Rumen fluid handling affects measurements of its enzymatic activity and in vitro digestibility

Raffrenato, E.; Badenhorst, Marelise; Shipandeni, M. N. T.; Zyl, Willem H. van

doi:10.1016/j.anifeedsci.2021.115060

Cited by 6 publications

(3 citation statements)

References 32 publications

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“…The relative proportion of proteins in the abomasum obtained from feed and microbial proteins synthesised in the rumen, however, depends on the physiological status of lambs and on diet composition. GIN infection can therefore probably also affect the specific activities of enzymes of the ruminal microorganisms, especially amylase, which is associated with the particulate fraction [53]. Diet in our study, however, may also have contributed to the increase in amylase activity in the ruminal microorganisms in CON.…”

Section: Discussionmentioning

confidence: 52%

Growth Performance, Nutrient Composition, Ruminal Microbiota, and Fermentation In Vivo and In Vitro Associated with Endoparasites of Lambs

Mikulová¹,

Petrič²,

Komáromyová³

et al. 2023

Preprint

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We conducted an experiment on lambs to determine the effect of infection with the gastrointestinal nematode (GIN) Haemonchus contortus on growth performance, nutrient composition, microbiota, and ruminal fermentation in vivo and in vitro. Twelve female lambs were equally divided into two groups: uninfected animals (CON) and animals infected with approximately 5000 third-stage ISE larvae derived from the MHCo1 strain of H. contortus (INF). Two lambs per group were killed on days 48, 49, and 50 after infection and ruminal content was collected separately from each lamb. Batch cultures of ruminal fluid from CON and INF were incubated in vitro with 0.25 g meadow hay, a plant mixture and chicory used as substrates. The ruminal populations of protozoa, bacteria, total Archaea, and the archaean orders Methanobacteriales and Methanomicrobiales were significantly higher in INF than CON. In vivo concentrations of n-butyrate, iso-butyrate, and iso-valerate were significantly lower in INF than CON. In vitro values after 24 h of ruminal fermentation varied within the substrate. GIN infection by H. contortus was associated with increased methanogenesis. In conclusion, chicory was a promising substrate for mitigating methane production and may modulate the ruminal microbiome of GIN-infected lambs due to its chemical and phytochemical composition.

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Section: Discussionmentioning

confidence: 52%

Growth Performance, Nutrient Composition, Ruminal Microbiota, and Fermentation In Vivo and In Vitro Associated with Endoparasites of Lambs

Mikulová¹,

Petrič²,

Komáromyová³

et al. 2023

Preprint

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“…In Experiment 1, GIN infection also affected the specific activities of enzymes of the ruminal microorganisms, especially amylase (Table 4), which is associated with the particulate fraction [53]. Xylanase was higher (not significantly) in GIN-infected lambs because rumen protozoa, fungi, and bacteria are associated with xylanase activities that accelerate the biodegradation of xylan to SCFAs and gases during the processing of lignocellulosic biomass in the rumen [54].…”

Section: Discussionmentioning

confidence: 95%

Growth Performance and Ruminal Fermentation in Lambs with Endoparasites and In Vitro Effect of Medicinal Plants

Mikulová,

Petrič,

Komáromyová

et al. 2023

Agriculture

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We investigated growth performance and ruminal fermentation associated with gastrointestinal nematode (GIN) Haemonchus contortus in lambs and in vitro ruminal fermentation of mallow, chamomile, fumitory, wormwood (Herbmix), and chicory using inoculum from GIN-infected lambs. Twelve lambs were equally divided into two groups: uninfected animals (CON) and animals infected (INF) with approximately 5000 third-stage larvae derived from the MHCo1 strain of GIN H. contortus. Two lambs per group were killed on days 48, 49, and 50 after infection and ruminal content was collected separately from each lamb. Batch cultures of ruminal fluid from CON and INF were incubated for 24 h in vitro with 0.25 g meadow hay, Herbmix, and chicory using an in vitro gas production technique. Daily weight gain was relatively lower in the INF than the CON group, but not significantly (72.6 vs. 130.1 g/day). The ruminal populations of protozoa, bacteria, total Archaea, Methanobacteriales, and Methanomicrobiales were significantly higher in the INF than in the CON group. The substrates affected the concentrations of n-butyrate, iso-butyrate, n-valerate, iso-valerate, ammonia-N, total gas, and methane (p < 0.001) in vitro. GIN infection affected fermentation and microbial population in the rumens of the lambs, and chicory was a promising substrate to modulate ruminal fermentation in vitro.

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“…Enzymatic activity of amylase, endoglucanase, lichenase, and xylanase were assessed colorimetrically by measuring the amount of reducing sugars released using dinitrosalicylic acid (Miller, 1959;Raffrenato et al, 2021). Amylolytic activity was determined using 0.2% (wt/vol) cooked maize starch (S4126, Sigma-Aldrich) resuspended in 0.05 M citrate buffer (pH 6.0).…”

Section: Determination Of Enzymatic Activitiesmentioning

confidence: 99%

The diurnal patterns of ruminal enzymatic activity and in vitro digestibility of starch, neutral detergent fiber, and protein

Raffrenato

Badenhorst

Harvatine

et al. 2022

Journal of Dairy Science

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The objective of this study was to determine whether diurnal patterns in starch, neutral detergent fiber (NDF) and protein digestibilities and amylolytic, fibrolytic, and proteolytic activities exist in dairy cows. Rumen fluid was collected from 4 ruminally cannulated Holstein dairy cows before the morning feeding and subsequently every 4 h for a 24-h period. Two of the cows were restricted from feed for 8 h overnight, and the other 2 continued to receive their feed ad libitum, to isolate and quantify the effects of changes in feeding behavior at night. After 2 runs the cows were crossed over between night feeding treatments. Rumen fluid was analyzed for enzymatic activity and in vitro starch, NDF, and nitrogen digestibility. Circadian rhythm analyses of enzymatic activity and in vitro digestibility were conducted by fitting the linear form of a cosine function with a 24-h period. Patterns were observed in activity for amylase, lichenase, endoglucanase, and xylanase, with the highest activities observed at the time points subsequent to milking and feed delivery. Protease activity was unaffected by either feeding treatment or possible feeding behavior. When fitted to a cosine function, all the parameters tested followed a daily pattern that was sensitive to the overnight availability of feed, although the parameters responded differently to the feeding treatment. The patterns displayed by in vitro digestibility results of starch, NDF, and nitrogen, across the various fluid collection time points, were highly variable. The time at peak (acrophase) observed in the enzymatic analysis did not correspond to those observed in the in vitro analysis. These results suggest that different interpretations should be given to enzymatic activities and in vitro digestibility values, and the time of rumen fluid collection relative to feeding time should be considered and reported when rumen fluid is used for research or commercial purposes. Maximum digestibility appears in fact to be reached around 4 to 5 h after the main ration delivery for NDF and starch and around ration delivery for protein.

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Rumen fluid handling affects measurements of its enzymatic activity and in vitro digestibility

Cited by 6 publications

References 32 publications

Growth Performance, Nutrient Composition, Ruminal Microbiota, and Fermentation In Vivo and In Vitro Associated with Endoparasites of Lambs

Growth Performance, Nutrient Composition, Ruminal Microbiota, and Fermentation In Vivo and In Vitro Associated with Endoparasites of Lambs

Growth Performance and Ruminal Fermentation in Lambs with Endoparasites and In Vitro Effect of Medicinal Plants

The diurnal patterns of ruminal enzymatic activity and in vitro digestibility of starch, neutral detergent fiber, and protein

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