Abstract
This study developed and evaluated an in vitro rumen incubation (IVRI) method to describe the exsheathment kinetics of Haemonchus contortus third-stage infective larvae (L3) in ruminal liquor (RL). The specific objectives were (i) to standardize the IVRI method to facilitate the contact between L3 and RL as well as the larval recovery, and (ii) to apply the IVRI method to describe the exsheathment kinetics of H. contortus and to select the best fitting nonlinear model. Incubation devices containing H. contortus larvae were incubated according to the IVRI technique in cattle RL or PBS. The incubation conditions included RL mixed with a nitrogen-rich media, maintained at 39 °C, with pH = 7.0, vented with CO2 and manual agitation. The larvae were recovered after 0, 1, 3, 6, 9, 12, and 24 h. The exsheathed and ensheathed larvae were counted to estimate the exsheathment (%) in RL or PBS. Exsheathment in RL was analyzed with nonlinear regression models: Exponential, Gompertz, Logistic, Log-Logistic, and Weibull. The models’ fit was compared to select the one that best described the exsheathment kinetics. The exsheathment in RL reached 6.52%, 20.65%, 58.22%, 69.24%, 73.08%, and 77.20% in 1, 3, 6, 9, 12, and 24 h, respectively. Although the Gompertz, Weibull, and Logistic models were adequate to describe the observed exsheathment, the Log-Logistic model had the best fit. The IVRI method using bovine RL represents a suitable tool for the study of the in vitro exsheathment kinetics of H. contortus L3.
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Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We would like to thank the staff of the nutrition area and the Animal Nutrition Laboratory at FMVZ-UADY for all the support provided. We also thank the team in charge of the L3 larvae production at the FMVZ-UADY. C.G. Marin-Tun. received a doctoral scholarship from CONACYT, México.
Funding
This study was supported by the Consejo Nacional de Ciencia y Tecnología (CONACYT, México): Cátedras CONACYT Project No. 692, and doctoral grant No. 784280.
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All authors contributed to the study’s conception and design. Material preparation and data collection were performed by Cindy G. Marin-Tun and Concepción M. Capetillo-Leal. Data analyses were performed by Cindy G. Marin-Tun and Carlos A. Sandoval Castro. The first draft of the manuscript was written by Cindy G. Marin-Tun, María G. Mancilla-Montelongo, and Juan F.J. Torres-Acosta. All the figures were prepared by Cindy G. Marin-Tun. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Supplementary information
Supplementary 1
Online resource 1 Haemonchus contortus L3 larvae: sheath (a), larva during the exsheathment process (b) and exsheathed larva (b).
Supplementary 2
Online resource 2 Complete equations for the nonlinear models considered to select the model that best described the in vitro exsheathment kinetics of Haemonchus contortus L3 using the in vitro rumen incubation technique with ruminal liquid.
Supplementary 3
Online resource 3 Log-logistic equations using the original exsheathment values and the data at 30 or 50% of the original data of in vitro exsheathment from H. contortus L3 larvae.
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Marin-Tun, C.G., Mancilla-Montelongo, M.G., Torres-Acosta, J.F. et al. An in vitro rumen incubation method to study exsheathment kinetics of Haemonchus contortus third-stage infective larvae. Parasitol Res 122, 833–845 (2023). https://doi.org/10.1007/s00436-023-07780-z
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DOI: https://doi.org/10.1007/s00436-023-07780-z