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| Natura: | Artículo científico |
| Lingua: | en |
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Universidad de Costa Rica
2007
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| Accesso online: | https://www.redalyc.org/articulo.oa?id=44955113 |
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| author | Roger I. Rodríguez-Vivas |
| author_facet | Roger I. Rodríguez-Vivas |
| contents | Aislamiento de una cepa de campo de Babesia bigemina (Piroplasma: Babesiidae) y establecimiento del cultivo in vitro para la producción de antígenos Roger I. Rodríguez-Vivas Franklin J. Quiñones-Avila Gale Wagner Genny T. Ramírez-Cruz David Cruz Biología antigen isolation percentage Babesia bigemina in vitro culture Isolation of a field strain of Babesia bigemina (Piroplasma: Babesiidae) and establishment of in vitroculture for antigen production. Bovine babesiosis, caused by Babesia bigemina, is a barrier for livestock development;it results in high economic loss to Mexican livestock. Control requires adequate antigens for diagnosisand vaccination programs. However, because of antigenic variation among Babesia strains, it is necessary to useantigens prepared from local strains. The purpose of the present study was to isolate a local field strain and toestablish the in vitro culture of B. bigemina by the evaluation of the constituents concentration of culture media.Thirty engorged female Boophilus microplus were collected from cattle suffering clinical babesiosis (B. bigemina)in Yucatan state, Mexico. These ticks were sent to the laboratory for detection of Babesia sp. vermicules. Eggs werekept at 83-85 % humidity and 27 ºC until hatching. Larvae were transferred to an esplenectomized calf (B-1). Theresulting nymphs were transferred to an esplenectomized calf (B-2). Twelve days later, B. bigemina (local strain)was detected in calf B-2 and its infected blood was frozen in liquid nitrogen to initiate the in vitro culture. TheMicroaerophilus Stationary Phase (MASP) in vitro culture method was used to reactivate the parasite. Three differentconcentrations of culture media (70, 60 and 50 %), serum (30, 40 and 50 %) and uninfected red blood cells (5,10 and 15 %) were used in order to know the convenient concentrations to obtain the highest percentage of infectedred blood cells (PEI). The cultured strain was used to prepare antigens for the Immunofluorescence Antibody Test(IFAT) and several concentrations of serum and conjugate were tested. Strain isolation was successful; 30 days wereneeded to obtain a PEI of 1.5 %. The isolated strain was frozen in liquid nitrogen and the parasites were reactivatedwith the in vitro culture MASP method. The concentration of culture media that produced the highest PEI (14 %)(p<0.05) was 30 % serum, 70 % M199 and 5 %. Uninfected Red Blood cells antigens were successfully used in theIFAT and the best dilutions to differentiate between positive and negative controls were serum 1:80 and conjugate1:80. The isolated B. bigemina local strain requires particular conditions of in vitro culture by the MASP method toreach high numbers of infected red blood cells, needed to prepare and provide high quality antigens for serologicaldiagnosis of B. bigemina. Rev. Biol. Trop. 55 (1): 127-133. Epub 2007 March. 31. 2007 artículo científico 0034-7744 https://www.redalyc.org/articulo.oa?id=44955113 en http://www.redalyc.org/revista.oa?id=449 Revista de Biología Tropical application/pdf Universidad de Costa Rica Revista de Biología Tropical (Costa Rica) Num.1 Vol.55 |
| format | Artículo científico |
| id | redalyc_44955113 |
| language | en |
| publishDate | 2007 |
| publisher | Universidad de Costa Rica |
| spellingShingle | Aislamiento de una cepa de campo de Babesia bigemina (Piroplasma: Babesiidae) y establecimiento del cultivo in vitro para la producción de antígenos Roger I. Rodríguez-Vivas Biología antigen isolation percentage Babesia bigemina in vitro culture Aislamiento de una cepa de campo de Babesia bigemina (Piroplasma: Babesiidae) y establecimiento del cultivo in vitro para la producción de antígenos Roger I. Rodríguez-Vivas Franklin J. Quiñones-Avila Gale Wagner Genny T. Ramírez-Cruz David Cruz Biología antigen isolation percentage Babesia bigemina in vitro culture Isolation of a field strain of Babesia bigemina (Piroplasma: Babesiidae) and establishment of in vitroculture for antigen production. Bovine babesiosis, caused by Babesia bigemina, is a barrier for livestock development;it results in high economic loss to Mexican livestock. Control requires adequate antigens for diagnosisand vaccination programs. However, because of antigenic variation among Babesia strains, it is necessary to useantigens prepared from local strains. The purpose of the present study was to isolate a local field strain and toestablish the in vitro culture of B. bigemina by the evaluation of the constituents concentration of culture media.Thirty engorged female Boophilus microplus were collected from cattle suffering clinical babesiosis (B. bigemina)in Yucatan state, Mexico. These ticks were sent to the laboratory for detection of Babesia sp. vermicules. Eggs werekept at 83-85 % humidity and 27 ºC until hatching. Larvae were transferred to an esplenectomized calf (B-1). Theresulting nymphs were transferred to an esplenectomized calf (B-2). Twelve days later, B. bigemina (local strain)was detected in calf B-2 and its infected blood was frozen in liquid nitrogen to initiate the in vitro culture. TheMicroaerophilus Stationary Phase (MASP) in vitro culture method was used to reactivate the parasite. Three differentconcentrations of culture media (70, 60 and 50 %), serum (30, 40 and 50 %) and uninfected red blood cells (5,10 and 15 %) were used in order to know the convenient concentrations to obtain the highest percentage of infectedred blood cells (PEI). The cultured strain was used to prepare antigens for the Immunofluorescence Antibody Test(IFAT) and several concentrations of serum and conjugate were tested. Strain isolation was successful; 30 days wereneeded to obtain a PEI of 1.5 %. The isolated strain was frozen in liquid nitrogen and the parasites were reactivatedwith the in vitro culture MASP method. The concentration of culture media that produced the highest PEI (14 %)(p<0.05) was 30 % serum, 70 % M199 and 5 %. Uninfected Red Blood cells antigens were successfully used in theIFAT and the best dilutions to differentiate between positive and negative controls were serum 1:80 and conjugate1:80. The isolated B. bigemina local strain requires particular conditions of in vitro culture by the MASP method toreach high numbers of infected red blood cells, needed to prepare and provide high quality antigens for serologicaldiagnosis of B. bigemina. Rev. Biol. Trop. 55 (1): 127-133. Epub 2007 March. 31. 2007 artículo científico 0034-7744 https://www.redalyc.org/articulo.oa?id=44955113 en http://www.redalyc.org/revista.oa?id=449 Revista de Biología Tropical application/pdf Universidad de Costa Rica Revista de Biología Tropical (Costa Rica) Num.1 Vol.55 |
| title | Aislamiento de una cepa de campo de Babesia bigemina (Piroplasma: Babesiidae) y establecimiento del cultivo in vitro para la producción de antígenos |
| topic | Biología antigen isolation percentage Babesia bigemina in vitro culture |
| url | https://www.redalyc.org/articulo.oa?id=44955113 |