Commercial mastitis vaccines - effects on mastitis rates and somatic cell counts in Saanen goats

Background: Staphylococcus spp. are the most frequently isolated microorganisms in mastitis cases of small ruminants. The virulence factors of Staphylococcus spp. are critical in the treatment. Therefore, preventive medicine and mastitis control programs, especially herd vaccinations are of great importance in the prevention of mastitis. However, it is not always easy to obtain these vaccines under field conditions. This study, it was aimed to compare the effects of different commercial Staphylococcus spp. vaccines licensed for bovines and species-specific mastitis vaccines on mastitis rates and somatic cell count (SCC) on Saanen goats on field conditions. Materials, Methods & Results: The animal material consisted of 115 (230 udder halves) nulliparous Saanen goats. Goats were randomly grouped as bovine vaccine 1 (BV1 , n = 58), bovine vaccine 2 (BV2 , n = 58), small ruminant vaccine (SRV, n = 56), and control (n = 56). Vaccines were administered to goats in 2 doses according to the label regimen. First milk samples were collected between 0-5 days in milk (DIM) for microbiological analysis and 25-35 DIM for SCC. The other milk samples were collected at 25-35 (1st month) DIM, 60-65 (2nd month) DIM, 85-95 (3rd month) DIM, 115-125 (4th month) DIM, 145-155 (5th month) DIM for microbiological analysis and SCC. Non-aureus staphylococci (NAS) and Staphylococcus aureus were the most frequently isolated microorganism. It was found that the total mastitis rate decreased in vaccine groups compared to the control group. A significant difference was found only in the BV2 and SRV groups. The significant difference in S. aureus infection was found only in the SRV group. Mastitis vaccines used in this study decreased the NAS mastitis rate, but no significant difference was observed. It was found that the clinical mastitis incidence decreased in all vaccine groups compared to the control group, and a significant difference was found between the BV2 and SRV groups compared to the control group (P < 0.05). Somatic cell count was lower in the SRV and BV2 groups compared to the control group (P < 0.05). Discussion: In this study, compatible with the previous reports NAS and S. aureus were the most frequently isolated microorganism. The diversity of virulence factors of Staphylococcus spp. also plays an important role in its high incidence. In some countries, mastitis vaccines used in cows are also administered to small ruminants for reducing infection rates. Similarly, in this study, it was found that the mastitis rate decreased in all vaccine groups compared to the control group. A significant difference was found only in the BV2 and SRV groups. It is thought that the reason for the statistical difference may be due to the biofilm antigen in the BV2 and SRV. In addition, J5 strain in the BV2 is estimated to be effective in reducing the prevalence of gram-negative mastitis. It was observed that the infection rates decreased in the vaccine groups, especially due to S. aureus and NAS. Spontaneous treatment rates were very close to each other between the groups. The reason for the high rate of spontaneous treatment in this study can be explained by the fact that the animals were young and in their 1st lactation. SCC was lower in all vaccine groups compared to the control group. This situation is associated with the decrease in infection rates related to the use of vaccines. It was observed that SCC was lower in the vaccine groups. In addition, SCC was found to be lower in this study compared to similar studies. However, it is evident that the use of species-specific vaccines in the SRV group significantly reduced the rates of total S. aureus mastitis, subclinical NAS mastitis, and new infections by NAS compared to other vaccines. Furthermore, the species-specific vaccine significantly increased the rate of spontaneous treatment for S. aureus mastitis.

Comparação entre a imunidade induzida em bovinos vacinados com bacterinas polivalentes comerciais e uma monovalente experimental / Comparison between immunity induced in cattle vaccinated with commercial polyvalent bacterins and one experimental monovalent

O presente estudo avaliou a indução da produção de anticorpos contra Leptospira spp.por dez bacterinas, sendo nove polivalentes e uma monovalente experimental para a sorovariedade Hardjo amostra Norma. A concentração celular foi controlada e utilizou-se adjuvante de emulsão óleo em água. Um ensaio imunoenzimático (ELISA) indireto foi desenvolvido utilizando-se conjugado anti-IgG total para mensurar os níveis de anticorpos da classe IgG conferido pelas bacterinas utilizando três amostras diferentes: Hardjoprajitino, Norma e Hardjo-bovis. Paralelamente foi utilizado também o Teste de Soroaglutinação Microscópica (SAM) para mensurar os níveis de anticorpos contra as mesmas amostras. Encontraram-se títulos variáveis entre as bacterinas de acordo com o teste ELISA. Os títulos no SAM foram de pouca intensidade e de curta duração indicando a necessidade de controle celular para uma posterior padronização destes produtos. Com base nos resultados encontrados no presente estudo, a bacterina monovalente foi a que apresentou melhor desempenho.

The study evaluated the induction of antibody production against ten bacterins, nine polyvalent and one experimental monovalent to serovar Hardjo strain Norma. An indirect enzyme linked immunosorbent assay (ELISA) was developed using anti-IgG conjugate to measure total levels of IgG class antibodies conferred by bacterins using three different strains: Hardjoprajiitino, Norma and Hardjo-bovis. Microscopic Agglutination Test (MAT) was also used to measure immunoglobulin levels of the same strains. Variable ELISA titers were induced by the tested bacterins. The MAT titers found showed lower intensity and shorter duration, indicating the need to cellular control in further standardization of these vaccines. Based on results of this study, the monovalent bacterin showed best performance.

Comparação entre a imunidade induzida em bovinos vacinados com bacterinas polivalentes comerciais e uma monovalente experimental / Comparison between immunity induced in cattle vaccinated with commercial polyvalent bacterins and one experimental monovalent

O presente estudo avaliou a indução da produção de anticorpos contra Leptospira spp.por dez bacterinas, sendo nove polivalentes e uma monovalente experimental para a sorovariedade Hardjo amostra Norma. A concentração celular foi controlada e utilizou-se adjuvante de emulsão óleo em água. Um ensaio imunoenzimático (ELISA) indireto foi desenvolvido utilizando-se conjugado anti-IgG total para mensurar os níveis de anticorpos da classe IgG conferido pelas bacterinas utilizando três amostras diferentes: Hardjoprajitino, Norma e Hardjo-bovis. Paralelamente foi utilizado também o Teste de Soroaglutinação Microscópica (SAM) para mensurar os níveis de anticorpos contra as mesmas amostras. Encontraram-se títulos variáveis entre as bacterinas de acordo com o teste ELISA. Ostítulos no SAM foram de pouca intensidade e de curta duração indicando a necessidade de controle celular para uma posterior padronização destes produtos. Com base nos resultados encontrados no presente estudo, a bacterina monovalente foi a que apresentou melhor desempenho.(AU)

The study evaluated the induction of antibody production against ten bacterins, nine polyvalent and one experimental monovalent to serovar Hardjo strain Norma. An indirect enzyme linked immunosorbent assay (ELISA) was developed using anti-IgG conjugate to measure total levels of IgG class antibodies conferred by bacterins using three different strains: Hardjoprajiitino, Norma and Hardjo-bovis. Microscopic Agglutination Test (MAT) was also used to measure immunoglobulin levels of the same strains. Variable ELISA titers were induced by the tested bacterins. The MAT titers found showed lower intensity and shorter duration, indicating the need to cellular control in further standardization of these vaccines. Based on results of this study, the monovalent bacterin showed best performance.(AU)

Evaluation of the flocculation limit technique for quality control of Clostridial vaccines / Avaliação da técnica limite de floculação para o controle de qualidade de vacinas clostridiais

This study was carried out to assess and standardize the 'Limit of Flocculation' in vitro test to be used in the quality control of vaccine against enterotoxemy. This study used experimental vaccines against enterotoxemy and toxicoid produced from the epsilon toxin of Clostridium perfringens type D, previously evaluated by serum neutralization in mice and TCP test. The 'Limit of Flocculation' technique was performed by mixing variable amounts of antitoxin and a fixed value of toxicoid to each tube. It was proposed, in the first tube to flocculate, that its Fl would be the unit of flocculation of the tested sample. The main results indicated that the lower degree of antitoxin detectable both in rabbit and in sheep serum through the Limit of Flocculation' technique is equivalent to 10 UI/mL. The TCP and 'Limit of Flocculation' correlation ratio was 99.97%. In conclusion, the Limit of Flocculation is suitable to replace in vivo methods for the analysis of epsilon. Nevertheless, the referred technique is less effective to be used for the potency test of vaccines against enterotoxemy in substitution to serum neutralization in mice.

Este trabalho foi desenvolvido com o objetivo de avaliar e padronizar o teste in vitro Limite de Floculação, para ser utilizado no controle de qualidade de vacinas contra enterotoxemia. Foram utilizadas vacinas experimentais contra enterotoxemia e toxóides produzidos pelo Clostridium perfringens tipo D, previamente avaliados por métodos in vivo, empregando-se os testes de soroneutralização em camundongo TCP. A técnica Limite de Floculação consistiu em misturar volumes variados de antitoxina com volumes fixos de toxóide em tubos de ensaio. O primeiro tubo a flocular foi dito como sendo o que continha a quantidade mais próxima de antitoxina equivalente à quantidade de antígeno na amostra e se propôs que o Lf contido no tubo foi unidade de Lf da amostra testada. Os principais resultados obtidos indicaram que o nível mínimo de antitoxina detectável em soros de coelhos e carneiros pela técnica limite de floculação é 10 UI/mL. O coeficiente de correlação, obtido entre os títulos de toxóide pelos métodos TCP e Limite de floculação foi 99,97%. Conclui-se que o Limite de Floculação é satisfatório para substituir os tradicionais métodos in vivo utilizados na análise de toxóide epsilon. No entanto, a referida técnica é pouco eficiente para ser utilizada no teste de potência de vacinas contra enterotoxemia, em substituição à soroneutralização em camundongos.

Enzyme linked immunosorbent assay for potency testing of vaccines containing Clostridium perfringens type D epsilon-toxoid.

For ethical, economic and technical reasons in vivo assays need to be replaced in routine laboratory procedures. Based on a method which is already accepted by the British authorities, an indirect ELISA has been developed and evaluated for Clostridium perfringens type D-containing vaccines. Individual and pooled sera of vaccinated rabbits were tested at a single dilution level, the results transferred into IU/ml, and compared with the conventional toxin neutralization test in mice. The ELISA was found to give reproducible estimates of antitoxin levels and showed good correlation with the conventional in vivo test in mice.

Quantification of bovine albumin fraction V in cellular antigens of antileptospirosical Cuban vaccine vaxSpiral.

A solid phase immunoradiometric assay (IRMA) for quantification of bovine albumin fraction V (Bov.Alb.FV) in antileptospirosical Cuban vaccine vaxSpiral is described in the present work. Anti-Bov.Alb.FV IgG raised against rabbit purified by affinity chromatography was used as first antibody. Anti-rabbit IgG labeled by Chloromine-T reaction was used as a tracer and the method has demonstrated to be sensitive with high intra- and inter-assay reproducibility. Eight lots of vaccinal antigens were evaluated and in all of the cases, the bovine albumin fraction V concentration was lower than 1 microgram/mL, as the World Health Organization (WHO) establishes. This IRMA is a simple and sensitive assay and could be used as control method for all human vaccines that use Bov.Alb.FV in their production process, even cellular vaccines.

Analysis of vaccines prepared from armadillo-derived M. leprae; results of an inter-laboratory study coordinated by the World Health Organization. Immunology of Mycobacterial Diseases (IMMYC) Steering Committee, World Health Organization.

Preparations of armadillo-derived Mycobacterium leprae used in vaccine trials were analyzed using a combination of morphological, chemical and immunological criteria. When compared to more recent preparations, vaccine lots prepared in 1984 and 1985 were found to contain fewer intact bacilli and lower amounts of M. leprae antigens. These differences may be characteristic of the original preparations, or alternatively, may have arisen during prolonged storage. The early vaccine lots were those used in the recently published Venezuela trial.

Valor terapéutico de las vacunas bacterianas polivalentes en niños de 2do año de vida portadores de infecciones recurrentes

Se realiza un estudio clínico e inmunológico en 52 niños de 2do año de vida de los círculos infantiles "Juana de la Torre" y "Mis hermanitos y yo" de Holguín durante un período comprendido desde enero hasta junio de 1987, con la finalidad de valorar terapéuticamente las vacunas bacterianas polivalentes por vía intradérmica en las infecciones recurrentes, demostrándose la mejoría clínica, así como un incremento significativo de la IgA y de la respuesta cútanea a la Candidina y el Toxoide Tetánico. No se encontaron reacciones secundarias al tratamiento