Geographic variation of individual venom profile of Crotalus durissus snakes

South American rattlesnakes are represented in Brazil by a single species, Crotalus durissus, which has public health importance due to the severity of its envenomation and to its wide geographical distribution. The species is subdivided into several subspecies, but the current classification is controversial. In Brazil, the venoms of C. d. terrificus and C. d. collilineatus are used for hyperimmunization of horses for antivenom production, even though the distinction of these two subspecies are mostly by their geographical distribution. In this context, we described a comparative compositional and functional characterization of individual C. d. collilineatus and C. d. terrificus venoms from three Brazilian states. Methods: We compared the compositional patterns of C. d. terrificus and C. d. collilineatus individual venoms by 1-DE and RP-HPLC. For functional analyzes, the enzymatic activities of PLA2, LAAO, and coagulant activity were evaluated. Finally, the immunorecognition of venom toxins by the crotalic antivenom produced at Butantan Institute was evaluated using Western blotting. Results: The protein profile of individual venoms from C. d. collilineatus and C. d. terrificus showed a comparable overall composition, despite some intraspecific variation, especially regarding crotamine and LAAO. Interestingly, HPLC analysis showed a geographic pattern concerning PLA2. In addition, a remarkable intraspecific variation was also observed in PLA2, LAAO and coagulant activities. The immunorecognition pattern of individual venoms from C. d. collilineatus and C. d. terrificus by crotalic antivenom produced at Butantan Institute was similar. Conclusions: The results highlighted the individual variability among the venoms of C. durissus ssp. specimens. Importantly, our data point to a geographical variation of C. durissus ssp. venom profile, regardless of the subspecies, as evidenced by PLA2 isoforms complexity, which may explain the increase in venom neurotoxicity from Northeastern through Southern Brazil reported for the species.(AU)

Geographic variation of individual venom profile of Crotalus durissus snakes

South American rattlesnakes are represented in Brazil by a single species, Crotalus durissus, which has public health importance due to the severity of its envenomation and to its wide geographical distribution. The species is subdivided into several subspecies, but the current classification is controversial. In Brazil, the venoms of C. d. terrificus and C. d. collilineatus are used for hyperimmunization of horses for antivenom production, even though the distinction of these two subspecies are mostly by their geographical distribution. In this context, we described a comparative compositional and functional characterization of individual C. d. collilineatus and C. d. terrificus venoms from three Brazilian states. Methods: We compared the compositional patterns of C. d. terrificus and C. d. collilineatus individual venoms by 1-DE and RP-HPLC. For functional analyzes, the enzymatic activities of PLA2, LAAO, and coagulant activity were evaluated. Finally, the immunorecognition of venom toxins by the crotalic antivenom produced at Butantan Institute was evaluated using Western blotting. Results: The protein profile of individual venoms from C. d. collilineatus and C. d. terrificus showed a comparable overall composition, despite some intraspecific variation, especially regarding crotamine and LAAO. Interestingly, HPLC analysis showed a geographic pattern concerning PLA2. In addition, a remarkable intraspecific variation was also observed in PLA2, LAAO and coagulant activities. The immunorecognition pattern of individual venoms from C. d. collilineatus and C. d. terrificus by crotalic antivenom produced at Butantan Institute was similar. Conclusions: The results highlighted the individual variability among the venoms of C. durissus ssp. specimens. Importantly, our data point to a geographical variation of C. durissus ssp. venom profile, regardless of the subspecies, as evidenced by PLA2 isoforms complexity, which may explain the increase in venom neurotoxicity from Northeastern through Southern Brazil reported for the species.(AU)

PERFIL HEMATOLÓGICO, BIOQUÍMICO, URINÁRIO E ULTRASSONOGRÁFICO DE EQUINOS SUBMETIDOS À INOCULAÇÃO DO VENENO CROTÁLICO

Os acidentes ofídicos representam um problema de saúde pública nos países tropicais devido à alta incidência e morbimortalidade. No tratamento é utilizado infusão do soro antiofídico que é produzido a partir da hiperimunização de cavalos com a mistura de venenos característicos da região. O objetivo desta pesquisa foi avaliar a função hepática e renal dos equinos submetidos à produção do soro antiofídico com o protocolo de imunização empregado pelo Instituto Vital Brazil (IVB), para um melhor entendimento da resposta imune do equino, visando protocolos que minimizem os efeitos deletérios à saúde dos mesmos. O estudo foi realizado na fazenda do IVB durante o ano de 2019.Os animais foram examinados antes do início dos protocolos para o grupo controle. Depois foram realizados exames hematológicos e perfil bioquímico até 48hs após a inoculação do veneno e semanalmente, avaliação ultrassonográfica hepática e renal até 48hs após a inoculação do veneno e a cada 15 dias, exames de urinálise a cada 15 dias e avaliação clínica dos animais semanalmente após a inoculação do veneno. Os exames foram separados em quatro grupos de acordo com os protocolos de inoculação do veneno: C (grupo controle), IB (imunização de base), PR (primeira reimunização) e SR (segunda reimunização). Foram encontradas diferenças estatísticas nas variáveis hematimetria, hemoglobina, VG, VGM, CHGM, proteína, fibrinogênio, segmentado e linfócito no perfil hematológico; nas variáveis: ureia, creatinina, GGT, albumina e globulina no perfil bioquímico; e nas variáveis: proteína, pH e na relação proteína: creatinina na urinálise. No exame ultrassonográfico do fígado foi encontrada diferença na variável dimensão e no rim nas variáveis: relação córticomedular, ecogenicidade e espessura da cortical. Concluímos que a função hepática e renal dos equinos foi afetada pela inoculação do veneno crotálico. Desta forma devemos ponderar a utilização de suplementações e medicações que minimizem o processo inflamatório e anêmico estabelecido nos equinos inoculados com o veneno. E sugerimos mais pesquisas relacionadas ao assunto para proporcionar bem-estar aos animais submetidos ao processo de inoculação do veneno, que é uma etapa primordial para a produção dos soros antiofídicos.

Ophidian accidents represent a public health problem in tropical countries due to the high incidence and morbimortality. An infusion of antiophidic serum is used for treatment, which is produced from the hyperimmunization of equines with a mixture of poisons characteristic of the region. The objective of this research was to evaluate liver and kidney function of equines submitted to production of antiophidic serum with immunization protocol used by Vital Brazil Institute (IVB), as we expect a better understanding on immune response of equines, aiming at protocols that minimize deleterious effects on their health. The study was carried out at IVB farm during 2019. Animals were examined before start of protocols for the control group. Complete Blood Count (CBC) and biochemistry profile, hepatic and renal ultrasound evaluation, urinalysis and clinical evaluation of the animals were performed up to 48 hours after inoculation of the venom, weekly (in the case of CBC, biochemistry profile and clinical evaluation) and every 15 days. (in the case of urinalysis and ultrasound exams).The exams were separated into four groups according to venom inoculation protocols: C (control group), IB (basic immunization), PR (first re-immunization) and SR (second re-immunization). Statistical differences were found in variables erythrocyte, hemoglobin, corpuscular volume (CV), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), protein, fibrinogen, segmented neutrophils and lymphocyte in CBC results; in urea, creatinine, GGT, albumin and globulin in the biochemistry profile; and in protein, pH, protein:creatinine ratio in urinalysis. In the hepatic ultrasound examination, a difference was found in variable dimension and in the kidneys in variables: corticomedullary ratio, echogenicity and cortical thickness. We conclude that liver and kidney function was affected in equines submitted to crotalic venom inoculation. Thus, we must consider the use of supplements and drugs that minimize the inflammatory and anemic process established in equines inoculated with the venom. And we suggest more research related to this matter to provide welfare to animals submitted to poison inoculation process, which is a primordial step for antiophidic serum production.

OBTENÇÃO E CARACTERIZAÇÃO DE IMUNÓGENO CONJUGADO DE LIPOPOLISSACARÍDEO DE PSEUDOMONAS AERUGINOSA E ALBUMINA BOVINA

ABSTRACT Pseudomonas aeruginosa is an etiologic agent of opportunistic infections mainly in immunocompromised patients. Its inherent feature of developing resistance to a wide range of antibacterial agents make it a critical point in infection control. In animals, problems with multidrug resistance occur in otitis, cystitis, uveo-conjunctivitis, endometritis and mastitis, and there is no commercially available vaccine. With the aim of improving its immunogenicity, the lipopolysaccharide (LPS) antigen was coupled to bovine serum albumin (BSA) with .-periodate as the reductive agent. The conjugation was evaluated by gel-permeation chromatography, by quantitating total sugar and protein, and both LPS and BSA were detected in similar proportions. The immunization of mice with LPS-BSA conjugate vaccine resulted in an agglutinating antibody response against P. aeruginosa lower than that obtained for a mixture of free LPS and BSA. They were 65% and 86% lower in the 6th and 10th weeks after the hyperimmunization procedure, respectively. This indicates that the conjugation reaction resulted in an immunogenic product, however its quality should be improved.

RESUMO A Pseudomonas aeruginosa é agente etiológico de infecções oportunistas, principalmente em pacientes imunocomprometidos. Suas características inerentes em desenvolver resistência aos mais variados tipos de antibacterianos a torna um ponto crítico no controle de infecções. Em animais, os problemas com multirresistência ocorrem principalmente em casos de otite, cistite, úveo-conjuntivite, endometrite e mastite, não havendo vacina comercialmente disponível. No intuito de melhorar a imunogenicidade desse antígeno, foi testada a técnica de conjugação do lipopolissacarídeo (LPS) de P. aeruginosa à albumina bovina (BSA) por aminação redutiva direta utilizando .-periodato de sódio. A conjugação foi avaliada por cromatografia de gel-permeação, dosando-se açúcar e proteína totais, e tanto o LPS quanto a BSA foram identificados em proporções semelhantes. A imunização de camundongos com a vacina conjugada LPS-BSA conferiu títulos de anticorpos aglutinantes contra P. aeruginosa inferiores aos obtidos com a mistura de LPS e BSA livres. Foram 65% e 86% menores na 6ª e na 10ª semanas após o procedimento de hiperimunização, respectivamente. Isto indica que a reação de conjugação resultou em um produto imunogênico, porém, sua qualidade precisará ser melhorada.

Obtenção e caracterização de imunógeno conjugado de lipopolissacarídeo de Pseudomonas aeruginosa e albumina bovina / Obtainment and characterization of an immunogenic conjugate of Pseudomonas aeruginosa lipopolysaccharide and bovine serum albumin

A Pseudomonas aeruginosa é agente etiológico de infecções oportunistas, principalmente em pacientes imunocomprometidos. Suas características inerentes em desenvolver resistência aos mais variados tipos de antibacterianos a torna um ponto crítico no controle de infecções. Em animais, os problemas com multirresistência ocorrem principalmente em casos de otite, cistite, úveo-conjuntivite, endometrite e mastite, não havendo vacina comercialmente disponível. No intuito de melhorar a imunogenicidade desse antígeno, foi testada a técnica de conjugação do lipopolissacarídeo (LPS) de P. aeruginosa à albumina bovina (BSA) por aminação redutiva direta utilizando .-periodato de sódio. A conjugação foi avaliada por cromatografia de gel-permeação, dosando-se açúcar e proteína totais, e tanto o LPS quanto a BSA foram identificados em proporções semelhantes. A imunização de camundongos com a vacina conjugada LPS-BSA conferiu títulos de anticorpos aglutinantes contra P. aeruginosa inferiores aos obtidos com a mistura de LPS e BSA livres. Foram 65% e 86% menores na 6ª e na 10ª semanas após o procedimento de hiperimunização, respectivamente. Isto indica que a reação de conjugação resultou em um produto imunogênico, porém, sua qualidade precisará ser melhorada.

Pseudomonas aeruginosa is an etiologic agent of opportunistic infections mainly in immunocompromised patients. Its inherent feature of developing resistance to a wide range of antibacterial agents make it a critical point in infection control. In animals, problems with multidrug resistance occur in otitis, cystitis, uveo-conjunctivitis, endometritis and mastitis, and there is no commercially available vaccine. With the aim of improving its immunogenicity, the lipopolysaccharide (LPS) antigen was coupled to bovine serum albumin (BSA) with .-periodate as the reductive agent. The conjugation was evaluated by gel-permeation chromatography, by quantitating total sugar and protein, and both LPS and BSA were detected in similar proportions. The immunization of mice with LPS-BSA conjugate vaccine resulted in an agglutinating antibody response against P. aeruginosa lower than that obtained for a mixture of free LPS and BSA. They were 65% and 86% lower in the 6th and 10th weeks after the hyperimmunization procedure, respectively. This indicates that the conjugation reaction resulted in an immunogenic product, however its quality should be improved.

Immune response and neutralization capacity of antibodies produced in young sheep immunized with Crotalus durissus terrificus native or Cobalt-60 irradiated venom / Resposta imune e capacidade de neutralização de anticorpos produzidos em ovinos jovens imunizados com veneno de Crotalus durissus terrificus nativo e irradiado com Cobalto 60

The ELISA technique was used to evaluate and compare young ovine humoral immune response during crotalic antiserum production. Animals were clinically evaluated throughout this process, and the neutralizing capacity of antisera raised against natural (NV) and Cobalt-60 irradiated (IrV) venoms of Crotalus durissus terrificus (C.d.t.) was verified by means of in vitro challenges. Three groups of six animals each were used: G1 received NV; G2 was inoculated with IrV; and G3 was used as control. Animals received six immunizations during 84 days at 14-day intervals. ELISA of antibody profile showed significant difference (p<5%) between experimental groups (G1

A técnica de Elisa foi utilizada para avaliar e comparar a resposta imune humoral de ovinos jovens para a produção de soro anticrotálico. Durante o processo de soroprodução, foi realizada a avaliação clínica dos animais. A capacidade de neutralização do soro produzido a partir de veneno de serpente Crotalus durissus terrificus, nativo (VN) e irradiado (VIr) com Cobalto–60 foi verificada por meio de desafios in vitro. Um grupo de seis animais recebeu veneno nativo, o segundo grupo recebeu veneno irradiado e o terceiro grupo foi o controle. Os animais receberam seis imunizações durante 84 dias com intervalo de14 dias. Houve diferença significativa (p<5%) no teste de ELISA do perfil de anticorpos produzidos pelos grupos experimentais (VN

Young ovine death during hyperimmunization: crotalic envenomation or copper toxicosis?

The unfavorable evolution of a young ovine during hyperimmunization process with Crotalus durissus terrificus venom was investigated in order to differentiate its origin between ophidic envenomation and copper toxicosis. Clinical, laboratory, necroscopic and histological exams as well as evaluation and measurement of heavy metals (copper) in the kidneys and in the liver were carried out. Blood counts revealed anemia and serological tests showed high levels of blood urea nitrogen, creatinine, aspartate aminotransferase, creatine phosphokinase, total bilirubin and indirect bilirubin; which indicates liver, kidney and skeletal muscle damages. At necropsy, the animal presented hepatopathy and nephropathy. Histological examination revealed renal and hepatic features that may imply copper intoxication. Copper levels were 237.8 µg/g in the liver and 51.2 µg/g in the kidneys. Although the amount of metal found in both organs was below the level that can cause death, according to the literature, anatomopathological signs were suggestive of copper intoxication. Therefore, the hypothesis of metal toxicosis during the hyperimmunization process became more consistent than the crotalic envenomation one.

Young ovine death during hyperimmunization: crotalic envenomation or copper toxicosis?

The unfavorable evolution of a young ovine during hyperimmunization process with Crotalus durissus terrificus venom was investigated in order to differentiate its origin between ophidic envenomation and copper toxicosis. Clinical, laboratory, necroscopic and histological exams as well as evaluation and measurement of heavy metals (copper) in the kidneys and in the liver were carried out. Blood counts revealed anemia and serological tests showed high levels of blood urea nitrogen, creatinine, aspartate aminotransferase, creatine phosphokinase, total bilirubin and indirect bilirubin; which indicates liver, kidney and skeletal muscle damages. At necropsy, the animal presented hepatopathy and nephropathy. Histological examination revealed renal and hepatic features that may imply copper intoxication. Copper levels were 237.8 µg/g in the liver and 51.2 µg/g in the kidneys. Although the amount of metal found in both organs was below the level that can cause death, according to the literature, anatomopathological signs were suggestive of copper intoxication. Therefore, the hypothesis of metal toxicosis during the hyperimmunization process became more consistent than the crotalic envenomation one.(AU)

Laboratory evaluation of young ovines inoculated with natural or 60co-irradiated Crotalus durissus terrificus venom during hyperimmunization process

Laboratory profile of young ovines was studied in order to evaluate and compare their antiserum production from natural and Cobalt-60 irradiated Crotalus durissus terrificus (C.d.t.) venoms. The parameters analyzed included complete blood count, and urea, creatinine, aspartate aminotransferase, total proteins, albumin and globulin serum measurements. Three groups of six animals each were used. Group 1 (G1) received natural C.d.t. venom; Group 2 (G2) received irradiated C.d.t. venom; and Group 3 (G3) was used as control and did not receive venom, only adjuvants, using seven venom inoculations. During the experimental period, animals were fortnightly weighed. According to clinical and weight evaluation, sheep in post-weaning phase showed no changes in their physiological profiles but had excellent weight gain. The parameters analyzed were not statistically different (p 5%) among the groups tested. The hyperimmunization process was successfully accomplished with the production of specific antibodies against Crotalus durissus terrificus venom. Results bring a new possibility of utilizing ovines in the commercial production of anticrotalic serum, which may be used to treat human and animal envenomation. Its production cost may be reduced by subsequent use of hyperimmunized sheep for human consumption.

Immunization with native and cobalt 60-irradiated Crotalus durissus terrificus venom in swiss mice: assessment of the neutralizing potency of antisera

ELISA was used to evaluate, follow, and compare the humoral immune response of Swiss mice during hyperimmunization with natural and Cobalt 60-irradiated (60Co) Crotalus durissus terrificus venom. Potency and neutralization were evaluated by in vitro challenges. After hyperimmunization, immunity was observed by "in vivo" challenge and the side effects were assessed. The animals immunization with one LD50 of the venom was on days one, 15, 21, 30, and 45, when blood samples were collected; the challenges occurred on the 60th day. Results showed that ELISA was efficient in evaluating, following, and comparing mouse immune response during hyperimmunization. Serum titers produced with natural venom were similar to those produced with irradiated venom. Immunogenic capacity was maintained after 60Co irradiation. Serum produced from Crotalus durissus terrificus irradiated venom showed higher potency and neutralization capacity than that from natural venom. All antibodies were able to neutralize five LD50 from these venoms. Clinical alterations were minimum during hyperimmunization with irradiated venom.

Immunological assessment of mice hyperimmunized with native and Cobalt-60-irradiated Bothrops venoms

ELISA was used to evaluate, accompany, and compare the humoral immune response of Swiss mice during hyperimmunization with native and Cobalt-60-irradiated (60Co) venoms of Bothrops jararaca, Bothrops jararacussu and Bothrops moojeni. Potency and neutralization were evaluated by in vitro challenges. After hyperimmunization, immunity was observed by in vivo challenge, and the side effects were assessed. The animals immunization with one LD50 of each venom occurred on days 1, 15, 21, 30, and 45, when blood samples were collected; challenges happened on the 60th day. Results showed that ELISA was efficient in evaluating, accompanying and comparing mouse immune response during hyperimmunization. Serum titers produced with natural venom were similar to those produced with irradiated venom. Immunogenic capacity was maintained after 60Co-irradiation. The sera produced with native venom showed neutralizing potency and capacity similar to those of the sera produced with irradiated venom. All antibodies were able to neutralize five LD50 from these venoms. Clinical alterations were minimum during hyperimmunization with irradiated venom, however, necrosis and death occurred in animals inoculated with native venom.

Model of sanitary management for the eradication of bovine herpesvirus 1 (HVB-1) in a dairy herd / Modelo de manejo sanitário para erradicação do herpesvírus bovino tipo 1 (HVB-1) em rebanho bovino leiteiro

This paper describes the efficacy of sanitary management on the eradication of HVB- 1 by means of hyperimmunization, that consists in infected animals vaccination in order to decrease the level of viral reactivation in carriers. Seroneutralization in microplates was the technique used for HVB-1 diagnosis in a dairy herd bred extensively. In the first year of the studies the occurrence of seroreagents older than six month old was 44.5% (166/373). After that, other evaluations were performed in non-reagents cows and calves. From the second year on, hyperimmunization was performed every six months on seropositive animals older than six months of age, using the inactivated monovalent vaccine with oily adjuvant. The greatest occurrence of seroreagents was observed in older animals in productive and reproductive phases. The rate of seroreagents decreased with the gradual culling of these animals, and three years after the establishment of the sanitary program, the number of seroreagents decreased to zero. In the first year, incidence was equal to 0.5% (2/371), in the second year, 0.8% (3/387), and was zero in the third year. Calves born from seroreagent cows became non-reagent after the disappearance of colostral immunity. Annual reduction in the number of reagent animals was due to the culling of seropositive females, substituted by non-reagent heifers. Fight against HVB-1 by monito

Avaliou-se a eficácia de manejo sanitário para erradicação do HVB-1 em rebanho leiteiro criado extensivamente por meio da hiperimunização, que consiste na vacinação de animais infectados pelo vírus visando diminuir a reativação viral de animais portadores. Utilizou-se a técnica de soroneutralização em microplacas para diagnóstico do HVB-1. A ocorrência de animais sororeagentes com idade superior a seis meses de idade no primeiro ano foi 44,5% (166/ 373). Posteriormente, outras avaliações foram realizadas em animais não reagentes e em bezerros. A partir do segundo ano, a hiperimunização foi realizada semestralmente nos animais soropositivos com idade superior a seis meses, aplicando-se vacina monovalente inativada com adjuvante oleoso. A maior ocorrência de animais sororeagentes foi observada nos animais mais velhos, em fase produtiva e reprodutiva. A taxa de sororeagentes decresceu com o descarte gradual destes animais e, três anos após a implantação das medidas sanitárias, chegou a zero. No primeiro ano, a incidência foi igual a 0,5% (2/371), no segundo, 0,8% (3/387) e zero no terceiro. Os bezerros filhos de mães sororeagentes tornaram-se não reagentes após o desaparecimento da imunidade colostral. A redução da ocorrência de ano para ano deveu-se ao descarte das matrizes sororeagentes e reposição com novilhas não reagentes. O combate do HVB-1 utilizando o monitoramento de indi

Model of sanitary management for the eradication of bovine herpesvirus 1 (HVB-1) in a dairy herd / Modelo de manejo sanitário para erradicação do herpesvírus bovino tipo 1 (HVB-1) em rebanho bovino leiteiro

This paper describes the efficacy of sanitary management on the eradication of HVB- 1 by means of hyperimmunization, that consists in infected animals vaccination in order to decrease the level of viral reactivation in carriers. Seroneutralization in microplates was the technique used for HVB-1 diagnosis in a dairy herd bred extensively. In the first year of the studies the occurrence of seroreagents older than six month old was 44.5% (166/373). After that, other evaluations were performed in non-reagents cows and calves. From the second year on, hyperimmunization was performed every six months on seropositive animals older than six months of age, using the inactivated monovalent vaccine with oily adjuvant. The greatest occurrence of seroreagents was observed in older animals in productive and reproductive phases. The rate of seroreagents decreased with the gradual culling of these animals, and three years after the establishment of the sanitary program, the number of seroreagents decreased to zero. In the first year, incidence was equal to 0.5% (2/371), in the second year, 0.8% (3/387), and was zero in the third year. Calves born from seroreagent cows became non-reagent after the disappearance of colostral immunity. Annual reduction in the number of reagent animals was due to the culling of seropositive females, substituted by non-reagent heifers. Fight against HVB-1 by monito

Avaliou-se a eficácia de manejo sanitário para erradicação do HVB-1 em rebanho leiteiro criado extensivamente por meio da hiperimunização, que consiste na vacinação de animais infectados pelo vírus visando diminuir a reativação viral de animais portadores. Utilizou-se a técnica de soroneutralização em microplacas para diagnóstico do HVB-1. A ocorrência de animais sororeagentes com idade superior a seis meses de idade no primeiro ano foi 44,5% (166/ 373). Posteriormente, outras avaliações foram realizadas em animais não reagentes e em bezerros. A partir do segundo ano, a hiperimunização foi realizada semestralmente nos animais soropositivos com idade superior a seis meses, aplicando-se vacina monovalente inativada com adjuvante oleoso. A maior ocorrência de animais sororeagentes foi observada nos animais mais velhos, em fase produtiva e reprodutiva. A taxa de sororeagentes decresceu com o descarte gradual destes animais e, três anos após a implantação das medidas sanitárias, chegou a zero. No primeiro ano, a incidência foi igual a 0,5% (2/371), no segundo, 0,8% (3/387) e zero no terceiro. Os bezerros filhos de mães sororeagentes tornaram-se não reagentes após o desaparecimento da imunidade colostral. A redução da ocorrência de ano para ano deveu-se ao descarte das matrizes sororeagentes e reposição com novilhas não reagentes. O combate do HVB-1 utilizando o monitoramento de indi

Variações da crase sangüínea durante a hiperimunização e após sangria e plasmaferese em eqüinos de produção de soro hiperimune anti-crotálico / Variation of blood profile during the hyperimmunization and after bleeding and plasmapherese in equine serum hyperimune anticrotalic\'s production

Para o estabelecimento do quadro sangüíneo de eqüinos durante a hiperimunização para produção de soro hiperimune anti-crotálico e após as sangrias de produção e a plasmaferese, foram utilizados 20 animais, obedecendo as normas do protocolo de hiperimunização do Instituto Butantan. As amostras foram colhidas antes das inoculações do antígeno (veneno crotálico), antes e depois das sangrias de produção, após as plasmafereses e 15, 30 e 45 dias após a última sangria, totalizando 340 colheitas, realizadas em 17 momentos. Nas amostras, foram determinados valores do hemograma (número de hemácias, volume globular, teor de hemoglobina, VCM, HCM, CHCM, número de leucócitos, contagem diferencial de leucócitos) e de alguns parâmetros de bioquímica sérica (ferro, transferrina e bilirrubinas). Significativas variações no quadro hematológico dos eqüinos foram observadas na fase de hiperimunização, caracterizando anemia normocítica, normocrômica, sem alterações significativas do leucograma. Além disso, verificou-se diminuição significativa da sideremia associada ao aumento da concentração de transferrina sérica, permitindo classificar-se a anemia, como do tipo ferropriva. Nas fases de sangrias, observou-se evidentes variações no quadro hematológico demonstrando uma anemia hipocitêmica normocítica hipercrômica pós-sangrias, sem variações nos valores do leucograma, mas com significativas variações nos constituintes bioquímicos (diminuição do ferro sérico e aumento da concentração de transferrina). No período de repouso foi evidente a eficácia da plasmaferese, com pronta , mas parcial recuperação do hemograma e da sideremia dos animais anêmicos, facilitando o rápido retorno para normalidade hematológica, tornando-os aptos a nova produção de soro hiperimune

Twenty animals were used to assess the blood profile of horses during hyperimmunization for anticrotalic hyperimmune serum production and after the bleedings of production and the plasmapherese, obeying the protocol schedule of Instituto Butantan. The samples were obtained before the antigen inoculations (crotalic poison), before and after the bleedings of production, right after the plasmapherese and 15, 30 and 45 days after the last one, making a total of 340 collections in 17 moments. Samples were analyzed for hemogram (number of erythrocytes, packed cell volume, hemoglobin, MCV, MCH, MCHC, white cell count, differential leukocytes count) and some serum biochemical components (iron, transferrin and bilirrubin). Significant variations were observed on the blood profile of the horses throughout the hyperimmunization, showing normocytic, normochromic anemia, without significant alterations on leukogram. Beside that, a significant reduction of the hemosiderin was observed in association with the elevation of the serum transferrin concentration, making possible its classification as an iron deficiency-like anemia. During the bleedings phases, evident variations were observed on the blood profile showing pos-sangrias hypocitemic normocytic, hyperchromic anemia without changing the leukogram values, but with significant variations on the biochemical components (reduction of serum iron and increase of transferrin concentrations). During rest periods, the efficacy of plasmapherese was evident with a rapid but partial recuperation of the hemogram and the hemosiderin of the anemic animals, making easier the fast return of hematologic normality and making them capable to a new hyperimmune serum production