Light Microscopic Observations of Infraciliature and Morphogenesis in Six Species of Rumen Ostracodinium Ciliates

Infraciliature and morphogenesis of six rumen ciliates, Ostracodinium mammosum, O. munham, O. dilobum, O. rugoloricatum, O. iwawoi, and O. tiete are described from pyridinated silver carbonate-impregnated specimens. These six Ostracodinium have a similar polybrachykinety arrangement, distinct from that of other ophryoscolecid ciliates and intermediate between those of O. gracile and 0. damaliscus. Buccal infraciliature is composed of three polybrachykineties, a kinety loop, and paralabial kineties. Small dorso-adoral polybrachykinety, slender vestibular polybrachykinety, and kinety loop are characteristic. Dorsal infraciliature consists of the dorsal polybrachykinety that extends laterally along the dorsal side of the body. Morphogenesis is different from that of O. gracile, because the right end of ventral primordium extends dorsally to grow into a right primordium without separation.     

Infraciliature and morphogenesis in three rumen Diplodinium ciliates, Diplodinium polygonale, Diplodinium leche, and Diplodinium nanum, observed by light microscopy

Infraciliature and morphogenesis of three rumen ophryoscolecid ciliates, Diplodinium polygonale Dogiel, 1925, Diplodinium leche Imai et al., 1992, and Diplodinium nanum Imai, 1988, are described from pyridinated silver carbonate-impregnated specimens. These three species have two polybrachykineties in the buccal area and a polybrachykinety in the dorsal ciliary zone. The vestibular polybrachykinety (VP) of D. polygonale and D. leche arises from the dorsal extremity of the adoral polybrachykinety (AP) as in Entodinium species, extending toward the left in D. polygonale and toward the left posterior in D. leche. The VP of D. nanum arises from the inner side of the AP, separate from its dorsal extremity, as in other Diplodinium species and extends toward the left posterior. These series of the polybrachykinety arrangements in D. polygonale, D. leche, and D. nanum can be regarded as transitional forms in the evolution of an Entodinium-like ancestor to Diplodinium. Morphogenesis of these three Diplodinium species is not different from that of other Diplodinium species.     

Light microscopical observation of infraciliary bands of Eodinium posterovesiculatum in comparison with Entodinium bursa and Diplodinium dentatum

Morphological features of the four morphotypes of the rumen ciliate, Eodinium posterovesiculatum, are described from pyridinated silver carbonate-impregnated specimens. Infraciliary bands are compared with those of Entodinium bursa and Diplodinium dentatum. In Entodinium bursa, the adoral polybrachykinety is "C" shaped and the vestibular polybrachykinety extends from the dorsal extremity of the adoral polybrachykinety as in other Entodinium species. In Diplodinium dentatum, the adoral polybrachykinety encircles most of the circumference of the vestibular opening and the vestibular polybrachykinety extends from the inner side of the adoral polybrachykinety as in other Diplodinium species. Infraciliary bands in Eodinium posterovesiculatum encircle the whole circumference of the vestibular opening and are, therefore, distinct from Entodinium, Diplodinium and other ciliates in the family Ophryoscolecidae. In Eodinium posterovesiculatum, the adoral polybrachykinety and the anterior part of the vestibular polybrachykinety encircle the vestibular opening. The wide and long vestibular polybrachykinety extends along the right wall of the tubular vestibulum and is bordered by a kinety. Kinetids in the central part of the vestibular polybrachykinety are randomly disposed. The genus Eodinium is valid because of this characteristic polybrachykinety arrangement in Eodinium posterovesiculatum.     

Diplodinium (Ostracodinium) minorum sp. n., Ciliate from the Rumen of Domestic Sheep*

SYNOPSIS. Diplodinium (Ostracodinium) minorum sp. n., observed in rumen contents from several domestic sheep, resembles Diplodinium (O.) ypsilon and Diplodinium (O.) magnum. It differs from the latter 2 species primarily in the size and proportions of the body, size of the rectum and anus, and in the presence of a short right caudal lobe in many individuals. In the samples studied, D. (O.) minorum constituted from 0.06 to 0.67% of the total rumen ciliate population.     

Ciliate Protozoa in the Rumen of Sassaby Antelope, Damaliscus lunatus lunatus, Including the Description of a New Species and Form

ABSTRACT. Composition of rumen ciliate fauna in five Zambian, sassaby antelopes was determined. Six genera, 18 species, and four forms were identified. One new species and form, belonging to the subfamily Diplodiniinae, were found, then labeled Ostracodinium damaliscus n. sp. and Diplodinium bubalidis f. aspinosum n. f., respectively. Only ophryoscolecid species were present while isotrichids were absent. Twelve of 18 total species are commmonly found in African antelopes. Three of those 12 species, Entodinium fyferi, Enoploplastron garstangi and Opisthotrichum janus , are only found in African antelopes. Percentage composition was low in the genera ml of rumen fluid, and the average number of ciliate species per host was 17.2.     

Rumen Ciliate Protozoa of the Blue Duiker (Cephalophus monticola), with Observations on Morphological Variation Lines Within the Species Entodinium dubardi

ABSTRACT. Protozoal concentrations were determined in rumen and cecal contents of 20 blue duikers ( Cephalophus monticola ). Ten animals of each sex were fed either a high concentrate or high roughage diet. Rumen protozoa were present in 19 of the 20 animals and concentrations ranged from 4.5 to 33.7 × 10⁶ per g of rumen contents. At the higher concentrations, protozoal cells equaled between 30–40% of the total rumen contents volume. No protozoa were found in cecal contents. Weight of rumen contents was higher in females than in males ( P < 0.01), and rumen protozoa concentrations were higher in males ( P < 0.05) and in those animals fed the high concentrate diet ( P < 0.05). All the protozoa were identified as belonging to a single species, Entodinium dubardi . However, an average of about 30% of the E. dubardi cells varied from the typical morphology of this species. These cells appeared to be on variation lines leading toward 7–10 other non-caudate species of Entodinium . The present data were used to evaluate and discuss the concept of variation lines within E. dubardi .     

Effects of fixation on routine,special and immunohistochemical stains: introduction to a symposium for the Biological Stain Commission

Orcein was first proposed as an elastic fiber stain by Taenzer in 1890, and has proved very useful for the purpose. It is an important constituent of the author's “Quad” stain. Unfortunately not all orcein samples have proved equally satisfactory, whether they are derived from the lichens from which the dye was originally prepared or have been manufactured by a synthetic process. At the present time several brands are available, and two of the brands of the synthetic product investigated have not proved to be the same thing; one of the latter proves only fair as an elastin stain, the other is one of the best samples the author has ever tried.     

Rumen Ciliate Protozoa in Ohio White-tailed Deer (Odocoileus virginianus)

ABSTRACT. Rumen contents were obtained from 23 white-tailed deer (Odocoileus virginianus), located in the eastern portion of central Ohio. Samples were taken during late fall and winter over a 4-yr period, 1986–1990. Protozoan numbers ranged from 0.002 to 7.25 × 10⁶ per ml of rumen contents, with a mean of 2.96 × 10⁶. Six deer had protozoan concentrations higher than any values previously reported in ruminants. In all 23 animals, Entodinium dubardi was the only ciliate protozoan species present.     

Rumen Ciliate Fauna of Alaskan Moose (Alces americana), Musk-Ox (Ovibos moschatus) and Dall Mountain Sheep (Ovis dalli)*

Total numbers, generic distribution and percentage species distribution were determined for the ciliate protozoa in rumen contents obtained from Alaskan moose (Alces americana), musk-ox (Ovibos moschatus) and Dall mountain sheep (Ovis dalli). The musk-ox has a fauna somewhat similar to that previously observed in reindeer and caribou. In contrast, only protozoa in the genus Entodinium were observed in moose, while Dall mountain sheep have a fauna unique among Alaskan ruminants studied to date. Other than Entodinium exiguum which was common to all animals, only 2 additional species of Entodinium, observed in the moose and musk-ox, occurred in more than one animal species. Four new species of protozoa are described, Entodinium dalli sp.n., Entodinium constrictum sp.n. and Polyplastron alaskum sp.n. from the Dall mountain sheep and Entodinium alces sp.n. from moose.     

Rumen microbes and fermentation of wild sika deer on the Shiretoko peninsula of Hokkaido Island,Japan

A total of 32 wild Hokkaido sika deer (Cervus nippon yesoensis) were shot (13 in summer, nine in autumn and 10 in winter) in the Syari district, Shiretoko Peninsula of Hokkaido Island, Japan. The ingested foods, rumen fermentation parameters and microbes were determined to evaluate digestive strategy and food availability in each season. Ingested foods and ruminal characteristics greatly varied by season. Rumen digesta mainly comprised of graminoids in summer, graminoids and agricultural products in autumn, and bark and twigs in winter. Rumen pH showed seasonal differences (P<0.05) and was lowest in summer, highest in winter, and intermediate in autumn, reflecting the seasonal differences in ruminal concentration of total volatile fatty acids which were significantly lower (P<0.05) in winter than in summer and autumn. Acetate proportions were significantly higher in winter than in other seasons (P<0.05), while the opposite trend was seen in proportions of propionate and butyrate. Rumen ammonia levels showed significant seasonal differences (P<0.05), decreasing from summer to autumn to winter. Rumen protozoa levels in autumn and winter decreased to 28 and 10% of the levels observed in summer, respectively (P<0.05 for both). The rumen bacteria level in winter was lower (P<0.05) than that in autumn, but no difference was seen for the other seasonal comparisons. Gram negative cocci were present in significantly higher proportions in winter than in other seasons (P<0.05), while Gram negative curved rods were less frequently observed in winter (P<0.05). Based on these results, wild sika deer in this area are shown to survive with rumen microbial populations altered with the dietary conditions that vary greatly by season.     

Isotricha jalaludinii N. Sp. Found from the Rumen of Lesser Mouse Deer, Tragulus javanicus, in Malaysia

Isotricha jalaludinii n. sp. found in the rumen of lesser mouse deer, Tragulus javanicus, in Malaysia was described and illustrated. This new species is characterized by the location and direction of the vestibulum, shape of the macronucleus, and absence of a dent at the vestibular opening. The presence of single peculiar isotrichid species in the rumen of mouse deer, which is recognized as one of the most primitive ruminants, suggests that the isotrichid ciliates similar to I. jalaludinii and Isotricha intestinalis were established at a fairly early period during the evolution of ruminants.     

Ciliate Protozoa in the Rumen of Kafue Lechwe, Kobus leche kafuensis, in Zambia, with the Description of Four New Species

ABSTRACT 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 times 10⁴, and the number of ciliate species per head of host was 10.8.     

Rumen ciliated protozoan fauna of the yak (Bos grunniens) in China with the description of Entodinium monuo n. sp

Rumen ciliate species composition was surveyed in domestic yaks kept in Tibet, Sichuan, and Inner Mongolia, China. Twelve genera including 36 species with 18 formae were identified. The species compositions were slightly different among the three areas: yaks in Tibet had the simplest fauna, in contrast, the fauna of yaks in Inner Mongolia were the most abundant and similar to those found in the cattle kept in the same area. This suggests that the rumen ciliate composition of yaks is affected by that of cattle kept together or in proximity. A new species belonging to the genus Entodinium, Entodinium monuo n. sp., was recognized from the yaks in all areas. This new species was common in the yaks but was not detected in the cattle fed near yaks in Inner Mongolia. There was a similar generic ciliate composition in yaks kept in respective areas. Entodinium was the most predominate ciliate (51.9-61.0%) with total ciliate densities estimated as 10(5)/ml per yak.     

Description of Opisthonecta Matiensis N. Sp. (Protozoa, Ciliophora), A New Peritrich Ciliate From Wastewater

Opisthonecta matiensis n. sp. was isolated from the inlet water of a wastewater treatment plant near Madrid, Spain, and studied in vivo, with silver methods, and using electronic and indirect immunofluorescence microscopy. This new species shows an amphora-like cell shape and has a size of 45-73 microns (x 58.2) x 25-40 microns (x 31.3). The oral infraciliature is formed by one haplokinety, three polykineties, and a short row of kinetosomes (epistomial membrane). The aboral infraciliature is made up of the trochal band and the scopula. From the trochal band arise three fibrillar systems: oral fibers, aboral fibers, and oblique fibers. The myoneme system is composed of a delicate peristomial ring, longitudinal branched fibers that reach the trochal band and of radial fibers extending from the scopula to the trochal band. The silverline system consists of an average of 147 lines. This new species is separated from other known forms by its smaller size, the presence of one single vacuole, and its higher number of silverlines.     

The Rumen Ciliate Fauna of Domestic Sheep (Ovis ammon aires) from the Turkish Republic of Northern Cyprus

ABSTRACT. 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 Ophryoscolex , 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. caudaquadricoronatum ), 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.     

Rumen protozoa are rich in polyunsaturated fatty acids due to the ingestion of chloroplasts

Within this study, we investigated whether the polyunsaturated fatty acids (PUFA)-rich nature of rumen protozoa is a consequence of ingestion of PUFA-rich chloroplasts. Four Hereford × Friesian steers were offered hay [low 18:3 (n-3) and low chlorophyll concentration] followed by freshly cut perennial ryegrass [high 18:3 (n-3) and high chlorophyll concentration] for 16 days. On the 14th and 16th days, rumen protozoa as well as attached and planktonic bacteria were fractionated 1 h before (−1 h), 2 and 6 h postfeeding, and their fatty acid concentrations determined. Protozoa fractionated from fresh grass-fed steers were richer ( P <0.05) in PUFA, except conjugated linoleic acid, for all time points compared with those from hay-fed steers. Protozoal density was higher ( P <0.05) for grass compared with hay. Entodinomorphid abundance was 3.4 times higher on fresh grass ( P <0.01) compared with hay. Confocal microscopy and transmission electron microscopy confirmed that Epidinium spp. were commonly saturated with intracellular cytoplasmic chloroplasts. These data suggest that engulfment of chloroplasts is a major contributor to the high 18:3 (n-3) concentration of protozoa.     

Diversity of the Formyltetrahydrofolate Synthetase Gene (fhs), a Key Enzyme for Reductive Acetogenesis,in the Bovine Rumen

A clone library of the partial formyltetrahydrofolate synthetase gene (fhs), a key enzyme in reductive acetogenesis, was constructed from the DNA of bovine rumen contents. Diverse sequences were recovered, the majority of which were clustered with the fhs of authentic acetogens. Low similarity values to known fhs were observed in all sequences, suggesting the presence of unknown acetogens.     

Fourteen morphotypes of Entodinium ovumrajae (Ophryoscolecidae, Entodiniomorphida) found in the Dromedary camel of Egypt

During a survey of the ciliate protozoal composition of the stomach contents of nine dromedary camels of Egypt, fourteen morphotypes of Entodinium ovumrajae, which has been considered as a species peculiar to camels, were found in six camels. Except for five morphotypes including one originally described as an independent species and its forms, these were newly detected. These morphotypes, divided into three groups, can be identified mainly by the morphology of their ectoplasmic processes. Each camel had on average, about five morphotypes of this species.     

Establishment of a spinated type of Diplodinium rangiferi by transfaunation of the rumen ciliates of Japanese sika deer (Cervus nippon centralis) to the rumen of two Japanese shorthorn calves (Bos taurus taurus)

One liter of rumen fluid containing 4.7 x 10(4) ciliates/ml, representing four genera including nine species of ciliates from a Japanese sika deer was inoculated into two unfaunated Japanese shorthorn calves. Two weeks after inoculation, all species originally present in the inoculum were subsequently detected in the rumen fluid of one or both calves. Ciliate densities ranged from 10(5)-10(6) cells/ml over the remainder of the 33-wk experiment. The inoculum contained Diplodinium rangiferi. which lacks caudal appendages, as is characteristic for the species. However, three weeks later, the rumen fluid of both calves contained D. rangiferi, which possesses caudal appendages varying from a single spine to multiple spines with a complicated furcate appearance. The caudal spines of D. rangiferi did not disappear during the experiment, even when the diet of the calves was switched to the ration of sika deer from which the inoculum was obtained.     

The effect of rumen ciliates on chitinolytic activity,chitin content and the number of fungal zoospores in the rumen fluid of sheep

The objective of this study was to investigate the effect of selected protozoa on the degradation and concentration of chitin and the numbers of fungal zoospores in the rumen fluid of sheep. Three adult ewes were fed a hay-concentrate diet, defaunated, then monofaunated with Entodinium caudatum or Diploplastron affine alone and refaunated with natural rumen fauna. The average density of the protozoa population varied from 6.1 · 10⁴ (D. affine) to 42.2 · 10⁴ cells/ml rumen fluid (natural rumen fauna). The inoculation of protozoa in the rumen of defaunated sheep increased the total activity of chitinolytic enzymes from 2.9 to 3.6 μmol N-acetylglucosamine/g dry matter (DM) of rumen fluid per min, the chitin concentration from 6.3 to 7.2 mg/g DM of rumen fluid and the number of fungal zoospores from 8.1 to 10.9 · 10⁵ cells/ml rumen fluid. All examined indices showed diurnal variations. Ciliate population density was highest immediately prior to feeding and lowest at 4 h thereafter. The opposite effects were observed for the numbers of fungal zoospores, the chitin concentration and chitinolytic activity. Furthermore, it was found that chitin from zoospores may account for up to 95% of total microbial chitin in the rumen fluid of sheep. In summary, the examined ciliate species showed the ability of chitin degradation as well as a positive influence on the development of the ruminal fungal population.