Evaluation of lugol solution used for counting rumen ciliates

Abstract

ABSTRACT. The present study proposes a modification of the technique described by PURSER & MOIR (1959) for the quantitative evaluation ofrumen ciliate based on an adaptation described by DEHORITY (1984). The modifYing process includes: the replacement of two drops of brilliant green dye, for at least four hours, by three drops oflugol solution, for at least 15 minutes. It was made acomparative evaluation ofthese stainings. It was concluded that Jugol solution can replace the brilliant green dye showing the following advantages: staining time reduction and subsequent speeding of sample processing; evidence of skeletal plates of entodiniomorphs making its identification easier; improved observation of small ciliates and inconspicuous structures; improved total counting and generic identitication of the ciliates. KEY WORDS. Rumen ciliate, quantitative evaluation, lugol solution, technique Among the techniques found in the literature for the quantitative evaluation of rumen ciliates are the one mentioned by OGIMOTO & IMAI (1981) which uses a plankton counting slide, the Fuchs-Rosenthal and the Sedgewick-Rafter counting chamber. DEHORITY (1984) described an adaptation of the procedure proposed by PURSER & MOIR (1959), using the Sedgewick-Rafter counting chamber. In order to improve the observation of ciliates and reduce the time of sample processing, the present work proposes the replacement of the brilliant green dye used by DEHORITY (1984) by the lugol solution. It also presents a comparison between these procedures. MATERIAL AND METHODSThirty samples of rumen contents were obtained from rumen-fistulate crossbred Holstein Friesian-Zebu cow. Each sample consisted of approximately 20 cm 3 of rumen content taken from the center of the rumen mass to which were added 20mI of rumen fluid obtained by aspiration. These 40 ml were fixed in an equal volume of 18.5% formalin and processed according to DEHORITY (1984). It involved the usage of I ml subsamp1es of the rumen content stained with two drops of brilliant green dye for at least four hours and subsequent dilution in 9 ml of 30% glycerol. The differential counting of the ciliates was performed twice using a Sedgewick-Rafter counting chamber with I ml of the diluted subsarnple. In another

Does virginiamycin supplementation affect the metabolism and performance of Nellore bulls grazing under low and high gain rates?

“…Five subsamples were obtained (liquid and solid) from different areas of the rumen (cranial dorsal, cranial ventral, central rumen, rumen ventral, and caudal dorsal) and pooled. Immediately after collection, 40 g of solid samples was mixed with 50 ml of formalin solution (1:1 of water: 370 ml/L formaldehyde; D'Agosto & Carneiro, ) and stored at −80°C until counting of protozoa.…”

“…Direct counts and protozoa identification were performed using a Sedgewick–Rafter counting chamber (Dehority, Tirabasso, & Grifo, ). Samples were diluted with 200 ml/L glycerol and stained using Lugol's solution before the cells were counted (D'Agosto & Carneiro, ). Ruminal bacteria were counted by vigorously stirring 20 g of solid and 5 ml of liquid samples plus 25 ml of phosphate‐buffered saline (pH 7.4) for 3 min, and the contents were filtered through four layers of cheesecloth.…”

Phylogenetic Analyses Support Validity of Genus Eodinium (Ciliophora, Entodiniomorphida, Ophryoscolecidae)

“…The identification of D. anisacanthum and E. posterovesiculatum morphotypes was acquired after morphometric analyses of 20 Lugol stained (Cedrola et al. ; D'Agosto and Carneiro ) specimens of each morphotype (Cedrola et al. unpubl.…”

Intestinal ciliates of Brazilian Capybara (Hydrochoerus hydrochaeris L.)

“…The average relative abundance was estimated from a sample of 300 cells (Ito et al 1994, Mishima et al 2009) and the density was determined in a Sedgewick-Rafter counting chamber as in methods of to Dehority (1984) andCarneiro (1999).…”