Bartonella sp. DGB1 from chicken mite and its application in preventing and treating chicken mite

Abstract

The application discloses a new Bartonella species DGB1 derived from chicken skin mite and application of the Bartonella species DGB1 in prevention and treatment of chicken skin mite. Bartonella sp. The application provides a new Bartonella species, specifically a Bartonella DGB1, which is registered and listed in the China General Microbiological Culture Collection Center with a registration number of CGMCC No. 36714. The application also provides a Bartonella inactivated bacteria solution and an inactivated vaccine prepared from the Bartonella. The Bartonella inactivated vaccine provided by the application can effectively prevent and control chicken skin mite.

Description

Technical Field

[0001] This invention relates to the field of microbiology, specifically to a new Bartonella species DGB1 derived from the chicken spiny mite and its application in the control of the chicken spiny mite. Background Technology

[0002] Chicken tussock mites are among the most serious blood-sucking ectoparasites in the poultry industry. They also infect various bird species, including pigeons, parrots, sparrows, and magpies. Chicken tussock mites bite chickens and feed on their blood, causing anemia, increased aggression, decreased egg production, and reduced egg quality. They are also carriers or vectors of various pathogens, including Salmonella, avian influenza, Newcastle disease, and avian leukosis virus, seriously endangering chicken health. Chicken tussock mites cause significant economic losses to the poultry industry; in Europe, the annual economic losses reach €231 million, and in my country, they cause tens of millions of yuan in economic losses annually. Chicken tussock mites can also bite humans (especially poultry farm workers), causing dermatitis and potentially triggering public health emergencies.

[0003] Currently, the control of chicken spiny mites mainly relies on chemical control, with commonly used pesticides including pyrethroids (such as permethrin), organophosphates (such as trichlorfon), isoxazoline pesticides (such as fluranal), carbamates, and organochlorine pesticides. However, chemical control has problems such as pesticide residues, environmental pollution, and pesticide resistance. New control methods for chicken spiny mites are urgently needed. Novel pest control methods often need to be based on their important physiological characteristics.

[0004] Symbiotic bacteria are often referred to as "multifunctional organs" in arthropods, participating in most of their life activities. They play a vital role in many physiological functions, such as nutrition, metabolism, and immunity, including food digestion, nutrient synthesis, energy intake from food, regulation of intestinal physiology and homeostasis, synthesis of important metabolites, promotion of immune system development and maturation, defense against pathogen invasion, assistance in interspecific and intraspecific communication, influencing mating and reproduction, and mediating drug resistance.

[0005] Eliminating and interfering with key symbiotic bacteria within arthropod pests to influence their vital physiological activities and thus control pests is a promising green control method. However, research on the isolation, identification, and functional studies of key symbiotic bacteria within the chicken cutworm mite, and their application in the control of the chicken cutworm mite, is scarce. Summary of the Invention

[0006] The purpose of this invention is to provide a new Bartonella species DGB1 derived from the chicken spiny mite and its application in the control of the chicken spiny mite.

[0007] Firstly, the present invention claims protection for a Bartonite.

[0008] The Bartonella species claimed in this invention is a new species of Bartonella derived from the chicken dermatophyte mite, specifically... Bartonella sp. DGB1 has the registration number CGMCC No.36714 at the China General Microbiological Culture Collection Center.

[0009] Secondly, the present invention claims protection for a Bartonella inactivated bacterial solution.

[0010] The Bartonella inactivated bacterial solution claimed in this invention is prepared by inactivating the Bartonella described in the first aspect above with formaldehyde.

[0011] Thirdly, the present invention claims protection for a Bartonella inactivated vaccine.

[0012] The Bartonella inactivated vaccine claimed in this invention consists of a vaccine antigen and an adjuvant;

[0013] The vaccine antigen is the Bartonella inactivated bacterial solution described in the second aspect above; The adjuvant is an aluminum colloid salt adjuvant.

[0014] In some embodiments of the present invention, the content of Bartonella in the vaccine antigen, after formaldehyde inactivation treatment, is 5 × 10⁻⁶. 9 CFU / mL (calculated based on the viable bacterial content before inactivation).

[0015] In some embodiments of the present invention, the aluminum colloid adjuvant is an aluminum hydroxide adjuvant, specifically a colloid formed by reacting 5% (i.e., 50 g / L) aluminum sulfate with 5% (i.e., 50 g / L) sodium hydroxide solution to obtain high-purity aluminum hydroxide.

[0016] In some embodiments of the present invention, the volume ratio of the vaccine antigen to the adjuvant in the Bartonella inactivated vaccine is 6:1.

[0017] Fourthly, the present invention claims a method for preparing a Bartonella inactivated vaccine.

[0018] The method for preparing a Bartonella inactivated vaccine claimed in this invention may include the following steps: (A1) Prepare the vaccine antigen by means of the following steps: inactivate Bartonella as described in the first aspect above with formaldehyde to obtain the Bartonella inactivated bacterial solution as described in the second aspect above, which is the vaccine antigen; (A2) The Bartonella inactivated vaccine is prepared by mixing the vaccine antigen and aluminum colloid adjuvant.

[0019] In some embodiments of the present invention, the content of Bartonella in the vaccine antigen, after formaldehyde inactivation treatment, is 5 × 10⁻⁶. 9CFU / mL (calculated based on the viable bacterial content before inactivation).

[0020] In some embodiments of the present invention, the aluminum colloid adjuvant is an aluminum hydroxide adjuvant, specifically a colloid formed by reacting 5% (i.e., 50 g / L) aluminum sulfate with 5% (i.e., 50 g / L) sodium hydroxide solution to obtain high-purity aluminum hydroxide.

[0021] In some embodiments of the present invention, the volume ratio of the vaccine antigen to the adjuvant in the Bartonella inactivated vaccine is 6:1.

[0022] In the second to fourth aspects mentioned above, in some embodiments of the present invention, the Bartonella inactivating bacterial solution is specifically prepared as follows: the bacterial solution concentration is 5 × 10⁻⁶. 9 Add formaldehyde solution to a final concentration of 0.2% (2 g / L) to a sterile PBS suspension of Bartonella cFU / mL, mix well, and inactivate at 37°C and 150 rpm for 24 h. Centrifuge the inactivated bacterial suspension (e.g., at 8000 rpm for 10 min), discard the supernatant, collect the bacterial precipitate, wash with sterile physiological saline (e.g., wash 3 times), and resuspend in sterile physiological saline to the original volume (i.e., the bacterial suspension concentration mentioned above is 5 × 10⁻⁶). 9 (The volume of the sterile PBS bacterial suspension of Bartonella (CFU / mL)).

[0023] Fifthly, the present invention claims the use of Bartonella as described in the first aspect above, or Bartonella inactivated bacterial solution as described in the second aspect above, or Bartonella inactivated vaccine as described in the third aspect above, in the preparation of veterinary biological products for the prevention and control of chicken cutaneous mites.

[0024] Sixthly, the present invention claims protection for a veterinary biological product for the control of chicken dermatophytes.

[0025] The effective ingredient of the veterinary biological product for the prevention and control of chicken dermatophytes claimed in this invention includes (or is) the Bartonella inactivated bacterial solution described in the second aspect above or the Bartonella inactivated vaccine described in the third aspect above.

[0026] Furthermore, the veterinary biological product may also include one or more of a pharmaceutically acceptable carrier, diluent, or preservative.

[0027] Seventhly, the present invention claims a non-therapeutic method for reducing the population density of chicken cutworms in a breeding environment.

[0028] The non-therapeutic method for reducing the population density of chicken cutworms in a breeding environment, as claimed in this invention, may include the following steps (B1) or (B2): (B1) Administer the Bartonella inactivated vaccine described in the third aspect above to the flock of chickens to induce the flock to produce specific antibodies against the Bartonella; after the blood of chickens carrying the specific antibodies is fed on by the chicken mites, the reproductive capacity of the chicken mites is reduced and / or the mortality rate is increased, thereby reducing the population density of chicken mites in the breeding environment. Principle: The specific antibodies against Bartonella carried in the blood of chickens cause a significant decrease in the abundance of Bartonella DGB1 parasites in the chicken's body after the blood is ingested by the chicken mites. This decrease in the abundance of Bartonella DGB1 leads to a reduction in the reproductive capacity and / or an increase in the mortality rate of the chicken mites.

[0029] (B2) When aminoglycoside antibiotics are administered to chickens, the mortality rate of chicken mites increases after the chickens' blood is sucked by the mites, thereby reducing the population density of chicken mites in the breeding environment.

[0030] Furthermore, the aminoglycoside antibiotic is tobramycin.

[0031] Principle: When antibiotics enter the bloodstream of chickens, the antibiotics enter the chicken mites' bodies along with the blood when they feed on the blood. This leads to a significant decrease in the abundance of Bartonella DGB1 parasites in the mites. As a result, the mortality rate of chicken mites increases due to the decrease in the abundance of Bartonella DGB1.

[0032] Eighthly, the present invention claims protection for a method of disease prevention and breeding of chicken flocks.

[0033] The disease prevention and breeding method for chicken flocks claimed in this invention may include the following steps (C1) or (C2): (C1) Apply the Bartonella inactivated vaccine described in the third aspect above to the flock to induce the flock to produce specific antibodies against Bartonella; after the blood of the chickens carrying the specific antibodies is fed on by the chicken mites, the reproductive capacity of the chicken mites decreases and / or the mortality rate increases, thereby reducing the population density of chicken mites in the breeding environment, thus achieving healthy breeding and disease prevention of the flock.

[0034] (C2) When aminoglycoside antibiotics are administered to chickens that are not infected with chicken mites, the mortality rate of chicken mites increases after the chickens' blood is sucked by the mites, which reduces the population density of chicken mites in the breeding environment, thereby achieving healthy breeding and disease prevention of chickens.

[0035] Furthermore, the aminoglycoside antibiotic is tobramycin.

[0036] In aspects seven and eight above (B1) and (C1), the administration of the Bartonella inactivated vaccine to the flock is by intramuscular injection of the Bartonella inactivated vaccine at a dose of 2 × 10⁻⁶. 9CFU was administered for a second immunization 14 days later, followed by a third and fourth immunization one week later. The administration method and dosage for the second to fourth immunizations were the same as for the first immunization.

[0037] In aspects 7 and 8 above (B2) and (C2), the administration of aminoglycoside antibiotics to chickens is by intramuscular injection of 35 mg / kg body weight of aminoglycoside antibiotics (such as tobramycin).

[0038] In the seventh and eighth aspects mentioned above, the flock of chickens referred to is a flock of chickens that is not infected with the chicken dermatophyte mite.

[0039] Compared with the prior art, the present invention has the following advantages: (1) The Bartonella DGB1 provided by the present invention is an important symbiotic bacterium in the chicken mites and has an important influence on the survival and reproduction of chicken mites.

[0040] (2) The Bartonella DGB1 provided by the present invention can be used as a target for the prevention and control of chicken bursal mites.

[0041] (3) Using aminoglycoside antibiotics to inhibit Bartonella DGB1 in the body of chicken mites can effectively prevent and control chicken mites.

[0042] (4) Using Bartonella DGB1 provided by the present invention to develop an inactivated vaccine can effectively prevent and control chicken dermatophytes.

[0043] This invention is of great significance for the prevention and control of chicken skin mites.

[0044] Preservation Instructions Reference biological material (strain): DGB1 Suggested category naming: Bartonella sp. Preservation Institution: China General Microbiological Culture Collection Center, China Microbiological Culture Collection Committee Abbreviation of depositary institution: CGMCC Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing Deposit date: November 14, 2025 CGMCC Registration Number: CGMCC No. 36714 Attached Figure Description Figure 1 Phylogenetic tree of the 16S rRNA gene of Bartonella DGB1.

[0045] Figure 2 The colony morphology of Bartonella DGB1.

[0046] Figure 3Gram staining results for Bartonella DGB1. Scale bar is 10 μm.

[0047] Figure 4 The ultrastructure of Bartonite DGB1 under transmission electron microscopy.

[0048] Figure 5 The results of genomic phylogenetic analysis of Bartonella DGB1.

[0049] Figure 6 The results of the average nucleotide identity analysis for Bartonella DGB1 are shown.

[0050] Figure 7 The results of DNA-DNA hybridization analysis of Bartonella DGB1 are shown.

[0051] Figure 8 The figures show changes in the reproduction, blood digestion, and mortality of chicken mites after tobramycin injection. In the figures, A represents bacterial abundance; B represents mite reproductive capacity; C represents mite blood digestion (scale bar at 500 μm); and D represents mite mortality. In the figures, CT represents the control group, and NN represents the tobramycin-treated group. This indicates a significant difference compared to the control group (P<0.01). ns indicates a highly significant difference from the control group (P<0.001); ns indicates no significant difference.

[0052] Figure 9 This figure shows the production of specific antibodies in chickens after immunization with the Bartonella inactivated vaccine. In the figure, CT represents the control group, and DGB1 vax represents the group vaccinated with the Bartonella DGB1 inactivated vaccine. This indicates a significant difference compared to the control group (P<0.05).

[0053] Figure 10 This figure shows the changes in the reproduction, blood digestion, and mortality of chicken mites after immunization with the Bartonella DGB1 inactivated vaccine. In the figure, A represents mite reproductive capacity; B represents mite mortality; and C represents mite blood digestion (scale bar is 500 μm). In the figure, CT represents the control group, and DGB1 vax represents the group vaccinated with the Bartonella DGB1 inactivated vaccine. ns indicates a highly significant difference from the control group (P<0.01); ns indicates no significant difference. Detailed Implementation

[0054] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0055] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0056] Example 1: Isolation and identification of Bartonella DGB1, a symbiotic bacterium of the chicken skin mite. I. Separation of the Barton body In this embodiment, Bartonella was isolated from the chicken mites using the following specific method: Chicken mites were isolated from a laboratory breeding system. Fifteen adult female mites were selected and soaked in 75% ethanol for 3 hours, followed by washing 3-4 times with sterile distilled water to remove microorganisms from their bodies. Under sterile conditions, 300 μL of sterile physiological saline was added, and the chicken mites were homogenized using a homogenizer. After a 10-fold serial dilution, 100 μL of the homogenate was spread onto TSA agar containing 10% sterile defibrinated sheep blood and incubated at 37°C in a 5% CO2 incubator for 5-7 days. Colonies suspected to be Bartonella were picked and purified to obtain pure cultures.

[0057] II. Identification of Bartonian Pure cultures were picked and thoroughly mixed by pipetting in a small amount of sterile physiological saline. Using the bacterial suspension as a template, bacterial PCR amplification was performed using the universal 16S primers 27F: 5'-AGAGTTTGATCMTGGCTCAG-3' (SEQ ID NO:1) and 1492R: 5'-TACGGYTACCTTGTTACGACTT-3' (SEQ ID NO:2). The amplified products were sequenced, and the sequencing results were analyzed by BLAST alignment and evolutionary analysis in the Genbank database. Figure 1 ). Figure 1 The results showed that a pure culture of a single colony clustered with a strain of the genus Bartonella on the phylogenetic tree, and this strain was named DGB1, thus confirming that DGB1 is a Bartonella.

[0058] Observe the morphology of Bartonella colony DGB1. On TSA blood agar plates, DGB1 colonies are round, with regular edges, and semi-transparent. Figure 2A single colony of Bartonella DGB1 was picked, thoroughly mixed by pipetting in a small amount of sterile physiological saline, and 10 μL of the bacterial suspension was evenly spread onto a glass slide. The colony was then stained with a Gram staining kit and observed under an oil immersion microscope. DGB1 is rod-shaped and stained red, indicating that it is a Gram-negative bacillus. Figure 3 ).

[0059] Under aseptic conditions, sterile distilled water was slowly added dropwise to a plate containing a pure culture of Bartonella DGB1 for in-situ elution. The suspended sample was then loaded onto an electron microscope grid. After washing away excess suspension, the sample was stained with 1% uranium acetate solution. After staining for 3 minutes, excess stain was aspirated. After drying, the morphology and structure of Bartonella DGB1 were observed using a transmission electron microscope. DGB1 is approximately 1.3 μm long and 0.5 μm wide, with a single flagellum. Numerous secretions form a capsule-like structure around the cell. Figure 4 ).

[0060] Whole-genome sequencing was performed on DGB1, and comparative genomic analysis was conducted on the sequencing results to further identify its species taxonomically. Figure 5 The results showed that Bartonella DGB1 was annotated as a new species; mean nucleotide identity ( ) Figure 6 Table 1 shows that the ANI value of DGB1 is below 95% of the species threshold; DNA-DNA hybridization analysis ( Figure 7 Table 1 shows that the DDH value of DGB1 is also lower than 70% of the species threshold classification standard.

[0061] Table 1. Comparison of ANI and DDH values ​​between DGB1 and reference strain genomes.

[0062] The above results indicate that DGB1 is a new species of the genus Bartonella. This strain was deposited on November 14, 2025, at the China General Microbiological Culture Collection Center (CGMCC), with the registration number CGMCC No. 36714. Hereinafter referred to as Bartonella DGB1.

[0063] The applicant also isolated and identified another new species of Bartonella, DGB2, from the chicken mites, and the relevant technical solution has been filed as a separate patent application.

[0064] Example 2: Controlling chicken mites by using antibiotics to inhibit Bartonella in the body of chicken mites. Female *Dermatophyte foetida* mites (with high abundance of Bartonella DGB1 parasites internally) were selected from a laboratory rearing system and placed in an intelligent artificial climate chamber at 30°C and 75% relative humidity for 5 days of starvation. Tobramycin (an antibiotic) was injected intramuscularly into the chickens at 35 mg / kg body weight. A control group without tobramycin was also included. Each group consisted of 3 chickens. Immediately after injection, the chickens were challenged with an external blood-feeding device, allowing the mites to feed on chicken blood containing tobramycin. Eight hours after the challenge, the external blood-feeding device was removed from the chickens. The engorged mites were then placed in an intelligent artificial climate chamber at 30°C and 75% relative humidity. Thirty chicken *Dendrobium chrysogenum* mites were collected on day 5 post-infection. Total DNA was extracted from the mites using a genomic DNA kit. The abundance changes of Bartonella DGB1 and other dominant bacteria (such as DGB2 and *Kocuria*) in the mites were detected by qPCR. The relevant primers are shown in Table 2. The *Dendrobium chrysogenum* β-actin gene was used as an internal control. -ΔΔCT The method involves data processing and statistical analysis.

[0065] Table 2. Primers for qPCR detection of symbiotic bacteria abundance in chicken skin mites

[0066] Five days after feeding on antibiotic-treated blood, all surviving chicken mites were collected and treated again using the same method, this time feeding on antibiotic-free blood. Eight hours after treatment, the external blood-feeding device was removed from the chickens' bodies. The blood-saturated mites were then collected, and their reproductive capacity, blood digestion, and mortality were observed and recorded over five days.

[0067] To observe reproductive capacity, *Dermatophyte gargarizans* were placed in 96-well plates, with one female mite in each well, and the plates were sealed with sealing film. Fifty mites constituted one replicate, with three replicates per group. The 96-well plates were placed in an artificial climate chamber, and the mite reproduction was observed and recorded daily for five days, including the number of mites laying eggs, the total number of eggs, and egg hatching status. The oviposition rate, reproductive capacity, and egg hatching rate were calculated using the following formulas.

[0068]

[0069] Morphological changes in mites were observed and photographed under a stereomicroscope over four days to evaluate the effect of antibiotics on the hemodigestion of *Dermatophytes gargarizans*. Mite mortality was also recorded, with 100 mites constituting one replicate and three replicates per group. Mites were placed in 9cm petri dishes, sealed with sealing film, and incubated in a climate chamber. Mite mortality and survival were observed and recorded daily for five days. Mites were considered dead if neither light nor mechanical stimulation elicited active movement of their legs. The mortality rate of *Dermatophytes gargarizans* was calculated using the following formula:

[0070] The experimental results showed that compared with the untreated group, the abundance of Bartonella DGB1 in the chicken mites injected with tobramycin was significantly decreased, while the abundance of the other two dominant symbiotic bacteria, Bartonella DGB2 and... Kocuria Abundance showed no significant change; mortality rate of chicken skin mites increased significantly, while reproductive capacity and blood digestion showed no significant changes. Figure 8 ).

[0071] The above studies confirm that aminoglycoside antibiotics such as tobramycin can be used to inhibit and kill Bartonella DGB1, a key symbiotic bacterium in the chicken mites, to prevent and control chicken mites.

[0072] Example 3: Developing a vaccine using Bartonella DGB1 to prevent and treat chicken cutaneous mites. Bartonella DGB1 isolated from the culture medium was inoculated onto TSA agar containing 10% sterile defibrinated sheep blood and incubated at 37°C in a 5% CO2 incubator for 5–7 days. Bartonella grown on the plates were collected, washed twice with sterile PBS, resuspended, and colony forming units (CFU) were determined. The bacterial concentration was adjusted to 5 × 10⁻⁶. 9 CFU / mL. Add formaldehyde solution to the bacterial culture to a final concentration of 0.2% (2 g / L), mix well, and inactivate at 37℃ and 150 rpm for 24 h. Centrifuge the inactivated bacterial culture at 8000 rpm for 10 min, discard the supernatant, collect the bacterial precipitate, wash three times with sterile physiological saline, and finally resuspend in sterile physiological saline to the original volume (i.e., the viable bacterial content before inactivation is 5 × 10⁻⁶ CFU / mL). 9Prepare a 5% (50 g / L) aluminum sulfate solution and a 5% (50 g / L) sodium hydroxide solution, and filter them through a 0.22 μm filter for sterilization. Mix the aluminum sulfate solution and sodium hydroxide solution at a volume ratio of 2.5:1 under vigorous stirring to form aluminum hydroxide precipitate. Centrifuge at 3000 r / min for 5 minutes, collect the precipitate, and wash it twice with sterile water to remove impurities and ensure purity. Finally, resuspend the precipitate to its original volume with sterile physiological saline to form a stable colloidal solution, which is the aluminum colloidal adjuvant. Take 0.4 mL of the inactivated DGB1 bacterial suspension and mix it with the aluminum colloidal adjuvant at a volume ratio of 6:1 to obtain the Bartonella DGB1 inactivated vaccine.

[0073] Chickens in the experimental group were intramuscularly injected with the Bartonella DGB1 inactivated vaccine prepared above, at a dose of 2 × 10⁻⁶. 9 CFU (calculated as live bacteria content before inactivation). On day 14 after the first immunization, a second immunization was administered using the same method (administration method and dosage as the first immunization), followed by third and fourth immunizations every week (administration method and dosage as the first immunization). A control group was given an intramuscular injection of an adjuvant (the only difference from the experimental group was that the inactivated vaccine was replaced with an equal amount of adjuvant). Three chickens were in each group. Blood was collected periodically from the immunized chickens, and serum was separated. Bartonella DGB1 was used as the coating antigen, and antibody levels in the chicken serum were detected using ELISA. The specific procedure was as follows: bacterial cells were collected from plates containing Bartonella DGB1, washed three times with PBS, and then diluted to 1×10⁻⁸ CFU with carbonate buffer (formulation: Na₂CO₃ 0.848g, NaHCO₃ 1.428g, distilled water to 500mL, pH adjusted to 9.6). 6 CFU / mL, 100 μL per well, incubated overnight at 4°C for coating; washed 3 times with PBST, then 200 μL of 5% skim milk was added to each well, and the mixture was blocked at 37°C for 2 hours; serum from immunized chickens was diluted 1:6400 as the primary antibody, incubated at 37°C for 1 hour, and washed 3 times with PBST; HRP-labeled goat anti-chicken IgG (bs-0310G-HRP, Beijing Bio-Sen Biotechnology) was diluted 1:5000 as the secondary antibody, incubated at 37°C for 1 hour, and washed 3 times with PBST; then 100 μL of TMB chromogenic solution (PR1200, Beijing Solarbio Science & Technology Co., Ltd.) was added to each well, and the mixture was incubated at 37°C in the dark for 15 minutes, and 50 μL of 2M sulfuric acid solution was added to each well to stop the reaction, and the absorbance at 450 nm was measured.

[0074] Female *Dermatophyte gargarizans* mites were selected from a laboratory rearing system and placed in an intelligent artificial climate chamber at 30°C and 75% relative humidity for 5 days of starvation. The starved mites were then placed in an external blood-feeding device, which was fixed to the skin of chicken wings. Vaccinated chickens on day 7 after their fourth immunization were challenged with the mites, allowing them to feed on the blood of the vaccinated chickens. Eight hours after challenge, the external blood-feeding device was removed from the chickens. Blood-saturated mites were then collected and placed in an intelligent artificial climate chamber at 30°C and 75% relative humidity. Their reproductive capacity, blood digestion, and mortality were observed and recorded over 5 days (see Example 2 for specific methods). The control efficacy of the Bartonella DGB1 inactivated vaccine against *Dermatophyte gargarizans* was evaluated.

[0075] Experimental results showed that immunization of chickens with Bartonella DGB1 inactivated vaccine induced the production of high levels of antibodies, which could be maintained for a relatively long period of time. Figure 9 After chickens immunized with Bartonella DGB1 inactivated vaccine fed on their blood, compared with the adjuvant control group, the mite reproductive capacity of the Bartonella DGB1 inactivated vaccine group was significantly reduced, and the mortality rate was significantly increased. However, blood digestion, egg production rate, and egg hatching rate were not significantly affected. Figure 10 The above results indicate that the Bartonella DGB1 inactivated vaccine achieves a "two-pronged" control of chicken mites by inducing high levels of antibodies in chickens—significantly increasing adult mortality to rapidly reduce the existing mite population, and reducing its reproductive capacity to weaken its population expansion ability, thereby rapidly, continuously, and stably reducing the mite population density and achieving highly efficient control of chicken mites.

[0076] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein.

Claims

1. A Bartonite, characterized by: The Bartonella species is a new species of Bartonella, specifically... Bartonella sp. DGB1 has the registration number CGMCCNo.36714 at the China General Microbiological Culture Collection Center.

2. A Bartonella inactivated bacterial solution, characterized in that: The Bartonella inactivated bacterial solution is prepared by inactivating the Bartonella as described in claim 1 with formaldehyde.

3. A Bartonella inactivated vaccine, characterized in that: The Bartonella inactivated vaccine consists of a vaccine antigen and an adjuvant. The vaccine antigen is the Bartonella inactivated bacterial solution as described in claim 2; The adjuvant is an aluminum colloid salt adjuvant.

4. The Bartonella inactivated vaccine according to claim 3, characterized in that: The content of Bartonella in the vaccine antigen, after formaldehyde inactivation treatment, is 5 × 10⁻⁶. 9 CFU / mL; and / or The aluminum colloid adjuvant is aluminum hydroxide adjuvant; and / or In the Bartonella inactivated vaccine, the volume ratio of the vaccine antigen to the adjuvant is 6:

1.

5. A method for preparing a Bartonella inactivated vaccine, comprising the following steps: (A1) Prepare the vaccine antigen by means of the following steps: inactivate the Bartonella as described in claim 1 with formaldehyde to obtain the Bartonella inactivated bacterial solution as described in claim 2, which is the vaccine antigen; (A2) The Bartonella inactivated vaccine is obtained by mixing the vaccine antigen and aluminum colloid adjuvant.

6. The method according to claim 5, characterized in that: The content of Bartonella in the vaccine antigen, after formaldehyde inactivation treatment, is 5 × 10⁻⁶. 9 CFU / mL; and / or The aluminum colloid adjuvant is aluminum hydroxide adjuvant; and / or In the Bartonella inactivated vaccine, the volume ratio of the vaccine antigen to the adjuvant is 6:

1.

7. The use of Bartonella as described in claim 1, or the Bartonella inactivated bacterial solution as described in claim 2, or the Bartonella inactivated vaccine as described in claim 3 or 4, in the preparation of veterinary biological products for the prevention and control of chicken cutaneous mites.

8. A veterinary biological product for controlling chicken cutaneous mites, characterized in that: The active ingredient of the veterinary biological product includes the Bartonella inactivated bacterial solution as described in claim 2 or the Bartonella inactivated vaccine as described in claim 3 or 4.

9. A non-therapeutic method for reducing the population density of chicken cutworms in a breeding environment, comprising the following steps (B1) or (B2): (B1) Apply the Bartonella inactivated vaccine of claim 3 or 4 to the flock of chickens to induce the flock to produce specific antibodies against Bartonella; after the blood of chickens carrying the specific antibodies is fed on by the chicken mites, the reproductive capacity of the chicken mites is reduced and / or the mortality rate is increased, thereby reducing the population density of chicken mites in the breeding environment. (B2) When aminoglycoside antibiotics are administered to chickens, the mortality rate of chicken mites increases after the chickens' blood is sucked by the mites, thereby reducing the population density of chicken mites in the breeding environment. Furthermore, the aminoglycoside antibiotic is tobramycin.

10. A method for disease prevention and control in chicken flocks, comprising the following steps (C1) or (C2): (C1) Apply the Bartonella inactivated vaccine of claim 3 or 4 to the flock of chickens to induce the flock to produce specific antibodies against the Bartonella; after the blood of chickens carrying the specific antibodies is fed on by the chicken mites, the reproductive capacity of the chicken mites is reduced and / or the mortality rate is increased, thereby reducing the population density of chicken mites in the breeding environment, thereby achieving healthy breeding and disease prevention of chickens. (C2) When aminoglycoside antibiotics are administered to chickens, the mortality rate of chicken mites increases after the chickens' blood is sucked by the mites, which reduces the population density of chicken mites in the breeding environment, thereby achieving healthy breeding and disease prevention in chickens. Furthermore, the aminoglycoside antibiotic is tobramycin.

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