Ciliate Protozoa in the Rumen of Brazilian Water Buffalo,Bubalus bubalis Linnaeus*

SYNOPSIS Total numbers and distribution of genera, subgenera and species were determined for the ciliate protozoa in rumen contents of 4 Brazilian water buffalo Bubalus bubalis Linnaeus. The fauna of one animal, housed in close proximity to European and zebu-type cattle, differed considerably from that of the remaining animals, which were somewhat isolated on a large ranch. Several of the protozoan species observed in the semi-isolated animals were first described in rumen contents from humped Indian cattle, and their subsequent occurrence in other hosts and geographic locations has been limited or absent. In all, 49 different species of protozoa were found, 8 of which have not been previously described. Three of the new species belong to the genus Entodinium: E. ciculum sp. n., E. spinonucleatum sp. n. and E. triangulum sp. n.: 4 to Diplodinium (Ostracodinium): D. (O.) brazili sp. n., D. (O.) esalqum sp. n., D. (O.) nucleolobum sp. n., and D. (O.) tiete sp. n.; and one to Diplodinium (Eudiplodinium): D. (E.) bubalus sp. n.     

Ciliate protozoa in the rumen of Brazilian water buffalo, Bubalus bubalis Linnaeus.

Total numbers and distribution of genera, subgenera and species were determined for the ciliate protozoa in rumen contents of 4 Brazilian water buffalo Bubalus bubalis Linnaeus. The fauna of one animal, housed in close proximity to European and zebu-type cattle, differed considerably from that of the remaining animals, which were somewhat isolated on a large ranch. Several of the protozoan species observed in the semi-isolated animals were first described in rumen contents from humped Indian cattle, and their subsequent occurrence in other hosts and geographic locations has been limited or absent. In all, 49 different species of protozoa were found, 8 of which have not been previously described. Three of the new species belong to the genus Entodinium: E. ciculum sp. n., E. spinonucleatum sp. n. and E. triangulum sp. n.; 4 to Diplodinium (Ostracodinium): D. (O.) brazili sp. n., D. (O.) esalqum sp. n., D. (O.) nucleolobum sp. n., and D. (O.) tiete sp. n.; and one to Diplodinium (Eudiplodinium): D. (E.) bubalus sp. n.     

Ciliate Protozoa in the Rumen of Kenyan Zebu Cattle,Bos taurus indicus,with the Description of Four New Species1

ABSTRACT. The composition of the fauna of rumen ciliates in zebu cattle in Kenya was surveyed and 13 genera containing 51 species with 19 formae were identified. Four new species were recognized, then described as Diplodinium africanum n. sp., Diplodinium nanum n. sp., Eudiplodinium kenyensis n. sp., and Ostracodinium iwawoi n. sp. In addition, Buetschlia triciliata and Ostracodinium stokyi were found for the second and third times and described as Hsiungia triciliata n. comb. and Enoploplastron stokyi n. stat., with two formae, respectively. The species composition was similar to that of Indian zebu but several species were considered to originate from African native ruminants. In the percentage composition of genera, the numbers of Entodinium, which normally predominate in rumens in other areas, were very low.     

Ciliate protozoa in the rumen of Kenyan zebu cattle, Bos taurus indicus, with the description of four new species

The composition of the fauna of rumen ciliates in zebu cattle in Kenya was surveyed and 13 genera containing 51 species with 19 formae were identified. Four new species were recognized, then described as Diplodinium africanum n. sp., Diplodinium namum n. sp., Eudiplodinium kenyensis n. sp., and Ostracodinium iwawoi n. sp. In addition, Buetschlia triciliata and Ostracodinium stokyi were found for the second and third times and described as Hsiungia triciliata n. comb. and Enoploplaston stokyi n. stat., with two formae, respectively. The species composition was similar to that of Indian zebu but several species were considered to originate from African native ruminants. In the percentage composition of genera, the numbers of Entodinium, which normally predominate in rumens in other areas, were very low.     

Protozoa and digestive tract parameters in Blue wildebeest (Connochaetes taurinus) and Black wildebeest (Connochaetes gnou), with description of Entodinium taurinus n. sp.

Rumen contents from four blue wildebeest (Connochaetes taurinus) and six black wildebeest (Connochaetes gnou) were collected from two locations during two winter culling seasons. A total of 16 species of protozoa were found in blue wildebeest with Dasytricha ruminantium, Opisthotrichum janus and Ostracodinium gracile occurring in all four animals. In black wildebeest, 23 species of protozoa were observed with only Diplodinium bubalidis (caudal spine morphotype) and Ostracodinium damaliscus being present in all animals. In the blue wildebeest, the total number of species in an individual animal varied from 9 to 11 and in the black wildebeest, the number ranged from 4 to 14. Concentrations of cells per ml ranged from 1110 to 5880 in the blue wildebeest and 3120 to 6240 in the black wildebeest. This study is the first report on protozoa species in the blue and black wildebeest. A new species of Entodinium is described, Entodinium taurinus n. sp., observed in the rumen contents of three blue wildebeest. Several physical parameters of the digestive tract were also measured, including distribution, pH and density of gastrointestinal contents in different sections of the total tract. In vitro gas production was estimated for rumen, cecum and colon contents.     

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.     

Ruminal ciliated protozoa in bison.

Ruminal contents from 79 slaughtered bison and 2 ruminally cannulated bison were collected to obtain information on total numbers and species distribution of ciliated protozoa. The bison originated from numerous herds throughout the Great Plains and were grouped into three dietary categories: (i) only forage; (ii) forage with moderate levels of supplementation; and (iii) feedlot concentrate-silage diet. Total ciliate counts were highest in bison receiving grain supplementation (210.1 x 10(4)/g) and lowest in bison consuming only forage (27.1 x 10(4)/g). All protozoan species found in bison have been reported in domestic livestock, although Ophryoscolex sp., a relatively common protozoan in cattle, was detected at low concentrations in only eight bison. The uncommon holotrich Microcetus lappus was present in five bison in concentrations reaching 8.4% of the total ciliate population. Charonina ventriculi, another infrequently observed species, was present in 18 bison, with the highest concentrations in forage-fed animals. Thirty bison possessed a type B protozoan population, characterized by Epidinium sp., Eudiplodinium maggii, and Eudiplodinium bovis. Thirty-eight bison possessed a mixed A-B population, characterized by Polyplastron sp. coexisting with low numbers of Eudiplodinium maggii or Epidinium sp. or both. Thirteen bison possessed populations lacking any remnant type B ciliate species. At least 29 of the bison possessing Polyplastron sp. were known to have been in contact with cattle, whereas all bison isolated from cattle had type B populations. The reduction of type B populations in bison becomes increasingly likely as bison production expands into areas inhabited by domestic livestock.     

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.     

Ruminal ciliated protozoa in bison

Ruminal contents from 79 slaughtered bison and 2 ruminally cannulated bison were collected to obtain information on total numbers and species distribution of ciliated protozoa. The bison originated from numerous herds throughout the Great Plains and were grouped into three dietary categories: (i) only forage; (ii) forage with moderate levels of supplementation; and (iii) feedlot concentrate-silage diet. Total ciliate counts were highest in bison receiving grain supplementation (210.1 x 10(4)/g) and lowest in bison consuming only forage (27.1 x 10(4)/g). All protozoan species found in bison have been reported in domestic livestock, although Ophryoscolex sp., a relatively common protozoan in cattle, was detected at low concentrations in only eight bison. The uncommon holotrich Microcetus lappus was present in five bison in concentrations reaching 8.4% of the total ciliate population. Charonina ventriculi, another infrequently observed species, was present in 18 bison, with the highest concentrations in forage-fed animals. Thirty bison possessed a type B protozoan population, characterized by Epidinium sp., Eudiplodinium maggii, and Eudiplodinium bovis. Thirty-eight bison possessed a mixed A-B population, characterized by Polyplastron sp. coexisting with low numbers of Eudiplodinium maggii or Epidinium sp. or both. Thirteen bison possessed populations lacking any remnant type B ciliate species. At least 29 of the bison possessing Polyplastron sp. were known to have been in contact with cattle, whereas all bison isolated from cattle had type B populations. The reduction of type B populations in bison becomes increasingly likely as bison production expands into areas inhabited by domestic livestock.     

Rumen ciliates from Tanzanian short horn zebu cattle,Bos taurus indicus,and the infraciliature of Entodinium palmare n.sp. and Enoploplastron stokyi ()

Rumen ciliates of ten Tanzanian short horn zebu cattle were examined. A total of 15 genera and 46 species were identified, including a new Entodinium species. The ciliate density was 22.2×10⁴ ml^?1. The number of species per host and the diversity index showed high values, 33.8 and 2.80, respectively. Rumen ciliates had a low percentage composition of the genus Entodinium (7.0–25.0%) and a slightly higher percentage composition of the genera, Eudiplodinium (19.3%), Diplodinium (14.1%), and Ostracodinium (13.1%). Entodinium palmare n.sp., Eudiplodinium kenyensis, and Enoploplastron stokyi were found in all cattle examined. The former two species have been found only in African zebu cattle. Entodinium palmare n. sp. has a characteristic right surface of the body like the “palm of a hand” because of the concave part on the postero-dorsal part of the body, and has the same pattern of infraciliary bands as in other Entodinium species. Enoploplastron stokyi has a characteristic pattern of infraciliary bands analogous to those of Epidinium ecaudatum and Ostracodinium mammosum; with the right side of the adoral polybrachykinety gradually tapering and a slender short vestibular polybrachykinety.     

Rumen Ciliates of White-tailed Deer (Odocoileus virginianus), Axis Deer (Axis axis), Sika Deer (Cervus nippon) and Fallow Deer (Dama dama) from Texas

Samples of rumen contents from 33 white-tailed deer (Odocoileus virginianus), 31 axis deer (Axis axis), 26 sika deer (Cervus nippon), and 25 fallow deer (Dama dama) were collected from four study areas in central Texas. The geometric mean concentration of total protozoa was 50.2 x 10(4) per ml, with no differences between species (P > 0.36). White-tailed deer had a higher percentage of Entodinium and lower percentage of Diplodiniinae (P < 0.01) than the other deer species, which were not different from each other. Occurrence of Epidinium, Isotricha, and Dasytricha was sporadic and did not differ among deer species. Numerous new host records of protozoan species were observed: white-tailed deer--four; axis deer--five; sika deer--five; fallow deer--four. This brings the total number of protozoan species identified in each deer species to: white-tailed--eight; axis--12; sika--15; fallow--16. For all species combined, protozoan concentration were 7.5 to 11-fold higher (P < 0.01) from Area 4, which differed from the other three areas by having a stream that allowed deer to have free access to water. Criteria used for identification of medium-size Eudiplodinium species were evaluated.     

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 Fauna of Some Brazilian Cattle: Occurrence of Several Ciliates New to the Rumen,Including the Cycloposthid Parentodinium africanum1

ABSTRACT. Parentodinium africanum was observed in rumen contents of four Brazilian cattle and constituted 4.4, 0.9, > 0.1, and 10.0% of the total ciliates. This appears to be the first observation of the family Cycloposthiidae in the rumen habitat. Blepharoconus krugerensis, previously observed in the intestines of the elephant, and an unknown species of ciliate with many characteristics similar to the family Paraisotrichidae were each observed in a single animal. A new subspecies, Diplodinium flabellum laterospinatum n. subsp., is also described. Total number of ciliates per ml of rumen contents ranged from 9.0 to 51.2 × 10⁴ and included an overall total of 55 species and 4 subspecies.     

The importance of methanogens associated with ciliate protozoa in ruminal methane production in vitro

The importance of methanogenic bacteria associated with ciliate protozoa was estimated either by removing protozoa from whole rumen fluid (using defaunated rumen fluid to correct for the effects of centrifugation on bacteria) or by isolating the protozoa. Rumen fluid was withdrawn from sheep inoculated with either Polyplastron multivesiculatum , a co-culture of Isotricha prostoma plus Entodinium spp. or a mixed type B fauna of Entodinium, Eudiplodinium and Epidinium spp. Methanogenesis was highest in rumen fluid containing a mixed protozoal population of the following genera: Entodinium, Eudiplodinium and Epidinium , was lower in defaunated rumen fluid and lowest in rumen fluid containing either I. prostoma plus Entodinium or P. multivesiculatum . Methanogenic bacteria associated with rumen ciliates were apparently responsible for between 9 and 25% of methanogenesis in rumen fluid.     

Rumen ciliates of domestic cattle (Bos taurus taurus) in Kastamonu,Turkey, with the description of a new species

Species composition and distribution of ciliates were investigated in the rumen contents of 25 domestic cattle (Bos taurus taurus L.) living in Kastamonu, Turkey. Forty-seven species and 37 morphotypes representing 15 genera were identified. Of them, a new species of Ostracodinium was recognized and described as Ostracodinium anatolicum n. sp. This new species has two caudal lobes. The dorsal lobe is small and rounded and the ventral lobe is triangular shaped and bent toward the dorsal side like a thick hook. Furthermore, the anterior end of the macronucleus (1/5 of the length) is bent toward the left like a hook. The density of rumen ciliates in cattle was 96.8 ± 43.3 × 10⁴ cells mL⁻¹ and the mean number of ciliate species per host was 14.2 ± 4.4. Entodinium longinucleatum, E. nanellum, E. simulans and Isotricha prostoma were the most abundant species, each with a prevalence of 88%. Entodinium chatterjeei, E. bifidum m. monospinosum, Hsiungia triciliata, Oligoisotricha bubali, Ostracodinium dogieli, O. mammosum and O. munham are new host records for cattle from Turkey.     

Suitability of different media for in vitro cultivation of the ruminal protozoa species Entodinium caudatum, Eudiplodinium maggii, and Epidinium ecaudatum

Three protozoal cultivation media were tested to determine the medium which best facilitated growth and viability of key B-type ciliates isolated from the sheep rumen. Entodinium caudatum and Eudiplodinium maggii were grown anaerobically in 50-ml flasks for 32 days in Caudatum-type (C), Kisidayova (K) or Dehority (M) medium. On day 32, in media K and M, E. caudatum cell counts were high with 5.6 × 10³ and 7.8 × 10³ mL⁻¹, respectively, and the proportion of dead cells was low with 0.6 and 1.4%, respectively. E. maggii concentrations when grown in medium M and C were 2.7 × 10³ and 2.4 × 10³ mL⁻¹, respectively, with 3.9 and 14.1% dead cells. Medium M, which favoured growth of both protozoa species, was tested again and Epidinium ecaudatum was included. Protozoa were grown for a 4-month period and samples were taken in the last two months on days 1, 7, 35 and 57. Average cell concentrations were 10.0, 0.8 and 0.5 × 10³ mL⁻¹ for E. caudatum, E. maggii, and E. ecaudatum, respectively. In conclusion, medium M would appear to be the best choice for cultivating these three species in one medium.     

Phylogenetic analysis of protozoa in the rumen contents of cow based on the 18S rDNA sequences

AIMS: To examine the diversity of protozoa in the rumen contents of cow. METHODS AND RESULTS: Protozoa that inhabit the rumen were detected by PCR using protozoan-specific primers. Libraries of protozoan rDNA sequences were constructed from rumen fluid, solid tissues and epithelium. Twenty-three clones isolated from rumen fluid fell into two genera identified as Entodinium (69.6% of clones) and Epidinium (31.4% of clones). Of the clones isolated from rumen fluid, a moderate number were unidentifiable (30.4%). CONCLUSIONS: The predominant protozoan genus identified in the whole rumen belonged to the Entodinium group (81.1%). Protozoa were not detected in the rumen epithelium. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings suggest that rumen fluid and solid tissues contain different protozoan populations that may play specific roles in rumen function. Quantitative PCR techniques and a more specific set of phylogenetic probes that distinguish between protozoan species are needed to determine the significance of newly identified groups and to determine the distribution of identified protozoan clusters in rumen microbial communities.     

In vitro determination of generation times for Entodinium exiguum, Ophryoscolex purkynjei and Eudiplodinium maggii

ABSTRACT. Most previously reported generation times for rumen ciliate protozoa are longer than would be required to prevent their being flushed out of the rumen. In an earlier study from this lab, using a sequential transfer procedure, generation times between 12 and 13 h were determined for both Epidinium caudatum and Entodinium caudatum . This would permit these species to be maintained in a rumen with a fluid volume turnover rate as rapid as twice a day. In this study, generation times were estimated for Entodinium exiguum (13.2 h), Eudiplodinium maggii (26.8 h), and Ophryoscolex purkynjei (29 h), by sequential transfer at both 12 and 24 h time periods. The generation time for E. exiguum is lower than reported for this and other Entodinium species as determined by logarithmic growth from a small inoculum, but similar to that obtained for Ent. caudatum using sequential transfer. Eudiplodinium maggii and O. purkynjei generation times are similar to previous estimates of 24- and 24–48 h, respectively. However, it was observed that after an adaptation period of 36 to 48 h (generally 3–4 transfers) cell concentrations decreased and generation times were markedly decreased, i.e. 12.2 h for Ent. exiguum , 15.0 h for E. maggii and 12.8 h for O. purkynjei . In a separate study, varying both the concentration of Epidinium and the quantity of substrate fed per cell had no effect on generation time.