Patent Publication Number: US-2022236268-A1

Title: NOVEL SECRETED ANTIGENS FOR DIAGNOSIS OF ACTIVE BABESIA MICROTI AND BABESIA DUNCANI INFECTION IN HUMANS AND ANIMALs

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     The present application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Application No. 62/937,645 filed Nov. 19, 2019, and No. 62/860,662 filed Jun. 12, 2019, all of which applications are hereby incorporated by reference in their entireties herein. 
    
    
     STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT 
     This invention was made with government support under AI136118, AI123321, AI138139 and GM110506 awarded by National Institutes of Health. The government has certain rights in the invention. 
    
    
     BACKGROUND OF THE INVENTION 
     Human babesiosis is an emerging tick-borne disease endemic in the United States and increasingly reported in other parts of the world including Asia and Europe. Three species,  Babesia microti, B. duncani  and  B. divergens  have been shown to cause infection in humans with  B. microti  accounting for the large majority of clinical cases reported worldwide. In susceptible patients,  B. microti  infection can result in severe symptoms including respiratory distress, splenic rupture and renal failure. The mortality rates associated with babesiosis infections range between 6 and 21%. Furthermore, severe infections as well as end-organ complications may develop in up to 57% of immunocompromised patients. There is a need in the art for compositions and methods useful for the detection and treatment of Babesiosis. The present disclosure addresses this need. 
     BRIEF SUMMARY OF THE INVENTION 
     In certain aspects, the present invention provides methods of detecting a  Babesia  infection in a subject. In certain embodiments, the method includes: detecting whether one or more secreted antigens selected from SEQ ID NOs: 1-62 are present in a biological sample collected from the subject; wherein detecting presence of one or more of the antigens in the biological sample indicates that the subject has a  Babesia  infection. In certain embodiments, the method includes: detecting one or more secreted antigens selected from SEQ ID NOs: 1-62 in a biological sample collected from the subject; wherein detecting one or more of the antigens indicates the presence of a  Babesia  infection. In some embodiments, the  Babesia  infection comprises  Babesia microti  or  Babesia duncani . In some embodiments, the biological sample comprises one or more selected from the group consisting of: a blood sample, an erythrocyte sample, a leukocyte sample, a plasma sample, a urine sample, a saliva sample, and/or one or more combinations thereof. In some embodiments, the one or more antigens further comprises BmGPI12. In some embodiments, the subject is a human. In some embodiments, the subject is a mammal known to carry a  Babesia  parasite. In some embodiments, the one or more antigens is detected by one or more antibody-based techniques selected from the group consisting of: Western blot, immunofluorescent assay, IEM, ELISA, PCR amplification-based immunoassay and immunoprecipitation. 
     In certain aspects the present invention provides a diagnostic tool for identifying or diagnosing a babesiosis infection. In certain embodiments, the tool comprises: an assay platform, an immunologic agent having specificity for one or more  Babesia  antigens selected from SEQ ID NOs: 1-62. In some embodiments, the babesiosis infection comprises a  Babesia microti  infection or a  Babesia duncani  infection. In some embodiments, the assay platform comprises one or more selected from the group consisting of: an enzyme-based assay, a radioimmunoassay, a PCR amplification-based immunoassay, a fluorogenic immunoassay, a chemiluminescence-based assay, immunoblotting assay, and combinations thereof. In some embodiments, the immunologic agent comprises one or more of antibodies or antibody fragments. 
     In certain aspects, the present invention provides a method of treating, ameliorating, and/or preventing a  Babesia  infection in a subject in need thereof. In certain embodiments, the method comprises: obtaining a first sample from a subject at a first time point; assaying the sample using a diagnostic tool contemplated herein to detect the presence or absence of an infection relative to a comparator control, administering one or more therapeutic agents to the subject; obtaining a second sample from the subject at a second time point, wherein the second time point comprises one or more time points after the first time point; and assaying the second sample obtained from the subject at the one or more second time points using the diagnostic tool contemplated herein to detect the presence or absence of an infection relative to a comparator control. In some embodiments, the sample comprises a blood sample. In some embodiments, the infection comprises a  Babesia microti  infection or a  Babesia duncani  infection. In some embodiments, the first time point is before administration of the therapeutic agent. In some embodiments, the second time point comprises an interval after a therapeutic agent is administered. 
     In certain embodiments the present invention provides a method of treating, ameliorating, and/or preventing a  Babesia  infection in a subject. In certain embodiments, the method comprises: detecting presence of one or more peptides selected from SEQ ID NOs: 1-62 in a biological sample collected from the subject; and administering to the subject at least one anti-protozoan therapeutic agent. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIGS. 1A-1B  demonstrate that BmGPI12 is secreted into the erythrocyte cytoplasm and subsequently into the extracellular environment of the  B. microti -infected erythrocyte.  FIG. 1A  depicts immunoblotting analysis using pre-immune (PI) and anti-BmGPI12 immune rabbit sera on fractions of uninfected erythrocytes (UI) or erythrocytes infected with  B. microti  strain LabS1. S: mouse plasma, H: hemolysate; (P) membrane fractions collected following saponin treatment of erythrocytes. In uninfected erythrocytes, the P fraction consists primarily of erythrocyte membrane. In  B. microti -infected erythrocytes, the P fraction includes both erythrocyte membrane and protein extracts from isolated parasites. The erythrocyte membrane protein TER-119 (52 kDa) was detected using an anti-TER-119 monoclonal antibody only in the P fractions from uninfected and  B. microti -infected red blood cells.  FIG. 1B  depicts immunofluorescence assay on mouse erythrocytes infected with the LabS1 strain of  B. microti . BmGPI12 was labelled with polyclonal anti-BmGPI12 antibodies and could be observed within the parasite cytoplasm, the parasite plasma membrane as well as in the erythrocyte cytoplasm and within individual vesicles (IV) and tubes of vesicles (TOVs) (indicated by arrowheads). Monoclonal antibodies against TER-119 were used to label the plasma membrane of the infected mouse erythrocytes. The DAPI staining was applied to verify the presence of parasites within the erythrocytes by labelling parasitic nuclear DNA. Staining of control uninfected red blood cells is shown. Scale bars: 3 μm. 
         FIGS. 2A-2C  demonstrate that  B. microti  develops an interlacement of vesicles (IOV) system in the cytoplasm of the infected erythrocytes.  FIG. 2A  depicts results indicating that giemsa-stained blood smears from  B. microti -infected erythrocytes with representative images of LabS1-infected erythrocytes revealed tubular structures within the erythrocyte cytoplasm (see arrowheads). Scale bar: 3 μm.  FIGS. 2B and 2C  depict analysis of blood smears from four infected mice over a 13-day period following infection with  B. microti  LabS1 strain.  FIG. 2B  depicts parasitemia levels in individual mice. A total of 5000 erythrocytes were analyzed per smear (Mean±SEM).  FIG. 2C  depicts a proportion of each morphological form detected in the blood smears at days 3, 5, 7, 10 and 13 post-infection. A total of 20 images were analyzed per smear at a given day (Mean±SEM). 
         FIGS. 3A-3C  demonstrate that  B. microti  develops an IOV system in the cytoplasm of the infected erythrocytes.  FIG. 3A  depicts that EPON-embedded, LabS1-infected erythrocytes revealed the presence of the IOV in the erythrocyte cytoplasm. The IOV contains the same electron dense structures as the parasite, indicating that these structures are of parasitic origin. Various structures of parasites and erythrocytes are shown by arrows.  FIGS. 3B and 3C  depict a comparison of ultrathin sections of EPON-embedded infected ( FIG. 3B ) and uninfected ( FIG. 3C ) erythrocytes. IV: individual vesicle, P: parasite, PPM: parasite plasma membrane, R: ribosomes, RBCC: red blood cell cytoplasm, RBCM: red blood cell membrane, TOV: tube of vesicles. 
         FIGS. 4A-4D  demonstrate that BmGPI12 is localized to the parasite plasma membrane and associated with vesicles and tubules.  FIGS. 4A and 4B  depict immunoelectron microscopic analysis of  B. microti  LabS1-infected mouse erythrocytes. Ultrathin sections of high pressure frozen and Durcupan resin-embedded infected erythrocytes were immunolabeled with anti-BmGPI12. IV: individual vesicle, P: parasite, PPM: parasite plasma membrane, RBCC: red blood cell cytoplasm, RBCM: red blood cell membrane, TOV: tube of vesicles.  FIG. 4C  depicts a schematic diagram showing the steps in the ultracentrifugation of plasma samples collected from  B. microti -infected mice.  FIG. 4D  depicts immunoblot analyses using pre-immune (PI) serum, and anti-BmGPI12 or anti-TER-119 antibodies on either intact plasma (PL) collected from mice infected with  B. microti  LabS1 strain or on 2 fractions (supernatant: Us, and pellet: Up) of plasma following ultracentrifugation at 120,000×g. 
         FIGS. 4E-4F  depict immunoelectron microscopic analysis of the plasma membrane fraction (Up) from mice infected with  B. microti  LabS1 using anti-BmGPI12 antibodies coupled to 10 nm gold particles. 
         FIGS. 5A-5E  demonstrate vesicle-mediated secretion of BmIPA48 antigen by  B. microti .  FIG. 5A  depicts distribution of BmIPA48 in the plasma (S), hemolysate (H) and membrane (P) fractions isolated from blood of uninfected or  B. microti  (LabS1)-infected erythrocytes was determined by western blotting using polyclonal antibodies against BmIPA48 (48 kDa). Pre-immune (PI) sera were used as a control.  FIG. 5B  depicts immunoblot analysis using pre-immune and anti-BmIPA48 sera on either intact plasma (PL) collected from mice infected with  B. microti  LabS1 strain or on 2 fractions (supernatant: Us, and pellet: Up) of plasma following ultracentrifugation at 120,000×g. BmIPA48 is associated primarily with the membrane fraction of plasma.  FIG. 5C  depicts immunofluorescence assay of BmIPA48 distribution in LabS1-infected erythrocytes. BmIPA48 was labelled with polyclonal antibodies and could be detected within the parasite and in discrete IV within the cytoplasm of the infected erythrocyte. Monoclonal antibodies against TER-119 were applied to label the erythrocyte plasma membrane and the DAPI labelling verified the presence of parasitic DNA in the otherwise enucleated host-erythrocytes. Staining of control uninfected red blood cells is shown.  FIGS. 5D and 5E  depict representative images of immunoelectron micrographs of  B. microti  LabS1-infected mouse erythrocytes. Ultrathin sections of high pressure frozen and Durcupan resin-embedded infected erythrocytes were immunolabeled with anti-BmIPA48 antibodies coupled to 10 nm gold particles. IV: individual vesicle, P: parasite, PPM: parasite plasma membrane, RBCC: red blood cell cytoplasm, RBCM: red blood cell membrane, TOV: tube of vesicles. 
         FIGS. 6A-6B  depicts a model of vesicular-mediated export of antigens by  B. microti.    
         FIG. 7  demonstrates BmGPI12 distribution in plasma and erythrocytes infected with the PRA99 strain of  B. microti . Immunoblotting analysis using pre-immune (PI) and anti-BmGPI12 polyclonal antibodies on fractions of uninfected erythrocytes (UI) or erythrocytes infected with  B. microti  strain PRA99. S: mouse plasma, H: hemolysate; (P) membrane fractions collected following saponin treatment of erythrocytes. In uninfected erythrocytes, the P fraction consists primarily of erythrocyte membrane. In  B. microti -infected erythrocytes, the P fraction includes both erythrocyte membrane and protein extracts from isolated parasites. The erythrocyte membrane protein TER-119 (52 kDa) was detected using an anti-TER-119 monoclonal antibody only in the P fractions from uninfected and  B. microti -infected red blood cells. 
         FIG. 8  demonstrates distribution of the apical end protein BmRON2 in  B. microti -infected cells. (A) Immunoblotting analysis using pre-immune (PI) and anti-BmRON2 polyclonal antibodies on fractions of uninfected erythrocytes (UI) or erythrocytes infected with  B. microti  strain LabS1. S: mouse plasma, H: hemolysate; (P) membrane fractions collected following saponin treatment of erythrocytes. Consistent with previous studies, BmRON2 (163 kDa) undergoes proteolytic degradation in infected cells. The 163 kDa band is found both in the P and S fractions but not in the H fraction consistent with the presence of BmRON2 on the surface of daughter parasites and its release to the plasma following rupture of the infected erythrocyte. No signal was detected when the pre-immune (PI) rabbit serum was used for immunodetection. 
         FIGS. 9A-9C  demonstrate electron microscopy evidence for IOV system emerging from the parasite plasma membrane. Ultrathin EPON-sections of LabS1  B. microti -infected mouse erythrocytes showing individual vesicles ( FIG. 9A ) as well as tubes of vesicles that are originating from the parasite ( FIGS. 9B and 9C ). IV: individual vesicle, P: parasite, PPM: parasite plasma membrane, RBCC: red blood cell cytoplasm, RBCM: red blood cell membrane, TOV: tube of vesicles. 
         FIGS. 10A-10B  depict Western blot and immunofluorescence analysis of  B. duncani -infected samples and analyzed using antibodies against BdGPI2.  FIG. 10A  depicts results from a preliminary immunoblot assay using anti-BdGPI2 peptide antibodies (raised in rabbits) on plasma (PL) from uninfected (U) or  B. duncani -infected (I) mice (in vivo) or on supernatant (Sup) or parasite fraction (P) from in vitro-cultured  B. duncani  in human red blood cells (I) or control uninfected human red blood cells (U).  FIG. 10B  depicts fluorescence microscopy imaging of uninfected or  B. duncani -infected human red blood cells using anti-BdGPI2 antibodies. A monoclonal antibody against human Band3 is used as a control to stain the surface of human red blood cells. DAPI is used to stain the nucleus of the parasites 
         FIGS. 11A-11B  depict Western blot and immunofluorescence analysis of  B. duncani -infected samples analyzed using antibodies against BdMGF3-1/HSP-70 precurser.  FIG. 11A  depicts preliminary immunoblot assay using anti-BdHsp70-2 peptide antibodies (raised in rabbits) on plasma (PL) from uninfected (U) or  B. duncani -infected (I) mice (in vivo) or on supernatant (Sup) or parasite fraction (P) from in vitro-cultured  B. duncani  in human red blood cells (I) or control uninfected human red blood cells (U).  FIG. 11B  depicts fluorescence microscopy on uninfected or  B. duncani -infected human red blood cells using anti-BdHsp70-2 antibodies. A monoclonal antibody against human Band3 is used as a control to stain the surface of human red blood cells. DAPI is used to stain the nucleus of the parasites. 
     
    
    
     DETAILED DESCRIPTION 
     Definitions 
     Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described. 
     As used herein, each of the following terms has the meaning associated with it in this section. 
     The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. 
     “About” as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20%, ±10%, ±5%, ±1%, or ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods. 
     As used herein, to “alleviate” a disease, defect, disorder or condition means reducing the severity of one or more symptoms of the disease, defect, disorder or condition. 
     The term “antigen” as used herein is defined as a molecule that provokes an immune response. This immune response may involve either antibody production, or the activation of specific immunologically-competent cells, or both. The skilled artisan will understand that any macromolecule, including virtually all proteins or peptides, can serve as an antigen. 
     Furthermore, antigens can be derived from recombinant or genomic DNA. A skilled artisan will understand that any DNA, which comprises a nucleotide sequences or a partial nucleotide sequence encoding a protein that elicits an immune response therefore encodes an “antigen” as that term is used herein. Furthermore, one skilled in the art will understand that an antigen need not be encoded solely by a full length nucleotide sequence of a gene. It is readily apparent that the present invention includes, but is not limited to, the use of partial nucleotide sequences of more than one gene and that these nucleotide sequences are arranged in various combinations to elicit the desired immune response. Moreover, a skilled artisan will understand that an antigen need not be encoded by a “gene” at all. It is readily apparent that an antigen can be generated synthesized or can be derived from a biological sample. Such a biological sample can include, but is not limited to a tissue sample, a tumor sample, a cell or a biological fluid. 
     The term “epitope” as used herein is defined as a small chemical molecule on an antigen that can elicit an immune response, inducing B and/or T cell responses. An antigen can have one or more epitopes. Most antigens have many epitopes; i.e., they are multivalent. In general, an epitope is roughly about 10 amino acids and/or sugars in size. Preferably, the epitope is about 4-18 amino acids, more preferably about 5-16 amino acids, and even more preferably 6-14 amino acids, more preferably about 7-12, and most preferably about 8-10 amino acids. One skilled in the art understands that generally the overall three-dimensional structure, rather than the specific linear sequence of the molecule, is the main criterion of antigenic specificity and therefore distinguishes one epitope from another. Based on the present disclosure, a peptide used in the present invention can be an epitope. 
     The term “immune response” as used herein is defined as a cellular response to an antigen that occurs when lymphocytes identify antigenic molecules as foreign and induce the formation of antibodies and/or activate lymphocytes to remove the antigen. 
     By the term “modified” as used herein, is meant a changed state or structure of a molecule or cell of the invention. Molecules may be modified in many ways, including chemically, structurally, and functionally. Cells may be modified through the introduction of nucleic acids. 
     By the term “modulating,” as used herein, is meant mediating a detectable increase or decrease in the level of a response in a subject compared with the level of a response in the subject in the absence of a treatment or compound, and/or compared with the level of a response in an otherwise identical but untreated subject. The term encompasses perturbing and/or affecting a native signal or response thereby mediating a beneficial therapeutic response in a subject, preferably, a human. 
     As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein&#39;s or peptide&#39;s sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof. 
     By the term “specifically binds,” as used herein with respect to an antibody, is meant an antibody which recognizes a specific antigen, but does not substantially recognize or bind other molecules in a sample. For example, an antibody that specifically binds to an antigen from one species may also bind to that antigen from one or more species. But, such cross-species reactivity does not itself alter the classification of an antibody as specific. In another example, an antibody that specifically binds to an antigen may also bind to different allelic forms of the antigen. However, such cross reactivity does not itself alter the classification of an antibody as specific. In some instances, the terms “specific binding” or “specifically binding,” can be used in reference to the interaction of an antibody, a protein, or a peptide with a second chemical species, to mean that the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody is specific for epitope “A”, the presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled “A” and the antibody, will reduce the amount of labeled A bound to the antibody. 
     As used herein, the terms “subject” and “patient” are used interchangeably. As used herein, a subject is preferably a mammal such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) and a primate (e.g., monkey and human), most preferably a human. 
     As used herein, to “treat” means reducing the frequency with which symptoms of a disease, defect, disorder, or adverse condition, and the like, are experienced by a patient. 
     The term “therapeutic” as used herein means a treatment and/or prophylaxis. A therapeutic effect is obtained by suppression, remission, or eradication of a disease state. 
     As used herein, a “therapeutically effective amount” is the amount of a composition of the invention sufficient to provide a beneficial effect to the individual to whom the composition is administered. 
     Ranges: throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range. 
     DESCRIPTION 
     The present invention relates to compositions and methods for detecting, diagnosing, screening for, treating and/or reducing  Babesia  infections such as babesiosis in a subject. The  Babesia  infection may include  Babesia microti  ( B. microti ) infections,  Babesia duncani  ( B. duncani ) infections, and/or combinations thereof. The subject may include a mammal. In certain embodiments, the subject is a human. 
     Compositions and Methods for Detecting and/or Diagnosing an Infection 
     In certain embodiments, the present invention provides compositions and methods for detecting an infection in a subject. The infection includes parasitic infections such as  Babesia  infections caused by one or more  Babesia  strains. The  Babesia  infection may include  Babesia microti  ( B. microti ) infections,  Babesia duncani  ( B. duncani ) infections, and/or combinations thereof. 
     In certain embodiments, the compositions includes one or more compositions for detecting one or more secreted antigens. The one or more antigens include one or more antigens generated by one or more cells infected with a  Babesia  infection (e.g.  B. duncani  infection,  B. microti  infection and/or combination thereof). The one or more antigens may include one or more peptides including one or more of SEQ ID NOs: 1-62. 
     Compositions for Detecting/Diagnosing a  Babesia  Infection 
     In certain aspects, the present invention provides compositions for detecting and/or diagnosing and infection in a subject. The compositions may include one or more immunologic agents for detecting one or more antigens or peptides generated by one or more cells. The one or more antigens or peptides may be detected in a sample obtained from a subject with an infection. 
     In various embodiments, the one or more immunologic agents include one or more immunologic agents that have specificity for one or more antigens generated by the one or more cells. For example the one or more immunologic agents may include one or more of an antibody, an antibody fragment, antisense peptide, synthetic peptide, synthetic antibody, synthetic antibody fragment, synthetic hybridized antibody or antibody fragment. The one or more immunologic agents detect, bind to, or interact with one or more peptides or antigens generated by one or more cells infected with one or more of  B. duncani, B. microti , or both. The one or more peptides of antigens may include one or more selected from SEQ. ID. NOs: 1-46, 47-62. In some embodiments, one or more antigens or peptides selected from SEQ. ID. NOs: 1-46 may be used to detect a  B. duncani  infection in a sample obtained from a subject. In some embodiments, one or more antigens or peptides selected from SEQ. ID. NOs: 47-62 may be used to detect a  B. microti  infection in a sample obtained from a subject. 
     In certain embodiments, the biological sample includes one or more of blood, urine, saliva, feces, lymph, bile and the like, and/or one or more combinations thereof. The sample may include one or more of a whole blood sample, a plasma sample, a serum sample, a hemolysate sample, and the like. In some embodiments, the one or more infected cells include one or more blood-derived cells such as one or more of erythrocyte, leukocyte, plasma cell, platelets, and the like. 
     Methods for Detecting/Diagnosing a  Babesia  Infection 
     In certain embodiments, the present invention relates to methods for detecting and/or diagnosing one or more  Babesia  infections including babesiosis,  B. duncani  infection,  B. microti  infection, and/or combinations thereof. 
     In certain embodiments, the methods include obtaining one or more biological samples obtained from a subject. The one or more biological samples may include one or more of blood, urine, saliva, feces, lymph, bile and the like, and/or one or more combinations thereof. The blood sample may include whole blood sample, a plasma sample, a serum sample, a hemolysate sample, and the like. The subject may include a human subject. The subject may have a known or suspected infection of one or more  Babesia  strains including one or more of  B. duncani, B. microti , and/or combinations thereof. 
     Embodiments of the methods include detecting one or more infections in one or more biological samples obtained from a subject. The one or more infections include one or more of babesiosis,  B. duncani  infection,  B. microti  infection, and/or combinations thereof. The infection may be detected using one or more immunologic agent having specificity for one or more antigens or peptides including one or more of SEQ. ID. NOs: 1-46, 47-62, and/or one or more combinations thereof. In certain embodiments, the one or more antigens or peptides are detected using one or more assay platforms including for example, an enzyme-based assay, a radioimmunoassay, a PCR amplification-based assay, a fluorogenic immunoassay, a chemiluminescence-based assay, immunoblotting assay, and combinations thereof. The assay may include one or more of Western blot, immunofluorescence, immune-electron microscopy, ELISA, immunoprecipitation, and the like. 
     In certain embodiments, the one or more antigens or peptides are detected relative to a comparator control. In certain embodiments, an infection is detected if the signal is greater than or less than a threshold value, a difference relative to a comparator control, or the like. 
     Embodiments of the methods include measuring one or more biological samples obtained from a subject in order to detect the presence of an infection. Embodiments of the methods include measuring one or more biological samples obtained from a subject in order to evaluate the efficacy of one or more therapeutic agents. In various embodiments, the sample is a blood sample, a sera sample, and/or one or more samples containing one or more other suitable bodily fluids. In various embodiments, the subject is a subject that is healthy, a subject that is confirmed to be infected, a subject that is suspected to be infected, a subject that has been treated for an infection and is believed to no longer be infected, and the like. The subject may be evaluated by assaying the one or more samples obtained from the subject. The one or more samples may be evaluated at one or more time points in order to evaluate the status of an infection and/or the efficacy of one or more treatments for the infection. In various embodiments, the one or more time points include prior to an intentional infection, such as about 24 hours prior to infection, from about 24 hours to about 12 hours prior to infection, from about 12 hours to about 8 hours prior to infection, from about 8 hours to about 4 hours prior to infection, from about 4 hours to about 1 hour prior to infection, less than about 1 hour prior to infection, and any and all intervals and increments therebetween. The one or more time points include at the time of infection, up to 1 hour post-infection, from about 1 hour to about 1 day post-infection, from about 1 day to about 2 days post infection, about 2 days to about 3 days post-infection, from about 3 days to about 4 days post-infection, from about 4 days to about 5 day post-infection, from about 5 days to about 6 days post-infection, from about 6 day to about 7 days post-infection, from about 1 week to about 2 weeks post-infection, from about 2 weeks to about 4 weeks post-infection, from about 4 weeks to about 6 weeks post-infection, and so on. In various embodiments, the one or more time points include at the time of infection, immediately prior to administering a therapeutic agents, at the time of administering a therapeutic agent, and at one or more time points after administration of a therapeutic agent. The one or more time points may include up to 24 hours prior to administration of a therapeutic agent, from about 24 hours to about 12 hours prior to administration of a therapeutic agent, from about 12 hours to about 8 hours prior to administration of a therapeutic agent, from about 8 hours to about 4 hours prior to administration of a therapeutic agent, from about 4 hours to about 1 hour prior to administration of a therapeutic agent, less than about 1 hour prior to administration of a therapeutic agent, and any and all increments and intervals therebetween. In various embodiments, the one or more time points include: up to 1 hour post-administration, from about 1 hour to about 12 hours post-administration, from about 12 hours to about 24 hours post-administration, from about 1 day to about 2 days post-administration, from about 2 day to about 5 days post-administration, from about 5 days to about 7 days post-administration, from about 7 days to about 14 days post-administration, from about 14 day to about 28 days post-administration, from about 28 days to about 42 days post-administration, and so on. The one or more time points may include from about 1 week to about 2 weeks post-administration, from about 2 weeks to about 4 weeks post-administration, from about 4 weeks to about 6 weeks post-administration, and so on. 
     The methods include detecting an infection in a subject using one or more techniques as described herein. The methods include administering to the subject an effective amount of at least one therapeutic agent including one or more anti-protazoan therapeutic agents. In various embodiments, the anti-protazoan therapeutic agent includes one or more anti-protazoan agents as understood in the art. 
     EXPERIMENTAL EXAMPLES 
     The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only, and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples, but rather, should be construed to encompass any and all variations which become evident as a result of the teaching provided herein. 
     Example 1 
     The apicomplexan parasite  Babesia microti  is the primary agent of human babesiosis, a malaria-like illness and potentially fatal tick-borne disease. Unlike its close relatives, the agents of human malaria,  B. microti  develops within human and mouse red blood cells in the absence of a parasitophorous vacuole, and its secreted antigens lack trafficking motifs found in malarial secreted antigens. Here, it is shows that after invasion of erythrocytes,  B. microti  undergoes a major morphogenic change during which it produces an interlacement of vesicles (IOV); the IOV system extends from the plasma membrane of the parasite into the cytoplasm of the host erythrocyte. 
     Methods 
     Parasite Strains 
       B. microti  isolates used in this study are LabS1 and PRA99. These strains were maintained in rag2−/− Knockout (B6.12956-Rag2tmlFwa N12) and SCID (severe combined immunodeficiency) C.B17 SCID C.B-Igh-1b/IcrTac-Prkdcscid and CB17/Icr-Prkdcscid/IcrIcoCrl mice as previously described. Parasitemia was determined by thin blood recombinant protein consisting of amino acids 1-302 of BmGPI12, BmIPA48 (Genebank ID # XP_021338473; EupathDB ID #: BMR1_03g00947) peptide CNKIKTDGGKVDSNS, BmRON2 peptide NKIKTDGGKVDSNS. Monoclonal anti-mouse TER-119 (INVITROGEN®) was used as a control. 
     Plasma Collection and Fractionation Studies 
     Blood from uninfected or  B. microti -infected animals was collected by cardiac puncture and stored in tubes containing K 2 EDTA (dipotassium ethylenediaminetetraacetate) solution. For plasma separation, samples were spun down at 1,300 rpm (200×g) for 20 minutes at room temperature. Plasma or supernatant was removed and added to new 1.7 ml microcentrifuge tubes. The remaining cell pellet was washed twice with PBS supplemented with 1% saponin, incubated on ice for 30 minutes and spun at 9,300×g for 10 minutes at 4° C. The resulting supernatant (hemolysate) was collected and the remaining pellet (uninfected) or parasite (infected) fractions were washed twice with PBS and spun at 9,300×g for 10 minutes at 4° C. Plasma (S), hemolysate (H) and pellet (P) fractions were mixed with Lammeli buffer, separated on SDS-PAGE and analyzed by immunoblotting. 
     Isolation of Vesicles from Plasma 
     IOVs were isolated from the plasma of uninfected mice or mice infected with  B. microti  by sequential centrifugations following the protocol for the isolation of exosomes with some modifications. Briefly, 400 μm plasma from animals was diluted with 5 ml PBS and centrifuged at 500×g for 30 minutes and then at 16,000×g for 45 min to remove microvesicles. IOVs were pelleted with ultracentrifugation (UC) at 120,000×g for 14 hours at 4° C. using a SORVALL™ MTX 150 Micro-Ultracentrifuge with a S52-ST Swinging-Bucket Rotor (THERMO FISHER SCIENTIFIC). The resulting pellet (P1) was collected and the supernatant was spun again using the same conditions. The resulting pellet (P2) and supernatant (Us) fractions were collected. 
     Immunoblot Analysis 
     Equal concentrations of plasma (prior to ultracentrifugation), supernatant (Us) and pellet (Up) fractions obtained after UC were analyzed by immunoblotting. To obtain equal concentrations, supernatant (Us) fractions were concentrated with 20% trichloroacetic acid. The pellet fractions (P1 and P2) were diluted further and combined as a single pellet (Up). All the samples were resuspended in Lammeli-buffer and loaded on 10% Mini-PROTEAN® TGX (BIO-RAD LABORATORIES®, Hercules, Calif.) and transferred to nitrocellulose membranes. Membranes were blocked in 5% milk and incubated with a rabbit anti-BmGPI12 serum (1:250) or pre-immune serum (1:250 dilution) overnight at 4° C. The next day, membranes were washed in TBS-T and incubated with ECL-horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (1:10,000 dilution) for 1 hour. Following additional washes, the membranes were incubated with ECL western blotting detection reagents (GE HEALTHCARE®, Amersham, UK) and exposed to X-ray film using KODAK® autoradiography. The same method was used for immunoblotting of parasite and erythrocyte membrane (P), erythrocyte cytoplasm or hemolysate (H) and plasma or supernatant (S) fractions. Polyclonal antibodies including rabbit anti-BmIPA48 (1:100) and rabbit anti-BmRON2 (1:100) and their corresponding pre-immune sera and monoclonal anti-mouse TER-119 antibody (1:500) were also analyzed by immunoblotting. 
     Immunofluorescence Assay (IFA) 
     Rabbit anti-BmGPI12 (BmSA-1) antibodies were used to assess the localization of BmGPI12 in  B. microti  LabS1 and PRA99. Rabbit anti-BmIPA48 serum was used to evaluate the localization of BmIPA48 in LabS1. Blood smears were prepared from retro orbital bleeding (after centrifugation at 1,500 rpm for 2 min (MINISPIN PLUS®, EPPENDORF™) on thin 22 mm×22 mm microscopy cover glasses (12-542-B; FISHERBRAND®) and immediately transferred into a 6-well plate (COSTAR®, CORNING®, Inc.) which was put on a cold metal plate on ice. Afterwards 6-well plates with the blood smears were stored at 4° C. before being further processed. To perform the IFA, thin blood smears were removed from the 4° C. storage and fixed in 1% formaldehyde (28908; THERMO SCIENTIFIC®) diluted in PBS (10010-023; GIBCO®) at 37° C. (PRECISION® mechanical convection incubator, model 6 LM) for 30 minutes followed by three brief rinses in PBS. Smears were then incubated for 1 hour in a blocking buffer (5% heat-inactivated fetal bovine serum (16000-044, GIBCO®) 5% normal goat serum (16210-072; GIBCO-BRL®) and 0.1% saponin (10% stock solution in PBS) (57900-100G; SIGMA®)) at 37° C. and then once rinsed in wash buffer (0.5% fetal bovine serum, 0.5% normal goat serum, 0.05% saponin). To target the mouse erythrocyte membrane, FITC anti-mouse TER-119/Erythroid Cells; Clone: TER-119 (116206; BIOLEGEND®) was used at a 1:1,000 dilution. The anti-BmGPI12 or anti-BmIPA48 antibodies were used at 1:1,000 dilution. 
     The smears were either incubated overnight at 4° C. while being rocked gently (Orbi-Blotter; BENCHMARK®) or for 1 hour at 37° C. The next day, slides were washed three times for 2 minutes each with wash buffer and incubated with secondary antibody, Goat anti-rabbit IgG (H+L) rhodamine conjugate (31670, Invitrogen) (1:1,000) at 37° C. for 1 hour and washed three times for 2 minutes each in wash buffer. After three brief rinses in PBS and one brief rinse in ddH 2 O, the coverslips were mounted on sandblasted single frosted pre-cleaned microscope slides (421-004T; THERMO SCIENTIFIC®) using PROLONG™ Gold antifade reagent supplemented with DAPI (P36935; INVITROGEN® by THERMO FISHER SCIENTIFIC®) and incubated at RT in the dark overnight, before they were examined with the LEICA® TCS SP8 STED 3× microscope (LEICA® Microsystems GmbH; Wetzlar, Germany). 
     Imaging on Leica TCS SP8 STED 3× 
     The confocal images were acquired with a LEICA® TCS SP8 STED 3× microscope. A HC PL APO CS2 100×/1.40 oil immersion objective was used for image acquisition. FITC anti-mouse TER-119/Erythroid Cells; Clone: TER-119 was excited at 488 nm (500 nm-571 nm, HyD3), rhodamine was excited at 550 nm (569 nm-650 nm, HyD3) and DAPI was excited at 405 nm (430 nm-470 nm, HyD1). The pinhole was set to 1 AU. The images were acquired in unidirectional confocal mode with 1,000 Hz scan speed (line average 6). The image size was image size was chosen to be 38.75 μm×38.75 μm (1,024×1,024 pixels). The PMT Trans was further activated to enable the acquisition of DIC images. 
     Sample Preparation for Electron Microscopy Cryosectioning 
     The sample pellet was fixed in 4% paraformaldehyde (PFA) in PBS for 30 minutes at room temperature followed by further fixation in 4% PFA at 4° C. for 1 hour. They were rinsed in PBS and re-suspended in 10% gelatin. Chilled blocks were trimmed and placed in 2.3 M sucrose overnight on a rotor at 4° C. They were transferred to aluminum pins and frozen rapidly in liquid nitrogen. The frozen blocks were cut on a LEICA® Cryo-EMUC6 UltraCut and 65 nm thick sections were collected using the Tokoyasu method (Tokuyasu, 1973) and placed on carbon/formvar-coated grids and floated in a dish of PBS for immunolabeling. 
     Immunolabeling of the Ultra-Thin Sections 
     Samples were processed according to the method described by Slot and Geuze (Slot and Geuze, Nature Protocols. 2007; 2(10):2480-2491.). The grids were incubated on rabbit anti-BmGPI12 (1:100) or rabbit anti-BmIPA48 (1:100) and their corresponding pre-immune sera (1:500). For secondary antibody, 10 nm Protein A gold (Utrecht Medical Center) was used. All grids were rinsed in PBS, fixed using 1% glutaraldehyde for 5 minutes, rinsed again and transferred to a UA/methylcellulose drop before being collected and dried. 
     High Pressure Freezing and Freeze Substitution Epon Section and Labeling 
     Samples fixed in 4% PFA were frozen using a LEICA® HMP100 at 2000 psi. The frozen samples were then freeze substituted using a LEICA® Freeze AFS unit starting at −95° C. using 1% osmium tetroxide, 1% glutaraldehyde and 1% water in acetone for 10 h, warmed to −20° C. for 12 hours and then to 4° C. for 2 hours. The samples were well rinsed in 100% acetone and infiltrated with DURCUPAN™ resin (ELECTRON MICROSCOPY SCIENCES®) and baked at 60° C. for 24 hours. Hardened blocks were cut using a LEICA® UltraCut UC7 and 60 nm sections were collected on formvar/carbon coated nickel grids. 
     Immuno-Labeling of Resin Sections Grids were placed section side down on drops of 1% hydrogen peroxide for 5 minutes, rinsed and blocked for non-specific binding on 3% bovine serum albumin in PBS containing 1% Triton-X for 30 minutes. Grids were incubated with a primary antibody rabbit anti-BmGPI12 or anti-BmIPA48 1:100 overnight, rinsed in buffer and then incubated with the secondary antibody 10 nm protein A gold (UtrechtUMC) for 30 minutes. The grids were rinsed and fixed using 1% glutaraldehyde for 5 minutes, rinsed well in distilled water, and contrast stained using 2% uranyl acetate and lead citrate. Grids were all viewed in a FEI Tencai Biotwin TEM at 80 kV. Images were taken using Morada CCD and iTEM (OLYMPUS®) software. 
     Image Processing 
     FIJI (imagej.net/Fiji) and MICROSOFT® POWERPOINT® (MICROSCOFT® Corporation) was used to analyze and prepare raw images of Giemsa smears, EM images and fluorescence images for presentation. Diameter and length of EM structures were determined via the line tool in FIJI. The Huygens Professional Software (Scientific Volume Imaging) was further used to deconvolve the fluorescence images acquired with the LEICA® TCS SP8 STED 3×. 
     Selected Results 
       B. microti  major secreted antigen localizes to vesicular structures associated with parasite morphogenesis. The immunodominant BmGPI12 of  B. microti  is encoded by a member of the bmn multigene family and one of the most highly expressed genes of the parasite during its development within red blood cells. Consistent with the secretion of BmGPI12 from the parasite into the red blood cytoplasm and subsequently into the host environment, immunoblot analyses using anti-BmGPI12 antibodies on blood collected from mice and fractionated to collect plasma (S), erythrocyte cytoplasm (H), and membrane (P) fractions showed the presence of BmGPI12 in all three fractions from animals infected with  B. microti  strains (LabS1 ( FIG. 1 ) or PRA99 ( FIG. 7 )) but not from uninfected animals ( FIG. 1A  and  FIG. 7 ). As a control, immunoblot analyses conducted using a monoclonal antibody against the mouse erythrocyte membrane protein, TER-119 (glycophorin A-associated protein (Ly-76)) identified this protein in the membrane (P) fractions of both uninfected and  B. microti -infected erythrocytes (Kina et al., 2000, Br J Haematol. 109:280-287) but not in the plasma or erythrocyte cytoplasm fractions ( FIG. 1B ). As expected, antibodies against the  B. microti  rhoptry neck protein BmRON2, a highly conserved protein among apicomplexan parasites that localizes to the parasite apical end (Ord et al., 2016, Infect Immun. 84:1574-1584), identified the protein in the membrane and plasma fractions, but not in the erythrocyte cytoplasm ( FIG. 8 ). This finding is consistent with the association of BmRON2 with the parasite during its intra-erythrocytic development and its release following the rupture of the infected erythrocyte and exit of daughter parasites (Ord et al., 2016, Infect Immun. 84:1574-1584). As a control, pre-immune sera were used to analyze the fractions from uninfected and  B. microti -infected erythrocytes and no signal could be detected ( FIGS. 7 and 8 ). 
     The localization of BmGPI12 was further examined by confocal microscopy. The analysis identified BmGPI12 in both the cytoplasm and plasma membrane of the parasite as well as in the cytoplasm of the infected erythrocyte in well-defined dendrite-like structures and distinct foci ( FIG. 1B ). These structures are reminiscent of membranous extensions often seen in Giemsa-stained blood smears of  B. microti -infected erythrocytes at different stages of parasite development (representative images of LabS1) ( FIG. 2A ). Blood smears prepared from four  B. microti -infected mice showed that the parasite undergoes major morphogenic changes throughout its intraerythrocytic life cycle that include ring-shaped forms, rings with dendrite-like tubovesicular structures (TOVs), dividing parasites (tetrads) and tetrads with TOVs ( FIGS. 2B and 2C ). The proportion of parasites with TOV structures was similar as parasite burden increased in infected animals suggesting that the morphogenic events are part of the intraerythrocytic life cycle of  B. microti  ( FIG. 2C ). 
     Vesicle-Mediated Export of BmGPI12 Antigen into the Cytoplasm of  B. microti -Erythrocytes and Host Plasma 
     To investigate at the ultrastructural level the nature of the TOVs, electron microscopy analyses of ultrathin sections of erythrocytes from  B. microti -infected mice were conducted. These analyses revealed tubes of connected vesicles as well as individual vesicles in the cytoplasm of infected cells ( FIGS. 3A and 3B , and  FIG. 9 ), but not in the uninfected cells ( FIG. 3C ). The measured diameter of individual vesicles (IV) is approximately 0.110 μm±0.0052 μm (Mean±SEM), whereas the length of tubes of vesicles (TOV) ranges between 0.405 μm±0.056 μm (Mean±SEM)) and 0.900 μm, depending on the sections. Interestingly, detailed analysis of these structures revealed that the TOVs emerge directly from the parasite plasma membrane and extend into the infected erythrocyte ( FIG. 3B  and  FIG. 9  (panels B and C)). Both the cytoplasm of the parasite and the content of the vesicles and tubules share the same electron density ( FIGS. 3A and 3B , and  FIG. 7 ), further demonstrating that the structures are of parasite origin. 
     To determine whether the IV and TOV structures identified by electron microscopy are the same BmGPI12-positive structures detected by confocal microscopy, immunoelectron microscopy (IEM) analyses were performed on  B. microti -infected murine erythrocytes using anti-BmGPI12 antibodies coupled to 10 nm gold particles. IEM analyses showed that BmGPI12 localizes to the parasite plasma membrane (PPM) as well as to the IVs and the TOVs ( FIGS. 4A and 4B ). 
     Export of BmGPI12-Containing Vesicles from  B. microti -Infected Erythrocytes 
     The finding that  B. microti  produces IVs and TOVs inside infected erythrocytes and that BmGPI12 is associated with these structures led us to further investigate whether this protein is secreted into the host environment via a vesicle-mediated secretory mechanism or as a free antigen. This was achieved by subjecting plasma samples from  B. microti -infected mice to ultracentrifugation at 120,000×g to separate fractions containing membrane-associated structures including vesicles and tubules (Up, ultracentrifuged pellet fraction) from those containing free proteins (Us, ultracentrifuged supernatant fraction) ( FIG. 4C ). As shown in  FIG. 4D , BmGPI12 was found in both the Up and Us fractions, suggesting that it is released into the erythrocyte environment as a membrane associated protein, but could also be found as a free protein, most likely due to cleavage of the GPI anchor by plasma enzymes. The host protein TER-119, which associates exclusively with the erythrocyte membrane, was not found in the Up or Us fractions ( FIG. 4D ), demonstrating that exported proteins are the main proteins found in these fractions. Analysis of the membrane fractions by immunoelectron microscopy identified BmGPI12 associated with vesicles and tubular structures with similar sizes as those observed inside the infected erythrocytes ( FIGS. 4E and 4F ). 
     Evidence for Vesicular-Mediated Export of the Immunodominant Antigen BmIPA48 in  B. microti    
     To determine whether the vesicular system used by  B. microti  for secretion of BmGPI12 is also used by the parasite to export other antigens, we examined the cellular distribution of another  B. microti  antigen BmIPA48, which was previously shown to trigger strong IgM and IgG response in infected outbred mice (Silva et al., 2016, Scientific reports, 6:35284). BmIPA48 encodes a 48-kDa antigen with an N-terminal signal peptide but no GPI-anchor motif or transmembrane domains (Silva et al., 2016, Scientific reports, 6:35284). As shown in  FIG. 5 , BmIPA48 is expressed in the parasite, secreted into the erythrocyte cytoplasm, and then released into the host plasma ( FIG. 5A ). Similar to BmGPI12, confocal microscopy shows association of the antigen with discrete foci in the infected red blood cell ( FIG. 5C ). However, unlike BmGPI12, analysis of the distribution of BmIPA48 following ultracentrifugation shows the presence of the protein exclusively in the vesicles-containing fraction (Up fraction) ( FIG. 5B ). Consistent with these findings, immunoelectron microscopy analyses of  B. microti -infected erythrocytes demonstrates the presence of BmIPA48 inside vesicles found both in the parasite cytoplasm as well as secreted by the parasite into the erythrocyte cytoplasm ( FIGS. 5D and 5E ). 
     This study reports the first evidence that the human pathogen  B. microti  employs a novel mechanism to export parasite proteins into the host erythrocyte and subsequently into the erythrocyte environment. This system employs a network of parasite made dendrite-like membrane branches consisting of connected vesicles. We show that at least two immunodominant antigens of  B. microti , BmGPI12 and BmIPA48, are exported by the parasite via this mechanism. 
     A recent study estimated that ˜398 proteins may be secreted by  B. microti  during its development within mammalian erythrocytes (Silva et al., 2016, Scientific reports, 6:35284). Some of these proteins may be exported into the erythrocyte cytoplasm or erythrocyte membrane where they may function to modulate the host cell cytoskeleton or to facilitate uptake of nutrients. Others might be further exported into the host plasma to modulate the host response or effect other changes beneficial to the parasite. Both BmGPI12 and BmIPA48 contain an N-terminal signal peptide but lack specific motifs such as the PEXEL motif found in other apicomplexan parasites and associated with secretion of proteins into the host (Cornillot et al., 2016, Transfusion, 56:2085-2099; de Koning-Ward et al., 2016, International journal for parasitology; Lanzer et al., 2006, International journal for parasitology. 36:23-36; Marti et al., 2005, J Cell Biology, 171:587-592; Pelle et al., 2015, Cell Microbiology. 17:1618-1639; Sherling and van Ooij, 2016, FEMS microbiology reviews, 40:701-721). In fact, all predicted secreted proteins of  B. microti  lack such a motif (Silva et al., 2016, Scientific reports, 6:35284), suggesting that this parasite might have evolved a novel mode of protein export. Interestingly, unlike  P. falciparum  where the PEXEL motif plays an important role in the trafficking of parasite proteins across the parasitophorous vacuole that separates the parasite from the erythrocyte cytoplasm,  B. microti  spends most of its intraerythrocytic development without a vacuole, thus eliminating the need for a translocon to export proteins into the host cytoplasm. Consistent with this model, analysis of the  B. microti  genome shows the lack of orthologs of most components of the malaria translocon (Cornillot et al., 2012, Transfusion, 56:2085-2099; Silva et al., 2016, Scientific reports, 6:35284). 
     The electron microscopy analyses of ultrathin sections of  B. microti -infected erythrocytes showed that the interlacement of vesicles consists of individual vesicles (IVs) and tubes of vesicles (TOVs) with a diameter of ˜0.110 μm±0.0052 μm. Giemsa staining showed that the TOVs can vary in length between infected erythrocytes with some extensions several μm in diameter. Further analysis of the blood smears showed the presence of TOVs throughout the life cycle of the parasite and suggest that production of filamentous forms represent a distinct morphogenic event in the development of the parasite. Interestingly, analysis of cryosections showed that the IOV system of  B. microti  is of a composition similar to that of the cytoplasm of the parasite. An enlargement of a section near the parasite plasma membrane in  FIG. 3B  showed a tubule consisting of two vesicles directly emerging out of the parasite membrane. This distinguishes this system from the TVM system previously described in  P. falciparum , which has been shown to emerge from the parasitophorous vacuolar membrane of the malaria parasite. 
     This study employed fractionation methods used to characterize the secreted vesicles produced by  B. microti . Using this approach, we found that BmGP12 could readily be detected in both the soluble and membrane fractions following centrifugation of plasma samples or culture medium of short-term cultured parasites, whereas BmIPA48 was found primarily in the vesicle-rich membrane fractions. The difference in the distribution of these two proteins could be due to their respective localization in the vesicles and tubules. Immunoelectron microscopy analyses showed that BmGPI12 is primarily found on the parasite plasma membrane and on the membranes of vesicles and tubules. Interestingly, the 10 nm gold particles could be found both inside and outside these vesicles and tubules. It is, therefore, likely that following vesicular release, BmGPI12 exposed outside the vesicles is cleaved and thus found in the soluble fraction whereas the proteins still residing inside the vesicles remain associated with the membrane fraction. On the other hand, immunoelectron microscopy analysis of BmIP48 showed the presence of the protein primarily inside the vesicles, consistent with their exclusive association with the membrane fraction. Interestingly, none of the vesicles could be detected on outer layer of the erythrocyte membrane of infected cells, suggesting that the vesicles are likely released into the host environment following rupture of  B. microti -infected erythrocytes as suggested in our model ( FIGS. 6A-6B ). 
     Example 2:  B. microti  Secreted Antigens Identified by NANOTRAP® Proteomic Approach 
       
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 List of 15 exported antigens of  B. microti  identified using  
               
               
                 a NANOTRAP ® proteomic approach 
               
            
           
           
               
               
               
               
            
               
                   
                   B. microti  Gene 
                 Annotated protein 
                 Protein length 
               
               
                   
                   
               
            
           
           
               
               
               
               
            
               
                   
                 BmR1_04g08775 
                 Putative HtpG 
                 712 
               
               
                   
                 BmR1_03g03490 
                 Putative EF-1 alpha subunit 
                 447 
               
               
                   
                 BmR1_04g05965 
                 Putative Enolase 
                 438 
               
               
                   
                 BMR1_03g01010 
                 Putative Ubiquitin family 
                 77 
               
               
                   
                 BMR1_03g01340 
                 Putative Ubiquitin C 
                 282 
               
               
                   
                 BMR1_03g02390 
                 Putative Actin 
                 376 
               
               
                   
                 BMR1_02g03540 
                 Putative RPL18 
                 187 
               
               
                   
                 BmR1_04g09800 
                 Multifunctional tryptophan 
                 425 
               
               
                   
                   
                 biosynthesis protein 
                   
               
               
                   
                 BMR1_01g02545 
                 Hsp70 
                 644 
               
               
                   
                 BMR1_02g03395 
                 Unknown function 
                 395 
               
               
                   
                 BMR1_03g04775 
                 S-adenosylmethionine  
                 404 
               
               
                   
                   
                 synthetase 
                   
               
               
                   
                 BMR1_03g03380 
                 Glutamate dehydrogenase 
                 467 
               
               
                   
                 BmR1_04g08050 
                 peptide chain release  
                 433 
               
               
                   
                   
                 factor eRF subunit 1 
                   
               
               
                   
                 BMR1_02g01430 
                 Unknown function 
                 1094 
               
               
                   
                 BmR1_04g08520 
                 Unknown function 
                 1024 
               
               
                   
                   
               
            
           
         
       
     
     Example 3:  B. duncani  Secreted Antigens Identified by NANOTRAP® Proteomic Approach 
       
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 List of 46 exported proteins of  B. duncani  identified  
               
               
                 by a NANOTRAP ®-based proteomic approach. 
               
            
           
           
               
               
               
            
               
                   B. dunani  Gene 
                 Protein Length 
                 Annotation 
               
               
                   
               
            
           
           
               
               
               
            
               
                 BdWA1_II1364 
                 229 
                 Hypothetical protein 
               
               
                 BdWA1_II1520 
                 644 
                 Hsp70 (BdHsp70-1) 
               
               
                 BdWA1_III2507 
                 1058 
                 Dynamin family, putative 
               
               
                 BdWA1_I0004 
                 183 
                 Hypothetical protein 
               
               
                 BdWA1_I0706 
                 376 
                 Peptidylprolyl isomerase, putative 
               
               
                 BdWA1_II1893 
                 160 
                 Hypothetical protein 
               
               
                 BdWA1_III3045 
                 132 
                 BdGPI6-Hypothetical protein 
               
               
                 BdWA1_I0760 
                 188 
                 RPL22p/L17e, putative 
               
               
                 BdWA1_III2950 
                 490 
                 Protein disulfide isomerase  
               
               
                   
                   
                 related protein 
               
               
                 BdWA1_II2303 
                 442 
                 Enolase 
               
               
                 BdWA1_I0910 
                 177 
                 Hypothetical protein 
               
               
                 BdWA1_I0555 
                 131 
                 Polyubiquitin, putative 
               
               
                 BdWA1_II1366 
                 229 
                 Hypothetical protein 
               
               
                 BdWA1_I0089 
                 797 
                 Metallo-beta-lactamase  
               
               
                   
                   
                 superfamily, putative 
               
               
                 BdWA1_III2562 
                 392 
                 Alpha/beta hydrolase family,  
               
               
                   
                   
                 putative 
               
               
                 BdWA1_I0812 
                 713 
                 Hsp90 (BdHsp90-1) 
               
               
                 BdWA1_III2693 
                 126 
                 Hypothetical protein 
               
               
                 BdWA1_II1642 
                 255 
                 14-3-3 protein, putative 
               
               
                 BdWA1_II2273 
                 742 
                 RAP domain/Core histone  
               
               
                   
                   
                 H2A/H2B/H3/H4, putative 
               
               
                 BdWA1_III2537 
                 400 
                 Rhoptry-associated protein 1  
               
               
                   
                   
                 (RAP-1), putative 
               
               
                 BdWA1_II1831 
                 242 
                 BdGPI8-Hypothetical protein 
               
               
                 BdWA1_II2038 
                 917 
                 RPL1p/L10e family, putative 
               
               
                 BdWA1_I0231 
                 618 
                 BdGPI17-Hypothetical protein 
               
               
                 BdWA1_II1370 
                 266 
                 Hypothetical protein 
               
               
                 BdWA1_I0253 
                 498 
                 Hypothetical protein 
               
               
                 BdWA1_III2944 
                 296 
                 phosphatidylinositol-4-phosphate  
               
               
                   
                   
                 5-kinase, putative 
               
               
                 BdWA1_I0708 
                 376 
                 Actin, putative 
               
               
                 BdWA1_II2269 
                 292 
                 EF-hand domain/EF-hand  
               
               
                   
                   
                 domain pair/EF hand, putative 
               
               
                 BdWA1_II2320 
                 476 
                 Sodium: dicarboxylate  
               
               
                   
                   
                 symporter family, putative 
               
               
                 BdWA1_III3035 
                 207 
                 Hypothetical protein 
               
               
                 BdWA1_I0924 
                 299 
                 Ethanolamine-phosphate  
               
               
                   
                   
                 cytidylyltransferase, putative 
               
               
                 BdWA1_III2663 
                 516 
                 MAC/Perforin domain  
               
               
                   
                   
                 containing protein, putative 
               
               
                 BdWA1_II2261 
                 1416 
                 RNA polymerase Rpb1,  
               
               
                   
                   
                 domain containing protein 
               
               
                 BdWA1_III3073 
                 333 
                 PP-loop family, putative 
               
               
                 BdWA1_II2013 
                 278 
                 5′-3′ exonuclease, putative 
               
               
                 BdWA1_I0563 
                 825 
                 Hypothetical protein 
               
               
                 BdWA1_II2256 
                 374 
                 Hypothetical protein 
               
               
                 BdWA1_II2291 
                 535 
                 Hsp70 (BdHsp70-2) 
               
               
                 BdWA1_II1496 
                 400 
                 elF4A-3 
               
               
                 BdWA1_II2200 
                 108 
                 L7Ae 
               
               
                 BdWA1_I0250 
                 418 
                 26S protease regulatory subunit 6A 
               
               
                 BdWA1_II1994 
                 524 
                 GMP synthase 
               
               
                 BdWA1_III2590 
                 126 
                 GTP-binding nuclear protein Ran 
               
               
                 BdWA1_I0810 
                 612 
                 Hsp70 (BdHsp70-3) 
               
               
                 BdWA1_II1634 
                 117 
                 Core histone H2A/H2B/H3/H4,  
               
               
                   
                   
                 putative 
               
               
                 BdWA1_II1773 
                 396 
                 26S protease regulatory subunit 4 
               
               
                   
               
            
           
         
       
     
     Example 4.  Babesia duncani  Secreted Antigens: SEQ ID NOs: 1-46 
       
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                   B. duncani  secreted antigens identified in the pellet fraction after  
               
               
                 ultracentrifugation of the culture supernatant: SEQ ID NOs: 1-23 
               
            
           
           
               
               
               
            
               
                   
                   
                 SEQ ID 
               
               
                 Antigen ID 
                 Annotated Sequence 
                 NO: 
               
               
                   
               
            
           
           
               
               
               
            
               
                 BdWA1_II1364, 
                 MKFLFGFFVILFLRLSKQEELVSLQLGDFHFDFEN 
                 1 
               
               
                 hypothetical protein 
                 GKYSTHEPFETCFIDVYNYRYDKTGPFLFNIFIKRT 
                   
               
               
                   
                 LENEYQSLFFKRENGKLVNFAPSHLSSPQTDNTGT 
                   
               
               
                   
                 YYSTKEPVVLESKNLSDIRNGIKKIGGNKLSSSGKI 
                   
               
               
                   
                 NWDTISTTLLLKTACGTYSTYSSGYEALLPVKDG 
                   
               
               
                   
                 NDTFCCCFSKAVLFTGYRFQMKKYEEVKPASNSQ 
                   
               
               
                   
                 STCKK 
                   
               
               
                   
               
               
                 BdWA1_II1520, 
                 MAATAIGIDLGTTYSCVAVYKDNNVEIIPNDQGN 
                 2 
               
               
                 heat shock protein 
                 RTTPSYVAFTDTERLVGDAAKNQEARNPENTVFD 
                   
               
               
                 70 precursor 
                 VKRLIGRRFDDPTVQSDMKHWPFKVNAGAGCKP 
                   
               
               
                   
                 TIEVTFEGQKKTFHPEEISSMVLIKMKEIAEAYLGR 
                   
               
               
                   
                 PVTDAVITVPAYFNDSQRQATKDAGTIAGLNVMR 
                   
               
               
                   
                 IINEPTAAAIAYGLDKKGSTEKNILIFDLGGGTFDV 
                   
               
               
                   
                 SILTIEDGIFEVKATTGDTHLGGEDFDNVLVEHCV 
                   
               
               
                   
                 RDFMRMNGGKNLATNKRALRRLRTHCERAKRV 
                   
               
               
                   
                 LSSSTQATIELDSLFEGIDYNTTISRARFEEMCNEK 
                   
               
               
                   
                 FRSTLIPVEKALRDADMDKRKINEVVLVGGSTRIP 
                   
               
               
                   
                 KIQQLIKDFFNGKEPSRSINPDEAVAYGAAVQAA 
                   
               
               
                   
                 VLSGNQSEKIQELLLLDVAPLSLGLETAGGVMTV 
                   
               
               
                   
                 LIKRNTTIPTKKTQIFTTNEDRQEGVLIQVFEGERA 
                   
               
               
                   
                 MTKDNNLLGKFHLSGIAPAPRGVPQIEVTFDIDAN 
                   
               
               
                   
                 G1LNVTAMDKSTGKSEQVTITNDKGRLSQTDIDR 
                   
               
               
                   
                 MVAEAEKFKEEDERRKCCIESKHKLENYLYSMRS 
                   
               
               
                   
                 TLNEDAVKQKLSTEELQNGLNTVEEAIKWVENN 
                   
               
               
                   
                 QLANQDEFEDKLKEVEKACAPLTAKMYQAAGG 
                   
               
               
                   
                 AGAGGMPGNFGGAAAPPSGGPTVEEVD 
                   
               
               
                   
               
               
                 BdWA1_III2507, 
                 MATIRKQRLEDLTDIHKEHLSTAHQLLDVIKSASD 
                 3 
               
               
                 Dynamin family 
                 PKLIYLHCYQLMKLGGLDAEVPRLVVFGQQSMG 
                   
               
               
                   
                 KTTLLDFIMGGPIGYSSTDTGTRQPVVIIMRPESAI 
                   
               
               
                   
                 DPRELELASGSVGSPSSTITSKKIWCKFNGKIMDIH 
                   
               
               
                   
                 NVQQNMRLHMQSLGERICSEELEVEVYVPDAITA 
                   
               
               
                   
                 IFVDLPGIKDDSKSGAEFTRRVVRNYVSNNPNDL 
                   
               
               
                   
                 YLLVKKSSDDPANWPWSLREFITAAAPNGLGLSP 
                   
               
               
                   
                 QQTMVVGTRAREFLINEKTDIRTQDQLYERVHKR 
                   
               
               
                   
                 AVLDSKGQALPLHLLELFSLSIQAKDKGDFLTNKE 
                   
               
               
                   
                 EMKRQIANGQVEVENMIRHGFEESNSINKEGRSV 
                   
               
               
                   
                 SEELLDTFSIRQFLRSLNSKFSQLLNGHLTNLERRL 
                   
               
               
                   
                 IRKKIDLERIVASLETKLQCFSPTTVRESIKQFIRQF 
                   
               
               
                   
                 MEIVHNMMMGNYTIMKLPIPPEQFLGIYGGSLRD 
                   
               
               
                   
                 NLEDGHELAQNLFPQPDMYESNFYTKITARTEEL 
                   
               
               
                   
                 YNKKLTMIDTVKPGRYVRYFTSKISVHMFGLIEPP 
                   
               
               
                   
                 RRVPPAVSGADLGPFDMMGQDYGADELINVEFIQ 
                   
               
               
                   
                 LNNNEENSLHKDIDRSKLSLLTPLASVSAEHLQVP 
                   
               
               
                   
                 LYAWHKTKEPKTGWVVVRPIVIDRLPPEILVQKS 
                   
               
               
                   
                 NYREVDVANKQISFRYLDVEFETKALGSGEEDQL 
                   
               
               
                   
                 ENSVETEISETPPVVEGVQNRCSNRYLYVTACSEI 
                   
               
               
                   
                 FLEEHRTAPYYASALEMVSGEHAEANLLNQLAVT 
                   
               
               
                   
                 NICHWLKFQIKHMEPEHVYSAEVLYQMLRSIDHV 
                   
               
               
                   
                 VDRADWEPLVADLVQSNVRGTLLHASRLAACAS 
                   
               
               
                   
                 AAALRRVLRASLAEAFRCIKLTDCDQTLYCLPDS 
                   
               
               
                   
                 LHFQEQIDHLSEEYCRQKAIDCANAMMNCIIEQT 
                   
               
               
                   
                 YSIQFDVAVDIFDGCRQFEKYFMGRAGHRSFMGD 
                   
               
               
                   
                 ALSSVKEDLALRKRRLAMTDIFEKSDAKTSIELIY 
                   
               
               
                   
                 EEVKVQFWATKLLLSTPLTTKLYTYFIKQVVDKA 
                   
               
               
                   
                 LPTTHSDPTASIKVEFEQFLIDNILYEQIDGTRTAK 
                   
               
               
                   
                 SNNRLSADYDLNNKYEQLVQQHNRNKRLLEYISC 
                   
               
               
                   
                 ALEGISRFKHHATADTDFLAHLD 
                   
               
               
                   
               
               
                 BdWA1_I0004, 
                 MISSPPSGFSLHGEDAKGTPRDVERQDSDRLDPG 
                 4 
               
               
                 hypothetical protein 
                 NFPPDKYWSPSIGLSTIEANRRVVWLLQEAISRYK 
                   
               
               
                   
                 IHLGYWNTTKSCLTIDILIGVLVLVLFILGAEPNLG 
                   
               
               
                   
                 VWHVVRPVLLLPWLVLNCYSKITVMRIKSSRACE 
                   
               
               
                   
                 RVRDVLESFSRELKTGGHTEVEQRRFSFDPPNSLL 
                   
               
               
                   
                 IPVYRDREWKRLPANVLLAGDVFKLQIGDYFPCN 
                   
               
               
                   
                 CRIILSCDREGKVQLDANLFNAGSVFKSSHLPSINT 
                   
               
               
                   
                 NDGGEWTDASFVAVTDSFVQSLETFLSADQGPQS 
                   
               
               
                   
                 RFMFFNERDWENCQKGPNTPPPTTCVWNFSDYT 
                   
               
               
                   
                 HFSKYGVDWIQLGIAFFISSIVTVLQIIWSGFQGWR 
                   
               
               
                   
                 RHVNVFATCIICLCHPAFDPFLNLADIWGNVKLQS 
                   
               
               
                   
                 LFQWHNEKRFTVDILPQQSSSSSSTFDSGSSEDSV 
                   
               
               
                   
                 DSEMAASRIPLLHQLRELNRVFKRGLDSEGSLLRT 
                   
               
               
                   
                 LCSVTLLCFVDDMGLLTEGCATPQELAVVDPSGG 
                   
               
               
                   
                 IGTQRRQQKESTAMFQSGNIDAIETINSTATSHKD 
                   
               
               
                   
                 SIGQTEQSDKGTIKAGPREGKQVVRKDPQGGERL 
                   
               
               
                   
                 VILDVFEDSSQYYKQYVKFNNDAERNCIPQVLSL 
                   
               
               
                   
                 SFAMSATQFPRVQPSLLQLKAAPDLIHTYMGMLA 
                   
               
               
                   
                 NGTLHDFTHCLCAFAGSVGLKRSYIRRFRLLRFIV 
                   
               
               
                   
                 VLDETLGTNGKMLIYFLRDPRKQIVQMLVKAKPE 
                   
               
               
                   
                 TVFDRSINYHDRARGAILPVSRLTKRKLRDLNMQ 
                   
               
               
                   
                 WVSSGLTPFAFIYKPIHLDEFNLIMAHLPGVAVFK 
                   
               
               
                   
                 VGHFLKLDRHQSVKYQESTTCQQDPQADYFGEG 
                   
               
               
                   
                 GKSDAKARRWYARLTDYDAISMRYGSMHTDLR 
                   
               
               
                   
                 QRILTNSIGGLVNSCLKNSILLGMCATKYQYPKEV 
                   
               
               
                   
                 PSRIQSFHEAGIRFVYFSKHDEKQTRIVGGLLGLET 
                   
               
               
                   
                 SWNSMISLVKSGRYSHVNQDGRVVLPSGIDNIRR 
                   
               
               
                   
                 HIRDVDDIPLQVSLFCDCTHTSTVEMMRILRENGE 
                   
               
               
                   
                 RIMCVGNGLRPSNFFVFCEAHSSVSVALGYHPTC 
                   
               
               
                   
                 RFCRGKRWSGIARAHAFEEATPEMKLSAFLTSLP 
                   
               
               
                   
                 CDLQTSKMYKVADPYFVMEMLHEVFKEARHMA 
                   
               
               
                   
                 TNIQDAAAFFKLASHSVAWMLMFQASLGFRRIL 
                   
               
               
                   
                 MPADLALLIFVYIPLMGSCLLSNVVAEGTMQQMP 
                   
               
               
                   
                 SRCTKDDAKVTIAAMCKTYPRLLLVATSLLVFYS 
                   
               
               
                   
                 FVLGQIQALLKIELERLYNFTLDDTQCSRFWRVAS 
                   
               
               
                   
                 YSCLQEHEQSIALLRTQVQSVIFRENTAAHLAEQT 
                   
               
               
                   
                 ASFAMAFLYSVSSASWIVRTGRIGAIASELLHSRW 
                   
               
               
                   
                 IISSAAILTIQGTILVARILAQPVPLASLNAIMPWGL 
                   
               
               
                   
                 VCGMLLGLSLAILAVDHLHKFYAARQHEMDQKN 
                   
               
               
                   
                 VESKVTSDLDIMDVTCICEPILSPSTQKISKRTSPPP 
                   
               
               
                   
                 IKQNVRGILQCKKRSYTSKSNRRRHPAIFNPDTTS 
                   
               
               
                   
                 AKDMGVLRTRQHYTSILLHFLQRGINGEQLQLVS 
                   
               
               
                   
                 LDIDQESFRLPLKKHSFGEEAAPAFLDVYVEEDIQ 
                   
               
               
                   
                 KIPVMFNLVIETKGDYTYRNYFFKRDGDKFINFD 
                   
               
               
                   
                 VPKLSDYYNVRTEGIATYYLTYKPIRVPREVLKRL 
                   
               
               
                   
                 RREFPDNNQLKVSNDPEDGPRSNEITLKLKLNSGS 
                   
               
               
                   
                 LVSYMGTYVPKTSSRGCRSKAPIFNKFRYKLTTD 
                   
               
               
                   
                 GQAIPVDEVEEEGEEEEEENENEKTEETDDQDRK 
                   
               
               
                   
                 VVTKEDDEEEEDEEVEEVEEELVEIKDEEEDKCYI 
                   
               
               
                   
                 EAWEKHSKNYSKVEVATSNKKHISDTGSLKPIDE 
                   
               
               
                   
                 LSGLFNALERKIATETALESDTTYTPKDNERWIID 
                   
               
               
                   
                 KCNSEESILDLHIKEDILTFDVPITQLPIQEKEIVDP 
                   
               
               
                   
                 CNSQRKSAFFNKIGSAGEHYLRAFGSNILSVIDIM 
                   
               
               
                   
                 KSDLNVSIKKVNSDEINEDLPTKEKLLAMEPIMFN 
                   
               
               
                   
                 SMDGEDNNNKNTDEEVNGTKTLMHSINSQSIERC 
                   
               
               
                   
                 EMNSAYNQGSFISIHDGTSDSFMDSDSFSISQKITQ 
                   
               
               
                   
                 DELLSKHDDKTLLNTDNALNDDSGLKTTEISHTV 
                   
               
               
                   
                 NDTVIPNLKLEKLLPQSKEQSAESTSSGEVGLIKK 
                   
               
               
                   
                 NKDVQSTKGKVGDLGNYVQINDDKEHPREIVTV 
                   
               
               
                   
                 EKFADETGIGTSTKKNEPATTSPLNEFDKIPNNDA 
                   
               
               
                   
                 AVNDSEKKTPADESRGNTNAKKTSDGNFKTISAN 
                   
               
               
                   
                 LKPLKPFRPDSLVTAMHISNDNKFLATGNENGEL 
                   
               
               
                   
                 YVWKFESAKMMSSPEISDNRINALPKMLNWEML 
                   
               
               
                   
                 SSTPTAGVQAHEFFIYSIHISKIQNRKHSGSVLVVT 
                   
               
               
                   
                 SGGNNYVRIWALTKSDKENVLILKGDRQFDDDIL 
                   
               
               
                   
                 TAFQLPISQHIAICGIDQVIEIWKFPPITTSVNDNPK 
                   
               
               
                   
                 NISSWNKTKERIDVELTIQSVSYSPSGMYIAIGSID 
                   
               
               
                   
                 GLLSLYSAETMKLISMAICRNEKGWYSNSASITGI 
                   
               
               
                   
                 VWNTKETLVCATTADSRIRLFSTNIENDNALIYAE 
                   
               
               
                   
                 KLKGHKYCGENVAARFTGLNDEYVICISKGGYIVI 
                   
               
               
                   
                 WKHSCTEERIFYDGNPIIKNTNYCKFKIIPKPYIGK 
                   
               
               
                   
                 LLGVFNPGTWGHLWPTYQEQSVYEPLNDKNSVG 
                   
               
               
                   
                 CLDRFLTKNTPGVKFSKANPKNAILLVASVDGSQ 
                   
               
               
                   
                 LLCTIIDASLLVFKKS 
                   
               
               
                   
               
               
                 BdWA1_I0706, 
                 MTESVDLTGDGGVLKTIIKPSKFFEQPEQGHEVEV 
                 5 
               
               
                 peptidylprolyl 
                 HYTGKLDTGFVFDSSHKRQTTFKFVLGDGNVIKG 
                   
               
               
                 isomerase 
                 WEVGVAAMNIGETALLVIKPEYGYGAAGSGSTIP 
                   
               
               
                   
                 PNATLHFEIELINSRIKPKQKWEMSMDEKIQAAAD 
                   
               
               
                   
                 AKVEGNAKFSQERILAAITYYEDGISYLSTRDEWP 
                   
               
               
                   
                 EDAIKASNVTKLQCHLNLANCYIKTEDFANAETH 
                   
               
               
                   
                 ATEALNIDPVNIKGLYRRGLARVKQGDFKEAIDD 
                   
               
               
                   
                 LSKLIKIEPTNAAGVSQLKLAKEKYTIFKQRERNV 
                   
               
               
                   
                 FGSVFKKMNLYDEKSGIRNLDTLPRVFLDISITGH 
                   
               
               
                   
                 VDVNRLVIALFEDTVPKTVENFKSLCDENAKLTF 
                   
               
               
                   
                 KGNKFHRLIKGFMIQGGDITNGDGTGGACIYGDQ 
                   
               
               
                   
                 FDDEGFKDSHSERGLLSMANCGPNTNNSQFFITFV 
                   
               
               
                   
                 PTKHLDGKHVVFGKIVEGMEFLDILENLSTGEND 
                   
               
               
                   
                 RPEADVTIEHCGTL 
                   
               
               
                   
               
               
                 BdWA1_II1893, 
                 MPSQGTVKAQTGTTEVDNADSSEEESQKSQPESK 
                 6 
               
               
                 hypothetical protein 
                 AVNGISGTESRNENSQSVDTNSSGDTNPSQSAGGS 
                   
               
               
                   
                 APTTGDSKNQQGKNVNAESSSNPNSEKSVATQDS 
                   
               
               
                   
                 STDQTGKTDSSTTHTTTGDSVEQKGDDNTETTDT 
                   
               
               
                   
                 AQNTATEEATTGSGTEGSADQTE 
                   
               
               
                   
               
               
                 BdWA1_III3045, 
                 MARFFSYKKLIAFAIVALASLKEVSFLGGCPYALA 
                 7 
               
               
                 hypothetical protein 
                 VATTTTTGTNGAATGTNGAATGTNGAGANDTSK 
                   
               
               
                   
                 NTSDPNTPATPPSSPESNKDNAAGGSDGQKPTGQ 
                   
               
               
                   
                 DPQKPNAGNGFAATSVIGAATIGLLTLAFN 
                   
               
               
                   
               
               
                 BdWA1_I0760, 
                 MTKYSQEPSNLAKSAKAYGAHLRVHFKNTYETG 
                 8 
               
               
                 Ribosomal protein 
                 RAIQGKMILEAKRYLNDVIEHKRCVPFRKFNGGV 
                   
               
               
                 L22p/L17e 
                 GRCAQAKAFKHTQGRWPEKSCRILLDLLTNLESN 
                   
               
               
                   
                 AEAKGLDVENMVIENVLVNRAPLGRRRSYRAHG 
                   
               
               
                   
                 RIIPFLSHPCHVALIAVEKDENVPRFTPEAAKTIKL 
                   
               
               
                   
                 NKRQIARMRLCNGKGVAK 
                   
               
               
                   
               
               
                 BdWA1_III2950, 
                 MISHYCLLLLAAATVFGGEATVTIEDAKSATLEAT 
                 9 
               
               
                 protein disulfide 
                 TGVPEVAEGDVSIPRGVVTLTADDLHKSIEKHEAI 
                   
               
               
                 isomerase related 
                 MIKFYATWCGHCKILAPEYIKAAKILEEENVNVV 
                   
               
               
                 protein 
                 LAEIDAVAHSDAVAEFEIKGYPTIKFFKRGIPIDYN 
                   
               
               
                   
                 SDRKAETIASWCKEMLNPALMETTNLEAEIASRK 
                   
               
               
                   
                 SKIALVAHGCNDKDELCVLFEKLAEVHRMDAHF 
                   
               
               
                   
                 FSVADSSSVWFEVRHVGDGTLKFNGLSPEELALF 
                   
               
               
                   
                 VKDETLPLLDEINPANYARYTSSGKSISWLCANTQ 
                   
               
               
                   
                 DYTKYRSSIVEVAKEMRSHTVFVWLDTEKFSAV 
                   
               
               
                   
                 NEAFAISKLPAIAHQTMKGRFILSPDAYDFTSKSA 
                   
               
               
                   
                 MLQFYTDVEQGKIPLSFRSEAEPQDATEGPVMLV 
                   
               
               
                   
                 VGKTLQQLFTQTDKAVLLMIHAPYCEHCRNFMP 
                   
               
               
                   
                 VFEDFAKTIDAQAPLIVAKLDGDANESPLDYVSW 
                   
               
               
                   
                 EAFPTVLLFKAGDKQPIPFKGTRTIEELTSFVQEHV 
                   
               
               
                   
                 TLAPVKTEL 
                   
               
               
                   
               
               
                 BdWA1_II2303, 
                 MVEELDGTKNEYGFCKSKLGANAILVVSMAAAR 
                 10 
               
               
                 Enolase, N-terminal 
                 AAAAHLNIPLYVHLANLAGKPTNKFILPVPCLNVI 
                   
               
               
                 domain/Enolase, C- 
                 NGGSHAGNMLAMQEFMILPIGAGSFREAIQMGSE 
                   
               
               
                 terminal TIM barrel 
                 VYHTLKKVISSKYGQDATNIGDEGGFAPNIKNAE 
                   
               
               
                 domain containing 
                 EALDLLLEAFRIAGVEGLFKIAMDVAASEFYDKN 
                   
               
               
                 protein 
                 TGMYNLGFKGKEPQNKTGEEMISYYKLLCAKYPI 
                   
               
               
                   
                 CSIEDPFDQDDFDSYTKLTAAIGEKVQIVGDDLLV 
                   
               
               
                   
                 TNPKRIEMALGKKACNALLLKVNQIGSVTESIDA 
                   
               
               
                   
                 CKMAHANKWGVMVSHRSGETEDTFIADLVVAL 
                   
               
               
                   
                 GTGQIKTGAPCRSERNAKYNQLLRIEEELGNKCE 
                   
               
               
                   
                 YAGHNFRTCGN 
                   
               
               
                   
               
               
                 BdWA1_I0910, 
                 MEEWYTYALRHVDLDLDNFFVEFGETILEDYTRI 
                 11 
               
               
                 hypothetical protein 
                 YPRSNVFVDNIRKGATLITLSTEHKGLLFVELRYP 
                   
               
               
                   
                 RPGKDSVMDIIYKNWEDEEVMFSLVWVFGDWVP 
                   
               
               
                   
                 QNFLYSGILDSGPEYVPVRRHDSR 
                   
               
               
                   
               
               
                 BdWA1_I0555, 
                 MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGI 
                 12 
               
               
                 polyubiquitin 
                 PPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVL 
                   
               
               
                   
                 RLRGGVIEPSLVILAQKYNCEKMVCRRCYARLPL 
                   
               
               
                   
                 RATNCRKKRCGRCSQLRPKKKIKGGN 
                   
               
               
                   
               
               
                 BdWA1_II1366, 
                 MGNPGLIFIALFKYSFNCYYTFLPFKFDNNHFDVL 
                 13 
               
               
                 hypothetical protein 
                 PHENFLPRATVPRFVDIYVSADDPRIPILVNFVTGG 
                   
               
               
                   
                 MTTSGPDTRPTNRNYFFKRDGNKLVNYKFTNKP 
                   
               
               
                   
                 NDTVENNDIVTRISRERVTYYVGTSNPEDIIVATK 
                   
               
               
                   
                 DPQNDLNQRENSMYASLATLDPKATIPRIGLKTK 
                   
               
               
                   
                 SKYNKVALYSDEHKEKIEISNADIDGVMHEKLILF 
                   
               
               
                   
                 RYRLVKEYRV 
                   
               
               
                   
               
               
                 BdWA1_I0089, 
                 MRNVKKRTVPFLWISCFVLYGIYNVEPIALYNKN 
                 14 
               
               
                 Metallo-beta- 
                 WVHKRVGFIANLKQPPCGITFQGLTHEKAKRGFK 
                   
               
               
                 lactamase 
                 RYAEQSSQVKEASVLFDTLSNTEVLDELDSNDAI 
                   
               
               
                 superfamily/Beta- 
                 QDTPDPLESETIIEVEVEDKGKKTFSDIHNKLKGSI 
                   
               
               
                 lactamase 
                 NKLPSTFAVMKDLVAFLDTMEQQGNEKAKSIAY 
                   
               
               
                 superfamily domain 
                 TSIKKKLITELNKNVEELKSDLPKTKKRLYISPFLK 
                   
               
               
                 containing protein 
                 NTVDPFFEDQYVNPVDFKNYFSLYDNYKQEYRKI 
                   
               
               
                   
                 AYLCMKTYIARLILGLKRKLKHGIVSTFLWPFAK 
                   
               
               
                   
                 WAKDNNVRYKEPKPSQLENFERLLKYYGHEFEN 
                   
               
               
                   
                 VYFEQNIAMVRPPQPKARPPNKWSLIFLGTGSRQP 
                   
               
               
                   
                 TDTRMTSTMAFTEHDGGRIWLFDCGEGTCACMQ 
                   
               
               
                   
                 KLNLNPKAVDRIFITHLHGDHCFGLFSFISNSARA 
                   
               
               
                   
                 LPITVYGPIGISKMLIDIMNFTTTSVLPKFVVHELV 
                   
               
               
                   
                 LHPDNEKHKTGWHVNYPFGGYIYPQEAGHYLV 
                   
               
               
                   
                 YENDTCKVMAAPLKHILPTVGYVIKEKSKNENDS 
                   
               
               
                   
                 TKTQRKIVICQDSCDSSKMVPISMNPNVLIHEATT 
                   
               
               
                   
                 STTSTIGSSLIMQLVYNFSKGKIDESLLSKINDIISQ 
                   
               
               
                   
                 EELRRSCFSVTFTTMAMKLRRKCYLIERSYSNLTK 
                   
               
               
                   
                 TLEQNELKATETESNNADMMTDIYNIVQLVHAAS 
                   
               
               
                   
                 KIKSCIAEAKKIESQLKELYDAPKHLGPRSTWLKS 
                   
               
               
                   
                 VYNHVRDVIENNGNTETSSSPTIQAMESLKNLLTK 
                   
               
               
                   
                 FWTTNKSMGLLFSLEINLPEAAPKDWFSLYSKSV 
                   
               
               
                   
                 RYSGHSTPWDAGKFAAKINAASLYLTHLSSVSLH 
                   
               
               
                   
                 VHFY 
                   
               
               
                   
               
               
                 BdWA1_III2562, 
                 MLLSCWFNLIACFICICSCYIRLNQTNCTNLRLINT 
                 15 
               
               
                 Alpha/beta 
                 NNSNIKMKATGKLSMSHFKNKQGLLIRTYAAEV 
                   
               
               
                 hydrolase family 
                 DKPKGSAILVHGNKSHFRADFTNYNVDFYKDKY 
                   
               
               
                   
                 GLESVDPNIVIREMHAIYPNVDHKIDFNNDYEFHY 
                   
               
               
                   
                 SKLDGKNALDITPRFTLNGSIVEYLNGLGYSAYGL 
                   
               
               
                   
                 DLQSQGMSQGHNGHRNYFKKFDDHVVDVIQFIDI 
                   
               
               
                   
                 IRRNKFHNVNEEWDPNVLGKNYSLNKCFLMGLS 
                   
               
               
                   
                 MGGNVVLRAAQIFKTLSDFKSNIVDGIVCFAPML 
                   
               
               
                   
                 DIDMHFSGAFNQLALAIAKMIVTFCHHSTFMINEK 
                   
               
               
                   
                 YEIDTLNSFLRVNDPYYITKTQTHKAIVSLLEATR 
                   
               
               
                   
                 TLEKNHSKYPVDMPTLVFHCKDDNVCDFKVFTC 
                   
               
               
                   
                 NEMI 
                   
               
               
                   
               
               
                 BdWA1_I0812, 
                 MADGMNQETYAFNADISQLLSLIINAFYSNKEIFL 
                 16 
               
               
                 heat shock protein 
                 RELISNASDALEKIRYEAIKDPSITEDQPEYFIKLYA 
                   
               
               
                 90 
                 DKNNNTLTIEDSGIGMTKADLINNLGTIAKSGTKA 
                   
               
               
                   
                 FMEAIQAGTDMSMIGQFGVGFYSAYLVADKVTV 
                   
               
               
                   
                 VSKNNNDEQYIWESNASGHFTITKDESGEQLKRG 
                   
               
               
                   
                 TRIILSLKDDQTEYLEERRLKELVKKHSEFIGFPIQ 
                   
               
               
                   
                 LSVEKTTETEVTDDEAEESADAEGEKDKIQDVTD 
                   
               
               
                   
                 KDETEAKEGEEGDKDKKKKKRKVQNVTREWEM 
                   
               
               
                   
                 LNKQKPIWMRSPNEVTNEEYASFYKNLSNDWED 
                   
               
               
                   
                 HLAVKHFSVEGQLEFRALLFVPKRAPFDMFESRK 
                   
               
               
                   
                 KKNNIKLYVRRVFIMDDCEELIPEWLGFIKGVVDS 
                   
               
               
                   
                 EDLPLNISREVLQQNKILKVIRKNLVKKCLELFNE 
                   
               
               
                   
                 LTEKKDDFKKFYEQFSKNLKLGIHEDNANRTKISE 
                   
               
               
                   
                 LLRFETTKSGDEAISLRDYVDRMKPEQKYIYYITG 
                   
               
               
                   
                 ESKQSVANSPFLETLRQRGMEVIYMTDPIDEYAV 
                   
               
               
                   
                 QQIKEFEGKKLKCCTKENLELDDDEEANKNFEKL 
                   
               
               
                   
                 KEEMEPLCKLIKEILHDKVEKVTCGRRFTESPCAL 
                   
               
               
                   
                 VTSEFGWSANMERIMKAQALRDPSITSYMVSKKT 
                   
               
               
                   
                 MELNPRHAIVRELRQRAESDKTDKTLKDLVWLL 
                   
               
               
                   
                 YDTALLTSGFNLDAPAEFGNRIYKMIKLGLSLDD 
                   
               
               
                   
                 DVAEASLDEVPALDEVPVDSRMEEVD 
                   
               
               
                   
               
               
                 BdWA1_III2693, 
                 MRTLSRLPPPGDPGHVASFQEIATFNIDEFLANIDK 
                 17 
               
               
                 hypothetical protein 
                 THDGVSQLQLPPNLLENLKQGAGSLLQHLAEQST 
                   
               
               
                   
                 GPFATHNQPSLIGASGMAVAAPQMQLPQVAMAP 
                   
               
               
                   
                 QVMAAPQPGMPPAVAMPQMTSQFAGAPGQALP 
                   
               
               
                   
                 VAAQAVPNMQAPAGPQLASVPISNVIRGNNIAFP 
                   
               
               
                   
                 GAVPNEELAMELVMKITWALLMHC 
                   
               
               
                   
               
               
                 BdWA1_II1642, 
                 MSEENSQRAQLTYSAKLAEQAERYDEMADAMK 
                 18 
               
               
                 14-3-3 protein 
                 LLVETCITDKDELTVEERNLLSVAYKNAVGSRRA 
                   
               
               
                   
                 SWRIVSSVEQKEASKSNSVHKTLAGEYRAKIEKE 
                   
               
               
                   
                 LNKICLCIIGLLDEKLIPATVDSESHVFYYKMKGD 
                   
               
               
                   
                 YYRYISEFSCDESKANASASARDSYQKATEIAESE 
                   
               
               
                   
                 LKSTHPIRLGLALNYSVFFYEILNRPQQACEMAKR 
                   
               
               
                   
                 AFDDAITEFDSVSEDSYKDSTLIMQLLRDNLTLWC 
                   
               
               
                   
                 SDVTSDAPDKQKQED 
                   
               
               
                   
               
               
                 BdWA1_II2273, 
                 MFGLLRPCLIRLSGAVATRQTFGNLADCLKIAESS 
                 19 
               
               
                 RAP domain/Core 
                 QSIATTDLYEAFKFISSSGQIRRQAIHDDRFVTLLD 
                   
               
               
                 histone 
                 QLDARISTLNCSYMGNFGIRLGLIIQSLGNLDRED 
                   
               
               
                 H2A/H2B/H3/H4 
                 PIVEKSVKVIERLCTEMMEKSGNIKEISQLAFAAA 
                   
               
               
                   
                 SAGLQHKFLDYAKQNLTLNIENADPDVLNLALLA 
                   
               
               
                   
                 SYKTKVHDKVFLALICEKLSELTDRFTANDVVST 
                   
               
               
                   
                 LRSLEKTSLMKGFLLRRLSMLIHDNLEQFTNEQLA 
                   
               
               
                   
                 QCCYRLSILKFQTPVQYSTILSLLEPKFQQLSIHLQI 
                   
               
               
                   
                 EVLASGCMCQCTDANERLVKLAKSITLTDKVDLA 
                   
               
               
                   
                 GLVNYIYSCVYLKLYKGDHLTGALEEALARSPFLI 
                   
               
               
                   
                 RKYALLFKEAYDTLSLECPNLTLELPEAWKMALE 
                   
               
               
                   
                 NYESAEHDRCIQTSIIAETGNILKTSAGDFETFSKV 
                   
               
               
                   
                 GPFTVAFADVARKLVILAETPNTLGGLALAQRSIK 
                   
               
               
                   
                 AMDYKVAIIKYWEWRRLKTEKSELSYAFKRFDK 
                   
               
               
                   
                 SRLGVLSHVQFVRLLGAIGIHLTRQELKFLQFEEDI 
                   
               
               
                   
                 RGGFTLEDLEALGRDFYNDEVIATKVLESLQEHF 
                   
               
               
                   
                 GPCNTLDKQELASVLMKLGASLGVAREELDTFLN 
                   
               
               
                   
                 FYSAHSNSISVEAFIRGAALGVLELCADHTVTILQ 
                   
               
               
                   
                 WETSRLQTKTSKMSGRGKGGKGLGKGGAKRHR 
                   
               
               
                   
                 KVLRDNIQGITKPAIRRLARRGGVKRISGLIYEEVR 
                   
               
               
                   
                 GVLKVFLENVIRDAVTYTEHARRKTVTAMDIVYS 
                   
               
               
                   
                 LKRQGRTLYGFGG 
                   
               
               
                   
               
               
                 BdWA1_III2537, 
                 MVLSPILQFSFLALPSMLLNNVFALRMSAPTPIESP 
                 20 
               
               
                 Rhoptry-associated 
                 VTKYGDSLNMLFLDLGNAFHENFSMFQVATSMS 
                   
               
               
                 protein 1 (RAP-1) 
                 NYAETNNDIVSRICERFESQKACFVLASKYINNCA 
                   
               
               
                   
                 KAKKCMQIEKFHLFQSPDMSIKLMNRAQLAAAIH 
                   
               
               
                   
                 VFRNSGVYEKNYLKRRFNKIFKRQPFGYSSYRTL 
                   
               
               
                   
                 LIPLLYSNASFNEYTTFSEIFITYYLNVATFMYATLI 
                   
               
               
                   
                 YRDTRLARFVNAFDKLNVLLIPLKKHLRNMVTGI 
                   
               
               
                   
                 ANASPVAFCEEDFEPILRIFGHYLSGFDKSLTPLAN 
                   
               
               
                   
                 RFAKLIRDVLKNELHKSESCILNRAGDFLKNAGR 
                   
               
               
                   
                 RAGKAVRDAGATIKERGMATYKGVKGGLAGAS 
                   
               
               
                   
                 DKIRNRFRSRNGNGDDASSYGLLDEDATGEATDD 
                   
               
               
                   
                 VKPEDDSTGDNEPKEDEITRL 
                   
               
               
                   
               
               
                 BdWA1_II1831, 
                 MNLKWLLGLALIGSKYALGGDPNDSEVDSGKER 
                 21 
               
               
                 hypothetical protein 
                 GPGKRMTFDELLDELKTAEASVLGIKAEINGGLN 
                   
               
               
                   
                 RLRYRIGNLDAITKSDYDEISDAIRDIITKRTEFAK 
                   
               
               
                   
                 AVNKRVQLEAIANKFSERTSMGNLEDIQFSTFWV 
                   
               
               
                   
                 KLEAITRVPDFQLKEDFVKMKDEIIDVKEKFIEKL 
                   
               
               
                   
                 KKAREATAEVIPETIVEDQEMKSDLHEEIKSHGDD 
                   
               
               
                   
                 DIFNDKSDKKQNSGFAATSSSLILLAMATIGYSLF 
                   
               
               
                   
               
               
                 BdWA1_II2038, 
                 MAKSALYILDSNVKSKKVDKESLKLIKKQSGKPK 
                 22 
               
               
                 Ribosomal protein 
                 KILRPVEHKAHNPSCLENAKVELDLDLLKKVIETL 
                   
               
               
                 L1p/L10e family 
                 KKRAEVVRESNTRDLLEDPSRNYVFIQIALTKVVT 
                   
               
               
                   
                 EVHVKPLQIKLKHPIYTDKEVCIFVKDPQKHWKEI 
                   
               
               
                   
                 IKRENVPQIKKVIGVTKLKKKYKQFEDRRKLCRSF 
                   
               
               
                   
                 DLFLCDKAVCCSLPSLLGKVFIQRKKMPVPISMSK 
                   
               
               
                   
                 GGLGNSMREAIQSTYYKLSTGNTCSVKVGICSMN 
                   
               
               
                   
                 TEQLIDNIKQVFQTIKKFHTEDPIFRNVISSIFLNWE 
                   
               
               
                   
                 GTESLMLYSRALADDDIQIPQSHVTSPSKPTAAKP 
                   
               
               
                   
                 TCKWFGTSQSDLVARIVATAKGGTTSLSVWRQFS 
                   
               
               
                   
                 KDVIETIDTLNIPDVYRILKCFSILRYRHDPLLNVIS 
                   
               
               
                   
                 YRIVESLDKIACKNLAEILKAYSKLECRNDFLLKT 
                   
               
               
                   
                 ALPTVARHLEFFTPSDLSSVFYSYCNLGFHDLNFI 
                   
               
               
                   
                 RQVEWRIFNTLGKLQSCDFALLFCGLTRLERINTR 
                   
               
               
                   
                 FVISLACQFCKSLDAIDEKHFSLCVNALGRLEFAE 
                   
               
               
                   
                 HPHLYGILVQHIYNEIKLHHLTSVSLALLVNGFSR 
                   
               
               
                   
                 AKPKDLKVFQMLSKQIESRLSEFDIHSLCLITAGY 
                   
               
               
                   
                 SRISNAMEQVYLFEKIAESVGRKSLQLYPMAITSL 
                   
               
               
                   
                 MYSFSRAGHVHGPLMFYGSQHLTKFAEHYNIVEL 
                   
               
               
                   
                 SMVSRAHCLLEIKNDDLMHCIAREIVKRFPNVIPT 
                   
               
               
                   
                 AADSPRVRRISQDSKDLEHVPQESEKCLILEAGTI 
                   
               
               
                   
                 NLLWIMQGFASFYIFDGNIRNAIMAICNEMCMRIV 
                   
               
               
                   
                 DLTPMLVSNFLHALATLRYRHETFLEILVRELEDP 
                   
               
               
                   
                 RLGVKFNQDELKLCYEALNTFGVHGPIYKVSKM 
                   
               
               
                   
                 ALKQAHEINGGDSTTTLRQVMAGELKTFQDDSEE 
                   
               
               
                   
                 LTQELKIPPPPKKKVYIHVPEVIRKHLQVPNAQVT 
                   
               
               
                   
                 HQYDFVTFQV 
                   
               
               
                   
               
               
                 BdWA1_I0231, 
                 MDVFSILLVFSAFYVNAIAADDVKTFLFKKDVES 
                 23 
               
               
                 hypothetical protein 
                 TVEIDANDDAVLVCPIASVLIIKKARWLPVTGGD 
                   
               
               
                   
                 MRVKDGFSRTTRIGWLCNGLENCAFRPVAHLSKI 
                   
               
               
                   
                 GDRYEFLGQPIETDIYKLTVTATCGNFMFKRPGR 
                   
               
               
                   
                 REMLCIPTSAKPDIVLGCKDNEAIELSYVRVGGKS 
                   
               
               
                   
                 KHQWRHRDYCAESIIKTAHPLCTGKKTCKIAHDV 
                   
               
               
                   
                 FLKNAKECIPREFNVEYYCAAPHKNSFYDPLDAV 
                   
               
               
                   
                 VVDGVSVATKYVLTAEDGARASAKTNAYQVLQ 
                   
               
               
                   
                 VDSALWESDGATERRDRLELVKFLCDGRAECVFS 
                   
               
               
                   
                 PTRSIIGPDERKCNDVVFGGMVKDTMSHFMLRAH 
                   
               
               
                   
                 FSLVPFDPKKYDEKEYHHVTIKSTEKKTLECPVN 
                   
               
               
                   
                 MSLTFYVALWGGKITDTSPLKGPKHFVEVDINGE 
                   
               
               
                   
                 KHRYSEIINIVGTQCFGKSKCEIEPLKLKPPRHEKD 
                   
               
               
                   
                 LKEFPTHEGVKKDDHQLELYYKCIDLQTLPSLVES 
                   
               
               
                   
                 LISDGPRYPREFITPIQLSPDMRIVVMLDIYGPTVL 
                   
               
               
                   
                 EVANALKLEIPVARTNEIKISWKDAKISQGIRLVK 
                   
               
               
                   
                 DTRNYVFEFVIGAEDYIHMTVNSFDNDGSPMSIPV 
                   
               
               
                   
                 EFEASKRILDFSRGIEDFVVATGEITNFRAFIKS 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                   B. duncani  secreted antigens identified in the supernatant fraction: SEQ ID NOs: 24- 
               
               
                 37 
               
            
           
           
               
               
               
            
               
                 Antigen ID 
                 Annotated Sequence 
                 SEQ ID NO: 
               
               
                   
               
               
                 BdWA1_II1370, 
                 MVTREMYIMGYANVISADIKKFKETLANEITTREE 
                 24 
               
               
                 hypothetical protein 
                 YEKLKADIASTRAIMGDLKSNVRVLTRLLNEVYIL 
                   
               
               
                   
                 KTLMREEDVRALGDLSMEAKTLVAGKKELQIKL 
                   
               
               
                   
                 ENEIREIKKKVDGLPLTKKILEENTEQLLEEISKIE 
                   
               
               
                   
                 MHVKKSAEAIEKNIDECNDKITQTNVMAEEEHEA 
                   
               
               
                   
                 LTNKLGITKIVLNGIKTGLLKLISISNRLKAATSGA 
                   
               
               
                   
                 TREENKILKETILNRIKFILAEGDVVFKRLEKDMT 
                   
               
               
                   
                 DIEKRIKRIPIEGTLPDLNTHGGYGTIETH 
                   
               
               
                   
               
               
                 BdWA1_I0253, 
                 MDFLWLLGFAIIYRKFVVGVGPDEDSDYPEVDVK 
                 25 
               
               
                 hypothetical protein 
                 SSKVNIGISATVDKFFDDMKLMEEDYKAYQDKIL 
                   
               
               
                   
                 GALNTVRRRLEKAKHLEATDLSDLANIMLDVNK 
                   
               
               
                   
                 ELTKMNSCVSRLAALRPYVEKSINLLHEDDKETA 
                   
               
               
                   
                 KKRLLNFMNPDEMVNVLHMLFSEYKELNVELVH 
                   
               
               
                   
                 VKKRGTAPSQSESLSKSKIESLSKSLSALNQEPGRI 
                   
               
               
                   
                 LEPALAIYDKIMNGGTEILEEMNKLNLKIQGKQT 
                   
               
               
                   
                 MSSMEYLYIVQNMLHAKEYVMESKQPLTSLGYL 
                   
               
               
                   
                 GTALDQMQFTYSPTEKSHVENIKKEVGEILNGIKD 
                   
               
               
                   
                 FQNKVKNSIDTVEKRVTTISVTDALPKERALEIISA 
                   
               
               
                   
                 VPAYVQIFKKQMDELKGAVLNDINELDKQLDEK 
                   
               
               
                   
                 KRIPSKEHEQMEAKIPILETQVQFFLEFIEAMKYFR 
                   
               
               
                   
                 VTCEMIPKMMGVDEKKQFRRELILCDMALKNEK 
                   
               
               
                   
                 QEYDFMIEQFKKVKERIVQTRPRASRFKRKHSGFS 
                   
               
               
                   
                 TMEPSSLLLVLPVIVSLFY 
                   
               
               
                   
               
               
                 BdWA1_III2944, 
                 MGNACCKSSPPAAVSDAKTADKPNFESLSTLSMK 
                 26 
               
               
                 phosphatidylinositol- 
                 NDGSHRNQEKAPSAKASPRKNVEFKFTDTGYDA 
                   
               
               
                 4-phosphate 5- 
                 TGAKKWDEKVISALLGGKPTNIEAAPGIDVGDGL 
                   
               
               
                 kinase 
                 VERGPVLLKDGSVYCGQWKGSVRHGRGQFFDV 
                   
               
               
                   
                 DGTQYIGNFSKGVFEGAGELRTWTGDKYQGLFK 
                   
               
               
                   
                 NGKYHGKGIFTQKNGDVYEGVFVDGMREGYGTE 
                   
               
               
                   
                 RYKDGSVYMGEFKGGKRMGNGELKMADGVLYE 
                   
               
               
                   
                 GEFNDEITGKGKMFWPTGECYVGSFLKGMKHGL 
                   
               
               
                   
                 GETTWKTGPMKSQRGKYENGKMCGTFENVMRD 
                   
               
               
                   
                 GKVVKGVYKDGILLQEITDTKPKVAPVVTQPPPV 
                   
               
               
                   
                 KEQATPTPTPKAAASPTTSTPTRAAASASPTLSRA 
                   
               
               
                   
                 PSTSSTPATATPPASTPTAAASTQAKPKAKSAKSS 
                   
               
               
                   
                 SAAKKKTSSKSSAR 
                   
               
               
                   
               
               
                 BdWA1_I0708, 
                 MADEEVTALVIDNGSGNVKAGVAGDDAPRCVFP 
                 27 
               
               
                 Actin 
                 SIVGRPKNPALMVGMDEKDTYVGDEAQSKRGIL 
                   
               
               
                   
                 TLKYPIEHGIVTNWEDMEKIWHHTFYNELRMAPE 
                   
               
               
                   
                 EHPVLLTEAPMNPKANREKMTTIMFETHNVPAM 
                   
               
               
                   
                 YVAIQAVLSLYSSGRTTGIVLDSGDGVTHTVPIYE 
                   
               
               
                   
                 GYALPHAMMRLDLAGRDLTDFMQKILAERGFSF 
                   
               
               
                   
                 TTTAEREIVRDIKEKLCYVALDFEEEMTNAESSSEI 
                   
               
               
                   
                 EKSYELPDGNIITVGNERFRCPEVLFQPSFIGMEAA 
                   
               
               
                   
                 GIHTTTFKSITKCDVDIRKDLYANVVLSGGTTMYE 
                   
               
               
                   
                 GIGQRMTKELNALVPSTMKIKVVAPPERKYSVWI 
                   
               
               
                   
                 GGSILSSLSTFQQMWITKEEFDESGPNIVHRKCF 
                   
               
               
                   
               
               
                 BdWA1_II2269, 
                 MAIWKLFVFGICGAGKVLGYRLDDVLQGDGEDF 
                 28 
               
               
                 EF-hand 
                 ALFGLGKEVIEARMDKLFSVIDLNNDGILDLEELA 
                   
               
               
                 domain/EF-hand 
                 AFHAKTFQTILDLQLNHEMELVDRNKDGFVDVEE 
                   
               
               
                 domain pair/EF 
                 LKVAFEREGTQDVDISTVEKGLQRRFVAADKDQ 
                   
               
               
                 hand 
                 DGKLNRQELGLLLNPGRDEELINIEIEEIMQTYDQ 
                   
               
               
                   
                 NGDGLVSLEEYSHGRSDQEGVSAEFKPFDSNADG 
                   
               
               
                   
                 FLSREEIRGVYVEENKNDLDSEHEDLFAITGKKPIT 
                   
               
               
                   
                 REVWNANLNKIAHTSLTDHGEMLRFPEDYHMDL 
                   
               
               
                   
                 GDIPRDRKDGAEERPSGEL 
                   
               
               
                   
               
               
                 BdWA1_II2320, 
                 MCGNGIINNLVYLKMDSIRESTSFHNDFYYITCPF 
                 29 
               
               
                 Sodium: dicarboxylate 
                 SIKGFLKEYWLSTITFFLTFAAVFIPSVFKLDLKDH 
                   
               
               
                 symporter family 
                 ADYVMLPANMFLRHIRGFVVLFMFFASAAKIRLF 
                   
               
               
                   
                 LQRKVDSLKTRIMIKYLIAGLLSLLVTLGLAALIIP 
                   
               
               
                   
                 LNTSLGGNTYFSPHSIKDYNPTTEFKNFINHLAVH 
                   
               
               
                   
                 DLPLNMATTRVDFVNEFGKDQLHEGLGDGYNAP 
                   
               
               
                   
                 GFVVYGLLFAFAIYTMDEHTDALCNIITAMHKCL 
                   
               
               
                   
                 LSVYWILVVYSPFAFFLAGLVTFDELKVKGGLGIA 
                   
               
               
                   
                 LMGYLYLTLAVLAVFVAWSFIVVPLLHFIRTARN 
                   
               
               
                   
                 PYPTIIKLLPYLPTAFCSGSSVLSGEKTKDFLKKRG 
                   
               
               
                   
                 FDPDDVENYLTFSTLVNFSGTTSGFTVCAILMLKL 
                   
               
               
                   
                 FGKSLDWVTVLKIIVTGLLVGFTIVEYIQGYLFGII 
                   
               
               
                   
                 FFLNNTMLPPGSIILLLQIDWLLDRFRIVSNVIDDA 
                   
               
               
                   
                 LTIDEITNVKSKLSSCPFHKV 
                   
               
               
                   
               
               
                 BdWA1_III3035, 
                 MGEGTNKTQYPFILGLHEEVVCAAKRLEICLLEKI 
                 30 
               
               
                 hypothetical protein 
                 EQLGHVDDKGWKVCVDSTIEGKLLGNHITCLSLC 
                   
               
               
                   
                 KVPNGSHGSEFLLAVGYKSGLIALARVDSQNEFHI 
                   
               
               
                   
                 LSHNDQNKSSISQIELKYVGESTAAYNLMSLRVY 
                   
               
               
                   
                 AFLDSGKIITFSWGDKIQTSIQSGEHVLGFAVNSQ 
                   
               
               
                   
                 NTTIAILQNTGIVCRSLDTETILRTLENVQIVNANR 
                   
               
               
                   
                 HYLSWHPYKNLLVFLDNKGISYTCHPKWDVYKF 
                   
               
               
                   
                 SQNSQHQELIRHVQFGICGDFVLLLTASFDKIIIWD 
                   
               
               
                   
                 FETESILYSHKGCDMVACGLIHLSNKRVRLAVFA 
                   
               
               
                   
                 NFASNEFWRCKRLIMSGDELMEEAKSELPQSHKT 
                   
               
               
                   
                 RRLKRHITELDDYHIRKMVDQEAVDEDNEEQDEE 
                   
               
               
                   
                 YKDEETYRDILDSYDNLDKDEMTHFDQSRVHVM 
                   
               
               
                   
                 HEISKLRKKVASLDNRVAERTTLVPGSCPAPDDA 
                   
               
               
                   
                 AVQWVLFWDEVGQITKQLISDVWCLHVHVFSGP 
                   
               
               
                   
                 LAGYKRKPDRYNCHTAALNQKYVVTGSEINVDG 
                   
               
               
                   
                 GHVHGILTFLDFENNTQWDRRFFNEYISAVAVGD 
                   
               
               
                   
                 SFVAAITADGILYILSLARSLMGVFQLKGSPIAIAA 
                   
               
               
                   
                 RGNVLATITEATTLTNVSSFGHVRMFWVNGLRGL 
                   
               
               
                   
                 AKNPASRIVDLYSDSLVLGPDKAISWISFSSQCTL 
                   
               
               
                   
                 WITDTSGQLMALLPAIESCYKVGGYSLEWIPFMN 
                   
               
               
                   
                 LENLVNESSDYEPSHTVFPLYVSDMKLNYILLKR 
                   
               
               
                   
                 GQSYPTCSQPLNFMGYTLKKACLRIDGCVGAYLP 
                   
               
               
                   
                 FQKFNGLVTKDPQLREVIGSDIISDVASIPWQQYD 
                   
               
               
                   
                 EMRHIQSLQACQNEYILKVFQNYNFWMQNSQGIS 
                   
               
               
                   
                 DDAISAFGNAERVHDKWTLRILRKTKDSKQDSGV 
                   
               
               
                   
                 LLDALWMLRFPKCLEAAFSILQNQLDAKQRQVL 
                   
               
               
                   
                 QEASLLLENVTPFENNVQPLDAQSQPTATVSIPKQ 
                   
               
               
                   
                 DLDPIPKKHAPLIQGPSNRLQEFVPLGNEPEESGPL 
                   
               
               
                   
                 FKNILE 
                   
               
               
                   
               
               
                 BdWA1_I0924, 
                 MVDFEPESCRKHVDKNTRIYVDGAFDLLHWGHL 
                 31 
               
               
                 ethanolamine- 
                 NALRQSYKLGGELIVGINGDVETFHAKGISPIYNQ 
                   
               
               
                 phosphate 
                 DERAELVKGCRWVNEVMVGTPYEVNLDFLVNIA 
                   
               
               
                 cytidylyltransferase 
                 KCDYIAHGDDIAIGASGKDAYDEPKKAGKFIFFRR 
                   
               
               
                   
                 SLGVSTSTTVGRLIDALESDHFSHLSKNSENKLQY 
                   
               
               
                   
                 GDFEKALQENEEQMINEGIIDDALFSGNNKHDNK 
                   
               
               
                   
                 NKKSTDVFPYPRFRLSTSLLGEFITPKPKPKGGKII 
                   
               
               
                   
                 YVDGSFDVFHVGHLRLLKRAREMGDYLIVGIYD 
                   
               
               
                   
                 DQTVRTLKGTPFPFSSLMNRALTILGMRYTDDVV 
                   
               
               
                   
                 LGAPYVPSRTYLENLGITTVVTGKQHDSKMINRD 
                   
               
               
                   
                 FDPYREARDMDILVEIDSGSDITTSDIIARVSSRMD 
                   
               
               
                   
                 QITANIRKRCAIEKTRKNIVCSL 
                   
               
               
                   
               
               
                 BdWA1_III2663, 
                 MLNNANTVKEVGFRKSVIYQPNVFKDADGVIKIK 
                 32 
               
               
                 MAC/Perforin 
                 TNNKNIWIRQNSKCWEFTSKTPITSVDDYYKELLS 
                   
               
               
                 domain containing 
                 DFNTDDSTSITLYNANVDLAKRGFNNFHEGYTKI 
                   
               
               
                 protein 
                 KKLYCSILEAGLMIPYSGMLSGDFMYAVVSLPDK 
                   
               
               
                   
                 LDITYCPIDNYMENPTKSECENINRWMEFFKMYG 
                   
               
               
                   
                 THVSTHIITGGKIFHEYKTLNITEYRKKSKFRQSTN 
                   
               
               
                   
                 IFEVTNINFDSAGQKSTKNTIVFGGNYVKGMESEA 
                   
               
               
                   
                 RHFYNEWSKTLESRSLPIKVTVKPLSIFMSYRNDI 
                   
               
               
                   
                 YKEALKFYRDVALLTTVGIQYSTKIDELLRESTTV 
                   
               
               
                   
                 VSDDGMAHCPTNQIVLAGFIMSKNPKVPIVNCEQ 
                   
               
               
                   
                 GKILCSNGTDKPASVYIICVKELNDIITTSTSMENV 
                   
               
               
                   
                 HICPNGNVTALGFAFRKQSESDDWTVVIPRIGKQ 
                   
               
               
                   
                 QMINYTKGLNMSWLLCVPIEIMFWHMEMIIDSAP 
                   
               
               
                   
                 KDDSRTVSCKTGWTILKGFKLMFLKKEDTTRVVL 
                   
               
               
                   
                 EECISHQQHCILNCNEECTKMYGTILCKKQHD 
                   
               
               
                   
               
               
                 BdWA1_II2261, 
                 MSTNGTVRWRTSSRSQLRDRRKNALTRRLRFEV 
                 33 
               
               
                 RNA polymerase 
                 GRLDTEAFETRTKYLREIFLDVDKRNSHDWTALE 
                   
               
               
                 Rpb1, domain 
                 KKILLLDWCQNPQKFEKWKPAATEDMQSRFFAN 
                   
               
               
                 containing protein 
                 LIFNYSHLAMINSGMCIVIVRLVPLVVNIKAPYQC 
                   
               
               
                   
                 VILLVRDNVNGMVIDRLKVGLIWRGKDTNTDVL 
                   
               
               
                   
                 GKSTRIFTPPLSTDFQFLSHGIGYNFKSLLPQTEFEP 
                   
               
               
                   
                 HVACSDCISRELESLCTLETPDIVLESPALGMEVTS 
                   
               
               
                   
                 SFVNGHKVYLVVTRGVDGIVRIHRVQELIQTSIDII 
                   
               
               
                   
                 RTWEQLRLKHLKRTMQSQQYAACTGCLVHKEEL 
                   
               
               
                   
                 QQLYHLKQCARSFAPECGPMQFVGLEMIDVARP 
                   
               
               
                   
                 RTCATKREACGLPLALGYTSVRNEQAVIPSLLMA 
                   
               
               
                   
                 MASNTVCIWSIYSYLKEWNLNPLGPIKVMTYPDG 
                   
               
               
                   
                 TLPTDISSTVSIAASLEALERLEFIHAPKFYTTDDR 
                   
               
               
                   
                 GHLSLWSLGSTGPEAQVTLDQNALCSCAVNRSYP 
                   
               
               
                   
                 HIVAVGLDVGKIKIYNTLTQGTTCIAQMEHPLLTK 
                   
               
               
                   
                 VQTLQSFLPDNVYDYQRWYHPVVKLEWISDVFIL 
                   
               
               
                   
                 AQYSEPIFTTESTNASAVAIWNVVKDIFDRDDALV 
                   
               
               
                   
                 CNKHWSLNSCNLYNTWHLASKLVCLYGGHFGCI 
                   
               
               
                   
                 CGVLSSDAKWSAEQGLLAVTIDSTGQLHVFKPGI 
                   
               
               
                   
                 WTWGDCDDPMAIARLTGDCEFYHSILARVQEQH 
                   
               
               
                   
                 RRLSVPVMEIDNSVDMGQGTRLRKTRAKFSTYIQ 
                   
               
               
                   
                 DAQTQLESVETASGLEEINSLEKLPMFCKKTLRLN 
                   
               
               
                   
                 SESEKYLKLVHRLVAEREHAEFLLSQGTPGKTGK 
                   
               
               
                   
                 RKCPLGAPNIKQEPLELIEAPQGPSSEAPVKSKNE 
                   
               
               
                   
                 YQNDAMEPESFNFSSVLTKFDTDVSMEPETELVE 
                   
               
               
                   
                 TALPVVQPTKKRLVEKGLEACCIEGVQFDIMGDV 
                   
               
               
                   
                 EIARCAELEVQKRELYYYLSSMPFPQGVLDLRLG 
                   
               
               
                   
                 CNRNDTKCETCGRALMECVGHWGYINLQLPVFH 
                   
               
               
                   
                 VGFFKYTIQLLYCICKRCSSLLLPFDTVAQLRDAR 
                   
               
               
                   
                 LRRSDDPLARAVIFKRILSSCRKVTKCPACGARQG 
                   
               
               
                   
                 VIRRIVKPTMDQFMKLRHVVKYKEGGKIVIVEDE 
                   
               
               
                   
                 LNPLSVLRLFEAIDPVHARILNIMDPQRLIISNLPVP 
                   
               
               
                   
                 PACIRPSVSLQGQGTTEDDLTCILSDIVELNNVMA 
                   
               
               
                   
                 TQMQQGFQTNQVIGNWQFLQLQCTRLINADAPA 
                   
               
               
                   
                 VSQLLAAKHISKPGRGICQRLKGKEGRFRGNLSG 
                   
               
               
                   
                 KRVDFSARTVISPDPNIGIDEIVIPEYIARRLTFPEK 
                   
               
               
                   
                 VTSANLGVLQKAVLNGVSKWPGACYVMKRDGV 
                   
               
               
                   
                 KCTLRFANPKQVAESLQIGDIVERHLWNGDVVLF 
                   
               
               
                   
                 NRQPSLHRMSIMAHKARVMPGSTFRFNECVCNP 
                   
               
               
                   
                 YNADFDGDEMNLHMPQTYEARAEALHLMGVLQ 
                   
               
               
                   
                 NITTPRNGDPLIAATQDFLSASYLLTSKDRFLSHQ 
                   
               
               
                   
                 EFCQLLCYVGDGILHAQVPAPAIVYPTFLWTGKQ 
                   
               
               
                   
                 VYTAILRQIGAVVNLECREREFQHPEPNLFGRMQ 
                   
               
               
                   
                 PNFMCIKDGHVIISNSELLCGALAKKTLGASKDGL 
                   
               
               
                   
                 FYQLLRRHGPHKAADVMLKVSKLTSRWVSDFG 
                   
               
               
                   
                 MTIGLDDTTPSPMLLATKQQLLSDGYAKVELAIA 
                   
               
               
                   
                 NAANIEPFPGCTRKETLELQVKGILDDLRNQAGK 
                   
               
               
                   
                 ACNKSLSANNKPMIMFNSGAKGALINIAQMIACV 
                   
               
               
                   
                 GQQNVMGQRIHHGFIGRTLPHFEVGCIDAKSRGF 
                   
               
               
                   
                 VSNSFFSGLDPAEFWFHTMSGREGLIDTAVKTSET 
                   
               
               
                   
                 GYMQRRLMKALEDLAICYDYTVRTSDGQIVQFIY 
                   
               
               
                   
                 GDDGLGASGAHATTPKALQETLAHVLTLSRFQRT 
                   
               
               
                   
                 PTILNSRAPGPNSKGDLRLPLPSGDGDDENFECGI 
                   
               
               
                   
                 VPKCKLAMPPKVKAKASARASWRTSQVGTPLDE 
                   
               
               
                   
                 ATSAQINRNMYAHWVVHVHLDPEHAKAPLFGDE 
                   
               
               
                   
                 VLDWVALLEPLCREPLPRCMQESVTSMDHTSEPR 
                   
               
               
                   
                 IHASSMEIDKRVNGCSMKATNTGPRFSAVLLDTIR 
                   
               
               
                   
                 QWAARLQTLDPEIQREYESSGMDLVQFRAFKIPP 
                   
               
               
                   
                 QDRISVLQIYEFLRCAWSDYLHGICEPGEAIGALG 
                   
               
               
                   
                 AQSIGEPGTQMTLKTFHFAGVASMNVTLGVPRIK 
                   
               
               
                   
                 EIINAASVIQTPIIEVPLAKKDSYDFAKSVRAKIER 
                   
               
               
                   
                 HTLEQVVHSIKQVYTPGATLLLVSLDSQYIQQNM 
                   
               
               
                   
                 LQLDASSVCAVIRNSNLITKFKLSKHNIEAPQKWL 
                   
               
               
                   
                 VSIRLVASEALLFQLNALVAALGQLVVCGVKDIK 
                   
               
               
                   
                 RCVVKREQKDVINSLTPGATFEYALAVEGYGLQQ 
                   
               
               
                   
                 VLGIFGVDAHRVISNHVAEVAKVLGIEAARLVIVT 
                   
               
               
                   
                 EIKKSMDAYGIDIDGRYMKLLGDVMTFRGEVVGI 
                   
               
               
                   
                 NRFGIQKMRASSLMLASFEETNEHLFQAAVHGRR 
                   
               
               
                   
                 DPIKGVSECIIMGKHIALGTGAFDLLYRE 
                   
               
               
                   
               
               
                 BdWA1_III3073, 
                 MANGGLYVFPLLLCMHSVYALQCPHAHRACFLK 
                 34 
               
               
                 PP-loop family 
                 YNGIYTSKANKLGVLDVVNVEDNALDLEDVVPL 
                   
               
               
                   
                 QEVYKFFNNYLMPFYHKRVSQNYGTNVPIIISCSG 
                   
               
               
                   
                 GVDSMALLHTFGLIKENSHTFIKKHPINYIDGSNA 
                   
               
               
                   
                 VIAETASNVLKYIFENVNVIYFDHKVRSDTKVDIEI 
                   
               
               
                   
                 IENACKKYNFNFNVQELDCNDSYFSNLEGGFQAN 
                   
               
               
                   
                 SRKWRRRELKKYVMELHSSKMLSNEGKNKIGIR 
                   
               
               
                   
                 NGSYNSITTNKQDGLNSNSIGIVFMGHHANDNIET 
                   
               
               
                   
                 FFMKFIRGTHLLQMSEINDQSFLDSKDRIPLIRPFIH 
                   
               
               
                   
                 LPKNALHQFMQDFRFQYNEDSTNVLFDYGRNQF 
                   
               
               
                   
                 RKLVMPNLIEYIMSINKCQNDKAIDLLDKRIHVLS 
                   
               
               
                   
                 KQASNFRNDIEFQIQMFKVYLKSKYGDLLPIKKK 
                   
               
               
                   
                 RYMNRMGEIYSDFYKRLYFNRYNTAIHSNIQELK 
                   
               
               
                   
                 YFHDLNIPIMDLFFVDEWLILESKLLREEVLYDFFS 
                   
               
               
                   
                 HHFRKPLFYNAFTKIVDRLEVNFSEGSIKQYCLSK 
                   
               
               
                   
                 DVSMSHQGSLLKVKHRPEKNEKVLIFKDDLCSFS 
                   
               
               
                   
                 VLDNLQLFVEKAEGYTRNVFHLLFKVPMYATTES 
                   
               
               
                   
                 IDFDIRLFRNDDVLPSKILWNREAGSLLTHMKIKHI 
                   
               
               
                   
                 IKDYIPVIAMAGTNKIVGFYGFNIIPPYHCRGRQEV 
                   
               
               
                   
                 TYSFVDVAHGSTEYREYSIRVTR 
                   
               
               
                   
               
               
                 BdWA1_II2013, 5′- 
                 MGFFIQSQLLSTLWILIVIPQNVQCLRNNSRNNSL 
                 35 
               
               
                 3′ exonuclease 
                 AFASGYAHSNVRNSLDSLNGLPNLINRKEPTCQD 
                   
               
               
                   
                 GFKLFGVPRMYGWMIENLGKINQSFDTCDFYED 
                   
               
               
                   
                 VDYFYIDMNAVIHSATHGNMSPIVEIEDEQRMRRI 
                   
               
               
                   
                 TSTLLKLFHMIKPKKVMYLAVDGVCPSAKINQQR 
                   
               
               
                   
                 TRRFRLAKKVEDLTARVQDICEMKPIEDYNIDKLP 
                   
               
               
                   
                 CGSYDNVTFNPNYISPGTEFMQMFDSEIKNWLAI 
                   
               
               
                   
                 KTLEKQWGDCLVIYDGANIPGEGEQKIYEFMRKL 
                   
               
               
                   
                 NESKVKSRNKNHLVYGLDADIMMLSLLTKMPNV 
                   
               
               
                   
                 CVLREKRDYTPHILSKIKPEPYFTPETGIVHYHAN 
                   
               
               
                   
                 DYIDLEAKHFDVVSMMDIRRALYNRCIKYVGLK 
                   
               
               
                   
                 YDLANIPFLNQENVPNRLADDFVLLSFLAGNDFLP 
                   
               
               
                   
                 HLPTVDLEFHTFSDMLNSYFFMLPRLHGFITRGYR 
                   
               
               
                   
                 IHMGRLQKLFRVLQRQEVHVFKEKAQHESVPEY 
                   
               
               
                   
                 RDVTKYAEHYYRVKCNINYKNRNQVTRMCQEYI 
                   
               
               
                   
                 KGLVWNLYYYYKGCPSWNWCYKYHYAPLVSDL 
                   
               
               
                   
                 SQTSGVFVSFKRGRPIKPLEHLLAVSPPNGNELLP 
                   
               
               
                   
                 EQYRKLSASPNGELAEFFPTDYEICEDGKVNEWE 
                   
               
               
                   
                 HVVKLPFLNTNKLVTHAEQANQHLKYNSLSKNK 
                   
               
               
                   
                 LGRVNVYKCRPHNVNKSGDNRLIFDDLTKKGLIV 
                   
               
               
                   
                 RDGMHYGATFSVYEERPGLAHSSCLIFARHGQTPI 
                   
               
               
                   
                 MIKDLVRWTRISQAANKRVLIITILYKTMNTSGWS 
                   
               
               
                   
                 SDFSSLSNSDKITGKDRINEQESISVPSGLHCGPAG 
                   
               
               
                   
                 RRTLIKWIEENYRVRKGMDDTNPGVVACRLWQD 
                   
               
               
                   
                 SQGFSDPVSKAAIDLLRLQGIPANVTYKTLFEQHL 
                   
               
               
                   
                 SKMLSSLINQSRNSQARLYSIAEKMIGMFQVAEIQ 
                   
               
               
                   
                 PLICDVLDQLDEIPQFALPKLLDDAASANNFYKIC 
                   
               
               
                   
                 NDGLKRKIWATSAVRLYEEMLPLFQEAVLCIQLG 
                   
               
               
                   
                 VLEPRMHHASFIQSCRERYEIVIEQICQMIGDCKS 
                   
               
               
                   
                 QASRQVFRLTMRIIKVLYVKSILGDKGDFQIYFTP 
                   
               
               
                   
                 HDHLDTALKLNWRDLAARSVGICFKSLQLEELDE 
                   
               
               
                   
                 AHDSFSSTDHSSFCALAHSIISRHQDMHKITSSDM 
                   
               
               
                   
                 QDLESFYTLWSLVDTVIASPCMSLDNEQIASVLN 
                   
               
               
                   
                 VVLGDLRIRDEIDLFDASFVLGHFEFLIKISKYIVT 
                   
               
               
                   
                 RSININEQLPEDQDTLGQWFSLASLGLSCGYIGAC 
                   
               
               
                   
                 QFLFRAQWNNDTCIWHFPTETEPGKPISLSKLLGY 
                   
               
               
                   
                 TFPYNRSDQMCLDTLPPFFYKIVHGQASPHLSETS 
                   
               
               
                   
                 PVRNPYEANKASLRNLLCLYLPMVDVWKFKRA 
                   
               
               
                   
                 MMDLKAFEQNGYGTLLNSFKRQTFQTEKLLFYM 
                   
               
               
                   
                 QDMAIRDFESDDYDWTIARKLYQNLERLQALKL 
                   
               
               
                   
                 ATPPHGTVMDPKSPQPHVDDGKRLMGTILIQGDI 
                   
               
               
                   
                 TCGFIRMMGVSMVEASDEMARRLENYWKNLLQ 
                   
               
               
                   
                 PYMCCMQFRLLRMLFSNWTKISSFSLDVLCKMV 
                   
               
               
                   
                 KGYPIVAFLPLFTKAPNCDISTALGRLSASLQGTLP 
                   
               
               
                   
                 NHSSKGWHRIVFTMYNKHS 
                   
               
               
                   
               
               
                 BdWA1_I0563, 
                 MKLQINSFNHIFFIKQNIRTNLFKLSQKPHPFNLKF 
                 36 
               
               
                 hypothetical protein 
                 FRQYSHHEIFTSIKLASNKSEMASVTQILLNRSNY 
                   
               
               
                   
                 KGVVTLDNSTVVSANDIVLYSEFNSKISFSLTLDT 
                   
               
               
                   
                 LKKLTKLQTPCKVVWNYVIDSILSDSSHFTPNEIA 
                   
               
               
                   
                 KILVYILNIKFNDIFIKSKIELASETLIEKLNGSLSLM 
                   
               
               
                   
                 VGFSVIDIKRAGLRNISDTIRHDAEFDKLDSMDFEI 
                   
               
               
                   
                 IWKLWYTLSKQRGNERIRGLLFEYLERGKDKVYK 
                   
               
               
                   
                 FDLPKVIRSMDIILSNSKSLDLLELKFLEKLFKRFL 
                   
               
               
                   
                 NLYNSDNIPTFGGSSNIYMTTIQAKKALYILGNICK 
                   
               
               
                   
                 RNPKIFENIVKHECAMLSLVNFLKQVKEITSRSLE 
                   
               
               
                   
                 VHCRLYNHLKNINVENTNMKHNVRIEAIKRSSNV 
                   
               
               
                   
                 HAKMFVYCVEFLNNFDLITKANYNNASSYSKATS 
                   
               
               
                   
                 DNKRSTDLDHTIFDICTNLLRCTAQSSFLIHNVTM 
                   
               
               
                   
                 YCRSIYQLLVLALYNTCTGINESFVKAALSSISNIQ 
                   
               
               
                   
                 ATTLQMDRDNWDISSVIRNLSYIDKGISVLADISY 
                   
               
               
                   
                 SSIILEPIQNANAEIVSKCSSKYEEVLMGGEILQLD 
                   
               
               
                   
                 KLVYCTLLNKYNIIDGTMLDVLLQFIKANELTSRE 
                   
               
               
                   
                 LCMFCDLLLKTREMLMDDCDKRNNVLAKLLPLA 
                   
               
               
                   
                 LELLKSIDFDFNKRTLKDLLLILECLGAFGFSCAD 
                   
               
               
                   
                 DLSKRILEHGAKLTISQLMKLYSIVNDDYKRRIRL 
                   
               
               
                   
                 LLPGILKQRGKINITAILKVLNVLDIDYNLLETLEH 
                   
               
               
                   
                 NLGDDLYLVDKAYFARKRLLHQRNSSNPRKKTK 
                   
               
               
                   
                 VDYNNSGSHHMFIIDSSVAKAFIKCNKGTHSSSKL 
                   
               
               
                   
                 YKILVSKYQSSSSSMAL 
                   
               
               
                   
               
               
                 BdWA1_II2256, 
                 MQGGWLLVCIVGFLACFGASTKPNDKKATNEEL 
                 37 
               
               
                 hypothetical protein 
                 TCPAVNDLQGTPIDLQYTFDRLDMTSLSRLLAIQN 
                   
               
               
                   
                 MYSKRNPRNIRIPKFWNTDVAHGIWSRFRIYGTN 
                   
               
               
                   
                 LVYKTSRGDGSCLFWSVSDSLRLGGFTIKKIKENL 
                   
               
               
                   
                 DHYGTKNRGIYEAIMSLPNDDEYLTMQDIQRIATI 
                   
               
               
                   
                 GFVGFDPDVEASVKQWDREPILSHLETVKSLKTA 
                   
               
               
                   
                 YMGDVIPFDLGCHRFTSFRAVDDFYNGIANDATC 
                   
               
               
                   
                 IKAGRDLFNHTIKYLASGAWGYEIDIYAIETALNL 
                   
               
               
                   
                 KIVLISYNTGSLVCYYWSEDYVPQSMILLYYHDS 
                   
               
               
                   
                 RHFDVAGLVDMFKVPNVRNPRAKVLTSFHIREM 
                   
               
               
                   
                 PMALAIILKDDCRITKRNERFSFGNEDLTHLY 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                   B. duncani  secreted antigens identified in the hemolysate fraction: SEQ ID NOs: 38- 
               
               
                 46 
               
            
           
           
               
               
               
            
               
                 Antigen ID 
                 Annotated Sequence 
                 SEQ ID NO: 
               
               
                   
               
               
                 BdWA1_II2291, 
                 MQMFNRFLKASVALLAVASFGIQYIFAKGSNSGK 
                 38 
               
               
                 heat shock protein 
                 IEGPIIGIDLGTTYSCVGIYKNGRVEIIANEMGNRIT 
                   
               
               
                 70 precursor 
                 PSYVSFVEGTQKVGEAAKSEATINTESTVFDVKR 
                   
               
               
                   
                 LIGRKFTDRDVQEDMKLLPYKIINKSTRPYISLHD 
                   
               
               
                   
                 GKEQRTFAPEEISAMVLKKMKQVAESYLGKEVK 
                   
               
               
                   
                 KAIITVPAYFNDSQRQSTKDAGAIAGLDVVRIINEP 
                   
               
               
                   
                 TAAAIAYGLDKANAESNILVYDLGGGTFDVSVLT 
                   
               
               
                   
                 LDSGVFEVIATGGDTHLGGEDFDRRVMDHFIDIF 
                   
               
               
                   
                 KKKHKVNIRDNKQSLQKLRKEVEAAKRTLSSTTE 
                   
               
               
                   
                 VLVEVENLINGIDFSEKLTRAKFESLNAELFEKTL 
                   
               
               
                   
                 ATVKKVVEDADIPIRDINQVVLVGGSTRIPRIREMI 
                   
               
               
                   
                 KEYFGKEPDYGINPDEAVAFGAAMQGGILSGESS 
                   
               
               
                   
                 DNLLLLDVCPLSLGIETLGEVMSVIIPRNTMIPAHK 
                   
               
               
                   
                 SQVFSTSVDNQPMVTIKVYQGERKLTKDNVILGK 
                   
               
               
                   
                 FDLSGIPPAPRGVPQIEVTFDIDTNGILSVSAEEKG 
                   
               
               
                   
                 SGNKHNIVITPDKGRLSPEEIERMIKDAEMNAEKD 
                   
               
               
                   
                 KEVFNRVQARQALEGYIDSMTKTINDDKTGKKLE 
                   
               
               
                   
                 DDEKEKIRDALDEGTKWLASNPEVGADEISAKQH 
                   
               
               
                   
                 EIEAICNPIISKLYGSGEDSDDSGYSDEL 
                   
               
               
                   
               
               
                 BdWA1_II1496, 
                 MAIPDNNNNTQSNGFDTLESNYDEVVDSFEALKL 
                 39 
               
               
                 eukaryotic initiation 
                 NEDLLRGIYSYGFERPSAIQQRGIKPIIENYDTIGQ 
                   
               
               
                 factor 4A-3 (eIF4A- 
                 AQSGTGKTATFSIAALQIINYNIMSCQTLILAPTRE 
                   
               
               
                 3) 
                 LAQQIQKVVLALGDYLKVQCHACVGGTVVRDDI 
                   
               
               
                   
                 HKLKAGVHMVVGTPGRVYDMIDKKALLTDKIRL 
                   
               
               
                   
                 FILDEADEMLSRGFKGQIHEVFKRMPPDVQVALF 
                   
               
               
                   
                 SATMPNEILELTTKFMRSPKLILVKKDELTLEGIK 
                   
               
               
                   
                 QFFVMIDKEDYKFDTLCDLYESVTITQAIIYCNTR 
                   
               
               
                   
                 RKVDMLTNKMQEKDFTVSSMHGDMGQKERDLI 
                   
               
               
                   
                 MREFRSGSTRVLITTDLLARGIDVQQVSLVINYDL 
                   
               
               
                   
                 PMSPDNYIHRIGRSGRFGRKGVAINFLTPLDMDA 
                   
               
               
                   
                 MKSIENYYNTQIEEMPADIAAYM 
                   
               
               
                   
               
               
                 BdWA1_II2200, 
                 MSKKLKTKGPENINHSLQLVMKSGKVCLGFKSTR 
                 40 
               
               
                 ribosomal protein 
                 AALRSGKAWMIILSNNIPALRRSEIEYYAMLAKCS 
                   
               
               
                 L7Ae containing 
                 VYRYSGDNNDLGTACGKYFRVGCMAVLDAGDS 
                   
               
               
                 protein 
                 DILRNIE 
                   
               
               
                   
               
               
                 BdWA1_I0250, 
                 MTSVNSDVDISEVSQMSDADIRVRINLIDSEIKILR 
                 41 
               
               
                 26S protease 
                 SEHTRLKSRQKTLQDRIKDNLEKIQLNKQLPYLV 
                   
               
               
                 regulatory subunit 
                 ANVVELLDFTSDDEQDDGLTPSPSQKKSKSLVIKT 
                   
               
               
                 6A 
                 STRQTIFLPVIGLIPASELHPGELVGVNKDSYLVLD 
                   
               
               
                   
                 KLPPEYDNRVKAMEVCEKPIEDYSDVGGLDKQIQ 
                   
               
               
                   
                 ELVEAIVLPITHQERFKKIGIKPPKGVLMHGPPGTG 
                   
               
               
                   
                 KTLLARACAAQTKATFLKLAGPQLVQMFIGDGA 
                   
               
               
                   
                 KMVRDAFSLAKEKAPTIIFIDEIDAIGTKRFDSELS 
                   
               
               
                   
                 GDREVQRTMLELLNQLDGFASDDRVKVIAATNR 
                   
               
               
                   
                 PDTLDPALLRSGRLDRKIELPHPNEQARCHILQIHS 
                   
               
               
                   
                 RRMNVNPDVNFKELARSTDDFNGAQLKAVCIEA 
                   
               
               
                   
                 GMVALRRDASELEHEDFVEGISMVQAKKKNTLN 
                   
               
               
                   
                 YLN 
                   
               
               
                   
               
               
                 BdWA1_II1994, 
                 MAGHAALILNFGSCFSGVLVRVLRDVGINCVLES 
                 42 
               
               
                 GMP synthase 
                 AEKALDALNTNSTVKVAILCGGLDSVYDETSLTV 
                   
               
               
                   
                 PEEFIKACEEKNVKILAISHAFYALCKTLGARLMN 
                   
               
               
                   
                 GKGNDYIIDTVTVQRPMVLFNNVGKHFKAKINPI 
                   
               
               
                   
                 NGVEVLPAGFESLATFGNGHYAAIGDEKRGIFGV 
                   
               
               
                   
                 AFHPESDDTENGLVILKNFCLEQGACPIEWSMEQ 
                   
               
               
                   
                 YLKDELARCIAQCGDTKVVVAGLSGGVDSTVCA 
                   
               
               
                   
                 AIVHKAIGNRFHGVMINTGLMRLDETKKCAERLK 
                   
               
               
                   
                 KEIPGIQLHIRESADVFFGELKGILDPEQKRKIIGKV 
                   
               
               
                   
                 YIDEFERAIKDLGFDKSNCLLLQGTIYPDILESELN 
                   
               
               
                   
                 RRNQMPIKSHHNVGGLPKDLALELIEPVRLLFKEE 
                   
               
               
                   
                 VRKLGRLLGLSQESCERQPFPGPGLGVRVIGELNP 
                   
               
               
                   
                 RNLDLVRRADAWNQILDARGYRSKISQSGCILL 
                   
               
               
                   
                 ADVHNTGIRNSGRTYGHAVIIRIIITTDFVTAQWA 
                   
               
               
                   
                 RIIDTDCLAEISKTITDTVPEITRVCYDITDKPPACIE 
                   
               
               
                   
                 WE 
                   
               
               
                   
               
               
                 BdWA1_III2590, 
                 MAEEMPQFKLLLVGDGGVGKTTLVKRHLTGEFE 
                 43 
               
               
                 GTP-binding 
                 KKYIPTLGVEVHPLKFRTNCGGIQFNAWDTAGQE 
                   
               
               
                 nuclear protein Ran 
                 KYGGLRDGYYIKGECAIIMFDVTSRITYRNVPNW 
                   
               
               
                   
                 HRDIVRVCENIPMVLVGNKADVKERQVKAGHIQF 
                   
               
               
                   
                 HRKRNLQYYDLSARSNFNFERPFLWLSRRLLNQP 
                   
               
               
                   
                 QLVFVGECAKAPEIQIDPLLAQQSERDLEAAARV 
                   
               
               
                   
                 AIDDDGDL 
                   
               
               
                   
               
               
                 BdWA1_I0810, 
                 MADRFTGRNNREAVVAYPGWFSETQKQCLRAC 
                 44 
               
               
                 Hsp70 protein 
                 VTASGLSCLRVISHVHAMAMDYGVYRVKQLNDE 
                   
               
               
                   
                 TPTRVALVMIGHCHASAAIVDFYASHCSILSQVSR 
                   
               
               
                   
                 RNLGGRNLDMMLMKYMATEFSKKYHCDPLENN 
                   
               
               
                   
                 KTRLKVEAVAVKTRRVLSANAESSYSAECLMED 
                   
               
               
                   
                 NDMSGHITRTQFEEMCNAEFIPQLIEMLKECIEAS 
                   
               
               
                   
                 RTDLDSIFSVEIAGGSSRIPCIQQAISSIFNKVPSRTL 
                   
               
               
                   
                 NADECIARGCVLEAAIKSNHYRVREYKTRLTLPR 
                   
               
               
                   
                 SLTLGYFNGQEPMLLEAIAAGTPLGDPIRVTLQAQ 
                   
               
               
                   
                 APVCVRVALGDALDPRSQDALGTLDIARHISQEA 
                   
               
               
                   
                 QPAPVTTNDGAAIQTDEQDAEIQSESSPSGGISVTL 
                   
               
               
                   
                 GFDDCGQFVASPECCEYRWLPATILDIARLEAAEL 
                   
               
               
                   
                 EARGRDLKENSRLQALNDFETLLYTVRDKMQSS 
                   
               
               
                   
                 HRDFIDPQMIPAYESELDHWREWLYENSGASQET 
                   
               
               
                   
                 LQEGIDKVSSEWKRIDKYFKEHQNKLENLEPFLQ 
                   
               
               
                   
                 RLQERYNFCCEDNNPNWHGATPEERLNFAQELM 
                   
               
               
                   
                 DLDSRVRQMHQDESQRPRHMEPLFTMQQIQGEM 
                   
               
               
                   
                 QKLLVSISEFCQAKAAKAPAQEPPEQQPKEQQE 
                   
               
               
                   
               
               
                 BdWA1_II1634, 
                 MDAGGKIGGKIGGKVGGMGKGGKGKTGSGKGK 
                 45 
               
               
                 Core histone 
                 KAPMSRAARAGLQFPVGRVHRMLKSRISADGRV 
                   
               
               
                 H2A/H2B/H3/H4 
                 GSTAAVYASAILEYLTAEVLELAGNASKDLKVKR 
                   
               
               
                   
                 ITPRHLQLAIRGDEELDTLIKATIAGGGVIPHIHKA 
                   
               
               
                   
                 LMNKGPAQVLVKPPKRI 
                   
               
               
                   
               
               
                 BdWA1_II1773, 
                 MGIVTASIAPLHIGDDLYTRMKTLEKKLEICEIQE 
                 46 
               
               
                 26S protease 
                 NYVREEYRNLKLELIRAREEIKRIQSVPLVIGQFLD 
                   
               
               
                 regulatory subunit 4 
                 MIDKNYGIVSSTAGSNYYVRILSTINRELLTPNSSV 
                   
               
               
                   
                 ALHRHSHSVVDLLPPEADSSIQLMQVSEKPDVTY 
                   
               
               
                   
                 ADIGGLDIQKQEIREAVELPLTCPELYRQIGIDPPV 
                   
               
               
                   
                 GVLLYGPPGTGKTMLAKAVANQTDAKFIRVVGS 
                   
               
               
                   
                 EFVQKYLGEGPRMVRDIFRLARENAPAILFIDEVD 
                   
               
               
                   
                 AIATKRFDAQTGADREVQRILLELLNQMDGFDQN 
                   
               
               
                   
                 ATVKVIMATNRADTLDPALLRPGRLDRKIEFPLPD 
                   
               
               
                   
                 RRQRRLIFQTITSNMNLAADVDLETFVARPEKVS 
                   
               
               
                   
                 AADIAAICQEAGIQAIRKNRYVVTTKDFERGWKR 
                   
               
               
                   
                 HIRKHERDYGFYGV 
               
               
                   
               
            
           
         
       
     
     Example 5.  Babesia microti  Secreted Antigens: SEQ ID NOs: 47-63 
       
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                   B. microti  secreted antigens identified in the supernatant fraction: SEQ ID NOs: 47- 
               
               
                 54 
               
            
           
           
               
               
               
            
               
                 Antigen ID 
                 Annotated Sequence 
                 SEQ ID NO: 
               
               
                   
               
               
                 BmR1_04g08775 
                 MSSQETFEFNADISQLLSLIINAFYSNKEIFLRELIS 
                 47 
               
               
                   
                 NASDALEKIRYELLRDGTKVSDESEFSIKISADKS 
                   
               
               
                   
                 NNTLTIEDSGIGMTKADLINNLGTIAKSGTKAFME 
                   
               
               
                   
                 AMQSGCDMSMIGQFGVGFYSAYLVAEKVTVVSK 
                   
               
               
                   
                 HNSDEQYIWESSASGVFTITKDETTEKMKRGTRLI 
                   
               
               
                   
                 LQLKEDQTEYLEERRLKELVKKHSEFISFPIHLLCE 
                   
               
               
                   
                 KTKEEEVTASDDEGDKKEDDKKEDDEKEDDKKG 
                   
               
               
                   
                 EDEKVEDVSEDKKKTKKVSTVTKEWEVLNKQKP 
                   
               
               
                   
                 IWMRQPNEVTNEEYANFYKNLTNDWEDHLAVK 
                   
               
               
                   
                 HFSVEGQLEFRAILFIPKRAPFDMFENRKKKNNIK 
                   
               
               
                   
                 LYVRRVFIMDDCEELIPEWLSFVKGVVDSEDLPL 
                   
               
               
                   
                 NISRETLQQNKILKVIRKNLVKKCLELFSELTEKK 
                   
               
               
                   
                 DDFKKFYEQFNKNLKLGIHEDSANRNKISELLRFE 
                   
               
               
                   
                 TTKSGDEAISLREYVDRMKPNQKYIYYITGESIQA 
                   
               
               
                   
                 VSNAPFLEKLKDKNIEVIYMTDPIDEYAVQQIKEF 
                   
               
               
                   
                 DGKKLRCCTKEGLDIDDEKDEEEEKRFEQVKQE 
                   
               
               
                   
                 MEPLCKTIKEVLHDKVEKVTCGKRFTTSPLALVT 
                   
               
               
                   
                 SEFGWSANMERIMRAQALRNSSITSYMVSKKTM 
                   
               
               
                   
                 EINPYHSIMKALKERVAADKSDKTVKDLIWLLYE 
                   
               
               
                   
                 SALLISGFNLEEPTQFGNRIFRMIKLGLALEDDQPD 
                   
               
               
                   
                 DTDLPPLDEGVAVDGGDSKMEEVD 
                   
               
               
                   
               
               
                 BmR1_03g03490 
                 MPKEKTHINLVVIGHVDSGKSTTTGHLIYKLGGID 
                 48 
               
               
                   
                 KRTIEKFEKDSSEMGKSSFKYAWVLDKLKSERER 
                   
               
               
                   
                 GITIDITLWKFETQKYEYTVIDAPGHRDFIKNMITG 
                   
               
               
                   
                 TSQADVAMLVVPAESGGFEAAFSKEGQTREHAL 
                   
               
               
                   
                 LAFTLGVKQMIVAINKMDSCQYKEDRYMEIFKEV 
                   
               
               
                   
                 QQYLKKVGYKVESVPFVAISGFHGDNMVEKSTN 
                   
               
               
                   
                 MPWYKGKTLVEALDQMEPPKRPVEKPLRLPLQS 
                   
               
               
                   
                 VYKIGGIGTVPVGRVETGQLKAGMIITFAPTGLTT 
                   
               
               
                   
                 ECKSVEMHHEVVEVASPGDNVGFNVKNVSVKDI 
                   
               
               
                   
                 KRGNVASDSKNDPAKEATSFSAQVIVLNHPGTIK 
                   
               
               
                   
                 AGYSPVVDCHTAHIACKFESLDTRIDKRTGKTLEE 
                   
               
               
                   
                 NPKTIKNGDAAMVTMKPNKPMVVETFTDYAPLG 
                   
               
               
                   
                 RFAVRDMRQTVAVGIIKAVEKKDPSSAKVTKSAV 
                   
               
               
                   
                 KAGKK 
                   
               
               
                   
               
               
                 BmR1_04g05965 
                 MTKIISACGREVLDSRGNPTVECEVTTEGGKFRAI 
                 49 
               
               
                   
                 VPSGASTGIYEALELRDGDKTRYLGKGVQNAIKN 
                   
               
               
                   
                 MHNIICPGIQGFLCTEQEKLDNHMVKVLDGTQNE 
                   
               
               
                   
                 WGFSKSKLGANTILAVSMGAARAGAAAKGIPLY 
                   
               
               
                   
                 EHLAQLSGKPTDKFIMPVPCLNVINGGSHAGNAL 
                   
               
               
                   
                 AFQEFMILPTVADNFSNALRMGVEVYHTLKKVIN 
                   
               
               
                   
                 KKYGQDATNVGDEGGFAPNISTPQEALDLLVEAI 
                   
               
               
                   
                 AAAGYTGKIKIAMDVAASEFYQKDVKMYNLTFK 
                   
               
               
                   
                 SSSPDIKTSDQLVELYKELVNKYPIVSIEDPFDQDD 
                   
               
               
                   
                 WEAYAKLTAAIGDKIQIVGDDLLVTNPKRIEAAIQ 
                   
               
               
                   
                 KKACNALLLKVNQIGSVTESIQACKISQENGWGV 
                   
               
               
                   
                 MVSHRSGETEDVFISDLVVALGTGQIKTGAPCRSE 
                   
               
               
                   
                 RNAKYNQLLRIEQELGERATYSKVFNK 
                   
               
               
                   
               
               
                 BMR1_03g01010 
                 MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGI 
                 50 
               
               
                   
                 PPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVL 
                   
               
               
                   
                 RLRGGD 
                   
               
               
                   
               
               
                 BMR1_03g01340 
                 MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGI 
                 51 
               
               
                   
                 PPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVL 
                   
               
               
                   
                 RLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKI 
                   
               
               
                   
                 QDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKES 
                   
               
               
                   
                 TLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIE 
                   
               
               
                   
                 NVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDY 
                   
               
               
                   
                 NIQKESTLHLVLRLRGGVIDPSLALLAQKYNCNK 
                   
               
               
                   
                 MVCRKCYARLPPRATNCRKKRCGHCNDLRPKKK 
                   
               
               
                   
                 IKGN 
                   
               
               
                   
               
               
                 BMR1_03g02390 
                 MGDDDNAALVVDNGSGNVKAGIAGDDAPRCVF 
                 52 
               
               
                   
                 PSIVGRPKNPALMIGMDEKEVYVGDEAQSKRGIL 
                   
               
               
                   
                 TLKYPIEHGIVTNWDDMEKIWHHTFYNELRVNPE 
                   
               
               
                   
                 EHSVLLTEAPLNPKTNREKMATIMFETHNVPAMY 
                   
               
               
                   
                 VAIQAVLSLYSSGRTTGIVLDSGDGVSHTVPIYEG 
                   
               
               
                   
                 YAMPSAIMRLDLAGRDLTEYMQKILVERGFSFTT 
                   
               
               
                   
                 SAEKEIVRDIKEKLCYIALDFDEEMQAAETSSDLE 
                   
               
               
                   
                 KSYELPDGNIITVGNERFRCAEVLFQPSFIGKECH 
                   
               
               
                   
                 GLHKTTFDSIIKCDVDIRRDLYSNVVLSGGTTMLQ 
                   
               
               
                   
                 GIGERLTKELSCLAPSTMKIKVVAPPERKYSVWIG 
                   
               
               
                   
                 GSILSSLSTFQQMWITKDEFDESGPVIVHRKCF 
                   
               
               
                   
               
               
                 BMR1_02g03540 
                 MHNHSYYTILLTIALIHTTGCHGFLHRNIKFILHSM 
                 53 
               
               
                   
                 TVYGKPKRLHKTPPWAHLFEEKVEPSPLGEPWAK 
                   
               
               
                   
                 LSKDVANGERRVRLTVKKSHLQIYAAVVDDYKN 
                   
               
               
                   
                 QVICIASSNLPVLADVLGTVPTKDPTVRRNKGNN 
                   
               
               
                   
                 VKAAYEVGKHIGRLALAKGVAKVYFDRAGYKY 
                   
               
               
                   
                 HGRVEAVAIGARKVGLQL 
                   
               
               
                   
               
               
                 BmR1_04g09800 
                 MYQIDRMIDKHKDPIDPLQTILSVKGTMKCKLSE 
                 54 
               
               
                   
                 MLTRHSTEDRHSLSLVADLKRESPTHTDSRSGVR 
                   
               
               
                   
                 LSFLDAGEVVVTMANTGFDVVLVNTDDIAYKGT 
                   
               
               
                   
                 LDDLKTSICAAHAIGNRSRPAVVMKDIILHPLQLA 
                   
               
               
                   
                 QAAQLHADGVVLNSFYLGPALESMIDTSYNLGIE 
                   
               
               
                   
                 PIVEVHTLEDALYAIQLHTKILMINQWDRLTNKC 
                   
               
               
                   
                 HPNRALQIREIVPDGIITIACGGIKTLEQIEQLGLAG 
                   
               
               
                   
                 YDAVVLGKKLADTNIPSFVGSIKKWNAPGKGILAI 
                   
               
               
                   
                 SKPLFFTDEMDEKDVSGGGHVIRMKQNYRQTLQ 
                   
               
               
                   
                 EMCEFYQIAENNCVEAPRPKDVLLRFIGNDMPEK 
                   
               
               
                   
                 WIFKREKWMKDHAHEYPSEDHATAVYDFNLAVE 
                   
               
               
                   
                 LNTTLECNKKLLSQHFKREMVDKLEEAIKNYVKK 
                   
               
               
                   
                 CYINLKRFTNKISCS 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                   B. microti  secreted antigens identified in the hemolysate fraction: SEQ ID NOs: 55- 
               
               
                 62 
               
            
           
           
               
               
               
            
               
                 Antigen ID 
                 Annotated Sequence 
                 SEQ ID NO: 
               
               
                   
               
               
                 BMR1_01g02545 
                 MSQGPAIGIDLGTTYSCVGVWKNETVEIIANDQG 
                 55 
               
               
                   
                 NRTTPSYVAFTDVERLVGDAAKNQDARNPENTV 
                   
               
               
                   
                 FDAKRLIGRKINDPCIQSDIKHWPFTVAAGPNDKP 
                   
               
               
                   
                 VIKVQFQGETKSFHPEEISSMVLTKMKEIAESYLG 
                   
               
               
                   
                 KTISNAVITVPAYFNDSQRQATKDAGTIAGLNVM 
                   
               
               
                   
                 RIINEPTAAAIAYGMDKKGTSEKNVLIFDLGGGTF 
                   
               
               
                   
                 DVSILTIEDGIFEVKATQGDTHLGGEDFDNRLVNF 
                   
               
               
                   
                 CVDDFKRKNGGKNISTNRRALRRLRTQCERAKRT 
                   
               
               
                   
                 LSHSTQATIVVEAIFDGIDYSCNITRARFEELCAEM 
                   
               
               
                   
                 FKNTLIPVEKALADADMDKKQIHEVVLVGGSTRI 
                   
               
               
                   
                 PKIQQLIKDFFNGKEPCKSINPDEAVAYGAAVQA 
                   
               
               
                   
                 AILTGEQSSKVQDLLLLDVTPLSLGLETAGGVMT 
                   
               
               
                   
                 VLIPRNTTIPAKKEQEFTTNENNQTGVMIQVFEGE 
                   
               
               
                   
                 RSMTCDNNLLGKFHLTGIPPAPRGVPQIKVTFDID 
                   
               
               
                   
                 ANGILTVSAADKSTGKTEHVTITNDKGRLSQQDID 
                   
               
               
                   
                 RMVAEAEKFREDDEKKKRCVESKNELENYCYSM 
                   
               
               
                   
                 KNALEEEGVKSKLSSSELSEAQKLLQNTFSWIESN 
                   
               
               
                   
                 QLAEKEEFEAKLKEVQAVCTPLTAKLYQAGGGV 
                   
               
               
                   
                 PGGAAPGGFNAGGAAPSGPTVEEVD 
                   
               
               
                   
               
               
                 BMR1_02g03395 
                 MTAYIPGLLERLSAPIVSSRLDDDDLAYLGKYESE 
                 56 
               
               
                   
                 ISDSDNLQNIYSNDTIVALGDLDAKYKQQTYLSSL 
                   
               
               
                   
                 NSHIVDQDAQIAFYPVKYTNKIIRNDSNLTYRRTT 
                   
               
               
                   
                 SSLSNPLVNYIQKSQHDTPDPVDSNIQNSSSEQYAI 
                   
               
               
                   
                 TTASSGSDKSKRLSHDNSKTDHDIHDKSGVVTAN 
                   
               
               
                   
                 NHSTDRFQNAGDLVWQPGFTRQLGTKGRRVLLD 
                   
               
               
                   
                 LIRKVYRGNPEYFKNILALKNPPTSISNLPFCNVT 
                   
               
               
                   
                 MLWELANDFGVFDQAIQIHYAHGKPGYPANYHN 
                   
               
               
                   
                 TANYGCNKTRKTNSGKSKKNKFYNYDYEAYDEY 
                   
               
               
                   
                 YEDEYISSFSNGRTSRGRKIVQPKRFDDYDHLLDS 
                   
               
               
                   
                 NYQVEYGYCSNRHLNKDGNTILYYKNKSHDYVN 
                   
               
               
                   
                 LGELKGHKIERFTSQLIC 
                   
               
               
                   
               
               
                 BMR1_03g04775 
                 MKNDFNSVELPGHFLFTSESVNEGHPDKLCDQIS 
                 57 
               
               
                   
                 DSILDACLEQDPESKVACEVCTKRGMVMVFGEIS 
                   
               
               
                   
                 TNAKVNYEEVVRNVVKNVGYDSEDKGIDYKTM 
                   
               
               
                   
                 DVIINLDQQSHEIAQAVHLGKDADNTCAGDQGIM 
                   
               
               
                   
                 FGYATDETPEYMPLSHSLATGLGKRLKDVRLSGL 
                   
               
               
                   
                 LPYLGPDGKTQITVEYIKEGYGSIKPIRVHTVLISQ 
                   
               
               
                   
                 QHSANVSNDKLREDLMTHVVKAVIPPHFLDDKT 
                   
               
               
                   
                 KYYLNPSGHFVVGGPSSDAGLTGRKVIVDTYGG 
                   
               
               
                   
                 WGAHGGGCFSGKDGTKVDRSAAYYARKVAKSL 
                   
               
               
                   
                 VANGFCRRALVQVSYSIGIRSPLSLHVDSYNTCIE 
                   
               
               
                   
                 GFTDLDLEQIAVRNFDFSVGNIIKELQLKKPIYSQT 
                   
               
               
                   
                 GVYGHFGKDNPEYLWENVKDLSHELTHKPKR 
                   
               
               
                   
               
               
                 BMR1_03g03380 
                 MDYTRSLFTLSGPATASEVEKHIQNAIEFVKRRDP 
                 58 
               
               
                   
                 DQVQFIQAFTEVANGLAPVFQTDLKYLEIFLSLSE 
                   
               
               
                   
                 PERVITFKVPWVNDAGKLMINRGFRVQFNSTLGP 
                   
               
               
                   
                 YKGGLRFHPSVNLSILKFLGFEQIFKNSLTTLAMG 
                   
               
               
                   
                 GGKGGSDFDPKGKSDNEVRSFCQSFMTELQRHIG 
                   
               
               
                   
                 PDTDVPAGDIGVGEREIGFMYGQYKRLSNSSTGT 
                   
               
               
                   
                 LTGKDPKWGGSFIRPQATGYGLVFFVQYILNDLH 
                   
               
               
                   
                 NGDSFKGKRVAISGSGNVAQYAADKVIDFGGIPIT 
                   
               
               
                   
                 FSDSSGYIYEPNGFTKEMVTVLMELKNIQRARVS 
                   
               
               
                   
                 EFLKYSNTAKFFPNKKAWDVDTNVNVALPCACE 
                   
               
               
                   
                 NELDKADAEMLVKKGCIIVGEGANMPTTPEAISV 
                   
               
               
                   
                 FKAAKVTVCPGKAANAGGVAVSGLEMSQNSQRE 
                   
               
               
                   
                 KWTSEKVLEKLQDIMKNMSKACQEAAAKYNVH 
                   
               
               
                   
                 GDIISGANIAGFLKVAHSYCDQGCV 
                   
               
               
                   
               
               
                 BmR1_04g08050 
                 MSYSAEETDASIEQWKILRLIRNLESAKGNGTSMI 
                 59 
               
               
                   
                 SLIIKPKDEIARINKMLADEFGTASNIKSRVNRLSV 
                   
               
               
                   
                 LSAITSTQQKLKLYRQTPPKGLVVYCGTILTEDGK 
                   
               
               
                   
                 EKKVSLDFEPFKPINTSLYLCDNKFHVEALKELLE 
                   
               
               
                   
                 SDEKFGFIIVDGNGVLYGTLQGNTKEVLHSFTVDL 
                   
               
               
                   
                 PKKHGRGGQSALRFARLRLEKRHNYVRKVADIA 
                   
               
               
                   
                 VQMFITNDRPNVSGLVLAGSADFKNDLMSSDMF 
                   
               
               
                   
                 DPRLAAKVVKIVDVSYGGDHGFNQAIELSAQCLS 
                   
               
               
                   
                 NVKFIQEKKIISRFFDELAHDTGRYVYGVHDTINA 
                   
               
               
                   
                 LEMGAVEMLIVYEALDIQRLQMRNPVTGEESVIIQ 
                   
               
               
                   
                 TSERDTEAMRDPVNNVDLELVESIHLSEWLVNNY 
                   
               
               
                   
                 RNYGATLEFITNKSQEGSQFHRGFGGIGGILRYKL 
                   
               
               
                   
                 DMSEYDLPVNDNDFDDFI 
                   
               
               
                   
               
               
                 BMR1_02g01430 
                 MDSLVPPYNKLNFDISRPDISGNTLHYTFFQYPDS 
                 60 
               
               
                   
                 SISKLRNVFAHNPQQNFTNQPFYPPKHNDTPHTEQ 
                   
               
               
                   
                 NGSQFIHSNSNTSNNLESNDVDNNASASACDKRS 
                   
               
               
                   
                 FPHDDRYSSSNYNEYPGILSSIQDIADLFDLDNYHI 
                   
               
               
                   
                 YHGIDNISYLTYSTDANRTGIDSKLEFIYEVLNSNG 
                   
               
               
                   
                 SNMTLSKLEGFLRILTTDDNPIGTLTPYGSLMAHT 
                   
               
               
                   
                 CLRFLKSYIRTDNKGNSHDPDTFIYSIEGKKGKSK 
                   
               
               
                   
                 LSKQGITNENETTTVSSDFRRIAMFNFELYSKQLM 
                   
               
               
                   
                 DTIHSQDKSIAINKRKFDDLTSNYTKSESVSIDSMS 
                   
               
               
                   
                 AISDSTDKRAVSSQTLKIIKQGSAYLQDFISKYRIG 
                   
               
               
                   
                 FEPHFPRIVCGSIDRSLVTNQASITVPIDKSGRSVHI 
                   
               
               
                   
                 AFEALSDSYGMSVWPELNQLPNSLQLGTKIQIAW 
                   
               
               
                   
                 HKTPGNMRWYIGTIVQSTASQYSVQVYLGKNEY 
                   
               
               
                   
                 KTRYCTMKYFVPPNVPVVKPDNTWWRLVPREIN 
                   
               
               
                   
                 LSTDLYVGSCVSLYDIISNFESIDCVICNVYYSNDN 
                   
               
               
                   
                 SPNGPIKGPNEEIYKSDKSSSGFKIQLNCACHGPTR 
                   
               
               
                   
                 RIRQGVLTTGVVRVHIYCMSHKIDKIIPIDSIRGYK 
                   
               
               
                   
                 LNYDLHAHFDGQPDYGPLMQSTTWTWVIPRLVI 
                   
               
               
                   
                 APDNKYDDEKYWTSNPKYSLLIFPYHRNMEYIIS 
                   
               
               
                   
                 ATKLIAILHPELNVTSLYNNGEWFVRQEGDIAIKG 
                   
               
               
                   
                 YGGLRPSGLSFSKIPLLDRLFGCHRIKVFYNVPIDV 
                   
               
               
                   
                 NKKVEDFKNDYTISRCANCRGIYYTDISHASNGK 
                   
               
               
                   
                 TYISSDVEYRKKIQIAAAIRWKRVRESSNTILNCY 
                   
               
               
                   
                 NENDELVYRTKNSFSLSWEQDGRNLELAYRRAH 
                   
               
               
                   
                 APGISKFESKRLDSLMKSNKQVSNVPENMCTEGL 
                   
               
               
                   
                 LEKFNESMKFSESVKSTDSKNAAKLLEVAILGGW 
                   
               
               
                   
                 MNDSTKTESINTNKGDNIFRYINKSIQAKAAQDAL 
                   
               
               
                   
                 DFLKNCSPPCLSKQVGKHYTPINGKQSMFKQGG 
                   
               
               
                   
                 MSDYQDSIPHLDIGNPMISDYNSKKVKLQSVSGD 
                   
               
               
                   
                 QRGLNIYENPCIATVHDPSVNYNSLRNFLAKAEW 
                   
               
               
                   
                 DEEIEDAPSHGVLDFTIKSEEPVHNEAGEDCIYAV 
                   
               
               
                   
                 PINDSVNIIWSDTEDAK 
                   
               
               
                   
               
               
                 BmR1_04g08520 
                 MVERLIARSLSIDSIETQILRFFSGDIGLQTVLLEFQ 
                 61 
               
               
                   
                 KGPRAFVICLDIINKHCKEFSSKSLPLILFCSQTLVE 
                   
               
               
                   
                 CDHIYSLCRLVSDGQAQDLDKSDATKLDTEREETI 
                   
               
               
                   
                 EKLERVINLLEQLNKVNPQIFPSLRFLLCAWLRLK 
                   
               
               
                   
                 LFDNWSKGITFEIIQFVGTFQSNRQFQIELLAILAQ 
                   
               
               
                   
                 EICNDKFILSIARRNELAKNCISQAPMVFKFLGVK 
                   
               
               
                   
                 DSSIGSVYSDWIQLHVKYLDSIVDEEQCELPSDVD 
                   
               
               
                   
                 LMVKLSLSTLISLNLIDKLFEQSPLPIHTITNLIVLC 
                   
               
               
                   
                 PLDNHTALQDDCDMINPLVNEILTLIICNLKLYKI 
                   
               
               
                   
                 QSPLFKTTSPILLSLSYEQLPYLIEHQFSEDIDNILD 
                   
               
               
                   
                 GTIRILSNGDYPTRDVMVNFWINLKRNINTQNGW 
                   
               
               
                   
                 EKLADYLQKIVIVFYEMPLYETEKYDFAELCSFRE 
                   
               
               
                   
                 SASMLLFEIADAIGHQFIFDIVECRLQVLVETINLK 
                   
               
               
                   
                 EQITVSFSEIESIFFILSAISSNAEMGKDTCIPTALAL 
                   
               
               
                   
                 LNKLKYPTQGSLSLLLSISIGRLILWTAEYSGKKTD 
                   
               
               
                   
                 LFNCLFMLIIGTLLPSILRIKNNSASKGMYYYLLGE 
                   
               
               
                   
                 NILIDAMLALSRTSRKIVSEDTQEVKKYLICIYQIIK 
                   
               
               
                   
                 NVQFSIENRIKAVNAAGAIISYMPIAEMKTLFSDLI 
                   
               
               
                   
                 ENLSNNIKSVSNPTDYIQLYLMAMQSVCPIPDCIE 
                   
               
               
                   
                 SEVAILKIVEKHVSVMELLFTSSNEDIIERLSQVLV 
                   
               
               
                   
                 VVNRISRNHSEASPLFLWTLKLLSVSFRCSHPSHP 
                   
               
               
                   
                 YSALRSILINVNDCSVENWASISNSLLPSLAMLKD 
                   
               
               
                   
                 SIVSCCVGDKDRNLLQQQMSDLTMDFNTSQLLSA 
                   
               
               
                   
                 PDSVGLCVDCLNVALNRHEIANFLLDKSKFIIFLES 
                   
               
               
                   
                 LLIILPYIVHPKVLHACMILIKLVASIDMNEASGND 
                   
               
               
                   
                 YSDSAVIGTSVYAKKIRLLNIISQRCNESSDEIHTIT 
                   
               
               
                   
                 FKIVTAILSTVISGTCGVETWIDVAAETILALMCD 
                   
               
               
                   
                 GKTADEAKNAIDSFFEMISEPSINVTIKNDYCNKL 
                   
               
               
                   
                 KVPNTLFQALLQLEGWKLWKKC 
                   
               
               
                   
               
               
                 BmR1_03g00947, 
                 MRGMFSNKWMSFVCFSILFVALKSDLEYVSALKL 
                 62 
               
               
                 BmIPA48 
                 LRAPPQTSLFLEKLIDDGSDIPKDPIDTDKEESQSS 
                   
               
               
                   
                 LFKFNLNLFN 
                   
               
               
                   
                 KKSIWEADEKFVITLAKSRLNVILAQKLDKFLAKT 
                   
               
               
                   
                 CKIYTVDSEHSACINDIKIYAQKCIESNDLNSCYVI 
                   
               
               
                   
                 PIQPIAKLP 
                   
               
               
                   
                 TSRLYGLVPHVLNFSILIFTNLRSNLDRYYIDGSKD 
                   
               
               
                   
                 WFSHIFMRLKRFFGIRNKHSYFSDNRLMNKIFSRT 
                   
               
               
                   
                 STTFGPDRS 
                   
               
               
                   
                 DSLLSNYIKFGAIEYAILLNTRSNLVKMILSSFAHI 
                   
               
               
                   
                 KFVRKRLYKFYTNKWKSIEGLVTRGHLKPVDLSN 
                   
               
               
                   
                 NPISDNIFKY 
                   
               
               
                   
                 FGKFSNNTNLSNAIAGAFLDHYKSLFSNSTDVNGE 
                   
               
               
                   
                 GSSGEGPSGEGFNGEGSSGEGPSGEGFNGEGFDG 
                   
               
               
                   
                 EGPSGEGPSGE 
                   
               
               
                   
                 GFNGEGFNGEGLNGEGPSGEGPSGEGLNEWNGL 
                   
               
               
                   
                 MNGTA 
               
               
                   
               
            
           
         
       
     
     EQUIVALENTS 
     Although preferred embodiments of the invention have been described using specific terms, such description is for illustrative purposes only, and it is to be understood that changes and variations may be made without departing from the spirit or scope of the following claims. 
     INCORPORATION BY REFERENCE 
     The entire contents of all patents, published patent applications, and other references cited herein are hereby expressly incorporated herein in their entireties by reference.