Hemagglutination inhibition (HI) is a serological assay used to detect antibodies against specific viruses in horses. This method is based on the principle that certain viruses can cause red blood cells to agglutinate, or clump together, and the presence of specific antibodies can inhibit this process. In equine research, the HI test is often utilized to assess the immune response to viral infections such as equine influenza. The assay provides information on the antibody levels in a horse's serum, which can be indicative of exposure to the virus or the effectiveness of a vaccination. This page compiles peer-reviewed research studies and scholarly articles that explore the methodology, application, and implications of hemagglutination inhibition in the context of equine viral diseases.
Kreutzfeldt N, Chambers TM, Reedy S, Spann KM, Pusterla N.Dexamethasone is routinely administered to horses but its effect on the antibody response to a commercial EIV/EHV vaccine is unclear. Objective: Horses receiving dexamethasone will have lower postvaccination antibody levels against EIV and EHV-1 than vaccinated controls. Methods: Fifty-five healthy adult research horses. Methods: Randomized cohort study. Control (no vaccine, group 1), vaccination only (EIV/EHV-1/EHV-4, Prestige 2, Merck Animal Health, group 2), vaccination and concurrent single intravenous dose of dexamethasone (approximately .05 mg/kg, group 3), vaccination and 3 intravenou...
Lim SI, Kim MJ, Kim MJ, Lee SK, Yang HS, Kwon M, Lim EH, Ouh IO, Kim EJ, Hyun BH, Lee YH.Equine influenza virus (EIV) causes acute respiratory disease in horses and belongs to the influenza A virus family , genus . This virus may have severe financial implications for the horse industry owing to its highly contagious nature and rapid transmission. In the Republic of Korea, vaccination against EIV has been practiced with the active involvement of the Korea Racing Authority since 1974. In this study, we monitored the viral RNA for EIV using PCR, as well as the antibody levels against 'A/equine/South Africa/4/03 (H3N8, clade 1)', from 2020 to 2022. EIV was not detected using RT-PCR. ...
Spruit CM, Zhu X, Tomris I, Ríos-Carrasco M, Han AX, Broszeit F, van der Woude R, Bouwman KM, Luu MMT, Matsuno K, Sakoda Y, Russell CA, Wilson IA....Influenza A viruses (IAV) initiate infection by binding to glycans with terminal sialic acids on the cell surface. Hosts of IAV variably express two major forms of sialic acid, -acetylneuraminic acid (NeuAc) and -glycolylneuraminic acid (NeuGc). NeuGc is produced in most mammals, including horses and pigs, but is absent in humans, ferrets, and birds. The only known naturally occurring IAV that exclusively bind NeuGc are extinct highly pathogenic equine H7N7 viruses. We determined the crystal structure of a representative equine H7 hemagglutinin (HA) in complex with NeuGc and observed high simi...
Arthropod-borne viruses (arboviruses) cause morbidity and mortality in humans and domestic animals worldwide. The percentage of population immunity or susceptibility to these viruses in Ecuador is unknown. To investigate the proportion of Ecuadorian populations with IgG antibodies (Abs) (past exposure/immunity) and IgM Abs (current exposure) against flaviviruses and alphaviruses and to study the activity of these viruses in Ecuador. During 2009-2011, we conducted a serosurvey for selected arboviruses in humans (n=1,842), equines (n=149), and sentinel hamsters (n=84) at two coastal locations an...
Favaro PF, Reischak D, Brandao PE, Villalobos EMC, Cunha EMS, Lara MCC, Benvenga GU, Dias RA, Mori E, Richtzenhain LJ.The equine influenza virus (EIV) H3N8 subtype is responsible for all EIV outbreaks worldwide while the H7N7 subtype is less pathogenic and is considered extinct as it has not been confirmed in outbreaks since 1980. Although EIV is enzootic in Brazil, few reports describe the actual EIV antibody status in the country. The aims of this study were: - to evaluate the efficiency of different serum treatments described by the World Organisation for Animal Health (OIE) and the World Health Organization (WHO) to remove non-specific haemagglutination inhibitors for the haemagglutination inhibition (HI)...
Dilai M, Piro M, Fougerolle S, El Harrak M, Mahir W, El Mourid R, Legrand L, Paillot R, Fassi Fihri O.In order to evaluate the vaccination status against equine influenza (EI) in Moroccan racehorses, a serological investigation was carried out on 509 racehorses using three serological tests: an Enzyme-Linked Immunosorbent Assay (ELISA), the Hemagglutination Inhibition (HI) test and the Single Radial Haemolysis (SRH) assay. The serological analysis showed 56% of seropositivity by ELISA, 67% by HI and 89.4% by SRH (with 69.9% above the clinical protection threshold). Using the Kappa test, the SRH and HI assays showed a strong agreement, the SRH and ELISA assays had a moderate agreement and the H...
Kang BK, Hwang JM, Moon H, Han SY, Kim JM, Yang DK, Park BK, Song D.The Japanese encephalitis virus (JEV) genotype circulating in Korea has changed from G3 to G1. Therefore, the purpose of this study was to compare the antigenic relationship between the two genotypes by using antibody tests. Methods: Blood samples from 42 sows and 216 horses were collected, and their seroprevalence was monitored using the hemagglutination inhibition and virus neutralization tests. Antisera against JEV G1 and G3 were isolated and prepared from guinea pigs. The cross-reactivity of these two viruses was then compared using the neutralizing antibody test. Results: We found that th...
Andoh K, Hattori S, Mahmoud HY, Takasugi M, Shimoda H, Bannai H, Tsujimura K, Matsumura T, Kondo T, Kirisawa R, Mochizuki M, Maeda K.Equine herpesvirus type 1 (EHV-1) has haemagglutination (HA) activity toward equine red blood cells (RBCs), but the identity of its haemagglutinin is unknown. To identify the haemagglutinin of EHV-1, the major glycoproteins of EHV-1 were expressed in 293T cells, and the cells or cell lysates were mixed with equine RBCs. The results showed that only EHV-1 glycoprotein C (gC)-producing cells adsorbed equine RBCs, and that the lysate of EHV-1 gC-expressing cells agglutinated equine RBCs. EHV-1 lacking gC did not show HA activity. HA activity was inhibited by monoclonal antibodies (MAbs) specific ...
Casseb Ado R, Nunes MR, Rodrigues SG, Travassos da Rosa ES, Casseb LM, Casseb SM, da Silva SP, Rodrigues ED, Vasconcelos PF.The Amazon as a whole is the largest reservoir of arboviruses worldwide, while the Brazilian Amazon hosts the largest variety of arboviruses isolated to date. In this study, the results of an indirect sandwich IgG ELISA, standardized for 19 arbovirustypes circulating among horses in Brazilian Amazon, were compared to results of the hemagglutination inhibition test. A screening test assessed the conditional probability distribution and a Pearson linear correlation test determined the correlation strength among the absorbance values recorded for viruses from the same family. Results: Sensitivity...
Karamendin K, Kydyrmanov A, Kasymbekov Y, Khan E, Daulbayeva K, Asanova S, Zhumatov K, Seidalina A, Sayatov M, Fereidouni SR.Equine influenza (EI) continues to be an important respiratory pathogen of horses worldwide. Since 2007 several outbreaks of EI have occurred in Central Asian countries, including Kazakhstan, western Mongolia, India and western China. Phylogenetic analysis showed that two H3N8 equine influenza virus (EIV) isolates from Kazakhstan, A/equine/Almaty/26/2007 and A/equine/South Kazakhstan/236/12, were related to Florida sublineage 2, with high similarity to EIVs circulating in the same period in neighbouring countries. New outbreaks of EI during 2011 and 2012 in Kazakhstan and other Central Asian c...
Sguazza GH, Fuentealba NA, Tizzano MA, Galosi CM, Pecoraro MR.Equine influenza virus is a leading cause of respiratory disease in horses worldwide. Disease prevention is by vaccination with inactivated whole virus vaccines. Most current influenza vaccines are generated in embryonated hens' eggs. Virions are harvested from allantoic fluid and chemically inactivated. Although this system has served well over the years, the use of eggs as the substrate for vaccine production has several well-recognized disadvantages (cost, egg supply, waste disposal and yield in eggs). The aim of this study was to evaluate a baculovirus system as a potential method for prod...
Thompson NN, Auguste AJ, Coombs D, Blitvich BJ, Carrington CV, da Rosa AP, Wang E, Chadee DD, Drebot MA, Tesh RB, Weaver SC, Adesiyun AA.Seroprevalence rates of selected arboviruses in animal populations in Trinidad were determined using serum samples collected between 2006 and 2009 from horses (n=506), cattle (n=163), sheep (n=198), goats (n=82), pigs (n=184), birds (n=140), rodents (n=116), and other vertebrates (n=23). The sera were screened for antibodies to West Nile virus (WNV), St. Louis encephalitis virus (SLEV), Ilheus virus (ILHV), Bussuquara virus (BSQV), Venezuelan equine encephalitis virus (VEEV), eastern equine encephalitis virus (EEEV), and western equine encephalitis virus (WEEV), using hemagglutination inhibiti...
Hirota J, Nishi H, Matsuda H, Tsunemitsu H, Shimiz S.Flavivirus-infected sera are known to show cross-reactions in serodiagnoses of heterologous flavivirus infections. Japanese encephalitis virus (JEV) is endemic in Asia and, in Japan, many horses are vaccinated against JEV. However, the cross-reactivity level of JEV-vaccinated horse sera in the serodiagnosis of West Nile virus (WNV) has not been clarified. The antibody cross-reactivity of JEV-vaccinated horse sera in WNV serological tests, such as the plaque reduction neutralization test (PRNT), IgG indirect ELISA (IgG-ELISA) and hemagglutination inhibition (HI) test, was examined. All JEV-vacc...
Ito M, Nagai M, Hayakawa Y, Komae H, Murakami N, Yotsuya S, Asakura S, Sakoda Y, Kida H.In August 2007, an outbreak of equine influenza occurred among vaccinated racehorses with Japanese commercial equine influenza vaccine at Kanazawa Racecourse in Ishikawa prefecture in Japan. Apparent symptoms were pyrexia (38.2-41.0 degrees C) and nasal discharge with or without coughing, although approximately half of the infected horses were subclinical. All horses had been shot with a vaccine that contained two inactivated H3N8 influenza virus strains [A/equine/La Plata/93 (La Plata/93) of American lineage and A/equine/Avesta/93 (Avesta/93) of European lineage] and an H7N7 strain (A/equine/...
Mancini DA, Mendonça RM, Dias AL, Mendonça RZ, Pinto JR.Trypsin is required in the hemagglutinin (HA) cleavage to in vitro influenza viruses activation. This HA cleavage is necessary for virus cell entry by receptor-mediated endocytosis. Bacteria in the respiratory tract are potential sources of proteases that could contribute to the cleavage of influenza virus in vivo. From 47 samples collected from horses, pigs, and from humans, influenza presence was confirmed in 13 and these samples demonstrated co-infection of influenza with flagellated bacteria, Stenotrophomonas maltophilia from the beginning of the experiments. Despite treatment with antibio...
Adeyefa CA, Hamblin C, Cullinane AA, McCauley JW.The objective of this work was to examine the incidence of equine influenza viruses in the equine population of an area of tropical Africa where equine influenza virus activity has recently been reported for the first time. A serological survey of sera from horses and donkeys from regions of Nigeria taken from 1990 to 1993 was carried out and the results obtained were com-pared with equine sera from Western Europe (Ireland). The sera were assayed for presence of antibodies by both haemagglutination inhibition (HI) and ELISA using a monoclonal antibody to the prototype H3 equine influenza virus...
Mall MP, Kumar A, Malik SV.A study on seroprevalence of Japanese Encephalitis (JE), West Nile (WN) and Dengue-2 (DN-2) was undertaken in and around Bareilly, Uttar Pradesh to explore the role of non-human hosts in the natural cycle of these infections. A total of 1449 animal sera samples collected from 104 dogs, 170 pigs, 170 horses, 333 buffaloes, 252 cattle, 168 sheep and 252 goats were screened for antibodies against JE, WN and DN-2 by Micro-haemagglutination inhibition (HI) test employing 4 to 16 HA units of JE, WN and DN-2 antigens. The HI positivity against JE was found in dogs (55.77%) followed by pigs (40%), hor...
Widjaja S, Soekotjo W, Hartati S, Jennings GB, Corwin AL.A study was conducted to measure the prevalence of hemagglutination-inhibition (HI) and neutralizing antibodies against two arboviruses (Chikungunya and Japanese encephalitis virus) in horses of Java, Indonesia. Blood specimens were collected from a sample of 112 horses at two stables: Pulo Mas, a racing track-horse complex, located in a residential area in North Jakarta, and Pamulang, a riding school, located in a rural environment of West Jaya. Sera were tested by the HI assay and plaque reduction neutralization test. JEV antibodies were detected by HI in 58 (52%) of the horses, while only 1...
Boussetta M, Chabchoub A, Ghram A, Jomaa I, Ghorbel A, Aouina T, Ben Amor H.Using the haemagglutination inhibition and immunodiffusion tests, a national serological survey was carried out to detect antibodies to equine influenza (EI) (A/equi/1/Prague 56 and A/equi/2/Miami 63) and equine infectious anaemia (EIA) in horse sera collected in northeastern Tunisia. 533 samples were analysed for EIA antibodies. All were negative. 13.6% of 433 equine sera tested for EI antibodies were positive. These results are discussed and compared with others obtained in Tunisia and bordering countries.
Aviles G, Bianchi TI, Daffner JF, Sabattini MS.It is shown here the WEE virus activity in ARgentina in 1983-1986 post-epizootic period. A surveillance system was established by the equine case notification and the sentinel animal method. Among the thirteen equine focus notified between September 1983-September 1985 in Córdoba and Santa Fe Provinces, 5 presumptive cases out of 16 sick horses were confirmed by the hemagglutination inhibition test for WEE epizootic virus. Twenty eight notified human cases were studied with negative results. The neutralizing antibody (Ac NT) prevalence among sentinel horses in Córdoba Province (4%) was lower...
Bogdan JR, Morley PS, Townsend HG, Haines DM.This study has tested the effect of using homologous or heterologous equine influenza A virus isolates to evaluate serum antibody levels to influenza A virus in vaccinated and naturally-infected horses. In addition, the potential effect of antigenic selection of virus variants in egg versus tissue culture propagation systems was studied. Serum antibody levels in samples from horses recently infected with a local influenza A virus isolate (A/equine 2/Saskatoon/1/90) or recently vaccinated with a prototype isolate (A/equine 2/Miami/1/63) were assessed by hemagglutination inhibition and by single...
Sentsui H, Kono Y.Horse erythrocytes treated with equine infectious anemia virus hemagglutinin were phagocytized by cultivated horse leukocytes (mainly macrophage-like cells and partly polymorphonuclear cells) after incubation with fresh horse serum but not with inactivated horse serum. The phagocytosis began as soon as the erythrocytes were added to the leukocyte cultures, and the majority of the reaction proceeded within 30 minutes. Addition of antiserum showed a slightly suppressing but no enhancing effect on the phagocytosis. Phagocytosis seemed to be caused by the recognition of the third complement compon...
Yamamoto K, Hashimoto K, Chiba J, Simizu B.To analyze the biological activities of the alphavirus glycoproteins, eight different monoclonal antibodies against the two glycoproteins of western equine encephalitis virus were isolated. Five of the eight monoclonal antibodies were shown to be specific for E1 and three for E2 protein by an enzyme-linked immunosorbent assay and by radioimmunoprecipitation. Three of the five anti-E1 and all of the anti-E2 monoclonal antibodies inhibited hemagglutination by purified virions. One anti-E1 and two anti-E2 monoclonal antibodies possessed high virus-neutralizing activity.
Smit H, Gaastra W, Kamerling JP, Vliegenthart JF, de Graaf FK.The erythrocyte receptor for Escherichia coli K99 fimbrial adhesin was isolated from equine erythrocytes and characterized as Neu5Gc-alpha(2----3)-Galp-beta(1----4)-GLcp-beta(1----1)-Ceramide. This glycolipid acted as the receptor for K99 by four different experimental approaches: inhibition of equine erythrocyte hemagglutination by preincubation of K99-positive bacteria or purified K99 fimbriae with the isolated glycolipid; inhibition of attachment of K99-positive bacteria to porcine intestinal epithelial cells in the presence of the isolated glycolipid; induction of binding of K99-positive b...
Nishimura M, Nakajima H.Using purified equine infectious anemia (EIA) virus labeled with 3H-glucosamine or 14C-protein hydrolysate, structural proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. As a result, 2 glycoproteins and 10 proteins with molecular weights (mol wt) ranging from 12,000 to 115,000 daltons were demonstrated. Of 12 structural proteins, 3 proteins, namely a glycoprotein with mol wt of 76,000 (gp76) and 2 proteins with mol wt of 25,000 (p25) and 12,000 (p12), respectively, had distinct antigenic activity from one another in immunodiffusion. Development of antibodies a...
Thorsen J, Willoughby RA, McDonell W, Valli VE, Viel L, Bignell W.Samples of mucus from the lower trachea were collected from 53 horses with chronic obstructive pulmonary disease and from 24 clinically normal horses. Serum samples were collected from 35 of the horses with chronic obstructive pulmonary disease and from the 24 normal horses. Samples were tested for inhibition of hemagglutination by influenza A equine 1 and 2 viruses. There were high levels of hemagglutination inhibiting activity against influenza A equine 1 in mucus samples from horses with chronic obstructive pulmonary disease.
Ryder E, Ryder S.Venezuelan equine encephalitis (VEE) virus has been implicated as producing alterations in glucose metabolism in animals. We performed oral glucose tolerance tests and measured serum immunoreactive insulin responses in 13 patients who were infected by VEE virus during an epidemic in 1969, in Zulia State, Venezuela. No significant alterations in the glucose tolerance test were found. Sera of 86 diabetic outpatients and 98 control individuals with normal glycemia at a local hospital were tested for antibodies to VEE virus by hemagglutination inhibition. No statistically significant difference wa...
Brundage-Anguish LJ, Holmes DF, Hosier NT, Murphy BR, Massicott JG, Appleyard G, Coggins L.Temperature-sensitive (ts) reassortants of an equine influenza virus, subtype A-1, were produced by mating a human influenza ts donor virus with an equine influenza A/Cornell/16/74 wild-type virus and by isolating a ts reassortant virus possessing the equine hemagglutinin and neuraminidase surface antigens. Two equine its reassortant clones, 8B1 and 71A1, were produced which had an in vitro shutoff temperature for plaque formation of 38 and 37 C, respectively. The human ts donor virus had ts mutation(s) on the polymerase 3 (P3) and nucleoprotein genes so that a ts equine reassortant virus coul...
Yamagishi H, Nagamine T, Shimoda K, Ide S, Igarashi Y, Yoshioka I, Matumoto M.Single radial hemolysis (SRH), neutralization (NT), and hemagglutination inhibition (HI) tests were carried out on sera from horses immunized against the Prague and Miami strains of equine influenza virus. The HI and NT tests demonstrated good sensitivity; the sensitivity of the SRH test was somewhat lower. The NT titers of individual sera were correlated very closely with the HI titers, although the NT titers were higher. SRH zone diameters of individual sera also showed significant correlation with the NT and NI titers. The SRH test appears to be suitable for large-scale serological surveys ...
Sentsui H, Kono Y.The serologic relationships between 6 strains of equine infectious anemia (EIA) viruses were investigated by hemagglutination-inhibition (HI) tests. Cross HI tests, using sera from horses in the early stage of infection, revealed that all strains were inhibited only by homologous strain antisera and that HI antibody was always detectable before virus-neutralizing antibody. In the later stages of infection, both homologous and heterologous HI antibodies were detected in a sera of most of the horses, and the order of appearance of heterologous HI antibodies was random in 2 horses inoculated with...
Smit H, Gaastra W, Kamerling JP, Vliegenthart JF, de Graaf FK.The erythrocyte receptor for Escherichia coli K99 fimbrial adhesin was isolated from equine erythrocytes and characterized as Neu5Gc-alpha(2----3)-Galp-beta(1----4)-GLcp-beta(1----1)-Ceramide. This glycolipid acted as the receptor for K99 by four different experimental approaches: inhibition of equine erythrocyte hemagglutination by preincubation of K99-positive bacteria or purified K99 fimbriae with the isolated glycolipid; inhibition of attachment of K99-positive bacteria to porcine intestinal epithelial cells in the presence of the isolated glycolipid; induction of binding of K99-positive b...
Hirota J, Nishi H, Matsuda H, Tsunemitsu H, Shimiz S.Flavivirus-infected sera are known to show cross-reactions in serodiagnoses of heterologous flavivirus infections. Japanese encephalitis virus (JEV) is endemic in Asia and, in Japan, many horses are vaccinated against JEV. However, the cross-reactivity level of JEV-vaccinated horse sera in the serodiagnosis of West Nile virus (WNV) has not been clarified. The antibody cross-reactivity of JEV-vaccinated horse sera in WNV serological tests, such as the plaque reduction neutralization test (PRNT), IgG indirect ELISA (IgG-ELISA) and hemagglutination inhibition (HI) test, was examined. All JEV-vacc...
Tumova B, Easterday BC.This study demonstrates relationships in envelope antigens of 4 human influenza A2 strains isolated during the period 1957-68 (including A2/Hong Kong/68), 2 strains of A/Equi-2/63 and 7 avian influenza viruses isolated in Europe, North America, and the Ukraine in the years 1960-67.Antigenic relationships among the strains were determined on the basis of haemagglutination-inhibition, virus-neutralization, strain-specific complement-fixation, and neuraminidase-inhibition tests.North American avian influenza strains, Turkey/California/64, Turkey/Massachusetts/65, Turkey/Wisconsin/66, Turkey/Ontar...
Ito M, Nagai M, Hayakawa Y, Komae H, Murakami N, Yotsuya S, Asakura S, Sakoda Y, Kida H.In August 2007, an outbreak of equine influenza occurred among vaccinated racehorses with Japanese commercial equine influenza vaccine at Kanazawa Racecourse in Ishikawa prefecture in Japan. Apparent symptoms were pyrexia (38.2-41.0 degrees C) and nasal discharge with or without coughing, although approximately half of the infected horses were subclinical. All horses had been shot with a vaccine that contained two inactivated H3N8 influenza virus strains [A/equine/La Plata/93 (La Plata/93) of American lineage and A/equine/Avesta/93 (Avesta/93) of European lineage] and an H7N7 strain (A/equine/...
Thompson NN, Auguste AJ, Coombs D, Blitvich BJ, Carrington CV, da Rosa AP, Wang E, Chadee DD, Drebot MA, Tesh RB, Weaver SC, Adesiyun AA.Seroprevalence rates of selected arboviruses in animal populations in Trinidad were determined using serum samples collected between 2006 and 2009 from horses (n=506), cattle (n=163), sheep (n=198), goats (n=82), pigs (n=184), birds (n=140), rodents (n=116), and other vertebrates (n=23). The sera were screened for antibodies to West Nile virus (WNV), St. Louis encephalitis virus (SLEV), Ilheus virus (ILHV), Bussuquara virus (BSQV), Venezuelan equine encephalitis virus (VEEV), eastern equine encephalitis virus (EEEV), and western equine encephalitis virus (WEEV), using hemagglutination inhibiti...
Mancini DA, Mendonça RM, Dias AL, Mendonça RZ, Pinto JR.Trypsin is required in the hemagglutinin (HA) cleavage to in vitro influenza viruses activation. This HA cleavage is necessary for virus cell entry by receptor-mediated endocytosis. Bacteria in the respiratory tract are potential sources of proteases that could contribute to the cleavage of influenza virus in vivo. From 47 samples collected from horses, pigs, and from humans, influenza presence was confirmed in 13 and these samples demonstrated co-infection of influenza with flagellated bacteria, Stenotrophomonas maltophilia from the beginning of the experiments. Despite treatment with antibio...
Karamendin K, Kydyrmanov A, Kasymbekov Y, Khan E, Daulbayeva K, Asanova S, Zhumatov K, Seidalina A, Sayatov M, Fereidouni SR.Equine influenza (EI) continues to be an important respiratory pathogen of horses worldwide. Since 2007 several outbreaks of EI have occurred in Central Asian countries, including Kazakhstan, western Mongolia, India and western China. Phylogenetic analysis showed that two H3N8 equine influenza virus (EIV) isolates from Kazakhstan, A/equine/Almaty/26/2007 and A/equine/South Kazakhstan/236/12, were related to Florida sublineage 2, with high similarity to EIVs circulating in the same period in neighbouring countries. New outbreaks of EI during 2011 and 2012 in Kazakhstan and other Central Asian c...
Mall MP, Kumar A, Malik SV.A study on seroprevalence of Japanese Encephalitis (JE), West Nile (WN) and Dengue-2 (DN-2) was undertaken in and around Bareilly, Uttar Pradesh to explore the role of non-human hosts in the natural cycle of these infections. A total of 1449 animal sera samples collected from 104 dogs, 170 pigs, 170 horses, 333 buffaloes, 252 cattle, 168 sheep and 252 goats were screened for antibodies against JE, WN and DN-2 by Micro-haemagglutination inhibition (HI) test employing 4 to 16 HA units of JE, WN and DN-2 antigens. The HI positivity against JE was found in dogs (55.77%) followed by pigs (40%), hor...
Yamagishi H, Nagamine T, Shimoda K, Ide S, Igarashi Y, Yoshioka I, Matumoto M.Single radial hemolysis (SRH), neutralization (NT), and hemagglutination inhibition (HI) tests were carried out on sera from horses immunized against the Prague and Miami strains of equine influenza virus. The HI and NT tests demonstrated good sensitivity; the sensitivity of the SRH test was somewhat lower. The NT titers of individual sera were correlated very closely with the HI titers, although the NT titers were higher. SRH zone diameters of individual sera also showed significant correlation with the NT and NI titers. The SRH test appears to be suitable for large-scale serological surveys ...
Hierholzer JC, Gamble WC, Quist KD, Chappell WA.Horses were immunized by a variety of inoculation procedures designed to determine the most efficient method of producing antisera to adenovirus types 25 to 31. The procedures evaluated included immunization by (i) direct intravenous (iv) injection, (ii) iv infusion, (iii) intramuscular (im) injection of virus with and without Freund's incomplete adjuvant, (iv) combined iv and im injections, and (v) combined iv infusion and im injection. The im schedule (no. 3) was superior to the others in terms of immunizing antigen and time required, and hemagglutination-inhibition (HI) and serum-neutralizi...
Sentsui H, Kono Y.Equine infectious anemia (EIA) virus which was propagated on an equine dermal cell line agglutinated guinea pig erythrocytes. Viral fluids containing about 10(7.5) mean tissue culture infective doses/ml showed hemagglutinating (HA) titers ranging from 16 to 32 units/0.05 ml. Results of cesium chloride equilibrium density gradient centrifugation revealed that the hemagglutinin was inseparable from the virus particles. The hemagglutination reaction persisted over a wide range of temperature and pH, and the absence of divalent cations did not decrease its activity. The HA activity was stable at 4...
Kang BK, Hwang JM, Moon H, Han SY, Kim JM, Yang DK, Park BK, Song D.The Japanese encephalitis virus (JEV) genotype circulating in Korea has changed from G3 to G1. Therefore, the purpose of this study was to compare the antigenic relationship between the two genotypes by using antibody tests. Methods: Blood samples from 42 sows and 216 horses were collected, and their seroprevalence was monitored using the hemagglutination inhibition and virus neutralization tests. Antisera against JEV G1 and G3 were isolated and prepared from guinea pigs. The cross-reactivity of these two viruses was then compared using the neutralizing antibody test. Results: We found that th...
Widjaja S, Soekotjo W, Hartati S, Jennings GB, Corwin AL.A study was conducted to measure the prevalence of hemagglutination-inhibition (HI) and neutralizing antibodies against two arboviruses (Chikungunya and Japanese encephalitis virus) in horses of Java, Indonesia. Blood specimens were collected from a sample of 112 horses at two stables: Pulo Mas, a racing track-horse complex, located in a residential area in North Jakarta, and Pamulang, a riding school, located in a rural environment of West Jaya. Sera were tested by the HI assay and plaque reduction neutralization test. JEV antibodies were detected by HI in 58 (52%) of the horses, while only 1...
Aviles G, Bianchi TI, Daffner JF, Sabattini MS.It is shown here the WEE virus activity in ARgentina in 1983-1986 post-epizootic period. A surveillance system was established by the equine case notification and the sentinel animal method. Among the thirteen equine focus notified between September 1983-September 1985 in Córdoba and Santa Fe Provinces, 5 presumptive cases out of 16 sick horses were confirmed by the hemagglutination inhibition test for WEE epizootic virus. Twenty eight notified human cases were studied with negative results. The neutralizing antibody (Ac NT) prevalence among sentinel horses in Córdoba Province (4%) was lower...
Anderson RS, Day NK, Good RA.Natural hemagglutinin activity against vertebrate erythrocytes is present in the hemolymph of the cockroach Blabarus craniifer. The hemagglutinin titer against rabbit erythrocytes is high, whereas sheep and horse red cells agglutinate weakly. Hemagglutinin activity was depressed by the complement inhibitor, cobra venom factor. Cockroach hemagglutinin is heat-labile; all activity is destroyed by heating at 56 C for 1 hr. A humoral factor similar to the complement component 3 proactivator is also present in cockroach hemolymph. The formation of the cobra venom factor-hemolymph "complex" is depen...
Sentsui H, Kono Y.Horse erythrocytes treated with equine infectious anemia virus hemagglutinin were phagocytized by cultivated horse leukocytes (mainly macrophage-like cells and partly polymorphonuclear cells) after incubation with fresh horse serum but not with inactivated horse serum. The phagocytosis began as soon as the erythrocytes were added to the leukocyte cultures, and the majority of the reaction proceeded within 30 minutes. Addition of antiserum showed a slightly suppressing but no enhancing effect on the phagocytosis. Phagocytosis seemed to be caused by the recognition of the third complement compon...
Spruit CM, Zhu X, Tomris I, Ríos-Carrasco M, Han AX, Broszeit F, van der Woude R, Bouwman KM, Luu MMT, Matsuno K, Sakoda Y, Russell CA, Wilson IA....Influenza A viruses (IAV) initiate infection by binding to glycans with terminal sialic acids on the cell surface. Hosts of IAV variably express two major forms of sialic acid, -acetylneuraminic acid (NeuAc) and -glycolylneuraminic acid (NeuGc). NeuGc is produced in most mammals, including horses and pigs, but is absent in humans, ferrets, and birds. The only known naturally occurring IAV that exclusively bind NeuGc are extinct highly pathogenic equine H7N7 viruses. We determined the crystal structure of a representative equine H7 hemagglutinin (HA) in complex with NeuGc and observed high simi...
Adeyefa CA, Hamblin C, Cullinane AA, McCauley JW.The objective of this work was to examine the incidence of equine influenza viruses in the equine population of an area of tropical Africa where equine influenza virus activity has recently been reported for the first time. A serological survey of sera from horses and donkeys from regions of Nigeria taken from 1990 to 1993 was carried out and the results obtained were com-pared with equine sera from Western Europe (Ireland). The sera were assayed for presence of antibodies by both haemagglutination inhibition (HI) and ELISA using a monoclonal antibody to the prototype H3 equine influenza virus...
Sentsui H, Kono Y.The serologic relationships between 6 strains of equine infectious anemia (EIA) viruses were investigated by hemagglutination-inhibition (HI) tests. Cross HI tests, using sera from horses in the early stage of infection, revealed that all strains were inhibited only by homologous strain antisera and that HI antibody was always detectable before virus-neutralizing antibody. In the later stages of infection, both homologous and heterologous HI antibodies were detected in a sera of most of the horses, and the order of appearance of heterologous HI antibodies was random in 2 horses inoculated with...
Sentsui H, Kono Y.Six strains of equine infectious anemia (EIA) virus propagated in equine leukocyte cultures were found to agglutinate horse erythrocytes. Concentrated virus material containing about 20 units of complement fixation (CF) titer showed hemagglutinating (HA) titers ranging from 4 to 8 units. The HA activity remained stable after ether treatment and was reduced by trypsin, formaldehyde and KIO4. Cesium chloride equilibrium density gradient centrifugation revealed two populations of hemagglutinin, one in the density range of 1.15-1.16 g/ml coinciding with a peak of CF antigen and the other at round ...
Nishimura M, Nakajima H.Using purified equine infectious anemia (EIA) virus labeled with 3H-glucosamine or 14C-protein hydrolysate, structural proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. As a result, 2 glycoproteins and 10 proteins with molecular weights (mol wt) ranging from 12,000 to 115,000 daltons were demonstrated. Of 12 structural proteins, 3 proteins, namely a glycoprotein with mol wt of 76,000 (gp76) and 2 proteins with mol wt of 25,000 (p25) and 12,000 (p12), respectively, had distinct antigenic activity from one another in immunodiffusion. Development of antibodies a...
Kucera CJ, Beckenhauer WH.An inactivated, aluminum hydroxide adjuvant equine influenza vaccine was tested in horses and guinea pigs to determine the levels of antigen that would elicit maximum serological responses. Vaccine containing serial twofold increments of A/Equi-1/Prague and A/Equi-2/Miami strains of equine influenza virus was administered to random groupings of both types of test animals. The hemagglutination inhibition antibody response for each group was then measured. Results in horses and guinea pigs were compared to determine if the equine serological values could be related to a potency test in laborator...
Goto H, Shimizu K.The first outbreak of equine influenza (EI) infection in Japan was recognized during the period December 1971 to January 1972 [1, 6]. No evidence of the disease had been found before then [2,6]. The etiological agent of this epizootic was identified by hemagglutination-inhibition (HI) and neutralization tests with chicken or ferret antiserum as the subtype 2 of EI virus (6, 7). However, the isolate, A/equine/Tokyo/71 (Tokyo) strain, was not completely identical to the prototypic A/equine/Miami /63 (Miami) strain of the subtype 2, since antibody responses of convalescent horses were 2 to 16 tim...
Andoh K, Hattori S, Mahmoud HY, Takasugi M, Shimoda H, Bannai H, Tsujimura K, Matsumura T, Kondo T, Kirisawa R, Mochizuki M, Maeda K.Equine herpesvirus type 1 (EHV-1) has haemagglutination (HA) activity toward equine red blood cells (RBCs), but the identity of its haemagglutinin is unknown. To identify the haemagglutinin of EHV-1, the major glycoproteins of EHV-1 were expressed in 293T cells, and the cells or cell lysates were mixed with equine RBCs. The results showed that only EHV-1 glycoprotein C (gC)-producing cells adsorbed equine RBCs, and that the lysate of EHV-1 gC-expressing cells agglutinated equine RBCs. EHV-1 lacking gC did not show HA activity. HA activity was inhibited by monoclonal antibodies (MAbs) specific ...
Ferguson JA, Reeves WC, Milby MM, Hardy JL.Of 359 horses vaccinated with attenuated Venezuelan equine encephalomyelitis (VEE) vaccine (strain TC-83), 87% developed hemagglutination-inhibition (HI) antibodies to VEE virus within 1 month. Blood from a subsample of 101 of the 359 horses was obtained over a 1-year period. Within 1 month after vaccination, 84% of the 101 horses had developed VEE HI antibodies, 87% had developed VEE-neutralizing (Nt) antibodies, and 78% had developed VEE complement-fixing (CF) antibodies. One year after vaccination, 58% of the horses had VEE HI antibodies and 73% had VEE Nt antibodies. The percentage of hors...
Yamamoto K, Hashimoto K, Chiba J, Simizu B.To analyze the biological activities of the alphavirus glycoproteins, eight different monoclonal antibodies against the two glycoproteins of western equine encephalitis virus were isolated. Five of the eight monoclonal antibodies were shown to be specific for E1 and three for E2 protein by an enzyme-linked immunosorbent assay and by radioimmunoprecipitation. Three of the five anti-E1 and all of the anti-E2 monoclonal antibodies inhibited hemagglutination by purified virions. One anti-E1 and two anti-E2 monoclonal antibodies possessed high virus-neutralizing activity.
Poskus E, Zakin MM, Fernámdez HN, Paladini AC.Peptide fragments, obtained from equine growth hormone by cyanogen bromide cleavage and further chemical treatment, were isolated and identified. Their immunological reactivities were tested by hemagglutination and complement fixation methods using rabbit antisera against native hormone. Antigenic determinants were detected in the fragments comprising amino acid sequences 5-72 and 73-123, this last one being predominant. Fragment 124-178 had very low reactivity. Nitration of peptide 73-123 did not modify its immunological properties,but oxidation diminished them. Comparison of the antigenicity...
