Family Ophryoscolecidae Research Articles

Endobiotic Ciliates from the Rumen of the European Bison Bison Bonasus (Linnaeus, 1758) from the Vologda Oblast of Russia

The fauna of endobiotic ciliates of the rumen of the European bison Bison bonasus from the Vologda oblast of Russia was investigated. In the studied bisons, 12 species of trichostomatids (Trichostomatia, Litostomatea) were found, ten of which were from the family Ophryoscolecidae and two species were from the family Isotrichidae. A high similarity of ciliate faunas in the rumen of different bison individuals in the studied population was noted. A comparative analysis of data on the species diversity and number of endobiotic ciliates in the rumen of various representatives of the genus Bison from different habitats was carried out.

Morphological Characterization Of Rumen Protozoa Isolated From Sweet Sorghum Hydrolysis Using Carabao (Bubalus bubalis Linn) Rumen Fluid

This study was conducted to characterize the morphology of rumen protozoa isolated in the batch type hydrolysis of sweet sorghum using 1% fresh carabao rumen fluid with duration of 0, 3, 6, 9, 12 and 15 days of the novel process hydrolysis. Hydrolysis was conducted using 5% chopped sweet sorghum bagasse. Nitrogen content was augmented with 0.35 g urea per 750 ml effective volume with low speed agitation of fermentation bottle at least 20 minutes twice daily and incubated at room temperature. Characterization was limited to microscopic evaluation of the morphological features of protozoa that was stained with methylene blue formalin saline solution. Protozoon population of 2.61 x 10 4 cfu/ml at initial period had significantly lowered down at the 15 days hydrolysis to 0.62 x 104 cfu/ml. Evaluation of morphology for the types of protozoa at different durations showed the predominance of species types with different orientations of caudal spines, shapes of the macromolecules, size of the body, and presence of adoral ciliary zones were features of species types from genus Eodinium Entodinium of the family Ophryoscolecidae. Evaluation of the hydrolysis condition showed that duration had significant effect on the pH (P>0.05). Sweet sorghum hydrolysis has initial pH 7 that significantly declined to pH 5 at 15 days duration. The population of the protozoa in the rumen fluid hydrolysis was significantly affected by duration of the hydrolysis (P>0.05). Entodinium were isolated at all durations of the acidic hydrolysis of sweet sorghum bagasse using carabao rumen fluid. In conclusion, morphological characteristics of the rumen protozoan shows how diverse are the composition of rumen protozoa in the carabao rumen fluid hydrolysis. The presence of the rumen protozoa is a justification of the novel carabao rumen fluid hydrolysis conversion of lignocelluloses in sweet sorghum bagasse into soluble carbohydrates for bioethanol production. The information is vital for animal feed utilization and bioethanol production optimization.

Insights into the systematics of the family Ophryoscolecidae (Ciliophora, Entodiniomorphida).

The family Ophryoscolecidae currently comprises 225species of trichostomatid ciliates, subdivided into three subfamilies (Entodiniinae, Diplodiniinae, and Ophryoscolecinae). The last taxonomic review of the family was performed 55years ago, but recent morphological and molecular studies indicate the need for a profound review of the systematics of this taxon, since its current taxonomy is insufficient to organize the diversity of the group. Here, we briefly review the systematics of the family Ophryoscolecidae based on information recovered from the literature and new morphological and molecular data. We add four new 18S rDNA sequences of ophryoscolecids to molecular databases, which contributed to improving the comprehension of intrafamily relationships within this group. Finally, we discuss some systematic problems and suggest approaches to resolve such inconsistencies in the future.

Morphological Characterization of Rumen Protozoa Isolated from Sweet Sorghum Hydrolysis Using Carabao (Bubalus bubalis Linn) Rumen Fluid

This study was conducted to morphologically characterize rumen protozoa isolated in the batch type hydrolysis of sweet sorghum using 1% fresh carabao rumen fluid with duration of Day 0, 3, 6, 9, 12 and 15 days of the novel process hydrolysis. Hydrolysis was conducted using 5% chopped sweet sorghum bagasse. Nitrogen content was augmented using 0.35 g urea per 750 ml effective volume with low speed agitation of fermentation bottle at least 20 minutes twice daily and incubated at room temperature. Characterization was limited to microscopic evaluation of the morphological features of protozoa that was stained with methylene blue formalin saline solution. Initial population of 2.61 x 10 4 cfu/ml at DO had significantly lowest at 0.62 x 104 cfu/ml during the 15 days hydrolysis. Evaluation of morphology for the types of protozoa at different durations showed the predominance of species types with different orientations of caudal spines, shapes of the macromolecules and size of the body, adoral ciliary zones closely related to the features of species types from genus Entodinium of the family Ophryoscolecidae. Evaluation of the hydrolysis condition showed that duration had significant effect on the pH (P>0.05). Sweet sorghum hydrolysis pH 7 at D0 had significantly declined to pH 5 at 15 days duration. The population of the protozoa in the rumen fluid hydrolysis was significantly affected by duration of the hydrolysis (P>0.05). Entodinium were isolated at all durations of the acidic hydrolysis of sweet sorghum bagasse using carabao rumen fluid. In conclusion, the diversity in composition of rumen protozoa is a justification of the novel carabao rumen fluid hydrolysis potential as pre-treatment in the conversion of lignocelluloses in sweet sorghum bagasse into soluble carbohydrates for bioethanol production. The information is vital for animal feed utilization and bioethanol production optimization.

Systematic review of the genus Ostracodinium (Ciliophora, Entodiniomorphida, Ophryoscolecidae) and notes on the taxonomy of Ostracodinium rugoloricatum Kofoid amp; MacLennan, 1932.

The family Ophryoscolecidae (Ciliophora, Entodiniomorphida) constitutes a diverse and monophyletic group of symbiotic ciliates of herbivorous mammals. The family includes approximately 200 species, distributed in three subfamilies and sixteen genera. The subfamily Diplodiniinae is the most diverse group in Ophryoscolecidae and comprises the genus Ostracodinium, which includes species with two retractable ciliary zones in the anterior body portion, a broad skeletal plate covering almost all the right surface of the body and a variable number of contractile vacuoles. The genus currently comprises 28 species, classified according to body size and shape, position and shape of the nuclear apparatus, number and position of contractile vacuoles, and number and shape of caudal projections. The present study performs a systematic review of the genus Ostracodinium, based on morphological and molecular data, and provides data about geographic distribution and hosts of each species.

Three new Prototapirella species, Opisthotrichum janus, and Troglocorys cava add to Entodiniomorphida (Ciliophora, Trichostomatia) diversity in mountain gorillas in Rwanda

Recently, a high number of Entodiniomorphida species was discovered in Virunga mountain gorillas (Gorilla beringei beringei) in Rwanda compared to other primates. Thirteen species of five genera (Troglocorys, Gorilloflasca, Prototapirella, Troglodytella, and Opisthotrichum) were identified with 10 species in Gorilloflasca, Prototapirella, and Troglodytella being host-specific, to our current knowledge. The remaining three species have been described for other herbivorous mammals; Prototapirella gorillae for western lowland gorillas; Troglocorys cava for western lowland gorillas, chimpanzees, and guinea baboons; Opisthotrichum janus in the family Ophryoscolecidae is regarded as specific to African antelopes. Here, we show the first indication that wild herbivorous hindgut fermenting mammals might have rumen ophryoscolecids in the intestine sympatric with their native intestinal ciliates. We also describe three new Prototapirella species, P. sabyinyo, P. sulcata, and P. curiosa. The somatic ciliary zones of the three new species are atypical of Prototapirella while their buccal infraciliature is not different from that of other Prototapirella species.

Infraciliature of Opisthotrichum janus, Epidinium ecaudatum, and Ophryoscolex purkynjei (Ciliophora, Entodiniomorphida)

We described the infraciliature of Opisthotrichum janus, Epidinium ecaudatum, and Ophryoscolex purkynjei, belonging to the family Ophryoscolecidae, from pyridinated silver carbonate impregnated specimens. The adoral polybrachykinety is C-shaped in Opi. janus, whereas it is spiral-shaped in Epi. ecaudatum and Oph. purkynjei. The vestibular polybrachykinety is slender and short in Opi. janus, whereas slender and long in Epi. ecaudatum and Oph. purkynjei. The paralabial kineties are composed of several short kineties along the adoral polybrachykinety. The dorsal polybrachykinety is displaced posteriorly. This study provides information on the diagnostic features of the subfamily Ophryoscolecinae to which Opi. janus, Epi. ecaudatum, and Oph. purkynjei belong. A C-shaped or spiral-shaped adoral polybrachykinety and slender vestibular polybrachykinety are the diagnostic features of the subfamily Ophryoscolecinae in addition to their general features of a cylindrical body, tubular vestibulum, dorsal ciliary zone considerably displaced posteriorly, well developed skeletal plates, and macronucleus beneath the right body surface. Opi. janus, Epi. ecaudatum, and Oph. purkynjei show the basic form of morphogenesis of the ciliature, similar to that in the subfamily Diplodiniinae, with four primordia developing into the oral and dorsal ciliature.

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

The validity of genus Eodinium has been historically disputed due to morphological similarities with Diplodinium (absence of skeletal plates as well as adoral and dorsal ciliary zones at the same body level). To address this issue, the 18S rDNA of four Eodinium posterovesiculatum morphotypes and four Diplodinium anisacanthum morphotypes were sequenced and phylogenetically analyzed. The different inference methods suggest the existence of a last common ancestor of Eodinium and Ostracodinium that is not shared with Diplodinium, strongly supporting the validity of genus Eodinium. Since skeletal plates are present in all members of genus Ostracodinium, the most parsimonious is a secondary loss of skeletal plates in E.posterovesiculatum. This work represents a breakthrough in the taxonomy and phylogeny of the family Ophryoscolecidae indicating that the skeletal plates may not reflect evolutionary divergence within this group of ciliates as traditionally proposed.

Presence of carbohydrate-digesting enzymes throughout the digestive tract of sheep

The activity of enzymes that digest carbohydrates at various points in the gastrointestinal tract of sheep was examined. In addition, weight and contents of digestive tract segments were examined. The experiment was performed on six adult ewes fed an 85% hay diet. The morphometric measurements revealed that the alimentary tract constituted 32% of the sheep's body mass and wet rumen contents constituted 70% of wet mass of the total digestive tract content. The protozoal population in the rumen varied from 37.2 × 10 4 to 54.0 × 10 4 /mL rumen fluid. In the examined samples, ciliates were identified as belonging to the family Ophryoscolecidae, genera Entodinium, Diplodinium, and Ophryoscolex, and family Isotrichidae, genera Isotricha and Dasytricha. Degradation of various carbohydrates by digesta revealed that the fastest digestion rate for plant cell wall carbohydrates and inulin occurred in the reticulum, whereas for starch and chitin, rates of digestion to reducing sugars were the highest in the small intestine. The pH and redox potential at various points in the digestive tract varied from 2.8 to 7.7 and from -72.1 to -289.2 mV, respectively.

Molecular Phylogeny of the Family Ophryscolecidae (Ciliophora, Litostomatea) Inferred from 18S rDNA Sequences.

The 18S rDNA was used to infer oral ciliature patterns of evolution within the family Ophryoscolecidae, with the addition of five new sequences of ciliates from the genus Ostracodinium. Our data confirmed the monophyly of the subfamilies Entodiniinae and Ophryoscolecinae, but more analysis would be required for the definition of the status of the subfamily Diplodiniinae. The oral infraciliature patterns reflect evolutionary divergence in the family Ophryscolecidae, observing monophyly on Entodinium-type, Diplodinium-type, Ostracodinium-type, Epidinium-type, and Ophryoscolex-type. The ancestral infraciliature of Entodinium-type cannot be proven, however, the position of Entodinium-type showed closer of Diplodinium-type than Ophryoscolex-type, corroborating previous studies using morphological characters. The observed inconsistencies reflect the need to increase the number of 18S rDNA sequences to family Ophryoscolecidae and investigate the evolution of this group using other molecular markers.

Infraciliature of eight Triplumaria species (Ciliophora, Entodiniomorphida) from Asian elephants with the description of six new species

Intestinal ciliates excreted in the feces of Asian elephants were surveyed. Fourteen species in the order Entodiniomorphida were detected. Nine Triplumaria species in the family Cycloposthiidae were found. Using the silver impregnation, two known species, T. antis and T. dvoinosi, were redescribed and six new species, T. sukuna n. sp., T. zuze n. sp., T. solea n. sp., T. suwako n. sp., T. fulgora n. sp., and T. harpagonis n. sp., were described. T. sukuna, T. zuze, T. solea, and T. suwako have the perivestibular polybrachykinety along the vestibular opening. The buccal infraciliary bands of T. suwako are similar to those of T. selenica found from elephants and the buccal infraciliary bands of T. sukuna, T. zuze, and T. solea are similar to those of T. grypoclunis described from rhinoceroses. T. antis, T. dvoinosi, T. fulgora, and T. harpagonis have the vestibular polybrachykinety extending down inside the vestibulum as found in rumen ciliates in the family Ophryoscolecidae. The caudalial ciliary zones of T. dvoinosi and T. fulgora were retractable as found in rumen ophryoscolecids. Raabena bella and Pseudoentodinium elephantis showed high composition values over 30%. Ciliate densities in the three fecal samples were 0.15, 1.09, and 2.07×104/ml.

Protozoa in the digestive tract of wild herbivores in South Africa. I: Warthogs (Phacochoerus aethiopicus)

Seventeen warthogs were harvested from their natural habitat during the winter hunting seasons of May to July 2001 and May to June 2002. Samples (200ml each) were collected and weighed from the stomach, cecum and colon of each animal for protozoal counts. Telamodinium onyx was the only protozoa present in seven animals and the predominant species in all others. Megadinium aethiopicum was observed in eight animals, while Teratodinium sphaeredon was present in two warthogs. Several different species of protozoa were seen in a few animals, two of which belong to the family Ophryoscolecidae and are considered to be normal inhabitants in the rumen (Diplodinium dentatum and Ophryoscolex purkynjei).

Nutritive defaunation of the rumen in steers with subsequent refaunation using a cryopreserved monoculture ofEntodinium caudatum

This study evaluated a technique for the nutritive defaunation of the rumen of cattle with subsequent single species refaunation using a cryopreserved monoculture of Entodinium caudatum (family Ophryoscolecidae). Four mature steers were nutritionally defaunated in two periods using two steers in each period. A diet containing (dry matter basis) 68% ground wheat grain, 7% wheat bran, 8% soybean oil and 17% wheat straw was used to decrease the pH of ruminal contents and to eliminate rumen ciliate protozoa. Protozoa-free rumens were observed on day 8 and 9 in the first and second period, respectively, after the start of defaunation. A monoculture of E. caudatum (34/89/94) was transported from the Institute of Animal Physiology, Slovak Academy of Sciences in Kosice to the University of Kiel (Germany) in liquid nitrogen in October 1996. The inoculation was accomplished on day 15 in the second period by applying 30 ml culture medium with a monoculture of E. caudatum (34/89/94; average concentration of protozoal cells 2 650/ml) into the rumen of a defaunated steer via the ruminal fistula. The mono-faunated steer was successfully inoculated with an average concentration of E. caudatum cells at 4.1 x 10(3)/ml (SD = 0.2) on day 2 after the inoculation.

New entodiniomorphid ciliates from the intestine of the wild African white rhinoceros belong to a new family, the Gilchristidae

Gilchristia artemis n.g., n.sp. and Digilchristia draconis n.g., n.sp. in the order Entodiniomorphida are described from the large intestine of the African white rhinoceros, and a new family Gilchristidae is proposed to contain them. These new species have a C-shaped adoral polybrachykinety, a slender vestibular polybrachykinety, and paralabial kineties along the ventral side of the adoral polybrachykinety in their retractable adoral ciliary zone, showing the same arrangement as in the rumen ciliates in the family Ophryoscolecidae. G. artemis has two skeletal plates and D. draconis one plate. In both species the dorsal skeletal plate is bow-shaped, folded in half longitudinally, twisting in the anterior part, and lying along the dorsal left side of the macronucleus. The second plate of G. artemis is slender and lies along the ventral side of the macronucleus. G. artemis has three ciliary arches and D. draconis has four arches along the dorsal and ventral sides of the body. Their arches are long and non-retractable, closely resembling those of ciliates in the families, Spirodiniidae and Cycloposthiidae, and are not analogous to the single retractable ciliary arch of the rumen ciliates in the family Ophryoscolecidae.

The rumen ciliate fauna of domestic sheep (Ovis ammon aires) from the Turkish Republic of Northern Cyprus.

Concentration and composition of ciliate protozoa in the families Ophryoscolecidae and Isotrichidae were determined in rumen contents of domestic sheep (Ovis ammon aries) from Cyprus. A total of five genera of Ophryoscolecidae were identified, Metadinium, Enoploplastron, Polyplastron, Epidinium, and Ophrvoscolex, which included six species: Metadinium affine, Enoploplastron triloricatum, Polyplastron multivesiculatum, Epidinium ecaudatum, Epidinium graini, and Ophryoscolex purkynjei. Eight separate forms of Epidinium were identified (E. ecaudatum f. ecaudatum, E. e. f. caudatum, E. e. f. bicaudatum, E. e. f. tricaudatum, E. e. f. quadricaudatum, E. graini f. graini, E. g. f. caudatricoronatum, and E. g. f. caudaquadricoroniatum), along with five forms of Ophryoscolex purkynjei (O. p. f. purkynjei, O. p. f. bifidobicinctus, O. p. f. bifidoquadricinctus, O. p. f. bicoronatus, O. p. f. tricoronatus, and O. p. f. quadricoronatus). Three species of Isotrichidae were observed, Isotricha intestinalis, I. prostoma, and Dasytricha ruminantium. This study reports new host records for three forms of Epidinium graini and Ophryoscolex purkynjei f. bifidobicinctus. The rumen fauna in the family Ophryoscolecidae from Cypriote domestic sheep appear to have limited diversity compared to those from Turkish and Far Eastern (Chinese/Japanese) sheep, while they are more diverse than those found in Western European (Scottish) and North American (Canadian/Alaskan) sheep.

Phylogenetic Taxonomy of Rumen Ciliate Protozoa Based on Their Morphology and Distribution

Imai, S. 1998. Phylogenetic taxonomy of rumen ciliate protozoa based on their morphology and distribution. J. Appl. Anim. Res., 13: 17–36. The ciliates belonging to the family Ophryoscolecidae in the suborder Entodiniomorphina peculiarly inhabit the rumen of various ruminants. They possess specialized and complicated bodies which seem to have evolved with the evolution of their hosts. Comparative morphology of these ciliates would be interesting as a model of co-evolution between these parasites and their hosts. As a result of morphologic comparison of 14 genera of this family, a temporary phylogenetic tree containing a line connected continuously from simpler species to more complicated ones, and a line independently derived on the way from the genus Diplodinium can be created However, there is no evidence that the morphologic relation of ophryoscolecid genera expresses the correct phylogenetic relation of this family. Thus it is necessary to examine this problem by another approach. When the relation of the distribution of respective genera of Ophryoscolecidae, in which the geographic factor is excluded as far as possible, to an advanced phylogenetic tree of host mammals is examined, we can draw another temporary phylogenetic tree. As compared to this figure with the morphologic relation of ophryoscolecid ciliates, the phylogenetic relation of these ciliates seems to agree as a rule. Accumulation of more detailed data from various hosts and their careful comparison would be necessary in future. In such examinations the species which are interesting phylogenetically may be discovered from unexamined animals, especially wild ones.

Ciliate protozoa in the rumen of Kafue lechwe, Kobus leche kafuensis, in Zambia, with the description of four new species.

The composition of the rumen ciliate fauna in 76 Kafue lechwe inhabiting a limited area in Zambia was surveyed and five genera containing 24 species with 16 formae belonging to the family Ophryoscolecidae were identified. Four new species belonging to Diplodiniinae were recognized and described as Diplodinium lochinvarense n. sp., Diplodinium leche n. sp., Diplodinium zambiense n. sp., and Metadinium ossiculi n. sp. In addition, Ostracodinium gracile form fissilaminatum Dogiel, 1932 was found for the second time and described as Metadinium fissilaminatum n. comb. The species composition was fairly unusual. Seven of the species have been found only in African wild antelopes and these species were found more frequently than cosmopolitan species. There was no evidence of isotrichid species. The average density of ciliates per 1 ml of the rumen fluid was 25.7 x 10(4), and the number of ciliate species per head of host was 10.8.

Association of methanogenic bacteria with rumen ciliates.

In 11 species of rumen ciliates belonging to nine genera of the family Ophryoscolecidae (order Entodiniomorphida) an ectosymbiosis with methanogenic bacteria was found. The bacteria could be identified as methanogens on the basis of the presence of specific fluorescent coenzymes (F350 and F420). This somatic interaction may reflect a metabolic interaction in which efficient interspecies hydrogen transfer benefits both partners.

Geographical distribution and population structure of springbok <I>Antidorcas marsupialis</I> rumen protozoa in southern Africa

Springbok were sampled in Angola, South West Africa, Kalahari Gemsbok National Park, the Cape Province and Eastern Transvaal between July 1973 and July 1974. All the ciliates found in the 20 rumens investigated belonged to a single genus Entodinium (family Ophryoscolecidae) and they varied in number from 0,3 x 106 to 2,5 x 106 per cm3 of rumen fluid. The different species were found and a positive correlation is seen between total numbers and protein content of the feed. It has been concluded that the numbers of E. par-uum are closely related to the protein concentration of the feed.

Rumen Ciliate Fauna of the Springbok (Antidorcas Marsupialis) in Southern Africa

AbstractSpringbok were sampled in Angola, South West Africa, the Kalahari, the Cape Province and Eastern Transvaal between July 1973 and July 1974. All the ciliates found in the twenty rumens investigated belonged to a single genus, Entodinium (family Ophryoscolecidae). Eight species were identified and two new species are described, E. kalaharicus and E. lucii.