Repeatability of in vitro digestibility assays of forages when using fresh, frozen or freeze-dried cow faeces instead of sheep rumen liquor as sources of micro-organisms

1995 ◽  
Vol 1995 ◽  
pp. 110-110 ◽  
Author(s):  
S Akhter ◽  
E Owen ◽  
M K Theodorou ◽  
S L Tembo ◽  
E R Deaville
Keyword(s):  
Freeze Drying ◽  
Rumen Fluid ◽  
In Vitro Digestibility ◽  
Two Stage ◽  
Freeze Dried ◽  
Fresh Frozen ◽  
Sheep Rumen ◽  
Micro Organisms

Previous studies (El Shaer, Omed and Axford, 1987; Akhter, Owen, Fall, O'Donovan and Theodorou, 1994) with the two-stage in vitro procedure of Tilley and Terry (1963) have shown a high correlation between digestibilities of forages as determined using either sheep rumen liquor, sheep faeces or cow faeces as the microbial inoculum. In the first study of the of the present investigation one objective was to examine the repeatability of these digestibility measurements when made on different occasions. A second objective was to assess whether the correlations between faecal and rumen fluid based inocula could be improved if microorganisms were obtained from pairs rather than individual animals. The objective in the second study using forages of known in vivo digestibility, was to investigate the effect of freezing or freeze-drying of faeces on the repeatability of digestibilities of forages determined in vitro using micro-organisms from cow faeces.

Comparison of sheep rumen liquor and Rusitec fluid as inoculum for determining the in vitro digestibility of hays

2001 ◽  
Vol 2001 ◽  
pp. 132-132
Author(s):  
M.L. Tejido ◽  
M.J. Ranilla ◽  
M.D. Carro
Keyword(s):  
Rumen Fluid ◽  
In Vitro Digestibility ◽  
Detergent Solution ◽  
Sheep Rumen ◽  
Restrictive Legislation ◽  
Practical Disadvantage

One practical disadvantage of some in vitro methods used to estimate the in vivo digestibility of forages is the need for fistulated donor ruminants to provide the rumen liquor. These are subject to restrictive legislation in many countries and are costly to prepare and maintain. The aim of this study was to investigate whether rumen liquor in the in vitro digestibility technique of Van Soest et al. (1966) could be replaced by microorganisms derived from Rusitec. This technique involves the incubation of samples with buffered rumen fluid for 48 h followed by an extraction with a neutral-detergent solution.

Effects of chelating agents during freeze-drying of boar spermatozoa on DNA fragmentation and on developmental ability in vitro and in vivo after intracytoplasmic sperm head injection

Zygote ◽  
2007 ◽  
Vol 15 (1) ◽  
pp. 15-24 ◽  
Author(s):  
M. Nakai ◽  
N. Kashiwazaki ◽  
A. Takizawa ◽  
N. Maedomari ◽  
M. Ozawa ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Freeze Drying ◽  
Chelating Agents ◽  
Sperm Head ◽  
Control Group ◽  
Sperm Dna Fragmentation ◽  
Freeze Dried ◽  
Boar Sperm

SUMMARYSuccessful offspring production after intracytoplasmic injection of freeze-dried sperm has been reported in laboratory animals but not in domesticated livestock, including pigs. The integrity of the DNA in the freeze-dried sperm is reported to affect embryogenesis. Release of endonucleases from the sperm is one of the causes of induction of sperm DNA fragmentation. We examined the effects of chelating agents, which inhibit the activation of such enzymes, on DNA fragmentation in freeze-dried sperm and on the in vitro and in vivo developmental ability of porcine oocytes following boar sperm head injection. Boar ejaculated sperm were sonicated, suspended in buffer supplemented with (1) 50 mM EGTA, (2) 50 mM EDTA, (3) 10 mM EDTA, or (4) no chelating agent and freeze-dried. A fertilization medium (Pig-FM) was used as a control. The rehydrated spermatozoa in each group were then incubated in Pig-FM at room temperature. The rate of DNA fragmentation in the control group, as assessed by the TUNEL method, increased gradually as time after rehydration elapsed (2.8% at 0 min to 12.2% at 180 min). However, the rates in all experimental groups (1–4) did not increase, even at 180 min (0.7–4.1%), which were all significantly lower (p < 0.05) than that of the control group. The rate of blastocyst formation after the injection in the control group (6.0%) was significantly lower (p < 0.05) than those in the 50 mM EGTA (23.1%) and 10 mM EDTA (22.6%) groups incubated for 120–180 min. The average number of blastocyst cells in the 50 mM EGTA group (33.1 cells) was significantly higher (p < 0.05) than that in the 10 mM EDTA group (17.8 cells). Finally, we transferred oocytes from 50 mM EGTA or control groups incubated for 0–60 min into estrous-synchronized recipients. The two recipients of the control oocytes became pregnant and one miscarried two fetuses on day 39.The results suggested that fragmentation of DNA in freeze-dried boar sperm is one of the causes of decreased in vitro developmental ability of injected oocytes to the blastocyst stage. Supplementation with EGTA in a freeze-drying buffer improves this ability.

Differences between the in vitro digestibility of extrusa collected from oesophageal fistulated steers and the forage consumed

2009 ◽  
Vol 49 (7) ◽  
pp. 563 ◽  
Author(s):  
David B. Coates ◽  
Robert J. Mayer
Keyword(s):  
Dry Matter ◽  
Rumen Fluid ◽  
In Vitro Digestibility ◽  
In Vitro Techniques ◽  
Pasture Legumes ◽  
Dry Matter Digestibility ◽  
Significant Difference ◽  
C3 Grasses

In a study that included C4 tropical grasses, C3 temperate grasses and C3 pasture legumes, in vitro dry matter digestibility of extrusa, measured as in vitro dry matter loss (IVDML) during incubation, compared with that of the forage consumed, was greater for grass extrusa but not for legume extrusa. The increase in digestibility was not caused by mastication or by the freezing of extrusa samples during storage but by the action of saliva. Comparable increases in IVDML were achieved merely by mixing bovine saliva with ground forage samples. Differences were greater than could be explained by increases due to completely digestible salivary DM. There was no significant difference between animals in relation to the saliva effect on IVDML and, except for some minor differences, similar saliva effects on IVDML were measured using either the pepsin–cellulase or rumen fluid–pepsin in vitro techniques. For both C4 and C3 grasses the magnitude of the differences were inversely related to IVDML of the feed and there was little or no difference between extrusa and feed at high digestibilities (>70%) whereas differences of more than 10 percentage units were measured on low quality grass forages. The data did not suggest that the extrusa or saliva effect on digestibility was different for C3 grasses than for C4 grasses but data on C3 grasses were limited to few species and to high digestibility samples. For legume forages there was no saliva effect when the pepsin–cellulase method was used but there was a small but significant positive effect using the rumen fluid–pepsin method. It was concluded that when samples of extrusa are analysed using in vitro techniques, predicted in vivo digestibility of the feed consumed will often be overestimated, especially for low quality grass diets. The implications of overestimating in vivo digestibility and suggestions for overcoming such errors are discussed.

scholarly journals A comparative study of ruminal activity in Churra and Merino sheep offered alfalfa hay

1997 ◽  
Vol 65 (1) ◽  
pp. 121-128 ◽  
Author(s):  
M. J. Ranilla ◽  
M. D. Carro ◽  
C. Valdés ◽  
F. J. Giráldez ◽  
S. López
Keyword(s):  
Cell Wall ◽  
Rumen Fluid ◽  
Merino Sheep ◽  
Rumen Ph ◽  
Sheep Rumen ◽  
Fermentation Parameters ◽  
Micro Organisms ◽  
Kinetics Of

AbstractA study was carried out to compare the fermentation parameters and kinetics of digestion of a range of different foods in the rumen of two breeds of sheep (Churra and Merino). Ten mature sheep (five Churra and five Merino), each fitted with a rumen cannula, were used in this study. In situ rumen degradability of both dry matter (DM) and cell wall was greater in Churra than in Merino sheep, the breed differences being significant for most of the foods used in the study (P < 0·05). These differences were greater when the foods had a higher cell wall concentration and this could be related to differences in the ruminal environment. However, when the foods were incubated with rumen fluid their in vitro organic matter (OM) degradability was similar in both breeds. Rumen pH was higher (P < 0·05) and ammonia concentrations were lower (P < 0·05) in Churra than in Merino sheep. Rumen volatile fatty acid concentrations tended to be higher in Merino than in Churra sheep, though differences were only significant just before feeding (P < 0·05). The ratio acetate: propionate was higher in the Churra than Merino breed before and 12 h after feeding (P < 0·05). Protozoa numbers in rumen liquid were similar for both genotypes. The greater degradation of forages in the rumen of Churra sheep is discussed in relation to the possible higher activity of fibre-degrading micro-organisms and the greater buffering capacity of the rumen contents against fermentation acids, which could result in more favourable conditions for the microbial degradation of foods in the rumen.

scholarly journals Estimation of the degradability of dietary protein in the sheep rumen by in vivo and in vitro procedures

1985 ◽  
Vol 54 (2) ◽  
pp. 545-561 ◽  
Author(s):  
R. C Siddons ◽  
J Paradine ◽  
D. L. Gale ◽  
R. T. Evans
Keyword(s):  
Rumen Fluid ◽  
Flow Measurements ◽  
Fractional Rate ◽  
Outflow Rate ◽  
Bean Meal ◽  
Sheep Rumen ◽  
Soya Bean Meal ◽  
Microbial N

1. Estimates of degradability of nitrogen in the sheep rumen for a basal hay diet and for soya-bean meal (SBM), groundnut meal (GNM) and fish meal (FM), when given together with the hay, were determined from measurements of (1) duodenal N flow, (2) ammonia kinetics and (3) rumen N disappearance from polyester bags and rumen outflow rate. The ability of various in vitro procedures to predict in vivo N degradability was also examined.2. Four sheep were given a basal hay diet (800 g dry matter (DM) and 19 g N/d) either alone or supplemented with isonitrogenous amounts (15 g N/d) of SBM, GNM or FM. Duodenal non-ammonia-N flow (g/d) was increased more by FM (8.0) than by GNM (5.9) and SBM (5.8), whilst microbial N flow (g/d) was increased more by SBM (3.9) than by GNM (2.3) and FM (1.6). N degradability values calculated from these results were 0.88, 0.76 and 0.57 for the SBM, GNM and FM respectively. The corresponding value for hay was calculated to be 0.76.3. The irreversible loss of ammonia in the forestomachs (g N/d) was increased more by SBM (11.9) than by GNM (7.2) and FM (5.8) whilst ammonia outflow from the rumen (g N/d) was increased to a similar extent by all supplements ( I.1, 0.9 and 0.8 respectively), as was the amount of microbial N (g/d) synthesized from sources other than rumen ammonia (1.8, 2.0 and 1.9 respectively). N degradability values calculated from these results were 0.84, 0.54 and 0.45 for the SBM, GNM and FM respectively.4. The fractional rate of N disappearance (/h) when the feedstuffs were incubated in polyester bags in the rumen of sheep receiving the basal hay diet (800 g DM/d) was the highest for SBM (0,145) and lowest for FM (0.037), with the hay (0.082) and GNM (0.071) intermediate, whilst the fractional outflow rates from the rumen (/h) of the three supplements were similar (0.034, 0.038 and 0,030 for SBM, GNM and FM espectively). N degradability values calculated from these results were 0.82, 0.67 and 0.60 for the SBM, GNM and FM respectively; the value for the hay was 0.73.5. Of a number of in vitro procedures tested, only N solubility in sodium hydroxide and ammonia or total non-protein-N (NPN) production during incubation with rumen fluid in the absence of hydrazine sulphate ranked the supplements, although not the hay, in the same order as the in vivo degradability procedures. In terms of absolute values, N solubility in NaOH, at room temperature, gave estimates similar to those derived from the duodenal flow measurements; estimates derived from ammonia and total NPN production were lower.

Influence of saponins and sapogenins on the bacteriolytic activity of ciliate protozoa from the sheep rumen

1998 ◽  
Vol 1998 ◽  
pp. 88-88
Author(s):  
B. Teferedegne ◽  
P.O. Osuji ◽  
A. Odenyo ◽  
R. J. Wallace ◽  
C.J. Newbold
Keyword(s):  
Rumen Fluid ◽  
Sesbania Sesban ◽  
Leguminous Tree ◽  
Bacteriolytic Activity ◽  
Rumen Protozoa ◽  
Sheep Rumen ◽  
Possible Cause ◽  
Ciliate Protozoa

Foliage from the tropical leguminous tree, Sesbania sesban, is toxic to rumen protozoa in vitro, due to materials present in a saponins-containing extract of the foliage (Newbold et al. 1997). Suppression of protozoal numbers in vivo when S. sesban is added to the diet is either transient or non-existent, however, even though washed protozoa remain sensitive to S. sesban in vitro (Newbold et al. 1997, Odenyo et al. 1997). A possible reason is that saponins are metabolised in rumen fluid (Makkar and Becker 1997). The aims of this study were to determine if the antiprotozoal effect of different accessions of S. sesban was related to their saponins composition, and if conversion of saponins to their sapogenin derivatives was a possible cause of the loss of the antiprotozoal effect in vivo.

scholarly journals Bioactive Flavonoids, Antioxidant Behaviour, and Cytoprotective Effects of Dried Grapefruit Peels (Citrus paradisiMacf.)

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Lucia Castro-Vazquez ◽  
María Elena Alañón ◽  
Virginia Rodríguez-Robledo ◽  
María Soledad Pérez-Coello ◽  
Isidro Hermosín-Gutierrez ◽  
...  
Keyword(s):  
Freeze Drying ◽  
Neuroblastoma Cell ◽  
Therapeutic Strategies ◽  
Citrus Paradisi ◽  
Cytoprotective Effect ◽  
Freeze Dried ◽  
Grapefruit Peel ◽  
Neuroblastoma Cell Lines

Grapefruit (Citrus paradisiMacf.) is an important cultivar of theCitrusgenus which contains a number of nutrients beneficial to human health. The objective of the present study was to evaluate changes in bioactive flavonoids, antioxidant behaviour, andin vitrocytoprotective effect of processed white and pink peels after oven-drying (45°C–60°C) and freeze-drying treatments. Comparison with fresh grapefruit peels was also assessed. Significant increases in DPPH, FRAPS, and ABTS values were observed in dried grapefruit peel samples in comparison with fresh peels, indicating the suitability of the treatments for use as tools to greatly enhance the antioxidant potential of these natural byproducts. A total of thirteen flavonoids were quantified in grapefruit peel extracts by HPLC-MS/MS. It was found that naringin, followed by isonaringin, was the main flavonoid occurring in fresh, oven-dried, and freeze-dried grapefruit peels.In vivoassay revealed that fresh and oven-dried grapefruit peel extracts (45°C) exerted a strong cytoprotective effect on SH-SY5Y neuroblastoma cell lines at concentrations ranging within 0.1–0.25 mg/mL. Our data suggest that grapefruit (Citrus paradisiMacf.) peel has considerable potential as a source of natural bioactive flavonoids with outstanding antioxidant activity which can be used as agents in several therapeutic strategies.

Utilization of urea by sheep

1964 ◽  
Vol 63 (3) ◽  
pp. 289-296 ◽  
Author(s):  
H. M. Schwartz ◽  
C. A. Schoeman ◽  
M. S. Färber
Keyword(s):  
Rumen Fluid ◽  
Soluble Starch ◽  
Large Numbers ◽  
Food Particles ◽  
Freely Suspended ◽  
Micro Organisms ◽  
Strained Material

1.The breakdown of urea, soluble starch and glucose was followed in the rumen of fistulated sheep and in artificial rumens containing rumen ingesta or strained rumen fluid.2. Carbohydrate remained in the rumen two to three times as long when starch was dosed in vivo as when glucose was given.3. The relative rates of breakdown of urea and carbohydrate by strained rumen fluid in vitro differed markedly from those in vivo. This was due to the fact that straining removed large numbers of starch- and glucose-utilizing micro-organisms which were attached to the larger food particles, while the concentration of the urea-splitting organisms which were freely suspended in the strained liquor was apparently the same as that obtained in the strained material. Strained rumen fluid should therefore not be used for studies of the utilization of urea in vitro.

scholarly journals Assessment of three generations of mice derived by ICSI using freeze-dried sperm

Zygote ◽  
2009 ◽  
Vol 17 (3) ◽  
pp. 239-251 ◽  
Author(s):  
Ming-Wen Li ◽  
Brandon J. Willis ◽  
Stephen M. Griffey ◽  
Jimmy L. Spearow ◽  
K. C. Kent Lloyd
Keyword(s):  
Genomic Instability ◽  
Freeze Drying ◽  
Mouse Strains ◽  
Control Group ◽  
Three Generations ◽  
Phenotypic Abnormality ◽  
Freeze Dried ◽  
Two Generations

SummaryAlthough the derivation of mice by intracytoplasmic sperm injection (ICSI) using freeze-dried sperm has been demonstrated previously, a comprehensive analysis of their viability, health, and fertility has not. The purpose of the present study was to determine the extent to which ICSI using freeze-dried sperm stored at 4 °C for 1–2 months from mice on either an inbred (C57BL/6J) or hybrid (B6D2F1/J) genetic background results in genomic instability and/or phenotypic abnormality in mice and two generations of their progeny. Fertilization rates (number of 2-cells per injected oocytes) using ICSI of fresh and freeze-dried sperm were similar within and between mouse strains, although fewer freeze-dried sperm-derived embryos than fresh sperm-derived embryos developed to blastocystsin vitro(C57BL/6J and B6D2F1/J) and liveborn pupsin vivo(B6D2F1/J only). Nevertheless, once born, mice derived by ICSI using freeze-dried sperm in both mouse strains were healthy and reproductively sound. No major differences in litter size, weaning rate, and sex ratio were noted in the two generations of progeny (F2 and F3) of ICSI-derived offspring using freeze-dried sperm compared with that in the natural mating (control) group. Further, there was no evidence that either ICSI or freeze drying induced genomic instability, as determined by microsatellite analysis of the derived mice and subsequent generations when compared with both parental genotypes, nor were there differences in the number or types of pathological changes in any of the three generations of progeny. We conclude that viable, healthy and genomically stable mice can be derived by ICSI using freeze-dried mouse sperm stored in the refrigerator for at least 2 months. Further, because freeze drying is a simpler and more economical technique compared with embryo and sperm cryopreservation, the results of this study justify additional research to continue to develop and enhance the technique for the preservation, storage, and sharing of genetically altered mice.

Influence of saponins and sapogenins on the bacteriolytic activity of ciliate protozoa from the sheep rumen

1998 ◽  
Vol 1998 ◽  
pp. 88-88
Author(s):  
B. Teferedegne ◽  
P.O. Osuji ◽  
A. Odenyo ◽  
R. J. Wallace ◽  
C.J. Newbold
Keyword(s):  
Rumen Fluid ◽  
Sesbania Sesban ◽  
Leguminous Tree ◽  
Bacteriolytic Activity ◽  
Rumen Protozoa ◽  
Sheep Rumen ◽  
Possible Cause ◽  
Ciliate Protozoa

Foliage from the tropical leguminous tree, Sesbania sesban, is toxic to rumen protozoa in vitro, due to materials present in a saponins-containing extract of the foliage (Newbold et al. 1997). Suppression of protozoal numbers in vivo when S. sesban is added to the diet is either transient or non-existent, however, even though washed protozoa remain sensitive to S. sesban in vitro (Newbold et al. 1997, Odenyo et al. 1997). A possible reason is that saponins are metabolised in rumen fluid (Makkar and Becker 1997). The aims of this study were to determine if the antiprotozoal effect of different accessions of S. sesban was related to their saponins composition, and if conversion of saponins to their sapogenin derivatives was a possible cause of the loss of the antiprotozoal effect in vivo.

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