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1.
Braz J Microbiol ; 54(2): 1231-1237, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36897516

RESUMO

Water buffaloes (Bubalus bubalis) have been introduced in many regions of the world as a source of animal protein. In many instances, bubaline cattle are reared close to or mixed with bovine or zebuine cattle. However, little is known about infectious diseases of bubaline and the interactions that may arise involving the microbiota of those species. Alphaherpesviruses of ruminants (bovine alphaherpesviruses types 1 and 5, BoHV-1, BoHV-5; bubaline alphaherpesvirus 1, BuHV-1) are highly cross-reactive in serological assays performed with bovine or zebuine sera. However, the profile of reactivity of bubaline cattle sera to alphaherpesviruses remains unknown. As such, it is not known which virus strain (or strains) would be most appropriate to be used as the challenge virus in the laboratory in search for alphaherpesvirus-neutralizing antibodies. In this study, the profile of neutralizing antibodies to alphaherpesviruses in bubaline sera was determined against different types/subtypes of bovine and bubaline alphaherpesviruses. Sera (n=339) were screened in a 24-h serum neutralization test (SN) against 100 TCID50 of each of the challenge viruses. From those, 159 (46.9 %) neutralized at least one of the viruses assayed; 131 (38.6%) sera neutralized the three viral strains used for screening. The viral strain that was neutralized by the largest number of sera was BoHV-5b A663 (149/159; 93.7%). A few sera neutralized only one of the challenge viruses: four sera neutralized BoHV-1 LA only; another neutralized BoHV-5 A663 only and four others neutralized BuHV-1 b6 only. SN testing with two additional strains gave rise to similar results, where maximum sensitivity (defined here as the largest number of sera that neutralized the challenge viruses) was obtained by adding positive results attained with three of the challenge strains. Differences in neutralizing antibody titers were not significant to allow inferences on which would be the most likely virus that induced the antibody responses detected here.


Assuntos
Alphaherpesvirinae , Infecções por Herpesviridae , Herpesvirus Bovino 1 , Bovinos , Animais , Búfalos , Anticorpos Neutralizantes , Infecções por Herpesviridae/veterinária , Anticorpos Antivirais
2.
Mol Immunol ; 155: 165-174, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36812764

RESUMO

Previously, it was demonstrated that from the single chain fragment variable (scFv) 3F it is possible to generate variants capable of neutralizing the Cn2 and Css2 toxins, as well as their respective venoms (Centruroides noxius and Centruroides suffusus). Despite this success, it has not been easy to modify the recognition of this family of scFvs toward other dangerous scorpion toxins. The analysis of toxin-scFv interactions and in vitro maturation strategies allowed us to propose a new maturation pathway for scFv 3F to broaden recognition toward other Mexican scorpion toxins. From maturation processes against toxins CeII9 from C. elegans and Ct1a from C. tecomanus, the scFv RAS27 was developed. This scFv showed an increased affinity and cross-reactivity for at least 9 different toxins while maintaining recognition for its original target, the Cn2 toxin. In addition, it was confirmed that it can neutralize at least three different toxins. These results constitute an important advance since it was possible to improve the cross-reactivity and neutralizing capacity of the scFv 3F family of antibodies.


Assuntos
Venenos de Escorpião , Animais , Humanos , Sequência de Aminoácidos , Caenorhabditis elegans , Anticorpos Neutralizantes , Fragmentos de Imunoglobulinas
3.
Artigo em Inglês | MEDLINE | ID: mdl-35432492

RESUMO

Background: A new pit viper, Protobothrops kelomohy, has been recently discovered in northern and northwestern Thailand. Envenoming by the other Protobothrops species across several Asian countries has been a serious health problem since their venom is highly hematotoxic. However, the management of P. kelomohy bites is required as no specific antivenom is available. This study aimed to investigate the biochemical properties and proteomes of P. kelomohy venom (PKV), including the cross-neutralization to its lethality with antivenoms available in Thailand. Methods: PKV was evaluated for its neutralizing capacity (ER50), lethality (LD50), procoagulant and hemorrhagic effects with three monovalent antivenoms (TAAV, DSAV, and CRAV) and one polyvalent (HPAV) hematotoxic antivenom. The enzymatic activities were examined in comparison with venoms of Trimeresurus albolabris (TAV), Daboia siamensis (DSV), Calloselasma rhodostoma (CRV). Molecular mass was separated on SDS-PAGE, then the specific proteins were determined by western blotting. The venom protein classification was analyzed using mass spectrometry-based proteomics. Results: Intravenous LD50 of PKV was 0.67 µg/g. ER50 of HPAV, DSAV and TAAV neutralize PKV at 1.02, 0.36 and 0.12 mg/mL, respectively. PKV exhibited procoagulant effect with a minimal coagulation dose of 12.5 ± 0.016 µg/mL and hemorrhagic effect with a minimal hemorrhagic dose of 1.20 ± 0.71 µg/mouse. HPAV was significantly effective in neutralizing procoagulant and hemorrhagic effects of PKV than those of TAAV, DSAV and CRAV. All enzymatic activities among four venoms exhibited significant differences. PKV proteome revealed eleven classes of putative snake venom proteins, predominantly metalloproteinase (40.85%), serine protease (29.93%), and phospholipase A2 (15.49%). Conclusions: Enzymatic activities of PKV are similarly related to other viperid venoms in this study by quantitatively hematotoxic properties. Three major venom toxins were responsible for coagulopathy in PKV envenomation. The antivenom HPAV was considered effective in neutralizing the lethality, procoagulant and hemorrhagic effects of PKV.

4.
J. venom. anim. toxins incl. trop. dis ; 28: e20210080, 2022. graf, tab
Artigo em Inglês | VETINDEX | ID: biblio-1395757

RESUMO

Background: A new pit viper, Protobothrops kelomohy, has been recently discovered in northern and northwestern Thailand. Envenoming by the other Protobothrops species across several Asian countries has been a serious health problem since their venom is highly hematotoxic. However, the management of P. kelomohy bites is required as no specific antivenom is available. This study aimed to investigate the biochemical properties and proteomes of P. kelomohy venom (PKV), including the cross-neutralization to its lethality with antivenoms available in Thailand. Methods: PKV was evaluated for its neutralizing capacity (ER50), lethality (LD50), procoagulant and hemorrhagic effects with three monovalent antivenoms (TAAV, DSAV, and CRAV) and one polyvalent (HPAV) hematotoxic antivenom. The enzymatic activities were examined in comparison with venoms of Trimeresurus albolabris (TAV), Daboia siamensis (DSV), Calloselasma rhodostoma (CRV). Molecular mass was separated on SDS-PAGE, then the specific proteins were determined by western blotting. The venom protein classification was analyzed using mass spectrometry-based proteomics. Results: Intravenous LD50 of PKV was 0.67 µg/g. ER50 of HPAV, DSAV and TAAV neutralize PKV at 1.02, 0.36 and 0.12 mg/mL, respectively. PKV exhibited procoagulant effect with a minimal coagulation dose of 12.5 ± 0.016 µg/mL and hemorrhagic effect with a minimal hemorrhagic dose of 1.20 ± 0.71 µg/mouse. HPAV was significantly effective in neutralizing procoagulant and hemorrhagic effects of PKV than those of TAAV, DSAV and CRAV. All enzymatic activities among four venoms exhibited significant differences. PKV proteome revealed eleven classes of putative snake venom proteins, predominantly metalloproteinase (40.85%), serine protease (29.93%), and phospholipase A2 (15.49%). Conclusions: Enzymatic activities of PKV are similarly related to other viperid venoms in this study by quantitatively hematotoxic properties. Three major venom toxins were responsible for coagulopathy in PKV envenomation. The antivenom HPAV was considered effective in neutralizing the lethality, procoagulant and hemorrhagic effects of PKV.(AU)


Assuntos
Animais , Venenos de Víboras/análise , Fenômenos Bioquímicos/fisiologia , Proteômica/métodos , Tailândia , Antivenenos/análise
5.
Avian Dis ; 65(3): 456-462, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34699143

RESUMO

In this study we evaluated the effectiveness of adding serotype 793B vaccine to an immunization program in order to control the infectious bronchitis virus (IBV) GI-16 lineage. Therefore, two different experiments were performed. First, a virus cross-neutralization test was carried out, which indicated that neither the Massachusetts (Mass) nor 793B serotypes are antigenically related to the field isolate A13 (GI-16). We also performed a challenge trial to evaluate if the Mass/793B combination is more efficient than Mass/Connecticut (Conn) to protect chickens against the Argentinian variant A13. Thus, 40 chickens were organized in four groups. Chickens in Group A were vaccinated at 1 day of age with Mass serotype and then at 14 days old with Mass plus Conn serotypes. Chickens in Group B received Mass and 793B serotypes at 1 and 14 days old, respectively. Groups C and D remained unvaccinated. At 28 days of age, Groups A, B, and C were challenged with the A13 isolate, while Group D remained as the negative control. The statistical analysis of the ciliostasis evaluation, performed at 7 days postchallenge (dpch), indicated that the difference between Mass/793B and Mass/Conn was not significant (p > 0.05). However, the comparison against the negative control showed that only Group A was significantly different, suggesting a slightly better performance on blocking ciliostasis for the Mass/793B combination. On the other hand, no significant differences were observed in the viral load, quantified by reverse-transcription quantitative real-time PCR (RT-qPCR) in tracheal swabs and kidneys (at 3 and 7 dpch, respectively) between vaccinated groups. Furthermore, some amounts of the viral genome were found in both vaccinated groups that could indicate that neither the Mass/793B nor the Mass/Conn combinations totally inhibited the viral replication. Such viral replication in vaccinated chickens should seriously be taken into consideration because it could promote the selection of new variants in the future.


Nota de investigación­Evaluación de la eficacia de vacunas comerciales contra el virus de la bronquitis infecciosa (IBV) perteneciente al linaje GI-16 aislado durante un brote argentino. En este estudio se evaluó la efectividad de agregar la vacuna del serotipo 793B a un programa de inmunización para controlar al virus de la bronquitis infecciosa (con las siglas en inglés IBV) linaje GI-16. Por tanto, se realizaron dos experimentos diferentes. Primeramente, se llevó a cabo una prueba de neutralización cruzada de virus, que indicó que ni los serotipos Massachusetts (Mass) ni 793B están antigénicamente relacionados con el aislado de campo A13 (GI-16). También se realizó una prueba de desafío para evaluar si la combinación Massachussets/793B era más eficiente que Massachussets/Connecticut (Conn) para proteger a los pollos contra la variante argentina A13. De esta forma, 40 pollos se organizaron en cuatro grupos. Los pollos del Grupo A se vacunaron al día de edad con el serotipo Massachussets y luego a los 14 días con los serotipos Massachussets más Connecticut. Los pollos del Grupo B recibieron los serotipos Massachussets y 793B a los 1 y 14 días de edad, respectivamente. Los grupos C y D permanecieron sin vacunar. A los 28 días de edad, los Grupos A, B y C fueron desafiados con el aislado A13, mientras que el Grupo D permaneció como control negativo. El análisis estadístico de la evaluación de la ciliostasis, realizada a los 7 días después del desafío (dpch), indicó que la diferencia entre el tratamiento Massachussets/793B y Massachussets/Connecticut no fue significativa (P> 0.05). Sin embargo, la comparación con el control negativo mostró que solo el Grupo A fue significativamente diferente, lo que sugiere un desempeño ligeramente mejor en el bloqueo de la ciliostasis para la combinación Massachussets/793B. Por otro lado, no se observaron diferencias significativas (P> 0.05) en la carga viral, cuantificada mediante transcripción reversa y PCR cuantitativa en tiempo real de hisopos traqueales y riñones (a 3 y 7 días después del desafío, respectivamente) entre los grupos vacunados. Además, se encontraron algunas cantidades del genoma viral en ambos grupos vacunados que podrían indicar que ni las combinaciones Massachussets/793B ni Massachussets/Connecticut inhibieron totalmente la replicación viral. Esta replicación viral en pollos vacunados debe tenerse muy en cuenta porque podría promover la selección de nuevas variantes en el futuro.


Assuntos
Infecções por Coronavirus , Vírus da Bronquite Infecciosa , Doenças das Aves Domésticas , Vacinas Virais , Animais , Galinhas , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/veterinária , Surtos de Doenças/prevenção & controle , Surtos de Doenças/veterinária , Vírus da Bronquite Infecciosa/genética , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/prevenção & controle
6.
Biologicals ; 68: 40-45, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32928631

RESUMO

New world Coral snakes comprise 82 species of medical importance distributed from southeastern United States to Argentina. In Colombia, Micrurus mipartitus and M. dumerilii are responsible for most coral snakebite accidents. Although infrequent, the severity of these envenomings, as well as the limited information available on the neutralizing coverage of commercially available antivenoms, underscores the need to perform studies to assess the cross-neutralizing ability of these life-saving immunobiologicals. In the present work, we evaluated the cross-recognition and neutralization ability of two equine therapeutic antivenoms: PROBIOL and SAC-ICP. PROBIOL antivenom showed cross-recognition towards both M. mipartitus and M. dumerilii venoms, with a significantly higher binding to the latter in both whole-venom ELISA and fractionated-venom immunoprofiling. In contrast, SAC-ICP antivenom cross-recognized M. dumerilii venom, but not that of M. mipartitus. Lethality of M. dumerilii venom was neutralized by both antivenoms, with a slightly higher potency for the SAC-ICP antivenom. However, the lethality of M. mipartitus venom was not neutralized by any of the two antivenoms. Results uncover the need to include M. mipartitus venom, or its most relevant toxins, in the production of coral snake antivenoms to be used in Colombia, to assure the neutralizing coverage for this species.


Assuntos
Antivenenos/imunologia , Cobras Corais/imunologia , Venenos Elapídicos/imunologia , Cavalos/imunologia , Mordeduras de Serpentes/imunologia , Animais , Antivenenos/administração & dosagem , Colômbia , Cobras Corais/classificação , Reações Cruzadas/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Masculino , Camundongos , Testes de Neutralização/métodos , Mordeduras de Serpentes/prevenção & controle , Especificidade da Espécie
7.
Toxicon ; 179: 92-100, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32345455

RESUMO

Specimens of the Crotalus genus represent a potential snakebite problem in Mexico, and despite the great number of species of Crotalus present in this country, only a few of them are relevant from a medical point of view. Crotalus envenomed patients can present a range of signs and symptoms, depending on the species involved, and their treatment is indistinctly with either of the anti-viperid antivenoms available in the Mexican Public Health System. One of these antivenoms is produced by immunization of horses with a mixture of only two venoms: Crotalus basiliscus and Bothrops asper venoms. In light of the high variability found in Crotalus species venom composition, it is important to demonstrate the cross-neutralization of this antivenom against other Crotalus species. Therefore, in this work the toxic variability of eight medically important Crotalus venoms from Mexico and its neutralization by the Crotalus basiliscus/Bothrops asper antivenom were assessed. The present study evidenced the variability of toxic and enzymatic activities among the following Crotalus venoms: (1) Crotalus atrox, (2) Crotalus basiliscus, (3) Crotalus culminatus, (4) Crotalus simus, (5) Crotalus tzabcan, (6) Crotalus scutulatus salvini, (7) Crotalus scutulatus scutulatus-A, and (8) Crotalus scutulatus scutulatus-B. All venoms studied possess lethal and hemorrhagic activity on a murine model, although there are important variations among the species; in contrast, the PLA2 activity was similar for all venoms. Interestingly, only C. simus venom exhibited coagulant activity on human plasma under 100 µg. The antivenom neutralized the lethality and all the other assessed activities for all venoms tested. However, the dose required varied depending on the venom and the evaluated activity. Our preclinical data support the recommendation of using this antivenom to clinically manage Crotalus snakebites produced by the species assessed in this study. Nonetheless, only clinical trials could categorically validate these results.


Assuntos
Antivenenos , Venenos de Crotalídeos/toxicidade , Crotalus , Animais , Bothrops , Venenos de Crotalídeos/química , Hemorragia , Cavalos , Humanos , México , Testes de Neutralização , Mordeduras de Serpentes
8.
Avian Pathol ; 48(3): 270-277, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30761909

RESUMO

Infectious bronchitis virus (IBV) is a persistent sanitary problem for the South American poultry industry despite extensive vaccination. The IBV single-stranded RNA genome has high rates of mutation and recombination that generate a notorious virus variability. Since most IBV vaccines are type-specific, there is a need for constant surveillance of the circulating lineages and knowledge about their genetic and antigenic properties. Here we present an integrative analysis that provides the pattern of genetic variation of the South American IBV strains and information about their antigenic characteristics. The genetic analysis was performed using the S1 complete coding sequences of all available South American strains, including newly obtained Argentine and Uruguayan field samples. Our phylogenetic and phylodynamic analyses evidence that three main lineages (GI-1, GI-11 and GI-16) are extensively circulating in South American flocks. Strains of the GI-1 lineage (Massachusetts-type) were detected in Argentina, Brazil, Chile and Colombia. The GI-11 lineage is an exclusively South American lineage that emerged in the 1950s, and is the predominant lineage in Brazil and Uruguay at present. The GI-16 lineage emerged around 1979, and is currently circulating in most South American territories (Argentina, Chile, Uruguay, Colombia and Peru). The virus cross-neutralization test performed here reveals very low antigenic relatedness between GI-11 and GI-16 lineages (i.e. they are different serotypes). The results of this study extend our knowledge about the present and past IBV variability in South America and provide relevant elements to improve the control programmes by considering the genetic and antigenic attributes of IBV.


Assuntos
Galinhas/virologia , Infecções por Coronavirus/veterinária , Vírus da Bronquite Infecciosa/imunologia , Doenças das Aves Domésticas/prevenção & controle , Animais , Variação Antigênica/genética , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/virologia , Genótipo , Vírus da Bronquite Infecciosa/genética , Doenças das Aves Domésticas/virologia , América do Sul
9.
Toxins (Basel) ; 11(1)2019 01 10.
Artigo em Inglês | MEDLINE | ID: mdl-30634620

RESUMO

The recombinant antibody fragments generated against the toxic components of scorpion venoms are considered a promising alternative for obtaining new antivenoms for therapy. Using directed evolution and site-directed mutagenesis, it was possible to generate a human single-chain antibody fragment with a broad cross-reactivity that retained recognition for its original antigen. This variant is the first antibody fragment that neutralizes the effect of an estimated 13 neurotoxins present in the venom of nine species of Mexican scorpions. This single antibody fragment showed the properties of a polyvalent antivenom. These results represent a significant advance in the development of new antivenoms against scorpion stings, since the number of components would be minimized due to their broad cross-neutralization capacity, while at the same time bypassing animal immunization.


Assuntos
Anticorpos Neutralizantes/imunologia , Neurotoxinas/imunologia , Venenos de Escorpião/imunologia , Anticorpos de Cadeia Única/imunologia , México
10.
Vet Microbiol ; 168(2-4): 324-30, 2014 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-24388627

RESUMO

Llamas and alpacas are domesticated South American camelids (SACs) important to ancestral population in the Altiplano region, and to different communities where they have been introduced worldwide. These ungulates have shown to be susceptible to several livestock viral pathogens such as members of the Pestivirus genus and mainly to bovine viral diarrhea virus (BVDV). Seventeen Chilean BVDV isolates were analyzed by serum cross neutralization with samples obtained from five llama, six alpacas, three bovines, plus three reference strains belonging to different subgroups and genotypes. The objective was to describe antigenic differences and similarities among them. Antigenic comparison showed significant differences between different subgroups. Consequently, antigenic similarities were observed among isolates belonging to the same subgroup and also between isolates from different animal species belonging the same subgroup. Among the analyzed samples, one pair of 1b subgroup isolates showed significant antigenic differences. On the other hand, one pair of isolates from different subgroups (1b and 1j) shared antigenic similarities indicating antigenic relatedness. This study shows for the first time the presence of antigenic differences within BVDV 1b subgroup and antigenic similarities within 1j subgroup isolates, demonstrating that genetic differences within BVDV subgroups do not necessary corresponds to differences on antigenicity.


Assuntos
Variação Antigênica , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Camelídeos Americanos/virologia , Vírus da Diarreia Viral Bovina Tipo 1/imunologia , Vírus da Diarreia Viral Bovina Tipo 2/imunologia , Animais , Antígenos Virais/imunologia , Bovinos/virologia , Chile , Diarreia/veterinária , Diarreia/virologia , Vírus da Diarreia Viral Bovina Tipo 1/genética , Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Vírus da Diarreia Viral Bovina Tipo 2/genética , Vírus da Diarreia Viral Bovina Tipo 2/isolamento & purificação , Genótipo , Testes de Neutralização
11.
Pesqui. vet. bras ; 30(7): 515-522, 2010. ilus, tab
Artigo em Inglês | VETINDEX | ID: vti-14247

RESUMO

The serum neutralization (SN) test is the gold standard method to measure neutralizing antibodies to bovine herpesviruses. However, in view of the further subdivisions of bovine herpesviruses in types/subtypes, defining which virus to use at challenge in SN tests may be difficult. In view of that, this study was carried out to re-evaluate (SN) sensitivity with different types/subtypes of bovine herpesviruses types 1 (BoHV-1) and 5 (BoHV-5) as challenge viruses. Bovine sera (n=810) were collected from two distinct geographic regions and tested by SN with three type 1 viruses (BoHV-1.1 strains "Los Angeles" and "EVI123/98"; BoHV-1.2a strain "SV265/96") and three type 5 viruses (BoHV-5a strain "EVI88/95"; BoHV-5b strain "A663" and BoHV-5c "ISO97/95"). SN tests were performed with a 1 hour incubation of the serum-virus mixtures at 37ºC against 100 TCID50 of each of the viruses. SN sensitivity varied greatly depending on the challenge virus used in the test. The highest sensitivity (327 positive/810 total sera tested; 40.37 percent) was attained when the positive results to the six viruses were added together. No association could be found between any particular type or subtype of virus and the sensitivity of the test. When positive results to each single strain were considered, SN sensitivity varied from 41.7 percent to 81.7 percent, depending on the virus and the geographic region of origin of the sera. Variation was detected even when challenge viruses belonged to the same subtype, where disagreement between positive results reached 41 percent. These results indicate that one hour incubation SN tests against single viruses, as performed here, may display a significantly low sensitivity (p=0.05); performing SN tests against a number of different viruses may increase considerably SN sensitivity. Furthermore, the choice of virus used for challenge is critical in SN tests. In addition, sera from different geographic regions may give rise to disagreeing results with different strains of BoHV-1 and BoHV-5. This might be particularly relevant for control programs and in international trade, were maximum sensitivity should be targeted.(AU)


O teste de soroneutralização (SN) é o método padrão para a mensuração de anticorpos neutralizantes para herpesvírus bovinos. Entretanto, com as subdivisões propostas destes agentes em tipos e subtipos, a definição de qual amostra utilizar como virus de desafio à SN pode ser difícil. Em vista disso, este estudo foi realizado para re-avaliar a sensibilidade de testes de SN utilizando diferentes tipos e subtipos de herpesvírus bovinos tipos 1 (BoHV-1) e 5 (BoHV-5) como amostras de desafio. Soros bovinos (n=810) foram coletados de duas regiões geográficas distintas e testados frente a amostras do tipo 1 (BoHV-1.1: amostras "Los Angeles" e "EVI123/98", BoHV-1.2a: amostra "SV265/96") e três amostras do tipo 5 (BoHV-5a: "EVI88/95"; BoHV-5b: "A663" e BoHV-5c "ISO97/95"). Os testes de SN foram realizados com incubação de 1 hora a 37ºC da mistura soro-vírus, frente a 100 doses infectantes para 50 por cento dos cultivos celulares (DICC50) de cada um dos vírus. A sensibilidade da SN variou grandemente em função do vírus utilizado no teste. A maior sensibilidade (327 soros positivos/810 soros testados; 40.37 por cento) foi alcançada quando os resultados positivos frente aos seis diferentes vírus foram somados. Nenhuma associação foi detectada entre determinado tipo/subtipo de vírus e a sensibilidade do teste. Quando resultados positivos frente a cada vírus foram considerados isoladamente, a sensibilidade da SN variou entre 41,7 por cento a 81,7 por cento, dependendo do vírus de desafio e da região geográfica de origem das amostras de soro. Variação foi detectada mesmo quando as amostras de desafio pertenciam a um mesmo subtipo; a discrepância entre os resultados positivos atingiu até 41 por cento. Estes resultados indicam que testes de SN contra amostras isoladas de vírus podem apresentar uma sensibilidade notadamente baixa; o emprego de diferentes amostras de vírus de desafio pode aumentar consideravelmente a sensibilidade da prova. Além disso, a escolha da amostra de vírus para a realização do teste é crítica. Outro achado importante é que sorors de diferentes regiões geográficas podem dar resultados discordantes frente a diferentes amostras de BoHV-1 e BoHV-5. Estes achados são particularmente relevantes para programas de controle destas infecções e para o comércio internacional, onde a sensibilidade deve ser maximizada.(AU)


Assuntos
Animais , Bovinos , Testes de Neutralização/instrumentação , Herpesvirus Bovino 1 , Herpesvirus Bovino 5 , Controle de Doenças Transmissíveis
12.
Pesqui. vet. bras ; Pesqui. vet. bras;30(7): 515-522, July 2010. ilus, tab
Artigo em Inglês | LILACS | ID: lil-557293

RESUMO

The serum neutralization (SN) test is the gold standard method to measure neutralizing antibodies to bovine herpesviruses. However, in view of the further subdivisions of bovine herpesviruses in types/subtypes, defining which virus to use at challenge in SN tests may be difficult. In view of that, this study was carried out to re-evaluate (SN) sensitivity with different types/subtypes of bovine herpesviruses types 1 (BoHV-1) and 5 (BoHV-5) as challenge viruses. Bovine sera (n=810) were collected from two distinct geographic regions and tested by SN with three type 1 viruses (BoHV-1.1 strains "Los Angeles" and "EVI123/98"; BoHV-1.2a strain "SV265/96") and three type 5 viruses (BoHV-5a strain "EVI88/95"; BoHV-5b strain "A663" and BoHV-5c "ISO97/95"). SN tests were performed with a 1 hour incubation of the serum-virus mixtures at 37ºC against 100 TCID50 of each of the viruses. SN sensitivity varied greatly depending on the challenge virus used in the test. The highest sensitivity (327 positive/810 total sera tested; 40.37 percent) was attained when the positive results to the six viruses were added together. No association could be found between any particular type or subtype of virus and the sensitivity of the test. When positive results to each single strain were considered, SN sensitivity varied from 41.7 percent to 81.7 percent, depending on the virus and the geographic region of origin of the sera. Variation was detected even when challenge viruses belonged to the same subtype, where disagreement between positive results reached 41 percent...


O teste de soroneutralização (SN) é o método padrão para a mensuração de anticorpos neutralizantes para herpesvírus bovinos. Entretanto, com as subdivisões propostas destes agentes em tipos e subtipos, a definição de qual amostra utilizar como virus de desafio à SN pode ser difícil. Em vista disso, este estudo foi realizado para re-avaliar a sensibilidade de testes de SN utilizando diferentes tipos e subtipos de herpesvírus bovinos tipos 1 (BoHV-1) e 5 (BoHV-5) como amostras de desafio. Soros bovinos (n=810) foram coletados de duas regiões geográficas distintas e testados frente a amostras do tipo 1 (BoHV-1.1: amostras "Los Angeles" e "EVI123/98", BoHV-1.2a: amostra "SV265/96") e três amostras do tipo 5 (BoHV-5a: "EVI88/95"; BoHV-5b: "A663" e BoHV-5c "ISO97/95"). Os testes de SN foram realizados com incubação de 1 hora a 37ºC da mistura soro-vírus, frente a 100 doses infectantes para 50 por cento dos cultivos celulares (DICC50) de cada um dos vírus. A sensibilidade da SN variou grandemente em função do vírus utilizado no teste. A maior sensibilidade (327 soros positivos/810 soros testados; 40.37 por cento) foi alcançada quando os resultados positivos frente aos seis diferentes vírus foram somados. Nenhuma associação foi detectada entre determinado tipo/subtipo de vírus e a sensibilidade do teste. Quando resultados positivos frente a cada vírus foram considerados isoladamente, a sensibilidade da SN variou entre 41,7 por cento a 81,7 por cento, dependendo do vírus de desafio e da região geográfica de origem das amostras de soro. Variação foi detectada mesmo quando as amostras de desafio pertenciam a um mesmo subtipo; a discrepância entre os resultados positivos atingiu até 41 por cento. Estes resultados indicam que testes de SN contra amostras isoladas de vírus podem apresentar uma sensibilidade notadamente baixa; o emprego de diferentes amostras de vírus de desafio pode aumentar consideravelmente a sensibilidade da prova...


Assuntos
Animais , Bovinos , Herpesvirus Bovino 1 , Testes de Neutralização/instrumentação , Controle de Doenças Transmissíveis
13.
Ci. Rural ; 32(5)2002.
Artigo em Português | VETINDEX | ID: vti-704069

RESUMO

Calves were immunized with two vaccines against bovine herspevirus type 1 (BHV-1; vaccine A, n=28; B, n=28) or with a vaccine containing BHV-1 and BHV-5 antigens (vaccine C, n=32) and the serum neutralizing activity against BHV-1 and BHV-5 was measured after three vaccine administrations. Neutralizing activity to BHV-1 was detected in sera of 100% (n=88) of the animals (geometric mean titers [GMT] of 13.1; 14.8 and 34.3 for vaccines A, B and C, respectively) whereas 82 sera (93.2%) reacted to BHV-5 (GMTs: 10.6; 11.5 and 29.8). In all three groups, the mean BHV-1 titers did not differ from BHV-5 titers. However, comparison among the vaccines demonstrated that the mean titers to BHV-1 and BHV-5 were higher in animals receiving vaccine C (p 0.01). This vaccine also induced a higher proportion of reagents to BHV-5 (96.9%; against 85.7% for vaccine A and 92.9% for vaccine B). These results demonstrate that vaccine C induced higher neutralizing antibody titers against both viruses; and that antibodies induced by BHV-1 antigens (vaccines A and B) cross-reacted and displayed relevant neutralizing activity against BHV-5.


Bezerros foram imunizados com duas vacinas contra o herpesvírus bovino tipo 1 (BHV-1; vacina A, n=28; B, n=28) ou com uma vacina contendo antígenos do BHV-1 e BHV-5 (n=32) e a atividade neutralizante sérica antiBHV-1 e BHV-5 foi testada após três doses vacinais. Todos os animais (n=88) produziram anticorpos com atividade neutralizante antiBHV-1 (títulos médios geométricos [GMT] de 13,1; 14,8 e 34,3 para as vacinas A, B e C, respectivamente) e 82 animais (93,2%) desenvolveram atividade neutralizante antiBHV-5 (GMTs: 10,6; 11,5 e 29,8, respectivamente). Nos três grupos vacinais, os títulos médios antiBHV-1 não diferiram dos títulos antiBHV-5. No entanto, comparando-se os títulos médios entre as vacinas, os títulos neutralizantes para o BHV-1 e BHV-5 foram superiores nos animais do grupo C (p 0,01 ). Esse grupo também apresentou a maior proporção de reagentes ao BHV-5 (96,9%; contra 85,7% da vacina A e 92,9% da vacina B). Esses resultados demonstram que a vacina C induziu títulos neutralizantes superiores contra os dois vírus e que anticorpos induzidos por antígenos do BHV-1 (vacinas A e B) possuem atividade neutralizante relevante também contra o BHV-5.

14.
Artigo em Português | LILACS-Express | VETINDEX | ID: biblio-1475839

RESUMO

Calves were immunized with two vaccines against bovine herspevirus type 1 (BHV-1; vaccine A, n=28; B, n=28) or with a vaccine containing BHV-1 and BHV-5 antigens (vaccine C, n=32) and the serum neutralizing activity against BHV-1 and BHV-5 was measured after three vaccine administrations. Neutralizing activity to BHV-1 was detected in sera of 100% (n=88) of the animals (geometric mean titers [GMT] of 13.1; 14.8 and 34.3 for vaccines A, B and C, respectively) whereas 82 sera (93.2%) reacted to BHV-5 (GMTs: 10.6; 11.5 and 29.8). In all three groups, the mean BHV-1 titers did not differ from BHV-5 titers. However, comparison among the vaccines demonstrated that the mean titers to BHV-1 and BHV-5 were higher in animals receiving vaccine C (p 0.01). This vaccine also induced a higher proportion of reagents to BHV-5 (96.9%; against 85.7% for vaccine A and 92.9% for vaccine B). These results demonstrate that vaccine C induced higher neutralizing antibody titers against both viruses; and that antibodies induced by BHV-1 antigens (vaccines A and B) cross-reacted and displayed relevant neutralizing activity against BHV-5.


Bezerros foram imunizados com duas vacinas contra o herpesvírus bovino tipo 1 (BHV-1; vacina A, n=28; B, n=28) ou com uma vacina contendo antígenos do BHV-1 e BHV-5 (n=32) e a atividade neutralizante sérica antiBHV-1 e BHV-5 foi testada após três doses vacinais. Todos os animais (n=88) produziram anticorpos com atividade neutralizante antiBHV-1 (títulos médios geométricos [GMT] de 13,1; 14,8 e 34,3 para as vacinas A, B e C, respectivamente) e 82 animais (93,2%) desenvolveram atividade neutralizante antiBHV-5 (GMTs: 10,6; 11,5 e 29,8, respectivamente). Nos três grupos vacinais, os títulos médios antiBHV-1 não diferiram dos títulos antiBHV-5. No entanto, comparando-se os títulos médios entre as vacinas, os títulos neutralizantes para o BHV-1 e BHV-5 foram superiores nos animais do grupo C (p 0,01 ). Esse grupo também apresentou a maior proporção de reagentes ao BHV-5 (96,9%; contra 85,7% da vacina A e 92,9% da vacina B). Esses resultados demonstram que a vacina C induziu títulos neutralizantes superiores contra os dois vírus e que anticorpos induzidos por antígenos do BHV-1 (vacinas A e B) possuem atividade neutralizante relevante também contra o BHV-5.

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