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Topic:Immunology

The equine immune system is a complex network of cells, tissues, and organs that work collaboratively to defend against pathogens and maintain homeostasis. It consists of innate and adaptive components, each with distinct functions and mechanisms. The innate immune system provides the first line of defense through physical barriers, phagocytic cells, and the complement system. The adaptive immune system involves lymphocytes, such as B cells and T cells, which generate specific responses to antigens and provide immunological memory. Research in equine immunology explores the interactions between these components, the impact of genetic and environmental factors on immune function, and the development of vaccines and therapeutics. This page gathers peer-reviewed studies and scholarly articles focusing on the mechanisms, regulation, and clinical applications of the equine immune system in health and disease.
Restricted and selective tropism of a Venezuelan equine encephalitis virus-derived replicon vector for human dendritic cells.
Viral immunology    April 12, 2007   Volume 20, Issue 1 88-104 doi: 10.1089/vim.2006.0090
Nishimoto KP, Laust AK, Wang K, Kamrud KI, Hubby B, Smith JF, Nelson EL.Dendritic cells (DCs) consist of heterogeneous phenotypic populations and have diverse immunostimulatory functions dependent on both lineage and functional phenotype, but as exceptionally potent antigen-presenting cells, they are targets for generating effective antigen-specific immune responses. A promising replicon particle vector derived from Venezuelan equine encephalitis virus (VEE) has been reported to transduce murine footpad DCs. However, the receptive DC subset, the degree of restriction for this tropism, and the extent of conservation between rodents and humans have not been well cha...
Molecular epidemiology of Streptococcus zooepidemicus infection in naturally occurring equine respiratory disease.
Veterinary journal (London, England : 1997)    April 11, 2007   Volume 175, Issue 3 338-345 doi: 10.1016/j.tvjl.2007.02.018
Newton JR, Laxton R, Wood JL, Chanter N.The objective of the study was to characterise the molecular epidemiology of Streptococcus zooepidemicus infection among isolates collected sequentially from recently weaned, pasture maintained Welsh mountain ponies with naturally occurring respiratory disease. Weekly nasopharyngeal and tracheal lavage samplings over a 10-week period were conducted in 29 ponies. Two PCR typing methods based on characterisation of the M-protein hypervariable (HV) region and the 16S-23S rRNA gene intergenic spacer were then applied to isolates of S. zooepidemicus recovered from nasopharyngeal swab and tracheal w...
Use of appropriate antimicrobials in wound management.
Emergency medicine clinics of North America    April 3, 2007   Volume 25, Issue 1 159-176 doi: 10.1016/j.emc.2007.01.007
Nakamura Y, Daya M.The primary goal of wound management is to achieve a functional closure with minimal scarring. Preventing infection is important to facilitate the healing process. Most simple, uncomplicated wounds do not need systemic antibiotics but benefit from the use of topical antibiotics. Judicious use of antibiotics reduces unnecessary adverse events and helps reduce the development of resistance. Although antibiotics can help reduce infection risk and promote healing, they are not a substitute for good local wound care, in particular irrigation and surgical débridement. This article reviews the role ...
Classes and crossreactivity of proteinases in the excretory-secretory products of Caenorhabditis elegans.
Journal of helminthology    March 27, 2007   Volume 81, Issue 1 93-99 doi: 10.1017/S0022149X0724725X
Nic An Ultaigh S, Ryan MF.Proteinases released during the in vitro maintenance of asynchronous cultures of the free-living nematode Caenorhabditis elegans were characterized on the basis of subunit composition, fluorogenic substrate specificity, inhibitor sensitivity and pH optima. Cysteine proteinases are present in the excretory-secretory products (ESP) as indicated by the hydrolysis of cathepsin fluorogenic substrates and confirmed by immunoblotting. Serine proteinases were predominant as indicated by substrate gel analysis and inhibitor studies. The presence of metallo-proteinases was also indicated by inhibitor st...
Envelope-specific T-helper and cytotoxic T-lymphocyte responses associated with protective immunity to equine infectious anemia virus.
The Journal of general virology    March 22, 2007   Volume 88, Issue Pt 4 1324-1336 doi: 10.1099/vir.0.82391-0
Tagmyer TL, Craigo JK, Cook SJ, Issel CJ, Montelaro RC.Equine infectious anemia virus (EIAV) infection of horses provides a valuable model for examining the natural immunological control of lentivirus infection and disease and the mechanisms of protective and enhancing vaccine immunity. We have previously hypothesized that the EIAV envelope (Env) proteins gp90 and gp45 are major determinants of vaccine efficacy, and that the development of protective immunity by attenuated viral vaccines may be associated with the progressive redirection of immune responses from immunodominant, variable Env segments to immunorecessive, conserved Env sequences. Whi...
Burkholderia Hep_Hag autotransporter (BuHA) proteins elicit a strong antibody response during experimental glanders but not human melioidosis.
BMC microbiology    March 15, 2007   Volume 7 19 doi: 10.1186/1471-2180-7-19
Tiyawisutsri R, Holden MT, Tumapa S, Rengpipat S, Clarke SR, Foster SJ, Nierman WC, Day NP, Peacock SJ.The bacterial biothreat agents Burkholderia mallei and Burkholderia pseudomallei are the cause of glanders and melioidosis, respectively. Genomic and epidemiological studies have shown that B. mallei is a recently emerged, host restricted clone of B. pseudomallei. Results: Using bacteriophage-mediated immunoscreening we identified genes expressed in vivo during experimental equine glanders infection. A family of immunodominant antigens were identified that share protein domain architectures with hemagglutinins and invasins. These have been designated Burkholderia Hep_Hag autotransporter (BuHA)...
Red blood cell erythropoietin, not plasma erythropoietin, concentrations correlate with changes in hematological indices in horses receiving a single dose of recombinant human erythropoietin by subcutaneous injection.
Journal of veterinary pharmacology and therapeutics    March 14, 2007   Volume 30, Issue 2 175-178 doi: 10.1111/j.1365-2885.2007.00828.x
Singh AK, Gupta S, Barnes A, Carlson JM, Ayers JK.No abstract available
Avian titer development against West nile virus after extralabel use of an equine vaccine.
Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians    February 28, 2007   Volume 36, Issue 2 257-264 doi: 10.1638/03-094.1
Johnson S.West Nile virus affects many animals, but the highest prevalence of morbidity and mortality is observed in birds, horses, and humans. The purpose of this study was to determine a protocol in birds of prey and corvids, using a vaccine developed for horses. The birds were assigned to five groups. Groups 1-4 received 0.25 ml, 0.5 ml, 0.75 ml, and 1.0 ml, respectively, and group 5 served as a control group. The greatest percentage of seroconversion (58.3%) was observed in the vaccine group that received a dose of 1.0 ml administered thrice, 3 wk apart. This report demonstrates that a vaccine devel...
Correlation between the induction of Th1 cytokines by an attenuated equine infectious anemia virus vaccine and protection against disease progression.
The Journal of general virology    February 28, 2007   Volume 88, Issue Pt 3 998-1004 doi: 10.1099/vir.0.82416-0
Zhang X, Wang Y, Liang H, Wei L, Xiang W, Shen R, Shao Y.The equine infectious anemia virus (EIAV) donkey-leukocyte attenuated vaccine (DLV) has been used to protect against equine infectious anaemia (EIA) disease for several decades in China. The attenuated mechanism and immunological protective mechanisms remain to be elucidated. To identify responses that correlate with the protection against disease, we immunized horses with DLV, followed by challenge with an EIAV wild-type strain LN. All vaccinated horses were asymptomatic and had a low level of virus replication (<10 copies ml-1). The expression level of cytokines including gamma interferon...
Immune selection of equine infectious anemia virus env variants during the long-term inapparent stage of disease.
Virology    February 27, 2007   Volume 363, Issue 1 156-165 doi: 10.1016/j.virol.2007.01.037
Sponseller BA, Sparks WO, Wannemuehler Y, Li Y, Antons AK, Oaks JL, Carpenter S.The principal neutralizing domain (PND) of equine infectious anemia virus (EIAV) is located in the V3 region of SU. Genetic variation in the PND is considered to play an important role in immune escape and EIAV persistence; however, few studies have characterized genetic variation in SU during the inapparent stage of disease. To better understand the mechanisms of virus persistence, we undertook a longitudinal study of SU variation in a pony experimentally inoculated with the virulent EIAV(Wyo). Viral RNA isolated from the inoculum and from sequential sera samples was amplified by RT-PCR, clon...
A comparison of Freund’s Complete and Freund’s Modified Adjuvants used with a contraceptive vaccine in wild horses (Equus caballus).
Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians    February 23, 2007   Volume 36, Issue 4 610-616 doi: 10.1638/04104.1
Lyda RO, Hall JR, Kirkpatrick JF.Fifteen captive wild mares (Equus caballus) were treated with porcine zona pellucida contraceptive vaccine and either Freund's Complete Adjuvant (n = 7) or Freund's Modified Adjuvant (n = 8). All mares received a booster inoculation of porcine zona pellucida plus Freund's Incomplete Adjuvant a month later. Anti-porcine zona pellucida antibodies were measured over 10 mo following the initial inoculation. There were no significant differences in antibody titers at any point during the 10 mo, and seven of the eight mares in the Freund's Modified Adjuvant group were above the 60% level at the end ...
Telogen defluxion associated with hypersensitivity causing alopecia in a horse.
Australian veterinary journal    February 16, 2007   Volume 85, Issue 1-2 56-58 doi: 10.1111/j.1751-0813.2006.00086.x
Jubb TF, Graydon RJ.No abstract available
Cerebellar abscess due to Rhodococcus equi in an immunocompetent patient: case report and literature review.
Journal of neurosurgical sciences    February 8, 2007   Volume 50, Issue 4 127-129 
Ulivieri S, Oliveri G.Rhodococcus equi (R. equi) is a facultative, intracellular, nonmotile, non-spore-forming, gram-positive coccobacillus, primarily causes zoonotic infections that affect grazing animals, mainly horses and foals and is a rare opportunistic pathogen found in severely compromised patients, and most commonly in recent years, in human immunodeficiency virus (HIV)-infected persons. Early cases, most in patients receiving immunosuppressant therapy, were more likely to be successfully treated with antimicrobial agents than cases in AIDS patients, it is emerging as an important pathogen in patients with ...
Influx of neutrophils and persistence of cytokine expression in airways of horses after performing exercise while breathing cold air.
American journal of veterinary research    February 3, 2007   Volume 68, Issue 2 185-189 doi: 10.2460/ajvr.68.2.185
Davis MS, Williams CC, Meinkoth JH, Malayer JR, Royer CM, Williamson KK, McKenzie EC.To determine effects of exercise performed while breathing cold air on expression of cytokines and influx of neutrophils in airways of horses. Methods: 9 adult horses. Methods: In a crossover study, bronchoalveolar lavage fluid (BALF) was obtained 24 and 48 hours after each of 2 submaximal exercise sessions performed by horses while breathing warm (25 degrees C) or cold (-5 degrees C) air. Total and differential nucleated cell counts were determined for each BALF sample. Relative mRNA expression of cytokines in BALF cells was quantified by use of a reverse transcription-PCR assay. Results: Hor...
Equine zona protein synthesis and ZP structure during folliculogenesis, oocyte maturation, and embryogenesis.
Molecular reproduction and development    January 26, 2007   Volume 74, Issue 7 851-859 doi: 10.1002/mrd.20501
Kölle S, Dubois CS, Caillaud M, Lahuec C, Sinowatz F, Goudet G.In the equine, the zona pellucida (ZP) is the major barrier to successful in vitro fertilization. Therefore the aim of our studies was to analyze species-specific features of the equine ZP in regard to structure and glycoprotein ZPB and ZPC expression sites during oocyte development and embryogenesis. The equine ZP revealed high immunological cross-reactivity to porcine ZPB and ZPC. In the ovary, the distribution of ZPB and ZPC was co-localized and correlated with the developmental stage of the follicle. ZPB and ZPC expression started in the oocyte of the late primordial and primary follicle. ...
Live-attenuated recombinant equine herpesvirus type 1 (EHV-1) induces a neutralizing antibody response against West Nile virus (WNV).
Virus research    January 22, 2007   Volume 125, Issue 1 69-78 doi: 10.1016/j.virusres.2006.12.009
Rosas CT, Tischer BK, Perkins GA, Wagner B, Goodman LB, Osterrieder N.The immunogenicity in horses of a recombinant equine herpesvirus type 1 (EHV-1) vaccine expressing West Nile virus (WNV) prM and E proteins was studied. To construct the recombinant EHV-1, two-step en passant mutagenesis was employed for manipulation of a bacterial artificial chromosome (BAC) of vaccine strain RacH. Recombinant EHV-1 stably expressed the WNV prM and E proteins as demonstrated by indirect immunofluorescence and Western blotting. In addition, growth properties in vitro of the EHV-1/WNV recombinant were found to not be significantly different from those of the parental virus. To ...
Standardization and validation of an agar gel immunodiffusion test for the diagnosis of equine infectious anemia using a recombinant p26 antigen.
Veterinary microbiology    January 17, 2007   Volume 121, Issue 3-4 344-351 doi: 10.1016/j.vetmic.2007.01.007
Alvarez I, Gutierrez G, Vissani A, Rodriguez S, Barrandeguy M, Trono K.We developed and validated an agar gel immunodiffusion test (AGID) test for the diagnosis of equine infectious anemia (EIA) using as antigen the p26 protein of equine infectious anemia virus (EIAV) produced in the Escherichia coli expression system. The developed rp26-AGID test showed an excellent diagnostic relative sensitivity (100%) and specificity (100%) compared to a commercial AGID assay when 1855 field serum samples were analyzed. In addition, the rp26-AGID demonstrated to be a precise assay with excellent repeatability and reproducibility. In the analytical sensitivity trial, positive ...
Cloning and functional characterization of recombinant equine P-selectin.
Veterinary immunology and immunopathology    January 16, 2007   Volume 116, Issue 3-4 115-130 doi: 10.1016/j.vetimm.2007.01.004
Xu J, Cai J, Anderson B, Wagner B, Albrecht R, Peek SF, Suresh M, Darien BJ.The recent molecular characterization and sequencing of equine P-selectin (ePsel), and its glycoprotein ligand, P-selectin glycoprotein ligand-1 (PSGL-1), have provided the tools for further investigation into their role in leukocyte trafficking. Here, we report the generation of a genetically engineered chimeric protein (ePsel-IgG) in which the equine P-selectin lectin and epithelial growth factor (EGF) domains were covalently linked to the equine IgG1 heavy chain constant region. The soluble ePsel-IgG was observed to bind to equine monocytes by confocal microscopy and flow cytometry. Further...
Lactoferrin, a glycoprotein with immunomodulatory and mast cell stabilising properties, in skin of horses suffering from Culicoides hypersensitivity.
Research in veterinary science    January 11, 2007   Volume 83, Issue 2 165-170 doi: 10.1016/j.rvsc.2006.11.007
Kolm G, Knapp E, Wagner R, Klein D.Lactoferrin (LF), a glycogen of the transferrin family with anti-bacterial and immunomodulatory properties, is expressed in various secretions and tissues. Cutaneous LF serves as a mast cell stabilising compound, modulates T cell activity and is found during IgE-mediated late phase reactions at allergen challenged sites. Culicoides hypersensitivity (CHS) in horses is a common IgE-mediated allergic dermatitis, characterised by an early and late phase cutaneous reaction upon allergen challenge. The aim of the study presented here was to examine whether LF mRNA expression in skin biopsies from ho...
Mutation and virulence assessment of chromosomal genes of Rhodococcus equi 103. Pei Y, Parreira V, Nicholson VM, Prescott JF.Rhodococcus equi can cause severe or fatal pneumonia in foals as well as in immunocompromised animals and humans. Its ability to persist in macrophages is fundamental to how it causes disease, but the basis of this is poorly understood. To examine further the general application of a recently developed system of targeted gene mutation and to assess the importance of different genes in resistance to innate immune defenses, we disrupted the genes encoding high-temperature requirement A (htrA), nitrate reductase (narG), peptidase D (pepD), phosphoribosylaminoimidazole-succinocarboxamide synthase ...
Lung surfactant function and composition in neonatal foals and adult horses.
Journal of veterinary internal medicine    December 26, 2006   Volume 20, Issue 6 1402-1407 doi: 10.1892/0891-6640(2006)20[1402:lsfaci]2.0.co;2
Christmann U, Livesey LC, Taintor JS, Waldridge BM, Schumacher J, Grier BL, Hite RD.Lung surfactant function and composition are varied and adapted to the specific respiratory physiology of all mammalian species. Objective: Lung surfactant function and composition are different in neonatal foals as compared to adult horses. Methods: Six adult horses, 7 term foals (<24 hours old), and 4 premature foals were used. Animals were part of the Auburn University teaching herd except for 3 client-owned premature foals. Methods: Bronchoalveolar lavage fluid (BALF) was obtained from all animals. Ultracentrifugation of cell-free BALF separated surfactant into crude surfactant pellets ...
West Nile virus: recent trends in diagnosis and vaccine development.
Vaccine    December 22, 2006   Volume 25, Issue 30 5563-5576 doi: 10.1016/j.vaccine.2006.12.005
Dauphin G, Zientara S.West Nile virus (WNV) is a mosquito-borne flavivirus, native to Africa, Europe, and Western Asia. In many respects, WNV is an outstanding example of a zoonotic pathogen that has leaped geographical barriers and can cause severe disease in human and horse. Before the emergence of WNV in the USA, only few methods of diagnosis were available. Recently, many changes in the fields of WN diagnosis and prevention have happened. This paper will review all these new tools. After a description of the main concerns in WNV and West Nile (WN) disease in humans and animals, this review will present the main...
Host cell tropism of equine herpesviruses: glycoprotein D of EHV-1 enables EHV-4 to infect a non-permissive cell line.
Archives of virology    December 15, 2006   Volume 152, Issue 4 717-725 doi: 10.1007/s00705-006-0885-x
Whalley JM, Ruitenberg KM, Sullivan K, Seshadri L, Hansen K, Birch D, Gilkerson JR, Wellington JE.Equine herpesviruses 1 and 4 (EHV-1 and EHV-4) cause equine respiratory disease worldwide. However, only EHV-1 is a cause of abortion and neurological disease, despite the two viruses having all 76 genes in common. In addition EHV-1 has a broader host range in cell culture than EHV-4, as exemplified by the rabbit kidney (RK) cell line that is permissive for EHV-1, but not for EHV-4. Here we describe that when EHV-4 produced in equine cells was inoculated onto RK cells expressing glycoprotein D of EHV-1 (RKgD1), infection developed as clusters of rounded cells, and this infectivity could be pas...
Comparative serological diagnosis of toxoplasmosis in horses using locally isolated Toxoplasma gondii.
Veterinary parasitology    December 14, 2006   Volume 145, Issue 1-2 31-36 doi: 10.1016/j.vetpar.2006.11.010
Ghazy AA, Shaapan RM, Abdel-Rahman EH.A total of 420 serum samples collected from horses of different ages, sexes and breeds, located at some horse farms in Egypt, were used for serological studies. A crude antigen of the locally isolated Toxoplasma gondii tachyzoites from horse tissues (LA) was used for the detection of T. gondii antibodies in horses. It showed good diagnostic efficiency (38.1%) by Enzyme Linked Immunosorbent Assay (ELISA). To increase this efficiency, an affinity purification process was performed. Two fractions were obtained from LA by CNBr-Sepharose 4B affinity column chromatography named; unbound (LAunb) and ...
Development and registration of recombinant veterinary vaccines. The example of the canarypox vector platform.
Vaccine    December 8, 2006   Volume 25, Issue 30 5606-5612 doi: 10.1016/j.vaccine.2006.11.066
Poulet H, Minke J, Pardo MC, Juillard V, Nordgren B, Audonnet JC.The canarypox vaccine vector (ALVAC) technology has been used to develop and license several vaccines for companion animals and horses in the European Union and USA. ALVAC is a ubiquitous vector with high biosafety since it is non-replicative in mammalians, is genetically and physically stable, and able to induce both humoral and cell-mediated immune responses against the expressed transgene product. Specific rules apply for the development and registration of recombinant vector vaccines. The biology of the vector as well as the recombinant virus must be thoroughly documented to allow the risk...
Clenbuterol affects the expression of messenger RNA for interleukin 10 in peripheral leukocytes from horses challenged intrabronchially with lipopolysaccharides.
Veterinary research communications    December 2, 2006   Volume 30, Issue 8 921-928 doi: 10.1007/s11259-006-3383-4
van den Hoven R, Duvigneau JC, Hartl RT, Gemeiner M.On four occasions, four horses with heaves and four horses with small airway inflammatory diseases inhaled 0.9% saline based aerosol mixtures with or without lipopolysaccharides (LPS). Prior to the first saline and LPS inhalation, horses were untreated, while three and a half days prior to the third and forth inhalation horses had received 0.8 microg/kg clenbuterol intravenously twice daily. The messenger RNA (mRNA) expression of tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-4, IL-6, IL-8, IL-10 and interferon- gamma (IFN- gamma) was investigated by RT-PCR, all of which ...
Monoclonal antibody-based competitive enzyme-linked immunosorbent assay for detecting and quantifying West Nile virus-neutralizing antibodies in horse sera.
Clinical and vaccine immunology : CVI    November 29, 2006   Volume 14, Issue 2 134-138 doi: 10.1128/CVI.00322-06
Choi KS, Ko YJ, Nah JJ, Kim YJ, Kang SY, Yoon KJ, Joo YS.A rapid immunoassay for detecting and quantifying West Nile virus (WNV)-neutralizing antibodies in sera was developed as an alternative to the plaque reduction neutralization test (PRNT), the gold standard test for WNV. The assay is a competitive, enzyme-linked immunosorbent assay using neutralizing monoclonal antibody 5E8 (NT-ELISA). A cutoff percent inhibition (PI) value of 35% (mean PI plus 3 standard deviations), with a specificity of 99%, was established based on analysis of 246 serum samples from horses free of WNV. The NT-ELISA detected neutralizing antibodies in all sera collected 7 or...
Se18.9, an anti-phagocytic factor H binding protein of Streptococcus equi.
Veterinary microbiology    November 28, 2006   Volume 121, Issue 1-2 105-115 doi: 10.1016/j.vetmic.2006.11.023
Tiwari R, Qin A, Artiushin S, Timoney JF.Evasion of phagocytosis is an important virulence determinant of Streptococcus equi (S. equi subsp. equi), the cause of equine strangles and distinguishes it from the closely related but much less virulent S. zooepidemicus (S. equi subsp. zooepidemicus). We describe Se18.9, a novel H factor binding protein secreted by S. equi but not by S. zooepidemicus that reduces deposition of C3 on the bacterial surface and significantly reduces the bactericidal activity of equine neutrophils suspended in normal serum for both S. equi and S. zooepidemicus. Se18.9 is secreted abundantly by actively dividing...
The involvement of mast cells and mast cell proteinases in the intestinal response to equine cyathostomin infection.
Veterinary immunology and immunopathology    November 21, 2006   Volume 115, Issue 1-2 35-42 doi: 10.1016/j.vetimm.2006.10.015
du Toit N, McGorum BC, Pemberton AD, Brown J, Dacre KJ.Cyathostomins (Cyathostominae) are regarded as the most pathogenic equine nematode worldwide. These nematodes are difficult to control in equine populations due to emerging anthelmintic resistance and evasion of encysted larval cyathostomins to regular modern anthelmintics. Mast cells and their proteinases have been shown to play a role in the mammalian immune response to nematode infections. Involvement of mast cells and mast cell proteinases in the equine immune response to cyathostomin infection is proposed. A technique was established to perform immunohistochemical staining using polyclona...
Serodiagnosis of equine infectious anemia by agar gel immunodiffusion and ELISA using a recombinant p26 viral protein expressed in Escherichia coli as antigen.
Preventive veterinary medicine    November 15, 2006   Volume 78, Issue 3-4 239-245 doi: 10.1016/j.prevetmed.2006.10.009
Piza AS, Pereira AR, Terreran MT, Mozzer O, Tanuri A, Brandão PE, Richtzenhain LJ.We used a p26 recombinant protein (p26r) from equine infectious-anemia virus (EIAV) expressed in Escherichia coli as antigen to standardize an agar-gel immunodiffusion (AGIDp26r) test and an indirect ELISA (ELISAp26r) for the detection of antibodies against EIAV in 720 equine sera from Brazil. We evaluated the tests's relative diagnostic sensitivities (relSe) and relative diagnostic specificities (relSp) against a commercial AGID kit (Idexx, USA). We used three sera panels: panel A--196 AGID-negative sera from an AIE non-endemic controlled area; panel B--194 AGID-negative sera from an AIE ende...
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