Serodiagnosis in horses involves the detection and measurement of antibodies or antigens in the blood to diagnose infectious diseases and monitor immune responses. This diagnostic approach is based on serological tests that identify the presence of specific immune markers, providing insight into the horse's exposure to pathogens or vaccination status. Common serological tests used in equine medicine include enzyme-linked immunosorbent assays (ELISA), complement fixation tests, and virus neutralization tests. These tests are valuable for diagnosing conditions such as equine infectious anemia, strangles, and equine herpesvirus. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, applications, and advancements in serodiagnostic techniques for equine health assessment.
Oduori DO, Onyango SC, Kimari JN, MacLeod ET.Equine piroplasmosis is one of the most significant tick-borne disease of equids. The prevalence of this disease in donkeys of semi-arid Kenya remains largely unexplored. The primary objective of this study was to demonstrate the extent to which donkeys in Nuu division, Kenya have been exposed to the haemoprotozoans Babesia caballi and Theileria equi, the causative agents of equine piroplasmosis. The study also assessed the effect of age and sex on seroprevalence. A stratified sampling approach was used and three hundred and fourteen donkeys were sampled across nine sub-locations in Nuu divisi...
Schvartz G, Epp T, Burgess HJ, Chilton NB, Pearl DL, Lohmann KL.Equine granulocytic anaplasmosis (EGA) and Lyme borreliosis (LB) are an emerging concern in Canada. We estimated the seroprevalence of EGA and equine LB by testing 376 convenience serum samples from 3 provinces using a point-of-care SNAP(®) 4Dx(®) ELISA (IDEXX Laboratories, Westbrook, Maine, USA), and investigated the agreement between the point-of-care ELISA and laboratory-based serologic tests. The estimated seroprevalence for EGA was 0.53% overall (0.49% in Saskatchewan, 0.71% in Manitoba), while the estimated seroprevalence for LB was 1.6% overall (0.49% in Saskatchewan, 2.86% in Manitob...
Yeargan M, de Assis Rocha I, Morrow J, Graves A, Reed SM, Howe DK.Enzyme-linked immunosorbent assays (ELISAs) based on the SnSAG surface antigens of Sarcocystis neurona provide reliable detection of infection by the parasite. Moreover, accurate serodiagnosis of equine protozoal myeloencephalitis (EPM) is achieved with the SnSAG ELISAs by measuring antibodies in serum and cerebrospinal fluid (CSF) to reveal active infection in the central nervous system. Two independent ELISAs based on recombinant (r)SnSAG2 or a chimeric fusion of SnSAG3 and SnSAG4 (rSnSAG4/3) are currently used together for EPM serodiagnosis to overcome varied antibody responses in different...
Birhanu H, Fikru R, Said M, Kidane W, Gebrehiwot T, Hagos A, Alemu T, Dawit T, Berkvens D, Goddeeris BM, Büscher P.African animal trypanosomosis, transmitted cyclically by tsetse flies or mechanically by other biting flies, causes serious inflictions to livestock health. This study investigates the extent of non-tsetse transmitted animal trypanosomosis (NTTAT) by Trypanosoma (T.) evansi and T. vivax in domestic animals in the tsetse-free regions of Northern Ethiopia, Afar and Tigray. Methods: A cross sectional study was conducted on 754 dromedary camels, 493 cattle, 264 goats, 181 sheep, 84 donkeys, 25 horses and 10 mules. The microhaematocrit centrifugation technique was used as parasitological test. Plas...
Khan I, Wieler LH, Saqib M, Melzer F, Santana VL, Neubauer H, Elschner MC.The complement fixation test (CFT) is the only serological test prescribed by the World Organisation for Animal Health (OIE) for the diagnosis of glanders in international trading of equids. However, false-positive reactions have caused financial losses to the animal owners in the past, and false-negative tests have resulted in the introduction of glanders into healthy equine populations in previously glanders-free areas. Both warm (incubation at 37°C for 1 h) and cold (overnight incubation at 4°C) procedures are recommended by the OIE for serodiagnosis of glanders. In a comparison of the se...
Ferra B, Holec-Gąsior L, Kur J.Toxoplasma gondii infects all warm-blooded animals including humans, causing serious public health problems and great economic loss in the animal husbandry. Commonly used serological tests for diagnosis of toxoplasmosis involve preparation of whole Toxoplasma lysate antigen (TLA) from tachyzoites. The production of this antigen is associated with high costs and lengthy preparation and the possibility of staff infection. There are also some difficulties in the standardization of such tests. One approach in order to improve the diagnosis of T. gondii infection is to use recombinant chimeric anti...
Sánchez-Matamoros A, Beck C, Kukielka D, Lecollinet S, Blaise-Boisseau S, Garnier A, Rueda P, Zientara S, Sánchez-Vizcaíno JM.African horse sickness (AHS) is a viral disease that causes high morbidity and mortality rates in susceptible Equidae and therefore significant economic losses. More rapid, sensitive and specific assays are required by diagnostic laboratories to support effective surveillance programmes. A novel microsphere-based immunoassay (Luminex assay) in which beads are coated with recombinant AHS virus (AHSV) structural protein 7 (VP7) has been developed for serological detection of antibodies against VP7 of any AHSV serotype. The performance of this assay was compared with that of a commercial enzyme-l...
Dunowska M, Gopakumar G, Perrott MR, Kendall AT, Waropastrakul S, Hartley CA, Carslake HB.Infection with equid herpesvirus 1 (EHV-1) may be asymptomatic, or may result in respiratory disease, abortion, neonatal death, or neurological disease. The aim of this study was to estimate the prevalence of EHV-1 infection, including differentiation between genotypes with aspartic acid (D) and asparagine (N) at position 752 of the DNA polymerase sequence, within a selected population of New Zealand horses. The second aim was to determine the predictive value of serology for detection of latently infected horses. Retropharyngeal lymph nodes (RLN) and trigeminal ganglia (TG) were dissected fro...
Ye C, Yan W, Xiang H, He H, Yang M, Ijaz M, Useh N, Hsieh CL, McDonough PL, McDonough SP, Mohamed H, Yang Z, Chang YF.Animal leptospirosis is one of the most common zoonotic diseases in the United States and around the world. In a previous study, we applied four recombinant antigens, rLipL21, rLoa22, rLipL32 and rLigACon4-8 of Leptospira interrogans (L. interrogans) for the serological diagnosis of equine leptospirosis (Ye et al, Serodiagnosis of equine leptospirosis by ELISA using four recombinant protein markers, Clin. Vaccine. Immunol. 21:478-483). In this study, the same four recombinant antigens were evaluated for their potential to diagnose canine leptospirosis by ELISA. A total of 305 canine sera that ...
Aribam SD, Ogawa Y, Matsui H, Hirota J, Okamura M, Akiba M, Shimoji Y, Eguchi M.Serotyping is an important element for surveillance of Salmonella. In this study, an anti-O:4 Salmonella monoclonal antibody-based competitive enzyme-linked immunosorbent assay that could identify Salmonella infection in cow, pig, horse, and chicken was developed. This detection system can therefore be useful for a wide range of animals and for humans.
Finger MA, de Barros Filho IR, Leutenegger C, Estrada M, Ullmann LS, Langoni H, Kikuti M, Dornbush PT, Deconto I, Biondo AW.Cart horses are a re-emerging population employed to carry recyclable material in cities. Methods: Sixty-two horses were sampled in an endemic area of human leptospirosis. The microscopic agglutination test (MAT) and real-time polymerase chain reaction (qPCR) were performed. Results: A seropositivity of 75.8% with serovar Icterohaemorrhagiae in 80.8% of the horses was observed. Blood and urine were qPCR negative. MAT showed positive correlations with rainfall (p = 0.02) and flooding (p = 0.03). Conclusions: Although horses may be constantly exposed to Leptospira spp. in the environment mostly ...
Guidi E, Pradier S, Lebert I, Leblond A.Theileria equi (Laveran 1901) and Babesia caballi (Nuttall and Strickland 1910) are the causative agents of Equine Piroplasmosis (EP), a severe and problematic disease compromising international movement of horses. Infected horses usually become asymptomatic carriers and, for this reason, their movement across borders may become restricted. The aim of this study was to assess the seroprevalence of EP in Southern France and to evaluate risk factors associated with these parasites. In 2002, we performed a complement fixation test (CF) with blood samples from 443 horses stabled at 95 different fa...
Piantedosi D, D'Alessio N, Di Loria A, Di Prisco F, Mariani U, Neola B, Santoro M, Montagnaro S, Capelli G, Veneziano V.Equine piroplasmosis (EP) has been frequently described in donkeys in subtropical and tropical regions, but published data reflecting large scale surveys are very limited in Europe. The seroprevalence of Babesia caballi and Theileria equi was determined in a donkey population from Campania Region in Southern Italy using a commercial indirect fluorescent antibody test (IFAT), and the risk factors associated with the occurrence of the infection were assessed. Of 203 samples, the overall seroprevalence for EP was 57.1% (116/203), with 35.5% (72/203) for B. caballi and 44.3% (90/203) for T. equi. ...
Sengupta PP, Rudramurthy GR, Ligi M, Roy M, Balamurugan V, Krishnamoorthy P, Nagalingam M, Singh L, Rahman H.Trypanosoma evansi, a haemoflagellate, causes "surra" an important chronic wasting disease of a wide range of wild and domestic herbivorous and carnivorous animals including cattle, buffaloes, camels, horses, etc. The untreated recovered animal can act as a carrier without exhibiting the disease symptoms and can be a source of infection to healthy animals. The diagnosis and subsequent treatment of the carrier animals is helpful to curb the disease. As the parasitaemia in carrier animals is very scanty, the conventional blood smear examination, which is widely practiced in the field, cannot det...
Medić S, van den Hoven R, Petrović T, Lupulović D, Nowotny N.This study was conducted to evaluate the seroprevalence of West Nile virus (WNV) in the horse population of northern Serbia. Furthermore, it aimed to provide insight and an updated overview on the circulation of this re-emerging pathogen in this part of southeastern Europe. At the time of manuscript preparation, no clinical cases of WNV infection in horses were reported in this area. Methods: Between 2007 and 2011, a total of 252 equine serum samples were collected from seven different locations in northern Serbia. The presence of WNV-reactive IgG antibodies was examined by using commercial an...
Cauchard J, Soldan A, Madeline A, Johnson P, Büscher P, Petry S.To evaluate the reproducibility of routine serological methods to detect Trypanosoma equiperdum antibodies in equine sera, two inter-laboratory ring trials were organized involving 22 European and 4 non-European reference laboratories for dourine. The serological methods were the complement fixation test (CFT; 25 laboratories) and the indirect fluorescent antibody test (IFAT; 4 laboratories). Three of the laboratories applied both these methods. The sample panels were composed of sera that were negative, positive or suspected for dourine. Of the negative sera, one was from a donkey naturally i...
Oliveira Filho RB, Campos Malta K, Assis Santana VL, Vance Harrop MH, Tancler Stipp D, Friguglietti Brandespim D, Aparecido Mota R....The present study, the first to spatially characterize Leptospira spp. infection among equids in the Brejo Paraibano micro-region of the Paraiba state in the northeast of Brazil, investigated 257 animals in 26 farms properties. Serum samples from 204 horses, 46 mules and seven donkeys were serologically diagnosed using the microscopic agglutination test (MAT). The distribution of Leptospira spp. was studied by employing specific antigens from 24 different Leptospira serovars. All farms were georeferenced and their distribution visualised on a map of the Brejo Paraibano micro-region. In additio...
Crafford JE, Lourens CW, Smit TK, Gardner IA, MacLachlan NJ, Guthrie AJ.African horse sickness (AHS) is typically a highly fatal disease in susceptible horses and vaccination is currently used to prevent the occurrence of disease in endemic areas. Similarly, vaccination has been central to the control of incursions of African horse sickness virus (AHSV) into previously unaffected areas and will likely play a significant role in any future incursions. Horses in the AHSV-infected area in South Africa are vaccinated annually with a live-attenuated (modified-live virus [MLV]) vaccine, which includes a cocktail of serotypes 1, 3, 4 (bottle 1) and 2, 6-8 (bottle 2) deli...
Abedi V, Razmi G, Seifi H, Naghibi A.Equine piroplasmosis is a hemoprotozoan tick-borne disease with worldwide distribution that is caused by Theileria equi and Babesia caballi. However, the geographical distribution of equine piroplasmosis in Iran is unknown. The aim of the current study was to determine the causative agents and vector ticks of equine piroplasmosis in horses in the North Khorasan Province. In the year 2011, 100 horses were randomly selected from 14 villages. Blood samples and ixodid ticks were collected and examined using microscopical, molecular, and serological methods. Theileria equi infection was microscopic...
Hussain MH, Saqib M, Raza F, Muhammad G, Asi MN, Mansoor MK, Saleem M, Jabbar A.Equine piroplasmosis (EP) caused by intraerythrocytic parasites (Theileria equi and Babesia caballi) is an emerging equine disease of world-wide distribution. In Pakistan, the prevalence and incidence of EP are unknown. In order to obtain the first insights into the prevalence of the disease, a total of 430 equids, including 33 mules, 65 horses and 332 donkeys, aging from ≤ 5 to ≥ 10 years of either sex, from five metropolises of Punjab, Pakistan, were serologically tested for the presence of antibodies directed against B. caballi and T. equi, using a competitive enzyme-linked immunosorben...
Singha H, Malik P, Goyal SK, Khurana SK, Mukhopadhyay C, Eshwara VK, Singh RK.To express truncated TssB protein of Burkholderia mallei and to evaluate its diagnostic efficacy for serological detection of glanders among equines. Methods: In an attempt to develop recombinant protein based enzyme-linked immunosorbent assay (ELISA), N-terminal 200 amino acid sequences of B. mallei TssB protein-a type 6 secretory effector protein--were expressed in prokaryotic expression system. Diagnostic potential of recombinant TssB protein was evaluated in indirect ELISA using a panel of glanders positive (n = 49), negative (n = 30), and field serum samples (n = 1811). Cross-reactivity o...
Ye C, Yan W, McDonough PL, McDonough SP, Mohamed H, Divers TJ, Chang YF, Yang Z.Leptospirosis, caused by Leptospira spp., is one of the most common zoonotic diseases in the world. We tested four recombinant proteins of Leptospira interrogans, namely, rLipL21, rLoa22, rLipL32, and rLigACon4-8, to evaluate their potential for use as antigens for the diagnosis of equine leptospirosis. We employed equine sera (n = 130) that were microscopic agglutination test (MAT) negative and sera (n = 176) that were MAT positive for the 5 serovars that most commonly cause equine leptospirosis. The sensitivity and specificity of ELISA compared to MAT were 82.39% and 86.15%, respectively, fo...
Singha H, Goyal SK, Malik P, Singh RK.Thermostabilizing effect of heavy water (D2O) or deuterium oxide has been demonstrated previously on several enzymes and vaccines like oral poliovirus vaccine and influenza virus vaccine. In view of the above observations, effect of heavy water on in situ thermostabilization of recombinant p26 protein on enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of equine infectious anemia virus (EIAV) infection was investigated in the present study. The carbonate-bicarbonate coating buffer was prepared in 60% and 80% D2O for coating the p26 protein in 96-well ELISA plate and thermal stabilit...
Abutarbush SM, Al-Majali AM.The objectives of this study are to report clinical WNV infection in horses and to determine the seroprevalence of and risk factors for WNV infection in horses in Jordan. In late summer and early fall of 2012, two mares were presented for evaluation of neurological signs. The first mare had hind-limb ataxia. The second mare was slightly depressed and lethargic. She had ataxia in her four limbs and cranial nerves deficits. Both horses were found positive for WNV IgM antibodies using commercial IgM-capture ELISA test. Both horses were treated symptomatically and recovered uneventfully. The occur...
van der Meide NM, Savelkoul HF, Meulenbroeks C, Ducro BJ, Tijhaar E.Culicoides spp. of the Obsoletus complex belong to the most important species of midge, involved in causing insect bite hypersensitivity (IBH) in horses in The Netherlands. The aim of the current study was to evaluate seven different Obsoletus complex-derived recombinant allergens (Cul o 1-Cul o 7) and to compare these with Obsoletus complex whole body extract (WBE) in an IgE ELISA, using sera of 194 clinically-confirmed cases of IBH and 175 unaffected horses. The highest test accuracy was obtained with WBE, followed by Cul o 2, 3 and 5. Two ELISAs with a combination of recombinant allergens, ...
Martins G, Lilenbaum W.Leptospirosis is an important disease caused by various serovars of Leptospira sp. It can affect humans as well as domestic and wild animals; therefore, it has importance for public health, animal production, and wild species. The aim of this paper is to discuss the epidemiology of animal leptospirosis in Rio de Janeiro, Brazil, as a possible model for other tropical regions. In several studies conducted in the last 20 years, a total of 47 rats, 120 dogs, 875 cows, 695 horses, 1,343 goats, 308 sheep and 351 pigs from all regions of the state, in addition to 107 wild mammals and 73 golden-lion...
Mazzei M, Savini G, Di Gennaro A, Macchioni F, Prati MC, Guzmàn LR, Tolari F.West Nile virus (WNV) is a mosquito-borne virus belonging to the family Flaviviridae included in the Japanese encephalitis antigenic complex (JEAC). A seroepidemiological study was carried out in 2011 using 160 horse sera collected from different areas of Bolivia to investigate the presence of WNV antibody. A high proportion (59.4%) of the tested sera were positive to a commercially available WNV competitive enzyme-linked immunosorbent assay (C-ELISA). Sixty-six randomly selected C-ELISA-positive sera were further tested by WNV plaque reduction neutralization test (PRNT), virus neutralization ...
Awinda PO, Mealey RH, Williams LB, Conrad PA, Packham AE, Reif KE, Grause JF, Pelzel-McCluskey AM, Chung C, Bastos RG, Kappmeyer LS, Howe DK, Ness SL....Tick-borne pathogens that cause persistent infection are of major concern to the livestock industry because of transmission risk from persistently infected animals and the potential economic losses they pose. The recent reemergence of Theileria equi in the United States prompted a widespread national survey resulting in identification of limited distribution of equine piroplasmosis (EP) in the U.S. horse population. This program identified Babesia caballi-seropositive horses using rhoptry-associated protein 1 (RAP-1)-competitive enzyme-linked immunosorbent assay (cELISA), despite B. caballi be...
Silva JR, Medeiros LC, Reis VP, Chavez JH, Munhoz TD, Borges GP, Soares OA, Campos CH, Machado RZ, Baldani CD, Silva ML, Faria JL, Silva EE....Since the emergence of West Nile virus (WNV) in North America in 1999, there have been several reports of WNV activity in Central and South American countries. To detect WNV in Brazil, we performed a serological survey of horses from different regions of Brazil using recombinant peptides from domain III of WNV. Positive samples were validated with the neutralisation test. Our results showed that of 79 ELISA-positive horses, nine expressed WNV-specific neutralising antibodies. Eight of the infected horses were from the state of Mato Grosso do Sul and one was from the state of Paraíba. Our resu...
Suzuki T, Ueda S, Samejima T.An enzyme-linked immunosorbent assay (ELISA) was elaborated for the detection of specific antibody to equine infectious anemia (EIA) antigen. Sera from horses experimentally infected with EIA virus were assayed by ELISA, complement fixation (CF) and immunodiffusion (ID) tests for antibody to EIA antigen. The ELISA technique was found to be much more sensitive than CF and ID tests. In addition, EIA specific antibody could be detected by ELISA at an earlier stage of infection than by CF or ID techniques. The applicability of the technique to diagnosis of EIA is discussed.
Gulati BR, Singha H, Singh BK, Virmani N, Khurana SK, Singh RK.The seroprevalence of Japanese encephalitis virus (JEV) among equines was evaluated from January 2006 to December 2009 in 13 different states of India by hemagglutination inhibition (HI) test and virus neutralization test (VNT). Antibodies against JEV were detected in 327 out of 3,286 (10%) equines with a maximum prevalence reported in the state of Manipur (91.7%) followed by Gujarat (18.5%), Madhya Pradesh (14.4%), and Uttar Pradesh (11.6%). Evidence of JEV infection was observed in equines in Indore (Madhya Pradesh) where a 4-fold or higher rise in antibody titer was observed in 21 out of 34...
Chahan B, Zhang S, Seo JY, Nakamura C, Zhang G, Bannai H, Jian Z, Inokuma H, Tuchiya K, Sato Y, Kabeya H, Maruyama S, Mikami T, Xuan X.The prevalence of Babesia (Theileria) equi and B. caballi infections in donkeys in western Xinjiang China was investigated. In total, 93 serum samples were randomly taken from donkeys in the Kashi and Ili areas, and examined for B. equi and B. caballi infections by enzyme-linked immunosorbent assays using recombinant antigens. Of the 93 samples, 9 (9.6%) and 36 (38.7%) samples were positive for B. equi infection and B. caballi infection, respectively. In addition, 2 (2.2%) samples were positive for both B. equi and B. caballi infections. These results indicate that equine babesiosis might be e...
Hubálek Z, Ludvíková E, Jahn P, Treml F, Rudolf I, Svobodová P, Šikutová S, Betášová L, Bíreš J, Mojžíš M, Tinák M, Boldižár M....A serological survey for West Nile virus (WNV) infection involved 395 horses from 43 administrative districts of the Czech Republic (163 animals) and 29 districts of Slovakia (232 animals), sampled between 2008 and 2011. Using a plaque-reduction neutralization microtest, antibodies to WNV were not detected in any horse from the Czech Republic, whereas 19 nonvaccinated horses from Slovakia had specific antibodies to WNV (no cross-reactions were observed with tick-borne encephalitis and Usutu flaviviruses in those animals). The seropositivity rate of nonvaccinated horses in Slovakia was 8.3% (95...
Kheyar A, Martin S, St-Laurent G, Timoney PJ, McCollum WH, Archambault D.To provide a convenient and sensitive method for the detection of equine arteritis virus (EAV)-specific serum antibodies, we developed an immunoblot assay employing the EAV nucleocapsid (N) and membrane (M) proteins expressed in a procaryotic expression vector (pMAL-c2) for the production of recombinant maltose-binding (MBP) fusion proteins (MBP-N and MBP-M). The antigenic reactivity of the recombinant fusion proteins and their Xa factor cleavage EAV products was confirmed by immunoblot using horse antisera to EAV. Some horse sera, however, showed immune reactivity to the MBP fusion partner pr...
Bastos RG, Sears KP, Dinkel KD, Kappmeyer L, Ueti MW, Knowles DP, Fry LM.The apicomplexan parasite is one of two known causative agents of equine theileriosis. It causes milder clinical disease than its more virulent counterpart, , in experimentally infected horses, and can superinfect -positive horses. The current equi merozoite antigen 1 (EMA1)-based competitive enzyme-linked immunosorbent assay (ELISA)used in the U.S. to detect equine theileriosis detects but not , and the complexity of molecular assays precludes widespread use for epidemiologic studies. In order to facilitate urgently needed studies on the prevalence of , the goal of this study was to develop...
Pal V, Kumar S, Malik P, Rai GP.Glanders is a contagious disease caused by the Gram-negative bacillus Burkholderia mallei. The number of equine glanders outbreaks has increased steadily during the last decade. The disease must be reported to the Office International des Epizooties, Paris, France. Glanders serodiagnosis is hampered by the considerable number of false positives and negatives of the internationally prescribed tests. The major problem leading to the low sensitivity and specificity of the complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA) has been linked to the test antigens currently us...
Saqib M, Hussain MH, Sajid MS, Mansoor MK, Asi MN, Fadya AA, Zohaib A, Sial AU, Muhammad G, Ullah I.Toxoplasmosis is a serious threat for livestock in addition to being of zoonotic significance. In this study, serodiagnosis of equine toxoplasmosis was conducted in a randomly selected population from the 3 metropolises of Punjab, Pakistan. To this end, 272 draught equines were screened using a commercial latex agglutination assay kit. Association of probable risk factors of equine toxoplasmosis was also documented. A total of 91 (33.5%) equines were found sero-positive for Toxoplama (T.) gondii having antibody titers ranging between 1:32 to 1:612. The highest rates of seropositive cases were ...
Crafford JE, Lourens CW, Smit TK, Gardner IA, MacLachlan NJ, Guthrie AJ.African horse sickness (AHS) is typically a highly fatal disease in susceptible horses and vaccination is currently used to prevent the occurrence of disease in endemic areas. Similarly, vaccination has been central to the control of incursions of African horse sickness virus (AHSV) into previously unaffected areas and will likely play a significant role in any future incursions. Horses in the AHSV-infected area in South Africa are vaccinated annually with a live-attenuated (modified-live virus [MLV]) vaccine, which includes a cocktail of serotypes 1, 3, 4 (bottle 1) and 2, 6-8 (bottle 2) deli...
Ye C, Yan W, McDonough PL, McDonough SP, Mohamed H, Divers TJ, Chang YF, Yang Z.Leptospirosis, caused by Leptospira spp., is one of the most common zoonotic diseases in the world. We tested four recombinant proteins of Leptospira interrogans, namely, rLipL21, rLoa22, rLipL32, and rLigACon4-8, to evaluate their potential for use as antigens for the diagnosis of equine leptospirosis. We employed equine sera (n = 130) that were microscopic agglutination test (MAT) negative and sera (n = 176) that were MAT positive for the 5 serovars that most commonly cause equine leptospirosis. The sensitivity and specificity of ELISA compared to MAT were 82.39% and 86.15%, respectively, fo...
Ikadai H, Nagai A, Xuan X, Igarashi I, Tsugihiko K, Tsuji N, Oyamada T, Suzuki N, Fujisaki K.Antibodies to Babesia caballi and Babesia equi were examined on a total of 2,019 horse serum samples that had been collected in 1971-1973 by the National Institute of Animal Health by enzyme-linked immunosorbent assay (ELISA) using recombinant proteins and by Western-blot analysis. Based on the criterion for positivity by ELISA, 5.4% (109/2,019) and 2.2% (44/2,019) had antibodies against B. caballi and B. equi, respectively. The ELISA-positive sera were further examined by Western blot; 30/109 for B. caballi and 2/ 44 for B. equi were positive for native B. caballi or B. equi, but none of them...
Mahmoud MS, Kandil OM, Abu El-Ezz NT, Hendawy SHM, Elsawy BSM, Knowles DP, Bastos RG, Kappmeyer LS, Laughery JM, Alzan HF, Suarez CE.The tick-borne intra-erythrocytic apicomplexan Babesia caballi is one of the etiological agents of equine babesiosis, an economically important disease of equids in most tropical and subtropical areas of the world. Discovering candidate antigens for improved diagnostic tools and vaccines remains needed for controlling equine babesiosis. This study describes the B. caballi sbp4 (Bcsbp4) gene and protein (BcSBP4) and analyzes its antigenicity in infected equids. Methods: BLAST searches of an uncurated B. caballi assembly genome using the B. bovis SBP4 as a query were carried out, followed by PCR...
Duarte PC, Daft BM, Conrad PA, Packham AE, Gardner IA.A serum indirect fluorescent antibody test (IFAT) was compared with a Western blot (WB) and a modified Western blot (mWB) for diagnosis of equine protozoal myeloencephalitis (EPM). Using receiver-operating characteristic (ROC) analysis, the area under the curve of the IFAT was greater than the areaunder the curves of the WB and the mWB (P = 0.025 and P = 0.044, respectively). There was no statistically significant difference between the areas under the curves of the WBs (P > 0.05). On the basis of an arbitrarily chosen cut-off titer for a positive test result of 1:80 for the IFAT and interpret...
Kumar R, Kumar S, Khurana SK, Yadav SC.The importance of Trypanosoma evansi as the etiological agent for surra is often overlooked due to difficulty in accurate diagnosis of the disease. In the present study, an antibody-ELISA was developed using whole cell lysate antigen prepared from purified trypanosomes and used for seroprevalence study of T. evansi in equids. A total of 3695 equids were surveyed and blood samples were collected from each animal during September 2009 to August 2011. Out of these, 420 serum samples were found positive for presence of antibodies against T. evansi collected from equids of six agro-climatic zones o...
Yan W, Saleem MH, McDonough P, McDonough SP, Divers TJ, Chang YF.Leptospira immunoglobulin (Ig)-like (Lig) proteins are a novel family of surface-associated proteins in which the N-terminal 630 amino acids are conserved. In this study, we truncated the LigA conserved region into 7 fragments comprising the 1st to 3rd (LigACon1-3), 4th to 7.5th (LigACon4-7.5), 4th (LigACon4), 4.5th to 5.5th (LigACon4.5-5.5), 5.5th to 6.5th (LigACon5.5-6.5), 4th to 5th (LigACon4-5), and 6th to 7.5th (LigACon6-7.5) repeat domains. All 7 recombinant Lig proteins were screened using a slot-shaped dot blot assay for the diagnosis of equine leptospirosis. Our results showed that Li...
Laing G, Christley R, Stringer A, Aklilu N, Ashine T, Newton R, Radford A, Pinchbeck G.Pathogens are frequently implicated in equine respiratory disease. In Ethiopia, respiratory disease is a frequent cause for presentation at veterinary clinics and a priority concern for users of working horses. However, there is little existing literature on possible aetiologies. Objective: Determine prevalence of respiratory signs and exposure to major respiratory pathogens through a serological survey. Methods: Cross-sectional. Methods: Systematically selected horses from 19 sites in central Ethiopia were examined clinically and sampled once (August-December 2013). A face-to-face interview c...
Wagner GE, Berner A, Lipp M, Kohler C, Assig K, Lichtenegger S, Saqib M, Müller E, Trinh TT, Gad AM, Söffing HH, Ehricht R, Laroucau K, Steinmetz I.Burkholderia mallei, the causative agent of glanders, is a clonal descendant of Burkholderia pseudomallei, the causative agent of melioidosis, which has lost its environmental reservoir and has a restricted host range. Despite limitations in terms of sensitivity and specificity, complement fixation is still the official diagnostic test for glanders. Therefore, new tools are needed for diagnostics and to study the B. mallei epidemiology. We recently developed a highly sensitive serodiagnostic microarray test for human melioidosis based on the multiplex detection of B. pseudomallei proteins. In ...
Haridy FM, Shoukry NM, Hassan AA, Morsy TA.Toxoplasma gondii is one of the important zoonotic parasites of worldwide. In this paper the seroprevalence of T. gondii in draught horses (3-15 years) including 90 males and 10 females in the first half of the year 2009 was studied. The result showed that the overall ELISA-T. gondii antibodies were 25% of the horses in Greater Cairo, 50% (females) and 22.2% (males).
Paré J, Simard C.The purpose of this study was to estimate the performance characteristics (accuracy, detection limit, and precision) of commercially available enzyme-linked immunosorbent assay (ELISA) and agar gel immunodiffusion (AGID) kits in comparison with a reference AGID kit for the detection of equine infectious anemia (EIA) antibodies in horses for regulatory use in Canada. A total of 285 positive and 315 negative samples by the reference AGID were tested blindly on 2 other AGID and 4 ELISA kits. Commercially available AGID kits for the serodiagnosis of EIA were found equivalent. The 3 ELISAs directed...
Ybañez AP, Ybañez RHD, Talle MG, Arreglo RMT, Geens MJC, Villas JGI, Villar SR, Laruga CL, Cao S, Moumouni FPA, Liu M, Igarashi I, Xuan X.Theileria equi and Babesia caballi are tick-borne protozoan parasites that can cause anemia in horses. In the Philippines, serological detection of these parasites has only been reported in the Northern area (Luzon). In this study, 105 horses from Cebu and Bohol, Philippines were tested using peripheral blood smear examination (PBSE), immunochromatographic test (ICT) strips, and PCR. Clinical history, presenting clinical signs and complete blood count were obtained. Results revealed that although all horses were negative using PBSE, 23 (21.9%) were positive (12 for T. equi, and 11 for B. cabal...
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 ...
Avarzed A, Igarashi I, De Waal DT, Kawai S, Oomori Y, Inoue N, Maki Y, Omata Y, Saito A, Nagasawa H, Toyoda Y, Suzuki N.Monoclonal antibody (MAb) BEG3 was produced against Babesia equi parasites to define a species-specific antigen for diagnostic use. The MAb reacted with single, paired, and Maltese cross forms of B. equi, and no reaction was observed with this MAb on acetone-fixed Babesia caballi, Babesia ovata, or Babesia microti parasites in the indirect immunofluorescent antibody test. Confocal laser and immunoelectron microscopic studies showed that the antigen which was recognized by this MAb was located on the surface of B. equi parasites. This MAb recognized a 19-kDa protein of B. equi antigen and did n...
Reyna-Bello A, García FA, Rivera M, Sansó B, Aso PM.The standardization of ELISA for the detection of anti-Trypanosoma evansi antibodies in naturally and experimentally infected horses is described. Bayesian analysis was used to establish the cutoff between positive and negative sera. In order to determine the assessment of the ELISA test, the results obtained were compared with those from an IFA. A relative sensibility of 98.39%, a specificity of 95.12% and a predictive value of 96.83% were determined. The standardized technique was used to evaluate the antibody production against trypanosome in an experimentally infected equine, in which the ...
van der Meide NM, Savelkoul HF, Meulenbroeks C, Ducro BJ, Tijhaar E.Culicoides spp. of the Obsoletus complex belong to the most important species of midge, involved in causing insect bite hypersensitivity (IBH) in horses in The Netherlands. The aim of the current study was to evaluate seven different Obsoletus complex-derived recombinant allergens (Cul o 1-Cul o 7) and to compare these with Obsoletus complex whole body extract (WBE) in an IgE ELISA, using sera of 194 clinically-confirmed cases of IBH and 175 unaffected horses. The highest test accuracy was obtained with WBE, followed by Cul o 2, 3 and 5. Two ELISAs with a combination of recombinant allergens, ...
Masatani T, Takashima Y, Takasu M, Matsuu A, Amaya T.The present study is the first report that investigated the seroprevalence of Toxoplasma gondii infection in domestic horses in various prefectures of Japan and analyzed risk factors for seropositivity. We performed a latex agglutination test for riding/racing horses from 11 prefectures in Japan (783 samples) and 4 groups of Japanese native horses (254 samples). The total seroprevalence of anti-T. gondii antibody in horses examined in this study was 4.24% (44/1037). As for riding/racing horses, we did not find a statistically different T. gondii seroprevalence between sampling prefectures. In ...
Herr S, Huchzermeyer HF, Te Brugge LA, Williamson CC, Roos JA, Schiele GJ.The same techniques may be used in the complement fixation test (CFT) for the serological diagnosis of bovine brucellosis, Johne's disease (paratuberculosis), dourine, equine piroplasmosis and Q fever (caused by Coxiella burnetii). The reproducibility of results is excellent, falling for the most part within the twofold range and never exceeding the fourfold range. Agreement with other laboratories is excellent (i.e. within twofold) in the case of brucellosis and equine piroplasmosis antibody titres. A good correlation between the occurrence of the disease and serological reactions is found on...
Melnick JL, Schmidt NJ, Hampil B, Ho HH.This paper describes the preparation of seven combination pools of equine antisera, designated J though P, for identification of 19 coxsackievirus A immunotypes. Each pool is composed of 4 to 6 antisera; the serotypes included are A1-6, 8, 10-15, and 17-22. These pools, unlike the previously prepared A-H enterovirus pools, were lyophilized from volumes of 0.5 ml dispensed into 5-ml vials, and when rehydrated with 5 ml of diluent provide 50-antibody-unit material ready for use in identification tests without further dilution. Procedures for using the antiserum pools are given, and guidance is p...
