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A growth trail was carried out to determine the effect of feeding rations with different levels of concentrate: roughage ratio on rumen development through examining the rumen fermentation and the identification and density of rumen ciliate protozoa for sequence five months. Eighteen early weaned Balady male goats with an average live body weight about 7.58 kg at 60 days age were randomly allocated in three groups (6 lambs each) according to body weight. The three groups were fed starter pelleted consists of different concentrate: roughage ratios (T1; 90:10, T2; 80:20 and T3; 70:30). Results showedthat rumen parameters concentration and ruminal ciliate protozoa count were significantly increased (P≤0.01) from the age of 8 weeks till the age of 24 weeks. Also, the data indicated that ruminal pH values and ruminal ciliate protozoa count were higher (P≤0.01) before feeding then it decreased at 3 hours post feeding followed by gradually increased, although total volatile fatty acids, ammonia nitrogen, non-protein nitrogen, total nitrogen and true protein nitrogen concentrations reached the highest (P≤0.01) value at 3hr post-feeding then decreased gradually. Seven species of ruminal protozoa were identified in this study, Entodinum spp was the most dominant specie. Comparison among the three experimental treatments showed that T3 had the highest (P≤0.01) concentrations of rumen parameters and ruminal ciliate protozoa count during the whole period followed by T2, while T1 had the lowest (P≤0.01) values. So we recommended involving high roughage ratio in goat kids feeding.

Research Areas: Science & Technology - Other Topics This study investigated the associations between abundance of rumen ciliate protozoa and the proportion of the main bioactive fatty acids related to rumen biohydrogenation, as 18:0, t10-18:1, t11-18:1, c9,t11-18:2, 18:3n-3 and 18:2 n-6, in rumen and meat of growing lambs, using data derived from 3 production experiments. A global correlation analysis and a linear regression analysis considering the effect of the experiment were performed. Ten of the 86 lambs involved in the experiments did not present ciliate cells in rumen liquor and the remaining lambs presented an average of 1.35 x 106ciliates / ml rumen liquor. From the nine genera of ciliates identified, Entodinium was the most abundant, averaging 1.17 x 106 cells / ml of rumen liquor. A large variation among lambs was observed for both rumen concentration and community structure of ciliates. Rumen t11-18:1 (P< 0.001) and meat deposition of t11-18:1 (P < 0.001) and of c9,t11-18:2 (P < 0.001) increased linearly with total ciliates, whereas the t10/t11 ratio in rumen (P= 0.002) and in meat (P= 0.036) decreased linearly. Entodiniomorphids seems to be strongly related with meat deposition of t11-18:1 and c9,t11-18:2 and with the reduction of the trans-10 shifted pathway. Completeness of RBH decreased linearly with Holotrichs (P = 0.029), Entodiniomorphids (P = 0.029), Isotricha (P = 0.011) and Epidinium (P = 0.027) abundances. Rumen 18:0 also decreased linearly with increasing counts of total ciliates (P = 0.015), Holotrichs (P = 0.020), Entodiniomorphids (P= 0.010) and Isotricha (P= 0.014). Rumen protozoa were positively linked with the deposition of healthy bioactive FA and simultaneously negatively associated with the occurrence of trans-10 shift. info:eu-repo/semantics/publishedVersion

SUMMARY The effects of cassava wastewater on nutrient digestibility and rumen protozoa were evaluated in 32 Santa Inês sheep fed a diet with Tifton hay, ground corn cob, and concentrate, containing 0, 500, 1000, or 1500 mL cassava wastewater. Uncastrated sheep at an average age of 167 days and average weight of 25.3 kg were housed in individual stalls and distributed in a randomized complete design with four treatments and eight replicates. The experimental period was 70 days: 15 days for adaptation to the diets and 60 days to collect the ingredients and diets supplied. Prior to this stage, the animals were adapted to the consumption of cassava wastewater, in collective cages, for seven days. Apparent digestibility was determined by partial feces collection and by using the LIPE® external marker in 250-mg capsules. The apparent digestibility of nutrients did not differ (P>0.05) in relation to the cassava wastewater levels tested. Eleven genera of rumen protozoa were identified, and they formed a community that, except for Ophryoscolex, did not differ among the cassava wastewater treatments with respect to composition. However, Entodinium species were observed at a higher density (P0,05) em relação aos níveis de manipueira estudados. Onze gêneros de protozoários foram identificados, compondo uma comunidade que à exceção de Ophryoscolex, não diferiu entre os tratamentos em sua composição. Entretanto, as espécies de Entodinium apresentaram maior densidade (P

The methods used for culturing rumen protozoa were found to be unsatisfactory for growth of ciliate protozoa from the kangaroo forestomach. Based on published measurements of physical parameters in the marsupial forestomach, several modifications were incorporated into the procedure, i.e., an increase in % hydrogen in the gas phase, adjustment of initial pH of the medium to 6.9–7.0 range, feed only forage as a substrate and incubate at a lower temperature (33–36 °C). Only incubation at the lower temperature increased survival time of the kangaroo protozoa. Two species of Bitricha were still viable after 28 d in culture. Cultures had to be terminated at that time. One of the species differed considerably in size and shape from previously described species and based on 18S rRNA data, may represent a new species of Bitricha. The second species, present in low numbers was identified as Bitricha oblata. In a separate trial, Macropodinium yalanbense survived for 11 d, at which time these cultures also had to be terminated.

The flow of ciliate protozoa from the reticulo-rumen is significantly less than expected given the total density of rumen protozoa present. To maintain their numbers in the reticulo-rumen, protozoa can be selectively retained through association with feed particles and the rumen wall. Few mathematical models have been designed to model rumen protozoa in both the free-living and attached phases, and the data used in the models were acquired using classical techniques. It has therefore become necessary to provide an updated model that more accurately represents these microorganisms and incorporates the recent literature on distribution, sequestration, and generation times. This paper represents a novel approach to synthesizing experimental data on rumen microorganisms in a quantitative and structured manner. The development of a linear programming model of rumen protozoa in an approximate steady state will be described and applied to data from healthy ruminants consuming commonly fed diets. In the model, protozoa associated with the liquid phase and protozoa attached to particulate matter or sequestered against the rumen wall are distinguished. Growth, passage, death, and transfer of protozoa between both pools are represented. The results from the model application using the contrasting diets of increased forage content versus increased starch content indicate that the majority of rumen protozoa, 63 to 90%, are found in the attached phase, either attached to feed particles or sequestered on the rumen wall. A slightly greater proportion of protozoa are found in the attached phase in animals fed a hay diet compared with a starch diet. This suggests that experimental protocols that only sample protozoa from the rumen fluid could be significantly underestimating the size of the protozoal population of the rumen. Further data are required on the distribution of ciliate protozoa in the rumen of healthy animals to improve model development, but the model described herein does indicate that the attached protozoal population is a significant component of the total rumen protozoal community. Key words: linear programming , protozoa , reticulorumen , sequestration

In order to study the effect of pH on defaunation in the rumen, four rumen fistulated steers were fed a basal roughage diet for a 4-week adaptation period followed by 17 weeks of feeding with three diets and two feeding levels of high concentrate diet. Rumen outflow fluid rate was evaluated in both ration levels. Rumen protozoa population was monitored weekly and when animals became defaunated, protozoa were reinoculated with rumen contents from one of the faunated steers. At every two weeks, during all the experimental period, rumen pH was measured in all animals at 0, 4, 8 and 12 h after feeding. It was observed an individual animal influence on the establishment and maintenance of the rumen ciliate protozoa population. In all sampling times, mean rumen pH values were higher in faunated steers than in the defaunated ones. No differences were observed in rumen outflow fluid rates between the two ration levels. Extended periods of low rumen pH are probably more detrimental to the survival of ciliate protozoa in the rumen than other factors.

Made available in DSpace on 2013-08-12T18:13:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-04-01 Made available in DSpace on 2013-09-30T18:01:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-04-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:18:36Z No. of bitstreams: 0 Made available in DSpace on 2014-05-20T13:18:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-04-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The objective of this research was to evaluate the effect of the dry sugar cane yeast (DSCY) protein addition on the intake, digestibility and rumen microbial population (bacteria, protozoa, and fungi) in zebu steers fed cotton seed processing residue (CSPR). Six Nellore steers, with rumen and duodenal cannulae were used. Data were analyzed according to a randomized block design, each one with three treatments: 0.0, 4.0 and 8.0% of DSCY as a percentage of supplied dietary dry matter. Parameters evaluated were: intake, digestibility and ruminal protozoa populations. In addition, the most probable numbers of rumen fungal and bacteria populations were estimated. The DM, OM, EE, NDF and ADF intakes and digestibilities were not influenced by different levels of DSCY, possibly because of the high value of the nitrogen C fraction of the CSPR (44.5% DM). Low levels of undegradable protein fraction from yeast did not provide an expected optimized synchronization energy/nitrogen in order to enhance the degradation rate. The presence of 4.0% DSCY in the diet significantly reduced fungal rumen population. There were no significant changes in fungal populations between the 4.0% and the 8.0% treatments. However, bacterial populations showed an increase when submitted to 8.0% DSCY compared to 4.0% in the diet. Protozoal populations did not vary significantly among the treatments. It was concluded that supplementation of DSCY to zebu steers did not affect the intake, digestibility and microbiological rumen populations. (C) 2010 Elsevier B.V. All rights reserved. São Paulo State Univ, Dept Zootecnia, Fac Ciencias Agr & Vet, UNESP,Dept Anim Sci, BR-14884900 Jaboticabal, SP, Brazil São Paulo State Univ, Dept Zootecnia, Fac Ciencias Agr & Vet, UNESP,Dept Anim Sci, BR-14884900 Jaboticabal, SP, Brazil

Estudaram-se os efeitos de fontes de enxofre na dieta sobre os parâmetros ruminais de bovinos Nelore, utilizando-se oito novilhos canulados no rúmen e submetidos a quatro tratamentos, segundo a fonte de enxofre. Os animais do grupo-controle não receberam suplementação de enxofre. Os dos outros três grupos receberam flor de enxofre ou metionina ou carboquelatado, como suplementação. Os animais foram arraçoados com dieta total, utilizando cana-de-açúcar picada como volumoso. O período experimental foi de 76 dias, dividido em quatro subperíodos de 19 dias, dos quais 14 eram para adaptação à dieta e cinco para as colheitas. Não foram encontradas diferenças entre as fontes de enxofre na degradabilidade da matéria seca, proteína bruta e fibras em detergente ácido e neutro e no pH ruminal. O tratamento com carboquelatado resultou em maior número de protozoários ciliados e o tratamento metionina em menor contagem. O carboquelatado pode ser uma boa alternativa para a suplementação de enxofre para bovinos em confinamento. This research was carried out at FZEA/USP to compare the effects of different sulphur sources in diets of Nellore cattle on ruminal parameters. Eight steers were rumen cannulated and submitted to four treatments, using the following sulphur sources: control (no supplement), elemental sulphur, methionine, and carboquelated. Animals were fed a total ration using sugarcane as the roughage. The proportion roughage:concentrate was 40:60. The experimental period lasted 76 days, divided in four 19-days sub-periods. The first 14 days of each sub-period were used to adapt the animals to the diet and the last five days to collect the samples. During the sample period, ruminal liquid was sampled to protozoa count and pH determination. Also, nylon bags were incubated in rumen to determinate the degradability of dietary dry matter, crude protein, and acid and neutral detergent fiber. Treatments did not affect dietary dry matter, crude protein, acid and neutral detergent fiber degradability, and the ruminal pH. However, carboquelated provided the higher total amount of ciliate protozoa and methionine provided the lowest values. Carboquelated can be a good alternative to supplement sulphur to bovines in a feedlot.

This experiment assayed the influence of the inclusion of dried Azardirachta indica, Albizzia lebbek or Ailanthus excelsa leaves in pearl millet stover-based complete feed block diets on feed intake, nutrient utilization, rumen fermentation characteristics, ciliate protozoa population and blood biochemical constituents in adult Malpura sheep. Complete feed blocks were formulated to have roughage-to-concentrate ratio of 70:30. Pearl millet stover (PMS) was used as basal roughage; 30 parts of pearl millet stover was replaced with dried leaves either of Azardirachta indica (NL), Albizzia lebbek (SL) or Ailanthus excelsa (AL). Twelve hogget Malpura rams, divided into four equal groups, were offered one of the four dietary treatments. A feeding-cum-metabolic trial was conducted to assess nutrient utilization. Rumen liquor samples were collected at 0, 3, 6, 12, 18 and 24 h post-feeding to assess rumen fermentation pattern and ciliate protozoa population. Inclusion of dried leaves in PMS-based diets improved CP and DCP content. Dietary DCP was low (P

Moose rumen samples from Vermont, Alaska and Norway were investigated for methanogenic archaeal and protozoal density using real-time PCR, and diversity using high-throughput sequencing of the 16S and 18S rRNA genes. Vermont moose showed the highest protozoal and methanogen densities. Alaskan samples had the highest percentages of Methanobrevibacter smithii, followed by the Norwegian samples. One Norwegian sample contained 43 % Methanobrevibacter thaueri, whilst all other samples contained 70 % Polyplastron multivesiculatum and one contained >75 % Entodinium spp. Protozoa from Norwegian moose belonged predominantly (>50 %) to the genus Entodinium, especially Entodinium caudatum. Norwegian moose contained a large proportion of sequences (25–97 %) which could not be classified beyond family. Protozoa from Vermont samples were predominantly Eudiplodinium rostratum (>75 %), with up to 7 % Diploplastron affine. Four of the eight Vermont samples also contained 5–12 % Entodinium spp. Samples were heterogeneous based on AMOVA, principal coordinate analysis and UniFrac. This study gives the first insight into the methanogenic archaeal diversity in the moose rumen. The high percentage of rumen archaeal species associated with high starch diets found in Alaskan moose corresponds well to previous data suggesting that they feed on plants high in starch. Similarly, the higher percentage of species related to forage diets in Vermont moose also relates well to their higher intake of fibre.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T19:06:22Z No. of bitstreams: 1 S1516-35982002000300022.pdf: 211839 bytes, checksum: 16fbca738e79e73c0dc02b7bd681d9bb (MD5) Made available in DSpace on 2013-08-22T19:06:22Z (GMT). No. of bitstreams: 1 S1516-35982002000300022.pdf: 211839 bytes, checksum: 16fbca738e79e73c0dc02b7bd681d9bb (MD5) Previous issue date: 2002-01-01 Made available in DSpace on 2013-09-30T20:10:38Z (GMT). No. of bitstreams: 2 S1516-35982002000300022.pdf: 211839 bytes, checksum: 16fbca738e79e73c0dc02b7bd681d9bb (MD5) S1516-35982002000300022.pdf.txt: 34604 bytes, checksum: a61c776fd89b20bda9d37c3bde24aa36 (MD5) Previous issue date: 2002-01-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:18:00Z No. of bitstreams: 2 S1516-35982002000300022.pdf: 211839 bytes, checksum: 16fbca738e79e73c0dc02b7bd681d9bb (MD5) S1516-35982002000300022.pdf.txt: 34604 bytes, checksum: a61c776fd89b20bda9d37c3bde24aa36 (MD5) Made available in DSpace on 2014-05-20T15:18:00Z (GMT). No. of bitstreams: 2 S1516-35982002000300022.pdf: 211839 bytes, checksum: 16fbca738e79e73c0dc02b7bd681d9bb (MD5) S1516-35982002000300022.pdf.txt: 34604 bytes, checksum: a61c776fd89b20bda9d37c3bde24aa36 (MD5) Previous issue date: 2002-01-01 Quatro bubalinos e quatro bovinos, fistulados no rúmen, foram distribuídos aleatoriamente a dois quadrados latinos (4 x 4) para avaliar os efeitos de rações com quatro níveis de FDN (54, 60, 66 e 72%) e de dois tamanhos de poros em sacos de náilon (53 e 100 mm) sobre a população de protozoários ciliados no conteúdo ruminal e dentro dos sacos incubados em diversos tempos. Não houve diferença nos diversos gêneros estudados e no total de protozoários nas amostras dos sacos de náilon entre 53 e 100 mm. As concentrações de Entodinium e de total de protozoários do conteúdo ruminal elevaram-se até o nível de 66% de FDN, com queda acentuada em 72% nos bubalinos, enquanto nos bovinos houve redução com o aumento de FDN na ração. Houve diferenças nas concentrações dos protozoários da subfamília Diplodiniinae, Epidinium e Dasytricha entre bubalinos e bovinos, dentro das amostras incubadas em sacos com poros de 53 mm. A concentração de Diplodiniinae aumentou com 72% de FDN na dieta em ambas as espécies animais. Os bubalinos apresentaram valores médios de pH e taxa de passagem do líquido ruminal mais elevados e volumes ruminais menores que os bovinos. Four buffaloes and four cattle, rumen fistulated, were utilized in two Latin Square (4 x 4) to evaluate the effects of rations with increasing levels of NDF (54, 60, 66 and 72% in DM) and two porous size of the nylon bags (53 and 100 mm) on the population of ciliate protozoa in rumen content and inside the nylon bags suspended in the rumen for several times. There was no difference in the several genus and in the total of protozoa in the samples of the nylon bags between 53 and 100 mm. The concentration of Entodinium and total of protozoa in the rumen content increased in the level of 66% of NDF with drop accentuated in 72% in the buffaloes, while in the cattle there was significant decrease with the increase of NDF in the ration. Rumen protozoa concentration of Diplodiniinae subfamily, Epidinium and Dasytricha were different for buffaloes and cattle inside the rumen incubated nylon bags with 53 mm pore size. Diplodiniinae concentration in the rumen content increased with 72% of NDF in diet in both animal species. The buffaloes presented higher pH average values and rate of passage and lower ruminal volumes than cattle. UNESP Faculdade de Medicina Veterinária e Zootecnia USP Faculdade de Zootecnia e Engenharia de Alimentos UNESP Faculdade de Medicina Veterinária e Zootecnia

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T19:06:07Z No. of bitstreams: 1 S1516-35982000000500025.pdf: 29415 bytes, checksum: 5adb970e014d72068caa4577dc4cbc8c (MD5) Made available in DSpace on 2013-08-22T19:06:07Z (GMT). No. of bitstreams: 1 S1516-35982000000500025.pdf: 29415 bytes, checksum: 5adb970e014d72068caa4577dc4cbc8c (MD5) Previous issue date: 2000-10-01 Made available in DSpace on 2013-09-30T20:10:15Z (GMT). No. of bitstreams: 2 S1516-35982000000500025.pdf: 29415 bytes, checksum: 5adb970e014d72068caa4577dc4cbc8c (MD5) S1516-35982000000500025.pdf.txt: 23327 bytes, checksum: 967dddad985157fbb8953e2d0fc0620b (MD5) Previous issue date: 2000-10-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:17:53Z No. of bitstreams: 2 S1516-35982000000500025.pdf: 29415 bytes, checksum: 5adb970e014d72068caa4577dc4cbc8c (MD5) S1516-35982000000500025.pdf.txt: 23327 bytes, checksum: 967dddad985157fbb8953e2d0fc0620b (MD5) Made available in DSpace on 2014-05-20T15:17:53Z (GMT). No. of bitstreams: 2 S1516-35982000000500025.pdf: 29415 bytes, checksum: 5adb970e014d72068caa4577dc4cbc8c (MD5) S1516-35982000000500025.pdf.txt: 23327 bytes, checksum: 967dddad985157fbb8953e2d0fc0620b (MD5) Previous issue date: 2000-10-01 Oito ovinos machos com fístulas ruminais foram delineados em dois quadrados latinos, em quatro períodos de 35 dias cada e quatro tratamentos, compreendendo níveis crescentes de cana-de-açúcar (CA) em substituição à silagem de milho (SM): A) 100% SM; B) 67% SM e 33% CA; C) 33% SM e 67% CA; e D 100% CA, para avaliação da população de protozoários ciliados no rúmen. As concentrações totais de protozoários ciliados no rúmen e de Entodinium diminuíram e os valores de pH do conteúdo ruminal aumentaram linearmente com o incremento das quantidades de cana-de-açúcar na ração. Não houve diferenças entre tratamentos para o volume ruminal e a taxa de passagem da fase líquida do rúmen. Eight rumen fistulated rams were delineated in two Latin Squares in four periods of 35 days each and four treatments consisted of increasing levels of sugar cane (SC) in substitution to the corn silage (CS): A) 100% CS, B) 67% CS and 33% SC, C) 33% CS and 67% SC and D) 100% SC, to evaluate the population of ciliate protozoa in the rumen. The total concentrations of rumen ciliate protozoa and of Entodinium decreased and the values of pH of the ruminal content increased linearly as dietary sugar cane levels increased. No differences between treatments were observed for the rumen volume and the rumen liquid turnover rate. UNESP FMVZ USP Faculdade de Medicina Veterinária e Zootecnia USP Faculdade de Zootecnia e Engenharia de Alimentos UNESP FMVZ

An experiment was conducted to study the effect of presence or absence of rumen ciliate protozoa on nutrient intake, its utilization, rate of body weight gain and feed conversion efficiency of lambs in post-weaning phase of growth. Forty five weaned Malpura lambs, divided into three equal groups (G1, G2 and G3) were either defaunated by periodic dosing of sodium lauryl sulfate at the rate of 8 g/100 kg body weight (G1) or defaunated and refaunated (G2) or maintained faunated (G3). The lambs were fed for 90 days on 50:50 pala (Zizyphus nummularia) leaf and concentrate based complete feed. With care and segregation the defaunated lambs were maintained free of rumen protozoa for 47 days after which Entodinomorphid appeared while Holotrichs were not detected. The total and differential protozoa count was similar in refaunated and faunated lambs and the residual toxic effect of sodium lauryl sulfate if any was not detected. Even with similar dry matter intake in the three groups, body weight gain and feed conversion efficiency were higher in defaunated lambs. The digestibilities of cell and cell wall contents were generally poor in defaunated lambs while intake of digestible protein and energy were similar in the three groups. The lambs in the three groups were in positive nitrogen balance while percent N retention was higher in defaunated than in refaunated and faunated lambs. It was concluded from the study that with similar dry matter intake and plane of nutrition, the growth performance and feed conversion efficiency were better in defaunated than refaunated and faunated lambs. In spite of reduced digestibility of neutral detergent fibre, acid detergent fibre and cellulose, the metabolise energy availability for body synthetic activity was probably higher in defaunated lambs as evident from the observed production traits.

Adult rumen-fistulated sheep and goats (four of each) were used to study comparative nutrient utilization and diurnal variation of rumen ciliate protozoa under maintenance level of feeding. The animals were control-fed on roughage ( Cenchrus ciliaris ) and concentrate mixture in the ratio of 65 : 35 as per their maintenance requirement. Dry-matter intake was higher ( P 0.75 /day) than in goats (57.1 g/kgW 0.75 /day), whereas digestibility of OM, NDF and ADF was better in goats than in sheep. The rumen protozoa of sheep and goats were a B -type population with numbers varying between 0.31–0.45×10 6 cells/ml rumen liquor. The total protozoa, holotrichs, spirotrichs, Isotricha , Dasytricha , small and large spirotrichs were higher ( P Isotricha and large spirotrichs, although numerically fewer in number, collectively contributed 91 and 89% of protozoal cell mass in the rumen of sheep and goats, respectively. It is concluded from the study that under control feeding, dry-matter intake was lower while fibre digestibility was higher in goats compared to sheep, possibly due to higher number of ciliate protozoa as well as holotrichs and spirotrichs in their rumen medium.

Pal, D.T., Kamra, D.N., Pathak, N.N. and Bisht G.S. 1998. Influence of rumen ciliate protozoa and protein level on rumen fermentation, enzyme activities and blood metabolites in crossbred cattle calves. J. Appl. Anim. Res., 13: 153–159. The influence of ciliate protozoa in the rumen on fermentation and enzyme activities in rumen fluid and blood metabolites of crossbred (Bos taurus X Bos indicue) cattle calves was studied in animals fed at two levels of protein. Sixteen crossbred male calves, separated from their dams at 24h to 48h of birth, were divided into two groups—faunated and defaunated and given a diet of normal protein or lower protein (75% of the requirement). The calves were reared in isolation stalls to avoid contact with faunated calves. On 3 months of age ruminal contents from all calves were collected at 4h post feeding and analysed The pH, total volatile fatty acids (TVFA), carboxymethylcellulase (CMC), xylanase and protease activities were similar in both faunated and fauna-free c...

Species composition and concentration of rumen ciliate protozoa were investigated in the rumen contents of 14 domestic sheep and 1 goat living in Bishkek, Kyrgyzstan. This is the first report on rumen ciliates from ruminants living in Kyrgyzstan. In sheep 12 genera, 28 species and 12 morphotypes were detected, whereas in goat 8 genera, 12 species and 4 morphotypes were detected. The density of ciliates in sheep was (28.1 ± 20.0) × 10(4) cells mL(-1) and in goat was 37.0 × 10(4) cells mL(-1). Dasytricha ruminantium, Isotricha prostoma, Entodinium simulans and Ophryoscolex caudatus were major species (100%) in sheep, and for the first time, Diplodinium rangiferi was detected in a domestic goat.

Protozoa species were identified in rumen contents of four domestic sheep (Ovis aries) from South Africa. All animals were fed a forage diet which consisted of 50% lucerne and 50% teff hay. Ten new host records were identified, bringing the total number of species and forms observed in sheep in South Africa to 30. The occurrence and geographic distribution of ciliate protozoa in both domestic and wild sheep from around the world are summarised. It was found that 15 genera and 131 species occur in domestic sheep globally.

Changes in the microbial populations, their activities, and the ruminal fermentation were monitored for 50 d following the reintroduction of ciliate protozoa into four defaunated sheep. A protozoal population was reestablished successfully in each recipient, using a washed inoculum containing approximately 103 cells, although there were between-animal differences in the rates of recolonization and genus establishment. Entodinium spp. predominated in the initial stages of the refaunation period and had an apparent maximal generation time of 9–10 h. Bacterial and fungal numbers did not decline following the reintroduction of protozoa and a small transient increase in the numbers of amylolytic and xylanolytic bacteria and fungal zoospores occurred in the early stages of refaunation when the protozoal population was 5/g ruminal contents, but these subsequently declined as the protozoa established. Although the fibrolytic bacterial population was lowest in period 3 (> 105 protozoa/g), the in sacco ruminal digestion of Lolium perenne hay and polysaccharolytic enzyme activities in the solids-associated populations were either maintained or increased when protozoa were present confirming the important contribution of protozoa to fibre breakdown in the rumen. Significant changes in ruminal microbial activities occurred after protozoal reinoculation but before the rumen had refaunated completely. Arylamidase activities in the liquor-phase population and ruminal ammonia concentrations increased significantly within 48 h of transfaunation; the magnitude of the effects became more pronounced as the protozoal population developed. However, volatile fatty acid formation and ruminal pH were not affected after the reintroduction of protozoa.Key words: rumen, sheep, ciliate protozoa.

The present results are the last part of a series of experiments conducted in this laboratory on metabolism of major individual rumen ciliate protozoa species and their combinations in sheep. These species were inoculated into the rumen of fauna-free (FF) sheep to study digestibility of dietary fibre and duodenal flow of N. Sixteen rumen- and duodenum-cannulated FF sheep were divided into four equal groups and used in the present experiment. One group remained FF, while the other three were each inoculated intraruminally with the Holotrich protozoa sp. (HT), Entodinium sp. (EN) or total mixed fauna (TF). In comparison with the FF group, the HT did not affect (P.0·05) the duodenal flow of non-NH3-N and bacterial-N, while the flow was decreased (P,0·05) by both the EN and TF populations. The digestibility of fibre was increased (P,0·05) by both the EN and TF populations, but the results for the HT group were mostly similar to those for the FF sheep. The fibre digestibility in the stomach for TF was intermediate between that for EN and FF. It was concluded that except when fed high-carbohydrate diets, the presence in the rumen of the HT protozoa are of no significant value or major detriment to ruminant production. Holotrich protozoa: Entodinium: Rumen ciliate protozoa: Sheep It is well established that the presence of mixed protozoa species populations in the rumen have a negative effect on the utilisation of dietary N by ruminants (1) . In comparison with defaunated (all existing protozoa eliminated from the rumen) or fauna-free (FF; protozoa never existed in the rumen) ruminants a number of experiments have shown a decreased flow of non-NH3-N (NAN) components from the stomach to the intestinal tract when mixed protozoa populations were present in the rumen (2‐5) . However, the species of the mixed population that contribute negatively to N utilisation and to what extent they do was not known. Although a number of studies involved individual species of protozoa such as Entodinium (EN) and Isotricha (IS) (6‐9) , the effect of these species on the duodenal flow of NAN components has not been measured. Therefore, we conducted a series of experiments in this laboratory on N metabolism by individual or combinations of ciliate protozoa species (IS, Dasytricha (DS), Polyplastron (PP), Epidinium (EP), Eudiplodinium (EU), EN and total mixed fauna (TF)). These species were established in the rumen of naturally FF sheep which originated from a sheep flock free of rumen ciliate protozoa for several generations (10) . The present work concludes the

The aim of this work was to determine whether reductive acetogenesis can provide an alternative to methanogenesis in the rumen. Gnotobiotic lambs were inoculated with a functional rumen microbiota lacking methanogens and reared to maturity on a fibrous diet. Lambs with a methanogen-free rumen grew well, and the feed intake and ruminal volatile fatty acid concentrations for lambs lacking ruminal methanogens were lower but not markedly dissimilar from those for conventional lambs reared on the same diet. A high population density (10 7 to 10 8 cells g −1 ) of ruminal acetogens slowly developed in methanogen-free lambs. Sulfate- and fumarate-reducing bacteria were present, but their population densities were highly variable. In methanogen-free lambs, the hydrogen capture from fermentation was low (28 to 46%) in comparison with that in lambs containing ruminal methanogens (>90%). Reductive acetogenesis was not a significant part of ruminal fermentation in conventional lambs but contributed 21 to 25% to the fermentation in methanogen-free meroxenic animals. Ruminal H 2 utilization was lower in lambs lacking ruminal methanogens, but when a methanogen-free lamb was inoculated with a methanogen, the ruminal H 2 utilization was similar to that in conventional lambs. H 2 utilization in lambs containing a normal ruminal microflora was age dependent and increased with the animal age. The animal age effect was less marked in lambs lacking ruminal methanogens. Addition of fumarate to rumen contents from methanogen-free lambs increased H 2 utilization. These findings provide the first evidence from animal studies that reductive acetogens can sustain a functional rumen and replace methanogens as a sink for H 2 in the rumen.

Quatro novilhos Nelore, fistulados e canulados no rúmen, foram distribuídos em um delineamento quadrado latino 4 x 4, para avaliar o caroço de algodão e o sal de cálcio de ácidos graxos como fontes de gordura assim como o efeito da monensina em dietas com caroço de algodão, sobre a população de protozoários ciliados e o pH do rúmen. Os tratamentos experimentais foram: dieta controle (CTRL), dieta com sal de cálcio de ácidos graxos (SC), dieta com caroço de algodão (CA) e dieta com caroço de algodão sem monensina (CASM). Amostras do conteúdo ruminal foram colhidas às 0, 2, 4, 6 e 8 horas após a alimentação. Foi utilizada câmara de contagem de Sedgwick-Rafter com capacidade de 1mL e microscópio ótico comum provido de retículo com área de 0,4362 mm², para identificação dos gêneros de ciliados. Não houve diferença entre os tratamentos para o pH do rúmen. Os tratamentos com caroço de algodão reduziram todos os gêneros de protozoários ciliados, em especial de Entodinium, provavelmente pela gordura liberada. Os protozoários dos gêneros Diplodinium, Epidinium e Eudiplodinum e o número total de protozoários foram os únicos que sofreram influência do tempo de alimentação. Sais de cálcio de ácidos graxos foram suficientemente inertes no ambiente ruminal, não afetando o número de protozoários ciliados no rúmen de bovinos Nelore. A utilização de monensina em dietas contendo caroço de algodão e alto concentrado não influenciou o número de protozoários ciliados no rúmen de bovinos Nelore. Four ruminally and duodenally cannulated Nellore steers were assigned to a 4 x 4 latin square to study the effects of feeding monensin (M) and different sources of fat on the ruminal population of ciliate protozoa and ruminal pH. The experimental diets contained (DM basis) 19% of sugar cane and were supplemented with M [control (CTRL)], M plus calcium salts of fatty acids (CSFA), M plus whole cottonseed (MWCS), or only WCS. Ruminal contents were sampled at 0, 2, 4, 6, and 8 hours post-feeding. In order to identify the ciliate protozoa a Sedgwick-Rafter count cell and an optic microscope with slide area of 0.4362 mm² were used. No significant differences in ruminal pH were observed among diets. Both cottonseed treatments (MWCS and WCS) reduced the ruminal population of the ciliate protozoa investigated, mainly the Entodinium, possibly because of the slow release of fat. Ruminal counts of Diplodinium, Epidinium, Eudiplodinium, and total protozoa were all affected by time post-feeding. Calcium salts of fatty acids seemed to be inert in the ruminal environment because no significant changes were observed in the protozoa counts. Moreover, feeding CRTL and MWCS diets to Nellore steers did not change the ruminal population of ciliate protozoa in this trial.

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.

Em delineamento inteiramente ao acaso, estudou-se o efeito da defaunação em ovinos que receberam cana-de-açúcar e uréia sobre o consumo e digestibilidade aparente de nutrientes, dinâmica da fase sólida, parâmetros de fermentação ruminal e de degradação in situ. As taxas de passagem no rúmen foram, respectivamente, de 3,05 e 1,97%/h, para ovinos faunados e defaunados. Valores mais elevados para taxa (5,4%/h) e extensão (46,7%) da digestão ruminal e de degradabilidade efetiva da FDN da cana (31,5%) foram obtidos nos ovinos faunados. Seus consumos diários (57,7 e 32,3 g/kg0,75, respectivamente para MS e FDN) foram superiores (P

The effects of dietary variations on microbial N turnover and recycling related to protozoal activities in the rumen were examined using a previously described model of rumen fermentation processes. Simulations were performed with diets containing variable proportions of concentrate (between 0 and 1 kg/kg diet dry matter) at a rate of daily dry matter intake (DMI) by cattle of between 5·3 and 21·0 kg/day. The roughages examined were hay, fresh grass and maize silage, and the concentrate supplements included molasses, barley, maize grain, untreated and formaldehyde-treated soyabean meal. The simulated fractional turnover rate of microbial N varied between 0·416 and 1·556/day, and simulated microbial N recycling ranged from 34·9 to 76·2% of gross microbial N synthesis. High turnover rates and recycling were observed particularly for diets containing maize silage as roughage and molasses and maize grain as concentrate supplements. Increased DMI generally resulted in reduced turnover rates with high-roughage diets, but increased turnover rates with high-concentrate diets. These responses of microbial N turnover and recycling were related to substrate availability and the substrate preferences of protozoa and, consequently, protozoal activities in the rumen. The large impact of recycling on the amount of organic matter required per unit of microbial protein washed out of the rumen was demonstrated. Thus, by quantification of the interactions between microbial populations in the rumen, the model can help to interpret the differences in microbial N recycling observed in the experiments and provide an improved understanding of the efficiency of microbial protein synthesis.

Twelve male sheep were used, in a completely randomized design, to evaluate apparent digestibility and rumen ciliate protozoa population under different diets: A) corn silage supplemented with 0.050 kg of soybean meal/kg original roughage; B) 50% corn silage + 50% sugar cane supplemented with 0.075 kg soybean meal/kg original roughage; C)sugar cane supplemented with 0.100 kg soybean meal/kg original roughage. Dry matter intake and digestibility of Crude fiber were linearly affected (P < 0.01) by increasing the level of sugar cane in the diets, and they had direct influence in feed consumption and T.D.N. Total, rumen protozoal population was represented by a quadratic regression(P < 0.01), and the average values for total ciliates ml-1 in the rumen contents were 235.01, 196.49 and 514.07 (x103) for treatments A, B and C, respectively. Entodinium genus was always predominant, with 76.8, 62.2 and 89.4% respectively in treatments A, B and C. Doze ovinos foram utilizados em delineamento inteiramente casualizado para avaliar a digestabilidade aparente e a população de protozoários ciliados no rúmen, em diferentes dietas: A) silagem de milho suplementada com 0.05 kg de farelo de soja/kg de matéria original; B) 50% de silagem de milho + 50% de cana-de-açúcar suplementados com 0.075 kg de farelo de soja/kg de matérial original e C) cana-de-açúcar suplementada com 0.100 kg de farelo de soja/kg de matéria original. A matéria seca ingerida e a digestabilidade da fibra diminuíram linearmente (P

The composition of ciliate protozoa in the rumen of 30 Japanese beef black cattle (Bos taurus taurus) fed in Yoron Island, Kagoshima, was compared with that of 125 Holstein-Friesian cattle (Bos taurus taurus) fed in Hokkaido. Thirteen genera including 44 species with 32 formae detected from Japanese beef black cattle and 15 genera including 48 species with 25 formae from Holstein-Friesian cattle were identified. Thirty-six species with 19 formae were common to both hosts. A new species was recognized from Japanese beef black cattle and named Entodinium yunnense n. sp. This new species was classified into such four formae as yunnense n.f., spinonucleatum n.f., acutonucleatum n.f. and spinolobum n.f. A new forma of Entodinium bifidum having only one caudal spine was also recognized from Japanese beef black cattle and named monospinosum n.f. The average ciliate density in Japanese beef black cattle, 18.7 x 10(4)/ml, was lower than 40.3 x 10(4)/ml in Holstein-Friesian cattle. The average number of ciliate species per head in Japanese beef black cattle, 14.4 was also lower than 17.8 in Holstein-Friesian cattle. The average value of diversity index was 1.789 in Japanese beef black cattle and 1.718 in Holstein-Friesian cattle. Both incidence and composition ratio of Entodinium rectangulatum and E. ovinum were significantly higher in Japanese beef black cattle than in Holstein-Friesian cattle. On the other hand, Charonina ventriculi, E. caudatum, E. rostratum, E. parvum, E. bursa, Metadinium affine and Ostracodinium mammosum showed a reverse relation.

The aim of this experiment was to ascertain how different feeding strategies affect the rumen function of reindeer after nutritional deprivation. Rumen adaptation to various diets, after restricted feeding, was studied in 44 eight-month-old semi-domesticated female reindeer (Rangifer tarandus tarandus). All animals were initially fed a simulated winter diet based on lichens (lichen diet). A control group, continuously offered the lichen diet ad libitum, was compared to four groups of reindeer that were first restrictively fed (half the ad libitum ration) for eight days followed by one day without feed. The rumen content of restrictively fed animals had higher pH, lower dry matter content and volatile fatty acid (VFA) concentration, a changed composition of VFAs, and lower counts of bacteria compared to that of the control group. The effect was less dramatic than previously reported for reindeer starved for several days. On day 10, the four restrictively fed groups were introduced to different diets. One group was re-fed the lichen diet ad libitum and did not differ from the control group when the experiment ended after five weeks of feeding. Two groups were fed grain-based reindeer feed (pellets) combined with either lichens or grass silage, and one group was fed silage with a gradually increased addition of pellets. Diarrhoea and so called “wet belly” occurred initially in the three latter groups. After five weeks of feeding, the reindeer in the three pellet-fed groups had an altered composition of VFAs and higher counts of protozoa, and also tended to have higher total VFA concentration in the rumen, compared to the control animals and those re-fed the lichen diet. Only small changes were observed in the size of rumen papillae and these could be associated with energy intake. Protozoa decreased over time on the lichen diet. This study confirmed that rumen function was significantly affected by a relatively short period of restricted feed intake. The experiment also revealed a clear difference in rumen function between reindeer adapted to a lichen-based diet and those adjusted to basically grain-based diets. Bacteria that were utilising lichens were drastically reduced when the diet lacked lichens; consequently these bacteria may be regarded as a substrate-specific group.Abstract in Swedish / Sammandrag:Vomfunktionen hos ren (Rangifer tarandus tarandus) efter begränsat födointag och påföljande utfodring Ett försök utfördes med syfte att ta reda på hur olika utfodringsstrategier påverkar vomfunktionerna hos ren (Rangifer t. tarandus) efter en period med lågt näringsintag. Vommens anpassning till olika dieter studerades hos 44 åtta månader gamla honkalvar Alla renarna gavs initialt en simulerad vinterdiet baserad på lav (lavdiet). En kontrollgrupp, som fick äta fritt av lavdieten under hela försöket, jämfördes med fyra grupper renar, som först utfodrades restriktivt med lavdieten (halva fodergivan jämfört med fri utfodring) under åtta dagar och sedan var helt utan foder under en dag. Vominnehåll från de restriktivt utfodrade renarna hade högre pH, lägre torrsubstans, lägre koncentration av flyktiga fettsyror (VFA), förändrad sammansättning av VFA och mindre mängd bakterier än vad som uppmättes i kontrollgruppen. Effekten var inte så dramatisk som den som tidigare rapporterats för renar som svultit flera dagar. Dag 10 sattes de fyra restriktivt utfodrade grupperna på olika dieter. En grupp utfodrades åter med lavdieten i fri mängd, och skiljde sig inte från kontrollgruppen när försöket avslutades efter fem veckors utfodring. Två grupper gavs spannmålsbaserat renfoder (pellets) kombinerat med antingen lav eller ensilage gjort på gräs, och en grupp gavs först enbart ensilage och därefter pellets i gradvis ökande mängd. Flera renar i de tre senare grupperna drabbades av diarré och så kallad ”blöt buk”. Efter fem veckors utfodring hade renarna i de tre pelletsutfodrade grupperna en ändrad sammansättning av VFA och mer protozooer, och tenderade även att ha högre total VFA-koncentration, än de två grupper som fick lavdiet. Endast små förändringar observerades i storleken på vompapiller, och dessa kunde kopplas till energiintag. Protozooerna minskade med tiden på lavdieten. Denna undersökning bekräftar att vomfunktionerna påverkas signifikant av en relativt kort period med minskat födointag. Undersökningen visar också en klar skillnad i vomfunktion mellan renar anpassade till lavdiet och dem som är anpassade till en i huvudsak spannmålsbaserad diet. Bakterier som växte på lav minskade drastiskt när laven uteslöts ur dieten; dessa bakterier kunde följaktligen betraktas som en substrat-specifik grupp.

Summary Methods and application of defaunation in the growing ruminant Defaunation (selective destruction of protozoa in the rumen) is suitable for the evaluation of the effects resulting from the activity of the rumen ciliate protozoa and also for possible growth promotion in ruminant fattening. In scientific experiments defaunation should be practised with detergents, whereas nutritive methods seem to be of advantage for the practical application of defaunation. Ruminants reared without ciliates from birth by isolation show, in contrast, no advantage over faunated animals. The evaluation of experiments with defaunated animals showed clear effects on the digestive physiology. Bacterial counts increase up to a 2–3 fold value and the formation of ammonia and methane is reduced by about a third on average. The proportion of butyrate is lower, while the proportion of propionate increases. With regard to the higher total content of volatile fatty acids in artificially defaunated animals there is an increase in propionate content of rumen fluid. This is not valid for animals kept ciliate-free by isolation from birth. The apparent digestibilities of almost all nutrients decrease after defaunation by 1–2 percentage units because of the reduced fermentation activity in the rumen. Daily live-weight gain, N retention and food conversion efficiency are clearly improved in artifically defaunated animals, while isolation from birth leads to exactly contrary effects. An evaluation of the defaunation experiments in relation to the applied food composition showed that defaunation seems to be of advantage if rations contain protein of a high rumen degradability, especially if the ration crude protein content is low. On the other hand, the ciliates seem to be of some importance if rations rich in starch are fed, but in this case the effects are not so clear as with protein. Zusammenfassung Defaunierung (selektive Abtotung der Protozoen im Pansen der Wiederkauer) eignet sich sowohl zur Untersuchung von Auswirkungen der Protozoentatigkeit als auch mit Einschrankungen zur Leistungssteigerung bei wachsenden Wiederkauern. Fur wissenschaftliche Zwecke wird Defaunierung am besten mit Detergentien durchgefuhrt, fur praktische Anwendungsbereiche verspricht die Weiterentwicklung von nutritiven Methoden den grosten Erfolg. Protozoenfreie Aufzucht von Geburt an scheint dagegen nicht vorteilhaft zu sein. Die Auswertung von Defaunierungsexperimenten ergab deutliche verdauungsphysiologische Verschiebungen bei Abwesenheit der Protozoen. Durch Defaunierung steigt die Bakterienpopulation auf die 2–3fache Zahl an, wahrend die Bildung von Ammoniak und Methan im Durchschnitt um etwa ein Drittel zuruckgeht. Der Anteil der Buttersaure an der Gesamtsaure im Pansen verringert sich zugunsten der Propionsaure, so das sich bei kunstlicher Defaunierung in Kombination mit erhohten Gesamtsauregehalten ein Anstieg der Propionsaure im Pansensaft ergibt. Sind die Tiere jedoch von Geburt an protozoenfrei, so verandert sich der Propionsauregehalt nur wenig. Die reduzierte Fermentationstatigkeit nach Defaunierung fuhrt zu einem Abfall in der scheinbaren Verdaulichkeit der meisten Futterinhaltsstoffe um ein bis zwei Prozentpunkte. Durch artifizielle Defaunierung verbessern sich tagliche Zunahmen, N-Ansatz und Futterverwertung deutlich, wahrend sich bei zeitlebens isolierter Haltung genau umgekehrte Effekte ergeben. Die Auswertung der Defaunierungsversuche anhand der eingesetzten Rationszusammensetzung ergab, das die Abwesenheit der Protozoen vor allem dann vorteilhaft sein muste, wenn die Abbaubarkeit des Rationsproteins hoch ist, insbesondere auch, wenn der Rohproteingehalt im Futter niedrig ist. Dagegen scheinen die Ciliaten bei starkereichen Rationen nicht so leicht durch Bakterien zu ersetzen sein, wenngleich hier die Effekte nicht so deutlich wie beim Protein ausfallen.

SUMMARYThe distribution of carboxymethylcellulase (CMCase) between various rumen fractions has been determined at three times after feeding in sheep containing no ciliate protozoa or five different protozoal populations. The total rumen CMCase was lowest in defaunated animals and tended to be higher in animals containing the amylolyticEntodinium caudatumor a natural mixed protozoal population and highest in sheep containing single cellulolytic species. In animals containing the cellulolytic species,Epidinium ecaudatum caudatumorEudiplodinium maggii, 70% of the CMCase was associated with the protozoal fraction whereas less than 15% was present in a fraction prepared in the same way from sheep containing no protozoa or onlyEntodinium caudatum. The activity associated with the free bacteria was higher in sheep containing onlyEntodinium caudatumthan in defaunated animals or those containing the cellulolytic species. Similar results were obtained 1·5, 6·5 or 24 h after feeding. Almost no activity was present in the cell-free rumen liquor under any condition. A variable amount of CMCase (up to 25% of the total) was associated with the plant debris in the rumen and the specific activity of the enzyme released by sonication was often higherthan that released from the free bacteria under the same conditions.

ホルスタイン種成雌牛2頭および黒毛和種成雌牛2頭を供試し,給与飼料は,粗飼料として稲わら,青草(オーチャードグラス),乾草(オーチャードグラス)およびサイレージ(トウモロコシ茎葉)を,濃厚飼料は大麦挽砕を使用した.これらの飼料を粗飼料単味または濃厚飼料を加えて給与することにより,反芻胃内繊毛虫類に及ぼす影響を比較検討した.その結果次のことが認められた.1) 粗飼料単味の給与では,反芻胃内繊毛虫類の反芻胃内容液1ml中の総数は減少した．2) 粗飼料に濃厚飼料(大麦)を加えて給与したときに繊毛虫数は著しい増加を示した.3) 反芻胃内繊毛虫類に対する飼料の影響は貧毛虫類に顕著にあらわれ,特にEntodinium属は飼料組成に敏感に反応して増減した.すなわち,繊毛虫数の大多数を占めるEntodinium属は,大麦給与時に増加し,また,その中ではEntodinium simplexの数が最も顕著に増加した.4) 給与飼料をもとの飼料に戻すことにより,もとの繊毛虫集団構成に近くなったことから,給与する飼料の種類が反芻胃内繊毛虫数の重要な変動要因であると考えられた.

SUMMARYMeadow hay and mixed food containing 13·3 or 26·9% of concentrates were used for culturing ciliates taken from the rumen of water buffalo. The culture medium was the sterilized rumen fluid or a solution of mineral salts. The cultures were mixed every 24h and one-half of the contents of each culture flask was transferred to an equal volume of fresh medium containing the appropriate food.Anoplodinium denticulatumand seven species ofEntodiniumwere the only protozoa which survived until the end of the experiment, but to maintain them on food containing 26·9% of concentrates an alkaline or neutral medium was required. Using rumen fluid and mineral medium the total number of protozoa was on average, respectively: 2·7 x 103and 2·2 x 103/ml in cultures on hay only; 9·4 x 103and 17·5 x 103/ml in cultures on food containing 26·9% of concentrates; and 14·0 x 103and 17·;8 x 103/ml in cultures on food containing 13·3% of concentrates.Entodiniumformed 66·6 and 72·5%; 95·5 and 91·8%; and 96·1 and 97·7% of the total counts. In cultures on hay alone there were 33·4 and 27·5% ofAnoplodinium denticulatum, while they formed only 4·5 and 8·2% and 3·9 and 2·3% in cultures on food containing 26·9 and 13·3% of concentrates respectively.The relation between number of ciliates and composition of food only partly agreed with the relation found previously under natural conditions. Changes in pH of the medium would appear to be the chief cause.

The lethal concentrations of dioctyl sodium Sulphosuccinate (DSS) to nine species of rumen ciliate protozoa were determined. The quantity of DSS required to remove these protozoa from the rumen of sheep was approximately 30 times the lethal concentration. This was caused by interaction of the DSS with rumen particulate material, and the protozoa were killed only after the particulate fraction was saturated with DSS. Defaunation was conducted most efficiently after withdrawal of food for 24 h followed by a period (1–4 d) of reduced food intake by the host animal. In toxic concentrations of DSS the cilia of both holotrich and entodiniomorphid protozoa became detached, followed by leakage of the cell contents via the gullet or anus (in the entodiniomorphids) or at variable points in the cell wall (in the holotrichs). The population densities of most of the other rumen micro-organisms examined decreased during defaunation, but regained their original population density within 11–21 d. After this time the population densities of these organisms exceeded that shown before defaunation, with the exception of Oscillospira guilliermondii which was eliminated.

The effect of defaunation on the energy metabolism of adult wethers was investigated in two experiments. For that purpose three semi-synthetic diets containing two different levels of cellulose and starch and two different sources of starch (native or steamflaked) were employed. Food intake and crude protein content of all rations remained constant. Defaunation improved the digestibility of energy in the starch-free diet and lowered it in the diet rich in native starch, whereas defaunation had no influence on the digestibility when steamflaked maize starch was fed. The activity of the ciliate protozoa was shown to be responsible for higher digestibility coefficients in rations rich in native compared to steamflaked starch, because these differences vanished completely in defaunated animals. Energy loss with urine was insignificantly lower in ciliate-free animals. Defaunation reduced methane formation in two rations, but not when native starch was fed. Reduced gaseous exchange after defaunation was found in all diets and led to a lower heat production of about 5% on average. The results indicate that defaunation could be advantageous when certain types of rations are fed to ruminants.

Of the 11 ciliate protozoa present in the rumen of the sable antelope, two are holotrichs and nine entodiniomorphs. One new species Diplodinium (Eudiplodinium) sablei is described. The seven antelopes investigated gave an average total number of protozoa per cm3 rumen fluid of 1,79 ± 0,39 × 105, a relatively low figure typical of grazing wild ungulates.Van die 11 siliaat-protozoa teenwoordig in die rumen van die swartwitpens was twee holotriche en nege entodiniomorphe vorme. Een nuwe spesies Diplodinium (Eudiplodinium) sablei word beskryf. Die sewe swartwitpense wat ondersoek is, het gemiddeld 1,79 ± 0,39 × 10* protozoa per cm3 rumenvloeistof gehad, 'n relatief lae syfer tipies van weiende wildsbokke.

本邦の主要な反芻家畜であるウシ126頭,ヒッジ51頭,ヤギ10頭におけるルーメン内繊毛虫の分布調査と移植試験をおこない,以下の所見を得た.1) この調査で検出された繊毛虫は,ウシ44種,ヒッジ34種,ヤギ26種の計3科16属47種で,そのうち7属15種は従来本邦での存在は不明のものであった.2) しかし,これらすべてが各個体に棲息するのではなく,その平均棲息種数はいずれの家畜でもほぼ14種であった.3) またルーーメン内容中の繊毛虫密度はいずれの家畜においても平均約5×105/mlであった.4) Entodinium属,特にE. simplex, E. nanellumは各種家畜において多数の個体に出現し,その密度も高く,最も優勢に棲息するものであった.5) 各種繊毛虫の密度と検出率は相関し,ルーメン内で高密度に存在する繊毛虫はその分布が広い傾向があった.6) 供試した3種の家畜におけるルーメン内の繊毛虫の種類構成はおおむね類似したが,13種の繊毛虫はウシのみに分布し,また2種はヒッジのみから検出された.7) しかし,ウシ特有と思われた繊毛虫をヒッジルーメン内に移植すると,それは明らかに増殖定着した.これは,家畜による繊毛虫の分布の相違は宿主特異性によるよりはむしろ感染の機会の有無によることを示唆するものと思われた.

反芻動物における非蛋白態窒素(NPN)化合物の利用におよぼすルーメンプロトゾア存否の影響を明らかにする目的で,プロトゾアの存在するヤギの区(Fヤギ区)とプロトゾアの存在しないヤギの区(Dヤギ区)にそれぞれ尿素含有飼料を給与し,N消化率,N保留率を測定した.さらに尿素含有飼料給与後10日目の反芻胃内容液についてアンモニア,総低級脂肪酸(VFA)および乳酸濃度を経時的に調べ,また14日目の内容液についてin vitroで反芻胃内微生物によるアンモニア利用の程度を両区で比較を行い以下の結果を得た.1) 飼料中のN源の約半分が尿素の場合,大豆蛋白質の場合に比べN消化率はFヤギ区およびDヤギ区でともに高かった.2) 尿素飼料給与時におけるN保留率は全般的にFヤギ区に比べDヤギ区で高い傾向が示され,特に尿素飼料に切り換えた直後でその傾向は顕著であった.3) 反芻胃内容液のアンモニアおよび総VFA濃度はいずれもFヤギ区で高かった.4) in vitro培養実験における微生物蛋白質合成量はDヤギ区で高い値が示された.以上の結果より反芻胃内にプロトゾアの存在しないDヤギ区で尿素の利用性が高い傾向が示された.

Two promising methods to remove the ciliate protozoa of the stomach of ruminants by simple alterations in the feeding system were examined in vivo with lambs and heifers in the present investigation. The use of pelleted concentrates fed ad libitum and of milk, especially of milk fat (cream), turned out to be effective against protozoa when given after few hours of fasting. Milk led to the defaunation of a great proportion of the animals supplied all at once in an amount of 5-10% of rumen fluid volume. With cream the amount could be lower according to the higher fat content. Defaunation with concentrate was found to be not dangerous only for animals adapted to intensive feeding sufficiently. Additionally, a series of in vitro experiments were carried out to evaluate the reasons for the anti-protozoal effects of these methods. Concentrates rich in starch had toxic effects against protozoa, which mainly derived out of the depression in ruminal pH. The formed lactic acid had further effects irrespective of pH. The addition of crude fibre, especially of cellulose or lignin, reduced or prevented the anti-protozoal effects of concentrate fed ad lib. This was not so with the addition of buffers. With milk the included fat was shown to be responsible for the anti-protozoal effects, while the other main constituents of milk were of no importance.

SYNOPSIS. Six protozoa-free sheep, 3 fed alfalfa hay and 3 fed a concentrate diet, were inoculated with rumen contents from a steer fed the same alfalfa hay. All 24 species of protozoa in the inoculum became established in the sheep fed alfalfa hay, while only 9 species established in the sheep fed concentrate. Percentage species composition in the alfalfa-fed sheep was fairly similar to that of the inoculum. Rumen volumes of the alfalfa hay-fed sheep were significantly higher than those of the concentrate-fed sheep; however, fluid turnover rates were similar. Total protozoan numbers per ml of rumen contents were significantly higher in the concentrate-fed sheep, but after adjustment for rumen volume, there was no significant difference in the total number of protozoa in the rumen.

The rumen contents of 70 lactating Holstein cows fed five different rations were examined for ciliate protozoa. Species of eight genera of the family Ophryoscolecidae and two genera of the family Isotrichidae were observed, with Entodinium, Epidinium, Polyplastron, Ophryoscolex , and Isotricha predominating. Isotricha appeared frequently in the rumens of cows fed a ration containing cane molasses, whereas Polyplastron, Epidinium , and Ophryoscolex were frequently present in cows fed a ration containing hemicellulose extract. There were no differences in the frequency of appearance of most ciliate protozoa in the rumens of cows fed different quantities of calcium and phosphorus. Entodinium caudatum appeared frequently in the rumens of cows fed rations containing large quantities of grain starch. Polyplastron multivesiculatum or Ophryoscolex caudatus or both were not seen in cows containing Epidinium caudatum or Eudiplodinium maggii . However, O. caudatus was seen together with Epidinium tricaudatum. P. multivesiculatum was not seen together with Metadinium medium .

SUMMARYThe ruminal activity, daily live-weight gain and feed conversion efficiency of zebu calves inoculated with whole rumen contents from a mature cow were compared with those of calves maintained under the same conditions except that they were ciliate-free and isolated from other ruminants.A thriving mixed population of ciliate protozoa became established in the rumen of calves two weeks after inoculation.Entodiniumwas the first genus to appear and was followed byIsotricha, Polyplastron, Ophryoscolex, DiplodiniumandDasytricha.The ruminal volatile fatty acid and ammonia concentrations in the inoculated calves showed a pronounced rise above the values found in the uninoculated controls.The inoculated calves grew faster and had a higher feed conversion efficiency than the uninoculated controls.

Two experimental diets with or without diethylstilbestrol (DES) were fed to four fistulated dairy cows in a 4 × 4 Latin square design. A continuous feeding system was used to give each cow 13–15 g of diet every two minutes. The continuous feeding appeared to remove the usual variation in the numbers of protozoa in the rumen content. This facilitated the sampling for studies of the effect of the diet on the concentrations and varieties of protozoa. Numbers of protozoa among days were similar with no significant differences. Total protozoa numbers were significantly higher (P

SUMMARY: The limitations of methods previously used for counting rumen ciliates, and the value of developing a method of known accuracy, are discussed. Two counting-chambers, one based on the Sedgwick-Rafter cell, the other on the MacMaster eelworm-counting cell, were constructed and tested. A MacMaster-type cell was adopted. The preparations of suitable suspensions for counting organisms from washed suspensions and from rumen contents are described. The counting technique adopted was found to be reliable; its accuracy depends upon the number of organisms counted. In one sheep on a constant diet, day-to-day differences in rumen ciliate population were far greater than the differences found throughout the rumen at any one time, or the variations due to technique.

The total numbers of ciliate protozoa in the contents of the rumen and reticulum were calculated from counts made on representative samples. The samples were obtained when the entire contents were removed through the rumen fistula, weighed, mixed, sampled, and returned to the animal. Diurnal variation was found in the populations of both oligotrichs and holotrichs. The fluctuations in total numbers of Isotricha appeared to be related to the diet and dietary regime of the host. When the host was fed chaffed red clover hay for two hours daily a marked rise in isotrich numbers occurred during feeding. When the host was fed fresh red clover for two two-hours periods each day a fall in isotrich numbers occurred. Destruction of the organisms by bursting may have caused this decrease. It is suggested that, since the cell contents thus liberated can have strong foam-stabilising properties, bursting of isotrichs could contribute to the occurrence of bloat.

1. The levels of blood reducing sugars, haemoglobin, protein N, NPN, ammonia N, and urea N in addition to the nitrogen retention and digestibility coefficients of food constituents, were studied in lambs with and without ciliate protozoa.2. The inoculated lambs have higher blood haemoglobin, protein N, and lower reducing sugars, ammonia N, and urea N values than those of the uninoculated lambs.

Dioctyl sodium sulfosuccinate (Aerosol OT) has been used in a remarkably short period to obtain rumen ciliate-free cows. Two 30-g doses of Aerosol OT given on consecutive days appeared to effectively eliminate all types of rumen ciliate protozoa without harming the health of the host animal. Measurement of the rumen metabolic parameters of gas production, carbon dioxide to methane ratio, p H value, volatile fatty acids, ammonia, and in vitro cellulose digestion, along with total amylolytic streptococci counts of the rumen contents, showed that conditions in rumens of defaunated animals are normal 4 days after such treatment. It appears that such animals may be used in ruminal studies that require defaunated animals.

1. The dynamics of large ciliate (holotrich) protozoa (Isotricha and Dasytricha spp.) in the rumen of cattle given cut, fresh ryegrass (Lolium multiflorium Lam) were studied by means of a single intrarumen injection of14C-labelled protozoa prepared in vitro by adding [Me14C]choline to rurnen fluid containing protozoa and incubating at 39° for 2 h.2. An indication of the lysis rate of protozoa in the rumen was obtained from the radioactivity apparently lost through the methane pool.3. The turnover time of the holotrich protozoa indicates that these protozoa were extensively retained in the rumen and that only a small proportion of those produced in the rumen flowed out in the digesta. This was supported by the estimation of the rate of lysis which was approximately 85% of the turnover rate in the rurnen.4. The apparent production rate of the larger protozoa indicates that they contribute only about 9% of the predicted net microbial protein synthesis in the rumen.

In a first experiment 4 rams consumed over a period of 70 days 0; 82; 208 and 345 mg of monensin daily, respectively. All doses of monensin caused a marked decrease in total numbers of rumen protozoa in samples taken after this period. More than 90% of total protozoal numbers belonged to the genus Entodinium. The sensitivity of the genus Entodinium to monensin was found to be lower than that of genera Dasytricha and Isotricha. In the second experiment 24 lambs were fed hay and concentrates in the ratio 60:40% (groups 1 and 2) and 40:60% (groups 3 and 4). Groups 2 and 4 received 40 mg monensin per animal daily during 16 weeks. The decrease in protozoal numbers due to monensin in samples taken after this period was significant (P less than 0.001) in lambs fed the concentrate diet (groups 3 and 4) and also in lambs from groups 1 and 2 fed the roughage diet (P less than 0.05). The statistical evaluation of the inhibition of total protozoal numbers by monensin (%) in lambs fed both diets has shown that the antiprotozoal effect of monensin was significantly more intensive with the concentrate diet (P less than 0.025) than with the roughage diet.

Three Norwegian reindeer were first fed lichen ad libitum for 40 days, then they were starved for 3 days, and thereafter offered a commercially available mill «waste product» (AB-84) ad libitum. One animal refused to eat AB-84 and was excluded. When eating lichen average daily dry matter intake was 1.1 ± 0.4 kg, while when eating AB-84 after the starvation period it averaged 1.1 kg at day 1, increasing to 2.5 kg at day 7. After 12 days daily dry matter intake varied between 2 and 4 kg. The rumen dry matter percentage was 16 when eating lichen, 6 after 2 days of starvation, and 17 after eating AB-84 for 5 days. Ruminal pH was 6.4 ± 0.3 when eating lichen, increasing to 7.4 after 3 days of starvation and dropping to an all time low of 6.1, 12 hours after refeeding. The viable bacterial count was 4.7 x 1010 ± 2.7, 5.5 x 108 ± 2.0 and 3.1 x 1010 ± 1.7/ml rumen fluid when the animals were eating lichen, had starved for 3 days and had been eating AB-84 for 5 days, respectively. The rumen ciliate protozoa numbered 17.1 x 105 ± 2.7/ml rumen fluid, when eating lichen, dropping to 4.3 x 105 ± 0.6./ml after 3 days of starvation. A further drop to 1.8 x 10s ± 0.5/ml was observed 1 day after refeeding, but the rumen ciliate number was normalized at 15 x 105 ± 1/ml already 8 days after refeeding. These results indicate that the mill «waste product» (AB-84) is very well tolerated even by reindeer suffering from starvation. Since it is moreover commercially available at a favourable price it could be of interest to reindeer herders. In any case, it deserves to be tested under controlled conditions on a larger number of animals.En test på anvendeligheten av kornavrens (AB-84) som for til utsultet rein.Abstract in Norwegian / Sammendrag: Tre norske rein ble gitt lav ad libitum i 40 dager, sultet i 3 dager og deretter gitt kornavrens (AB-84) ad libitum. Ett av dyrene nektet å spise AB-84 og ble satt ut av forsøket. Daglig foropptak i kg tørrstoff var gjennomsnittlig 1.1 ± 0.4 kg under lavforing, mens opptaket av AB-84 var gjennomsnittlig 1.1 kg/dag den første dagen etter sulteperioden. Den syvende dagen var opptaket 2.5 kg/dag, og varierte mellom 2 og 4 kg/dag etter 12 dager. Prosent tørrstoff i vominnholdet var 16 under lavforing, 6 etter 2 dagers sult og 17 etter 5 dagers foring med AB-84. pH i vominnholdet ble målt til gjennomsnittlig 6.4 ± 0.3 under lavforing, 7.4 etter 3 dagers sult og falt bare til 6.1, som lavest målte verdi, etter 12 timers ad lib. tilgang på AB-84. Antall levende bakterier pr. ml vomsaft var henholdsvis 4.7 x 1010 ± 2.7, 5.5 x 10" ± 2.0 og 3.1 x 1010 ± 1.7 når dyrene spiste lav, hadde sultet i tre dager og hadde spist AB-84 ad lib. i fem dager. Antall eiliater var 17.1 x 105 ± 2.7 pr. ml vomsaft når dyrene spiste lav og 4.3 x IO5 ± 0.6 etter tre dagers sult. Etter en dags ad lib. tilgang på AB-84 var antallet 1.8 x 105 ± 0.5 pr. ml og etter fem dager 7.7 x 105 ± 1.8/ml. Disse resultatene indikerer at selv sultet rein tolererer kornavrens (AB-84) meget godt, og da foret er kommersielt tilgjengelig til en rimelig pris, kan det være av interesse for reindriftsutøvere. I alle tilfeller fortjener foret å bli utprøvet på et større antall dyr under kontrollerte betingelser.Koe viljaperkeen (AB-84) käyttökelpoisuudesta rehuna nälkiintyneelle porolle.Abstract in Finnish / Yhteenveto: Kolmea norjalaista poroa ruokittiin jäkälällä ad libitum 40 päivää, annettiin nähdä nälkää 3 päivää ja sen jälkeen ruokittiin taas viljaperkeellä (AB-84) ad libitum. Yksi eläimistä kieltäytyi syömästä AB-84 ja näin ollen jouduttiin poistamaan kokeilusta. Päivittäinen rehuntarve kuiva-ainekilossa oli keskimäärin 1.1 ± 0,4 kg jäkäläruokinnassa, kun taas AB-84 ruokinta oli keskimäärin 1.1 kg/päivässä ensimmäisenä päivänä nälkäkauden jälkeen. Seitsemäntenä rehunkulunki oli 2.5 kg päivässä, vaihdellen 2-4 kg päivässä 12 päivän jälkeen. Kuiva-ainesprosentti pötsinsisällössä oli 16 jäkäläruokinnassa, 6 kahden päivän nälän jälkeen ja 17 viisipäiväisen AB-84 ruokinnan jälkeen. Pötsinsisällön happamuus, pH mitattiin keskimäärin 6.4 ± 0.3 jäkäläruokinnassa, 7.4 kolmipäiväisen nälän jälkeen ja väheni vain 6.1:een, joka on alhaisin mitattu arvo, 12 tunnin AB-84 ruokinnan ad lib. jälkeen. Elävien bakteerien lukumäärä pötsinestemillilitraa kohti oli kyseelliset 4.7 x 1010 ± 2.7, 5.5 x 10" ± 2.0, ja 3.1 x 1010 ± 1.7 kun eläimet söivät jäkälää, olivat nälkiintyneet 3 päivää ja syöneet AB-84 ad üb. viisi päivää. Ciliatmäärä oli 17.1 x 105 ± 2.7 pötsinestemillilitraa kohti eläinten syötyä jäkälää ja 4.3 x 105 ± 0.6 kolmipäiväisen nälkiintymisen jälkeen. Yhden päivän AB-84 ruokinnan ad lib. jälkeen määrä oli 1.8 x 105 ± 0.5 millilitralta ja viiden päivän jälkeen 7.7 x 105 ± 1.8 millilitralta. Nämä tulokset osoittavat, että jopa nälkiintynyt poro sietää viljaperkettä (AB-84) erittäin hyvin, ja koska rehua on saatavana kohtuulliseen hintaan se voinee kiinnostaa poromiehiä. Joka tapauksessa rehua kannattaa kokeilla tarkkailtuin edellytyksin suuremmalle eläinryhmälle.