Effect of Defaunation on In Vitro Fermentation Characteristics and Methane Emission When Incubated with Forages

Qin, Wei-Ze,Choi, Seong-Ho,Lee, Seung-Uk,Lee, Sang-Suk,Song, Man-Kang The Korean Society of Grassland and Forage Science 2013 한국초지조사료학회지 Vol.33 No.3

An in vitro study was conducted to determine the effects of defaunation (removal of protozoa) and forage sources (rice straw, ryegrass and tall fescue) on ruminal fermentation characteristics, methane ($CH_4$) production and degradation by rumen microbes. Sodium lauryl sulfate, as a defaunation reagent, was added into the mixed culture solution to remove ruminal protozoa at a concentration of 0.375 mg/ml. Pure cellulose (0.64 g, Sigma, C8002) and three forage sources were incubated in the bottle of culture solution of mixed rumen microbes (faunation) or defaunation for up to 24 h. The concentration of ammonia-N was high under condition of defaunation compared to that from faunation in all incubations (p<0.001). Total VFA concentration was increased at 3, 6 and 12 h (p<0.05~p<0.01) but was decreased at 24 h incubation (p<0.001) under condition of defaunation. Defaunation decreased acetate (p<0.001) and butyrate (p<0.001) proportions at 6, 12 and 24 h incubation times, but increased propionate (p<0.001) proportion at all incubation times for forages. Effective degradability of dry matter was decreased by defaunation (p<0.001). Defaunation not only decreased total gas (p<0.001) and $CO_2$ (p<0.01~0.001) production at 12 and 24 h incubations, but reduced $CH_4$ production (p<0.001) at all incubation times for all forages. The $CH_4$ production, regardless of defaunation, in order of forage sources were rice straw > tall fescue > ryegrass > cellulose (p<0.001) up to 24 h incubation.

Effects of Rumen Protozoa of Brahman Heifers and Nitrate on Fermentation and In vitro Methane Production

Nguyen, S.H.,Li, L.,Hegarty, R.S. Asian Australasian Association of Animal Productio 2016 Animal Bioscience Vol.29 No.6

Two experiments were conducted assessing the effects of presence or absence of rumen protozoa and dietary nitrate addition on rumen fermentation characteristics and in vitro methane production in Brahman heifers. The first experiment assessed changes in rumen fermentation pattern and in vitro methane production post-refaunation and the second experiment investigated whether addition of nitrate to the incubation would give rise to methane mitigation additional to that contributed by defaunation. Ten Brahman heifers were progressively adapted to a diet containing 4.5% coconut oil distillate for 18 d and then all heifers were defaunated using sodium 1-(2-sulfonatooxyethoxy) dodecane (Empicol). After 15 d, the heifers were given a second dose of Empicol. Fifteen days after the second dosing, all heifers were allocated to defaunated or refaunated groups by stratified randomisation, and the experiment commenced (d 0). On d 0, an oral dose of rumen fluid collected from unrelated faunated cattle was used to inoculate 5 heifers and form a refaunated group so that the effects of re-establishment of protozoa on fermentation characteristics could be investigated. Samples of rumen fluid collected from each animal using oesophageal intubation before feeding on d 0, 7, 14, and 21 were incubated for in vitro methane production. On d 35, 2% nitrate (as $NaNO_3$) was included in in vitro incubations to test for additivity of nitrate and absence of protozoa effects on fermentation and methane production. It was concluded that increasing protozoal numbers were associated with increased methane production in refaunated heifers 7, 14, and 21 d after refaunation. Methane production rate was significantly higher from refaunated heifers than from defaunated heifers 35 d after refaunation. Concentration and proportions of major volatile fatty acids, however, were not affected by protozoal treatments. There is scope for further reducing methane output through combining defaunation and dietary nitrate as the addition of nitrate in the defaunated heifers resulted in 86% reduction in methane production in vitro.

Effects of Defaunation on Fermentation Characteristics, Degradation of Ryegrass Hay and Methane Production by Rumen Microbes In Vitro When Incubated with Plant Oils

Qin, Wei-Ze,Li, Cheng-Yun,Choi, Seong-Ho,Jugder, Shinekhuu,Kim, Hyun-Ju,Lee, Sang-Suk,Song, Man-Kang The Korean Society of Grassland and Forage Science 2014 한국초지조사료학회지 Vol.34 No.3

This study was conducted to examine the effects of defaunation (removal of live protozoa) on fermentation characteristics, degradation of ryegrass hay and $CH_4$ (methane) production by rumen microbes when incubated with plant oils (SO, sunflower oil and LO, linseed oil) in vitro. Sodium lauryl sulfate (0.000375 g/ml) as a defaunation reagent was added into the culture solution and incubated anaerobically up to 24 h at $39^{\circ}C$. pH from defaunation was increased for all treatments from 6 h incubation times (p<0.01-0.001) compared with those from fauantion. Concentration of ammonia-N from defaunation is higher than that from faunation at 3 h (p<0.001), 12 h (p<0.05) and 24 h (p<0.001) incubation times. Defaunation decreased (p<0.01-0.001) total volatile fatty acid concentration at all incubation times. Molar proportions of $C_2$ (acetate, p<0.05-0.001) and butyrate (p<0.01-0.001) were also decreased by defaunation at all incubation times. Molar proportion of $C_3$ (propionate), however, was increased by defaunation at all incubation times (p<0.001). Thus the rate of $C_2$ to $C_3$ was decreased by defaunation at all incubation times (p<0.001). Defaunation decreased ED (effective degradability) of dry matter (p<0.001) and ED of neutral detergent fiber (p<0.001) of ryegrass hay. Defaunation decreased total gas, $CH_4$ production, $CH_4$ % in total gas and $CH_4/CO_2$ at all incubation times (p<0.001). Oil supplementation decreased total gas (p<0.05-0.001), $CH_4$ production (p<0.001) and $CH_4$ % in total gas (p<0.001) compared with control at all incubation times. The result of this study showed that defaunation combined with oil supplementation may cause an alteration of microbial communities and further medicate the fermentation pattern, resulting in both reduction of degradation of ryegrass hay and $CH_4$ production. No difference, however, was observed in all the examinations between SO and LO.

Effects of Defaunation on Fermentation Characteristics and Methane Production by Rumen Microbes In vitro When Incubated with Starchy Feed Sources

Qin, W.Z.,Li, C.Y.,Kim, J.K.,Ju, J.G.,Song, Man-K. Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.10

An in vitro experiment was conducted to examine the effects of defaunation (removal of protozoa) on ruminal fermentation characteristics, $CH_4$ production and degradation by rumen microbes when incubated with cereal grains (corn, wheat and rye). Sodium lauryl sulfate as a defaunation reagent was added into the culture solution at a concentration of 0.000375 g/ml, and incubated anaerobically for up to 12 h at $39^{\circ}C$. Following defaunation, live protozoa in the culture solution were rarely observed by microscopic examination. A difference in pH was found among grains regardless of defaunation at all incubation times (p<0.01 to 0.001). Defaunation significantly decreased pH at 12 h (p<0.05) when rumen fluid was incubated with grains. Ammonia-N concentration was increased by defaunation for all grains at 6 h (p<0.05) and 12 h (p<0.05) incubation times. Total VFA concentration was increased by defaunation at 6 h (p<0.05) and 12 h (p<0.01) for all grains. Meanwhile, defaunation decreased acetate and butyrate proportions at 6 h (p<0.05, p<0.01) and 12 h (p<0.01, p<0.001), but increased the propionate proportion at 3 h, 6 h and 12 h incubation (p<0.01 to 0.001) for all grains. Defaunation increased in vitro effective degradability of DM (p<0.05). Production of total gas and $CO_2$ was decreased by defaunation for all grains at 1 h (p<0.05, p<0.05) and then increased at 6 h (p<0.05, p<0.05) and 12 h (p<0.05, p<0.05). $CH_4$ production was higher from faunation than from defaunation at all incubation times (p<0.05).

Influence of Controlling Protozoa on the Degradation and Utilization of Dietary Fibre and Protein in the Rumen and Nitrogenous Flow Entering the Duodenum of Sheep

Han, C.Y.,Lu, D.X.,Hu, M.,Tan, Z.L. Asian Australasian Association of Animal Productio 1999 Animal Bioscience Vol.12 No.8

Nine two-year old sheep fitted with rumen and duodenum cannulas were used to study the effect of controlling protozoa flora on the degradation and utilization of dietary fibre and protein in the rumen and on nitrogenous flow to the duodenum. There were three groups in this experiment: defaunation (DF); partial defaunation (PDF); faunation (F) as control. Results showed that: 1,There were no differences between treatments in dietary DM degradation in the rumen, but defaunation and partial defaunation increased the quantity of nitrogenous material in the rumen and the flow of N to duodenum. 2, partial defaunation and defaunation improved the degradabilities of dietary NDF, ADF and HC, but there were no differences between the defaunated and partially defaunated groups. 3, Partial defaunation decreased the degradability of dietary protein in the rumen. There was no difference between defaunated and faunated groups. 4, Defaunation and partial defaunation increased the quantity of total N (TN) and microbial N (MCN) in the rumen and the amounts entering the duodenum. The protozoa N (PN) flow in the faunated group was higher than that in the partially defaunated group, and the amino acid pattern in the digesta at the proximal duodenum in the defaunated group was closer to the ideal amino acid pattern. 5, There were differences in the mole percent of acetic, propionic, total-VFAs and the non-glucogenic to glucogenic VFAs ratio (NGR) value in the rumen fluids. The order was as follows: mole percent of acetate: F>PDF>DF; mole percent of propionate: DF>PDF>F; total-VFAs: PDF>F>DF; NGR: F>PDF>DF.

Defaunation Reduced Production of CLAby Rumen Microbes In VitroWhen Incubated with Ryegrass Hay and Plant Oils

Wei-Ze Qin,Man-Kang Song 충북대학교 동물생명과학연구소 2014 동물생명과학연구 Vol.6 No.-

An in vitro experiment was conducted to examine the effects of defaunation(removal of protozoa) on production of conjugated linoleic acid (CLA) when incubated with ground ryegrass hay and plant oils ((linoleic acid (C18:2)-rich sunflower oil (SO) and linolenic acid (C183)-rich linseed oil (LO)) in the form of emulsion with egg albumin in vitro. Sodium lauryl sulfate as a defaunation reagent was added into the culture solution at a concentration of 0.000375g/ml, and incubated anaerobically for up to24h at 39℃. Defaunation decreased (P<0.05-0.001) the concentration of cis9, trans11-CLA (P<0.001), trans10, cis12-CLA (P<0.01) and total CLAs (P<0.001) at 6 and 24 h incubations compared with faunation. The concentrations of cis9, trans11-CLA and total CLAs were higher (P<0.001) by SO supplementation than those by LO at all incubation times. Interactions between defaunation and plant oil were found in C18:0 (P<0.01-0.001)from6incubations, cis9, trans11-CLA (P<0.01-0.001) and total CLAs (P<0.01-0.001) at 3, 6 and 24 h incubations.

EFFECTS OF DEFAUNATION ON DEGRADABILITY AND PROTEIN SYNTHESIS IN THE RUMEN OF SHEEP

Petkov, A.,Todorov, N.,Enev, E.,Oblakov, N.,Demeyer, D. Asian Australasian Association of Animal Productio 1989 Animal Bioscience Vol.2 No.3

Rumen Manipulation to Improve Animal Productivity

Santra, A.,Karim, S.A. Asian Australasian Association of Animal Productio 2003 Animal Bioscience Vol.16 No.5

Anaerobic rumen microorganisms mainly bacteria, protozoa and fungi degrade ligno-cellulosic feeds consumed by the ruminants. The ruminants in developing countries are predominantly maintained on low grade roughage and grazing on degraded range land resulting in their poor nutrient utilization and productivity. Hence, manipulation of rumen fermentation was tried during last two decades to optimize ruminal fermentation for improving nutrient utilization and productivity of the animals. Modification of rumen microbial composition and their activity was attempted by using chemical additives those selectively effect rumen microbes, introduction of naturally occurring or genetically modified foreign microbes into the rumen and genetically manipulation of existing microbes in the rumen ecosystem. Accordingly, rumen protozoa were eliminated by defaunation for reducing ruminal methane production and increasing protein outflow in the intestine, resulting in improve growth and feed conversion efficiency of the animals. Further, Interspecies trans-inoculation of rumen microbes was also successfully used for annulment of dietary toxic factor. Additionally, probiotics of bacterial and yeast origin have been used in animal feeding to stabilize rumen fermentation, reduced incidence of diarrhoea and thus improving growth and feed conversion efficiency of young stalk. It is envisaged that genetic manipulation of rumen microorganisms has enormous research potential in developing countries. In view of feed resource availability more emphasis has to be given for manipulating rumen fermentation to increase cellulolytic activity for efficient utilization of low grade roughage.

Energy Metabolism and Methane Production in Faunated and Defaunated Sheep Fed Two Diets with Same Concentrate to Roughage Ratio (70:30) but Varying in Composition

Chandramoni, Chandramoni,Jadhao, S.B.,Tiwad, C.M.,Haque, N.,Murarilal, Murarilal,Khan, M.Y. Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.9

Two calorimetric experiments were performed to investigate the effects of two diets with same concentrate: roughage ratio (70:30) but varying in composition on energy metabolism and methane production in faunated (F) and defaunated (DF) Muzaffarnagari sheep. For experiment I, ten animals were divided equally into two groups of which one was kept normally F as such while other was DF using 10% sodium lauryl sulphate. All the animals were offered diet I which comprised of oat hay and concentrate mixture I (CM I) containing maize grain (93%) as a major ingredient in 70:30 ratio. Similarly, the experiment II was conducted for which four F and four DF sheep (same as used for experiment I) were switched to diet II that consisted of maize hay and CM II (maize grain 59% + molasses 36%). Through diet II, DM intake in DF sheep was significantly (p<0.05) lower. Intake of GE through both the diets was similar in F and DF sheep. Digestibility of DM, OM, CP and GE and also metabolisability (ME/GE) was similar in F and DF sheep on both the diets. Total urinary energy loss did not differ in F and DF on both the diets, but methane energy loss as a percent of GE in DF was significantly (p<0.05) lower on diet I (3.75 vs 2.48), while it did not differ on diet II (3.20 vs 3.60). Heat production was significantly (p<0.01) reduced in DF on both the diets. Although, efficiency of utilisation of ME for maintenance calculated as per ARC (1984) did not differ in F and DF on both the diets, efficiency for maintenance and growth was higher (0.60 vs 0.672) on diet I in DF. It was inferred that methane production in DF sheep reduces on good quality hay-based diet supplemented with slowly fermentable carbohydrate (maize grain) but supplementation of molasses (rapidly fermentable CHO) nullify this effect when sheep were fed diets with concentrate: roughage ratio of 70:30.

Practical Application of Defaunation of Cattle on Farms in Vietnam: Response of Young Cattle Fed Rice Straw and Grass to a Single Drench of Groundnut Oil

Nguyen, Thi Hong Nhan,Nguyen, Van Hon,Nguyen, Trong Ngu,Nguyen, Tien Von,Preston, T.R.,Leng, R.A. Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.4

Farmers in the centre of Vietnam have a tradition of dosing young cattle with groundnut oil before fattening them on a diet of rice straw and road-side grass. These farmers claim the cattle grow faster. It was hypothesized that the effect of the oil could be to eliminate the protozoa from the rumen. This is known to increase the net microbial growth efficiency in the rumen and increase the protein supply to the animal. To test this hypothesis, two experiments were undertaken; one on-station with four cattle fitted with rumen cannulae and the second a growth trial with 25 young cattle in smallholder farms. When the cannulated animals were drenched with groundnut oil, the protozoa were eliminated from the rumen and animals could be kept free of protozoa by isolation. The ammonia concentration in the rumen fluid was decreased when the protozoa were eliminated and there was an indication of improved rumen dry matter degradability of the forage components of the diet. In the practical condition on the smallholder farms, the growth rates of cattle drenched with groundnut oil were increased considerably (65%) compared with untreated control animals. The laboratory results when taken together with the on-farm results indicate that these resource-poor farmers had been able to defaunate their cattle and to maintain the fauna-free state by isolation of their animals from extraneous stock. This traditional practice in Central Vietnam, whereby one family keeps only one or two animals that are hand fed and tethered, has quite a large potential for all of those countries where animals are fed agro-industrial by-products, as it is highly economic. The use of 1 litre of oil compared with 1 kg of rice polishing per day (300 kg over 300 days), would be highly profitable in all countries of South-East Asia.