Isolation and establishment in in vitro culture of a Theileria annulata - infected cell line from Spain

Viseras, J.; García-Fernández, Pedro; Adroher, Francisco Javier

doi:10.1007/s004360050270

Cited by 13 publications

(6 citation statements)

References 5 publications

(7 reference statements)

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“…Among the different parasite stages in the host, the schizont stage is the symptomatic stage, based on which attenuated schizont vaccines were designed. The cell culture-based attenuated vaccines have been used in countries, like India (Gupta et al, 1998), China (Zhang, 1997), Russia (Stepanova and Zablotskii, 1989), Turkey (Sayin et al, 1997), Spain (Viseras et al, 1997), Israel (Pipano and Tsur, 1966), Iran (Hashemi-Fesharki, 1988), Tunisia (Darghouth, 2008), and Morocco (Ouhelli et al, 1997) for controlling the T. annulata infection. The attenuation of the vaccine line because of the long term passage results in loss of genotypes, decreasing the diversity of the parasites (Darghouth et al, 1996;Pipano and Shkap, 2006;Bilgic et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Population Genetic Analysis of the Theileria annulata Parasites Identified Limited Diversity and Multiplicity of Infection in the Vaccine From India

et al. 2021

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Background: Apicomplexan parasite Theileria annulata causes significant economic loss to the livestock industry in India and other tropical countries. In India, parasite control is mainly dependent on the live attenuated schizont vaccine and the drug buparvaquone. For effective disease control, it is essential to study the population structure and genetic diversity of the Theileria annulata field isolates and vaccine currently used in India.Methodology/Results: A total of 125 T. annulata isolates were genotyped using 10 microsatellite markers from four states belonging to different geographical locations of India. Limited genetic diversity was observed in the vaccine isolates when compared to the parasites in the field; a level of geographical substructuring was evident in India. The number of genotypes observed per infection was highest in India when compared to other endemic countries, suggesting high transmission intensity and abundance of ticks in the country. A reduced panel of four markers can be used for future studies in these for surveillance of the T. annulata parasites in India.Conclusion: High genetic variation between the parasite populations in the country suggests their successful spread in the field and could hamper the disease control programs. Our findings provide the baseline data for the diversity and population structure of T. annulata parasites from India. The low diversity in the vaccine advocates improving the current vaccine, possibly by increasing its heterozygosity. The reduced panel of the markers identified in this study will be helpful in monitoring parasite and its reintroduction after Theileria eradication.

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Section: Introductionmentioning

confidence: 99%

Population Genetic Analysis of the Theileria annulata Parasites Identified Limited Diversity and Multiplicity of Infection in the Vaccine From India

et al. 2021

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“…One isolate, designated Cádiz 28E, originated from Algar in the province of Cádiz, Southern Spain and was established in vitro as described elsewhere. 10 The second isolate, designated Cáceres, originated from the Extremadura region in Western Spain and was established in culture as described elsewhere. 11 The Sudanese parasite was isolated in Soba, Sudan.…”

Section: Methodsmentioning

confidence: 99%

“…EDTA‐blood samples were taken at regular intervals of two weeks and during high fever in an effort to reestablish T. annulata cultures in vitro as described elsewhere. 10 Giemsa‐stained blood smears were obtained three times a week. Twenty microscopic fields were examined in the case of positive results, and fifty fields for negative smears.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Attenuated Theileria annulata Vaccines from Spain and the Sudan

Gubbels

Viseras

Habela

et al. 2000

Annals of the New York Academy of Sciences

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Theileriosis caused by Theileria annulata can be effectively prevented by vaccination with attenuated, cultured schizonts. Although these attenuated vaccines have been applied for a long time, not much is known about the fate of the vaccie strain in the field. Here, two experimental Spanish vaccine strains originating in Cádiz and Cáceres, and one Sudanese strain are studied to address the development of a carrier status and the infectivity for Hyalomma ticks. Moreover, the heterogeneity of the merozoite surface protein, Tams1, was analyzed in search for an attenuation marker. Using the sensitive reverse line blot (RLB) hybridization, the development of a low level carrier status was demonstrated in the Cáceres and Sudanese line vaccinated calves. Although no signal was detected in the Cádiz line vaccinated calves, seroconversion against the schizont stage was observed, as it was in all other calves. The experimental transmission of T. annulata by Hyalomma ticks to naïve calves was unsuccessful for all cell line inoculated calves. Tams1 heterogeneity indicated a clonal selection of parasites during the process of attenuation, but the Tams1 sequence itself has no connection with the attenuation status. In conclusion, a carrier status develops in attenuated schizont culture vaccinated calves, but is not infective for Hyalomma ticks. Based on these data, the risk for spread of the vaccine strains in the field may be very low.

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“…Microscope examination of the Giemsa-stained smears and the serology study with the IFAT were performed by the usual methods (Burridge et al 1974;Callow and Pepper 1974;GarcõÂ a-FernaÂ ndez et al 1996;Viseras et al 1997). Results were analyzed statistically using the v 2 test.…”

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confidence: 99%

Nested polymerase chain reaction for detection of Theileria annulata and comparison with conventional diagnostic techniques: its use in epidemiology studies

Martı́n-Sánchez¹,

Viseras²,

Adroher³

et al. 1999

Parasitol Res

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In this work we studied the ability of a nested polymerase chain reaction (PCR) to detect Theileria annulata, the causative agent of Mediterranean theileriosis, in blood samples obtained from cattle on farms in different Spanish regions and its possible use in epidemiology studies. Of the 214 samples analyzed, 78.04%, 69.86%, and 62.26% were found to be positive by nested PCR, indirect immunofluorescent antibody test, and optical microscopy of Giemsa-stained smears, respectively. The three techniques were in agreement in 68.6% of the results. The observation that the prevalence of Mediterranean theileriosis estimated using nested PCR alone (70.3%) and that obtained using all three diagnostic techniques together (80.4%) did not significantly differ verifies the utility of this technique in epidemiology studies.

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Isolation and establishment in in vitro culture of a Theileria annulata - infected cell line from Spain

Cited by 13 publications

References 5 publications

Population Genetic Analysis of the Theileria annulata Parasites Identified Limited Diversity and Multiplicity of Infection in the Vaccine From India

Population Genetic Analysis of the Theileria annulata Parasites Identified Limited Diversity and Multiplicity of Infection in the Vaccine From India

Characterization of Attenuated Theileria annulata Vaccines from Spain and the Sudan

Nested polymerase chain reaction for detection of Theileria annulata and comparison with conventional diagnostic techniques: its use in epidemiology studies

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