of embodiment 1, the method of embodiment 2, 3 or 8, the kit of embodiment 4 or the use of embodiment 6, wherein the flying insect attractant comprises a flying insect nutrient source.

[0160] 10. The composition, method, kit or use of embodiment 9, wherein the flying insect attractant comprises a flying insect nutrient source in combination with a second substance.

[0161] 11. The composition of embodiment 1, 9 or 10, the method of embodiment 2, 3, 7, 8, 9 or 10, the kit of embodiment 4, 9 or 10, or the use of embodiment 5, 6, 9 or 10, wherein the bacteriophage and / or immunogenic agent is active against bacteria associated with a disease in a terrestrial animal or plant.

[0162] 12. The composition or kit according to any preceding embodiment, wherein the composition comprises two or more immunogenic agents and / or bacteriophages, preferably two or more bacteriophages.

[0163] 13. The method or use according to any preceding embodiment, wherein two or more immunogenic agents and / or bacteriophages are used.

[0164] 14. A composition or kit according to any preceding embodiment, comprising a bacteriophage, or a method or use according to any preceding embodiment, wherein a bacteriophage is used.

[0165] 15. The composition, method, kit or use according to embodiment 14, wherein the bacteriophage is stabilized by being immobilized to a substrate, preferably wherein the bacteriophage is immobilized by its head.

[0166] 16. The composition, method, kit or use according to any preceding embodiment, wherein the immunogenic agent comprises a denatured bacterial antigen.

[0167] 17. A composition, method, kit or use according to any preceding embodiment, wherein the flying insect is a fly, preferably an insect of the order Diptera, preferably wherein the flying insect is a face fly, an Australian bush fly or a buffalo fly, most preferably a face fly.

[0168] 18. The composition, method, kit or use according to any preceding embodiment, wherein the flying insect is a wasp, an oriental fruit fly or an olive fruit fly.

[0169] 19. A composition, method, kit or use according to any preceding embodiment, wherein the bacteriophage and / or immunogenic agent is active against bacteria associated with a disease in a terrestrial animal, preferably wherein the animal is a bovine or equine animal, more preferably a bovine animal.

[0170] 20. The composition, method, kit or use according to any preceding embodiment, wherein the bacteriophage and / or immunogenic agent is active against bacteria associated with a disease in a terrestrial plant, preferably wherein the plant is a banana plant or an olive tree.

[0171] 21. The composition, method, kit or use according to any preceding embodiment, wherein the bacteriophage and / or immunogenic agent is active against a pathogen involved in the etiology of infectious bovine keratoconjunctivitis.

[0172] 22. The composition, method, kit or use of any preceding embodiment, wherein the bacteriophage and / or immunogenic agent is active against Moraxella bovis.

[0173] 23. A composition, method, kit or use according to any preceding embodiment, wherein the bacteriophage and / or immunogenic agent is active against Moraxella catarrhalis, Moraxella bovis, Mycoplasma, Xanthomonas campestris, Ralstonia solanacearum, Xylella fastidiosa and / or Pseudomonas sawantii.

[0174] 24. A composition, method, kit or use according to any preceding embodiment, wherein a flying insect attractant is present, preferably wherein the attractant comprises one or more of: fecal matter, pheromones, volatile agents, odorous materials, materials that can reflect or emit light within the visible electromagnetic spectrum, materials that reflect or emit electromagnetic radiation outside the visible spectrum, or resting places to which flying insects are naturally attracted.

[0175] 25. A composition, method, kit or use according to any preceding embodiment, wherein the attractant comprises one or more natural and / or synthetic substances, preferably one or more substances selected from pheromones, volatile agents and odorous materials, preferably volatile agents.

[0176] 26. The composition, method, kit or use according to any preceding embodiment, wherein the attractant comprises a material that reflects or emits light within the visible electromagnetic spectrum or a material that reflects or emits electromagnetic radiation outside the visible spectrum.

[0177] 27. The composition, method, kit or use according to any preceding embodiment, wherein the flying insect attractant comprises a flying insect nutrient source, wherein the nutrient source is a sugar.

[0178] 28. The composition, method, kit or use according to any preceding embodiment, wherein the attractant and the bacteriophage and / or immunogenic agent are presented in solid form or in liquid form.

[0179] 29. The composition, method, kit or use according to any preceding embodiment, wherein the attractant comprises fecal material, preferably bovine fecal material.

[0180] 30. The composition, method, kit or use according to any preceding embodiment, wherein the bacteriophage and / or immunogenic agent is mixed with the attractant.

[0181] 31. An article comprising the composition according to any preceding embodiment.

[0182] 32. The article of embodiment 31, wherein the article is capable of being worn by a terrestrial animal.

[0183] 33. The article of embodiment 32, wherein the article is a cattle ear tag or a horse ear tag.

[0184] 34. A method wherein a composition according to any preceding embodiment is applied to the skin of a terrestrial animal.

[0185] This disclosure provides exemplary embodiments of the invention and is not intended to be limiting. It will be appreciated that various other modifications and variations of the invention are possible.

Claims

1. A method of transferring a bacteriophage and / or an immunogenic agent to a flying insect, the method comprising attracting the flying insect to the bacteriophage and / or the immunogenic agent, comprising: (i) combining the bacteriophage and / or immunogenic agent with a flying insect attractant; or (ii) placing the bacteriophage and / or immunogenic agent within the attraction radius of a flying insect attractant; Wherein the bacteriophage and / or immunogenic agent is transferred to the flying insect when the flying insect moves toward or comes into contact with the attractant.

2. Use of a flying insect in a method for transferring a bacteriophage and / or an immunogenic agent to a terrestrial animal or plant, wherein the flying insect acts as a carrier of the bacteriophage and / or immunogenic agent.

3. The use according to claim 2, wherein the bacteriophage and / or immunogenic agent has been transferred to the flying insect by the method according to claim 1.

4. A method of treating or preventing a disease in a terrestrial animal or plant, the method comprising transferring an effective amount of a bacteriophage and / or an immunogenic agent to the animal or the plant using a flying insect, wherein the flying insect acts as a carrier of the bacteriophage and / or immunogenic agent.

5. The method according to claim 4, wherein the bacteriophage and / or immunogenic agent has been transferred to the flying insect by the method according to claim 1.

6. A composition for treating or preventing bacterial diseases in terrestrial plants or animals, the composition comprising: (i) bacteriophage and / or immunogenic agent; and (ii) a flying insect attractant selected from fecal matter, preferably cattle fecal matter, pheromones, volatile agents, odorous materials, materials capable of reflecting or emitting light within the visible electromagnetic spectrum, materials reflecting or emitting electromagnetic radiation outside the visible spectrum, flying insect nutrient sources, and combinations thereof.

7. A kit for treating or preventing bacterial diseases in terrestrial plants or animals, the kit comprising: (i) bacteriophage and / or immunogenic agent; and (ii) a flying insect attractant selected from fecal matter, preferably cattle fecal matter, pheromones, volatile agents, odorous materials, materials capable of reflecting or emitting light within the visible electromagnetic spectrum, materials reflecting or emitting electromagnetic radiation outside the visible spectrum, resting places to which flying insects are naturally attracted, flying insect nutrition sources, and combinations thereof.

8. The composition of claim 6, the method of claim 1 or 6, the kit of claim 7 or the use of claim 2 or 3, wherein the flying insect attractant comprises a flying insect nutrient source, optionally wherein the nutrient source is a sugar, preferably wherein the flying insect attractant comprises a flight insect nutrient source in combination with a second substance.

9. A composition according to claim 6 or 8, a method according to claim 1, 5 or 6, a kit according to claim 7 or 8, or a use according to claim 2, 3 or 8, wherein the bacteriophage and / or immunogenic agent is active against bacteria associated with a disease in a terrestrial animal or plant.

10. A composition or kit according to any preceding claim, wherein the composition or kit comprises two or more immunogenic agents and / or bacteriophages, preferably one or more bacteriophages.

11. The method or use according to any preceding claim, wherein two or more immunogenic agents and / or one or more bacteriophages are used.

12. A composition or kit according to any preceding claim comprising a bacteriophage, or a method or use according to any preceding claim, wherein a bacteriophage is used, preferably wherein the bacteriophage is stabilized by immobilization to a substrate, preferably wherein the bacteriophage is immobilized by its head.

13. The composition, method, kit or use of any preceding claim, wherein the immunogenic agent comprises a bacterial antigen.

14. A composition, method, kit or use according to any preceding claim, wherein the flying insect is a fly, preferably an insect of the order Diptera, preferably wherein the flying insect is a face fly, Australian bush fly, buffalo fly, horn fly or stable fly, most preferably a face fly; or wherein the flying insect is a wasp, oriental fruit fly or olive fruit fly, hoverfly, blowfly or honey bee.

15. A composition, method, kit or use according to any preceding claim, wherein the bacteriophage and / or immunogenic agent is active against bacteria associated with a disease in terrestrial animals, preferably wherein the animal is a bovine or equine animal, more preferably a bovine animal; and / or the bacteriophage and / or immunogenic agent is active against bacteria associated with a disease in terrestrial plants, preferably wherein the plant is a fruiting plant, most preferably wherein the plant is a banana plant, an olive tree or a kiwi plant; and / or The bacteriophage and / or immunogenic agent is active against a pathogen associated with the cause of infectious bovine keratoconjunctivitis; and / or The bacteriophage and / or immunogenic agent is active against Moraxella bovis; and / or The bacteriophage and / or immunogenic agent is active against Moraxella catarrhalis, Moraxella bovis, Mycoplasma, Xanthomonas campestris, Ralstonia solanacearum, Xylella fastidiosa, Pseudomonas sawanii, and / or Pseudomonas syringae.

16. A method or use according to any preceding claim, wherein the flying insect attractant comprises one or more of: fecal matter, preferably cattle fecal matter, pheromones, volatile agents, odorous materials, materials capable of reflecting or emitting light within the visible electromagnetic spectrum, materials reflecting or emitting electromagnetic radiation outside the visible spectrum, resting places to which flying insects are naturally attracted, or a source of nutrition for flying insects.

17. A composition, method, kit or use according to any preceding claim, wherein the attractant and the bacteriophage and / or immunogenic agent are presented in solid form or in liquid form; and / or wherein the bacteriophage and / or immunogenic agent is mixed with the attractant.

18. An article comprising a composition according to any preceding claim, preferably wherein the article is capable of being worn by a terrestrial animal, most preferably wherein the article is an ear tag, optionally a cattle ear tag or a horse ear tag.

19. A method wherein a composition according to any preceding claim is applied to the skin of a terrestrial animal.

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