Methods for the cultivation of ciliated protozoa from the large intestine of horses.
Abstract: This paper describes cultivation methods for ciliates from the digestive tract of horses. Members of three different genera were successfully grown in vitro for short periods of time. However, only cells belonging to the genus Blepharocorys, which resides in the horse's large intestine, were maintained for longer periods. This Blepharocorys culture was successfully grown in vitro after inoculation of freshly excreted horse faeces in culture medium containing a population of bacteria. The ciliates survived for over six months, and the density of their population varied between 1.7 × 10(3) and 2.4 × 10(3) cells mL(-1). Favourable conditions for the prolonged cultivation of this ciliate were observed when the medium was prepared by mixing horse faeces and 'caudatum' salt solution in a 1:1 V/V ratio together with food (60% powdered meadow hay, 16% wheat gluten, 12% barley flour and 12% microcrystalline cellulose) supplied as 0.20 mg mL(-1) culture per day.
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Publication Date: 2015-12-17 PubMed ID: 26684192DOI: 10.1093/femsle/fnv233Google Scholar: Lookup
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Summary
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This research article focuses on the methods used to cultivate ciliated protozoa, specifically the genus Blepharocorys, from the large intestine of horses. The researchers were able to maintain this culture for over six months using a mixture of horse feces, ‘caudatum’ salt solution, and a specific food combination.
Overview of Research and Methodology
- The study centers on the cultivation of ciliated protozoa, a type of single-celled organism bearing tiny hairlike structures called cilia, from the digestive tract of horses.
- The researchers were interested in three different classes of ciliated protozoa, but were only successful in maintaining cells from the Blepharocorys genus for an extended time.
- This particular genus is found in the horse’s large intestine.
- The team managed to grow the Blepharocorys culture in vitro, i.e., in a controlled artificial environment outside of an organism, such as a test tube or petri dish.
Inoculation and Cultivation Process
- The in vitro growth started with the inoculation of freshly excreted horse feces in a culture medium containing a population of bacteria.
- The culture was able to survive for over six months, with the population density of the ciliates ranging between 1.7 × 10(3) and 2.4 × 10(3) cells per milliliter.
- The medium preparation was key to this success – it was achieved by mixing horse feces and ‘caudatum’ salt solution in a 1:1 volume/volume ratio.
Nutrition for the Culture
- The ciliates were provided with food that consisted of 60% powdered meadow hay, 16% wheat gluten, 12% barley flour, and 12% microcrystalline cellulose.
- This food mixture was supplied at a rate of 0.20 milligrams per milliliter of culture per day.
- The combination of these elements provided the right conditions for a long-term cultivation of the ciliates.
Significance of the Study
- This research is important as it could potentially open up new avenues for studying the biology of ciliates from the horse’s large intestine, ultimately contributing to a better understanding of the digestive physiology of horses.
- Furthermore, the ability to maintain such cultures for extended periods in vitro gives researchers a sustainable supply of these cells for future research and experimentation.
Cite This Article
APA
Bełżecki G, Miltko R, Michałowski T, McEwan NR.
(2015).
Methods for the cultivation of ciliated protozoa from the large intestine of horses.
FEMS Microbiol Lett, 363(2), fnv233.
https://doi.org/10.1093/femsle/fnv233 Publication
Researcher Affiliations
- The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jabłonna, Poland g.belzecki@ifzz.pl.
- The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jabłonna, Poland.
- The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05-110 Jabłonna, Poland.
- Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, SY23 3DA, UK.
MeSH Terms
- Animals
- Ciliophora / growth & development
- Ciliophora / metabolism
- Ciliophora Infections / parasitology
- Ciliophora Infections / veterinary
- Culture Media / chemistry
- Culture Media / metabolism
- Feces / parasitology
- Horse Diseases / parasitology
- Horses
- Intestine, Large / parasitology
- Parasitology / methods
Citations
This article has been cited 7 times.- Rocchigiani G, Ricci E, Navarro MA, Samol MA, Uzal FA. Leukocyte numbers and intestinal mucosal morphometrics in horses with no clinical intestinal disease. J Vet Diagn Invest 2022 May;34(3):389-395.
- Bampidis V, Azimonti G, Bastos ML, Christensen H, Dusemund B, Fašmon Durjava M, Kouba M, López-Alonso M, López Puente S, Marcon F, Mayo B, Pechová A, Petkova M, Ramos F, Sanz Y, Villa RE, Woutersen R, C뫝a F, Focks A, Tosti L, Anguita M, Brozzi R, Galobart J, Innocenti ML, López-Gálvez G, Sofianidis K, Vettori MV, Gregoretti L. Safety and efficacy of a feed additive consisting of copper chelate of ethylenediamine for all animal species (Zinpro Animal Nutrition (Europe), Inc.). EFSA J 2021 Apr;19(4):e06541.
- Bampidis V, Azimonti G, Bastos ML, Christensen H, Dusemund B, Fašmon Durjava M, Kouba M, López-Alonso M, López Puente S, Marcon F, Mayo B, Pechová A, Petkova M, Ramos F, Sanz Y, Villa RE, Woutersen R, C뫝a F, Focks A, Tosti L, Anguita M, Brozzi R, Galobart J, Innocenti ML, López-Gálvez G, Vettori MV, Gregoretti L. Safety and efficacy of a feed additive consisting of iron chelate of ethylenediamine for all animal species (Zinpro Animal Nutrition (Europe), Inc.). EFSA J 2021 Apr;19(4):e06540.
- Bampidis V, Azimonti G, Bastos ML, Christensen H, Dusemund B, Durjava MF, Kouba M, López-Alonso M, López Puente S, Marcon F, Mayo B, Pechová A, Petkova M, Ramos F, Sanz Y, Villa RE, Woutersen R, Bories G, C뫝a F, Focks A, Tosti L, Brozzi R, Galobart J, Gregoretti L, Innocenti ML, Vettori MV, López-Gálvez G. Safety and efficacy of a feed additive consisting of manganese chelate of ethylenediamine for all animal species (Zinpro Animal Nutrition (Europe), Inc.). EFSA J 2021 Mar;19(3):e06468.
- Bampidis V, Azimonti G, Bastos ML, Christensen H, Dusemund B, Durjava MF, Kouba M, López-Alonso M, López Puente S, Marcon F, Mayo B, Pechová A, Petkova M, Ramos F, Sanz Y, Villa RE, Woutersen R, Bories G, C뫝a F, Focks A, Tosti L, Brozzi R, Galobart J, Gregoretti L, Innocenti ML, Vettori MV, López-Gálvez G. Safety and efficacy of a feed additive consisting of zinc chelate of ethylenediamine for all animal species (Zinpro Animal Nutrition (Europe), Inc.). EFSA J 2021 Mar;19(3):e06467.
- Cedrola F, Senra MVX, Rossi MF, Fregulia P, D'Agosto M, Dias RJP. Trichostomatid Ciliates (Alveolata, Ciliophora, Trichostomatia) Systematics and Diversity: Past, Present, and Future. Front Microbiol 2019;10:2967.
- Zheng W, Wang C, Yan Y, Gao F, Doak TG, Song W. Insights into an Extensively Fragmented Eukaryotic Genome: De Novo Genome Sequencing of the Multinuclear Ciliate Uroleptopsis citrina. Genome Biol Evol 2018 Mar 1;10(3):883-894.
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