Methods for the isolation, culture and assessment of the status of anaerobic rumen chytrids in both in vitro and in vivo systems

Rofiuddin, Mohammad; Beakes, Gordon W.; Parker, David S.

doi:10.1017/s0953756204000917

Cited by 23 publications

(16 citation statements)

References 40 publications

(47 reference statements)

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“…FNG5 isolated from faeces of wild nil gai ( Baselophus tragocamelus ) (Paul et al., 2003). However, the enzyme activity profile was similar to the results reported by Rezaeian et al. (2004).…”

Section: Resultssupporting

confidence: 87%

“…FNG5 isolated from faeces of wild nil gai (Baselophus tragocamelus) (Paul et al, 2003). However, the enzyme activity profile was similar to the results reported by Rezaeian et al (2004). The dayto-day enzyme activity variations in this study were also reported in other fungal isolates (Rezaeian et al, 2005;Yue et al, 2009).…”

Section: Resultssupporting

confidence: 85%

“…Carboxymethyl cellulase activity peaked on day 9 for both substrates, and it was lower than those in literatures (Sijtsma and Tan, 1993; Rezaeian et al., 2004; Tripathi et al., 2007) and higher than that of Piromyces sp. FNG5 isolated from faeces of wild nil gai ( Baselophus tragocamelus ) (Paul et al., 2003).…”

Section: Resultscontrasting

confidence: 70%

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Enzymatic characteristics of crude feruloyl and acetyl esterases of rumen fungus Neocallimastix sp. YAK11 isolated from yak (Bos grunniens)

Cao

Yang

Zhang

2012

Animal Physiology Nutrition

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Summary Rumen fungus Neocallimastix sp. YAK11 was isolated from yak (Bos grunniens), and three consecutive 10‐day pure cultures were anaerobically performed at 39 °C in 20‐ml Hungate’s tubes to explore ferulic acid esterase (FAE) and acetyl esterase (AE) activity profiles of the fungus grown on whole hay fraction of Chinese wildrye grass (Leymus chinensis) (WHOcw, n = 4) and its neutral detergent fibre fraction (NDFcw, n = 4), respectively. An aliquot of 0.7‐ml culture was sampled daily using a sterile syringe, and 0.7‐ml fresh medium was immediately added to the tubes to compensate for the withdrawn samples. Peak esterase activity occurred for FAE on day 5 (p < 0.001) and for AE on day 6 (p < 0.001). The mean activities of FAE and AE in WHOcw were 2.07 and 1.29 times of those in NDFcw (p < 0.001). Both FAE and AE activities were positively correlated with xylanase (r > 0.65, p < 0.001) and carboxymethyl cellulase (r > 0.57, p < 0.001) activities. Total volatile fatty acid concentration was positively correlated with enzyme activities of AE (r > 0.87, p < 0.001), FAE (r > 0.82, p < 0.001) and xylanase (r > 0.56, p < 0.001). Crude enzyme solution was harvested for the fungus grown on WHOcw, and the pH optimum of FAE activity was 8.0 while the optimum for AE was 9.0. Both FAE and AE had a broad pH stability range. The optimal temperatures for FAE and AE activity were 40 and 50 °C. The Michaelis constant (Km) and maximum velocity (Vmax) for FAE against methyl ferulate at pH 6.0 and 39 °C were 0.078 mm and 2.93 mU, respectively. The Km and Vmax for AE against p‐nitrophenyl acetate at pH 7.0 and 39 °C were 2.73 mm and 666.67 mU, respectively. Both FAE and AE may have prospective advantages for the enzymatic degradation of roughages in ruminant animals.

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Section: Resultssupporting

confidence: 87%

Section: Resultssupporting

confidence: 85%

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Enzymatic characteristics of crude feruloyl and acetyl esterases of rumen fungus Neocallimastix sp. YAK11 isolated from yak (Bos grunniens)

Cao

Yang

Zhang

2012

Animal Physiology Nutrition

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“…After anaerobic digestion, the particle size of CS in the digestate decreased obviously, and the pulverized CS was degraded from 0.5-1 cm to indiscernible minced material. The SEM images in Figure 6 demonstrated that the compact structure of the fibers had been swollen and destroyed by the amounts of bacteria in association with fungus [59,60], indicating that many ruminal microbes were still viable by these stages. These procedures of adherence and degradation were similar to the metabolic system in rumen, further confirming the positive effect of rumen microorganisms on the pilot-scale anaerobic digestion [13].…”

Section: Physical Form and Sem Images Of Substratesmentioning

confidence: 99%

Application of Rumen Microorganisms for Enhancing Biogas Production of Corn Straw and Livestock Manure in a Pilot-Scale Anaerobic Digestion System: Performance and Microbial Community Analysis

2018

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This study aimed to assess the feasibility of rumen microorganisms inoculated in a modified pilot-scale system for enhancing biogas production of (1) solely corn straw (CS) and (2) CS with livestock manure under different solid contents and mixture ratios. The biogas liquid was proven to pretreat CS at this scale. The digestion system was started up within 32 days at a retention time of 20 days. The rumen culture was found to have a positive response to the impact on temperature and pH. The optimal solid content of CS was detected to be 3%, resulting in a stable biogas yield of 395 L kg −1 •total solid (TS) −1. A higher biogas yield of 400 L kg −1 •TS −1-420 L kg −1 •TS −1 was achieved at a solid content of 10% organic loading rate (OLR, 4.42 kg volatile solid (VS) m −3 •d −1) in co-digestion systems with CS and livestock manure. The methane content could be maintained at about 60%. Hydrogenotrophic methanogens were dominated by Methanobacterium in the solely CS digestion system, and two methanogenetic pathways, including hydrogenotrophic and acetoclastic methanogens by Methanosarcina and Methanobacterium, co-occurred for methane production during the co-digestion of CS with pig manure (PM). This study indicates that rumen microbes could be utilized in a pilot-scale digestion system and that they greatly promoted the biogas yield.

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“…A variety of materials have been utilized for this purpose, including gauze (Ríspoli et al, 2009;Trujillo et al, 2010), nylon (Muetzel et al, 2009), sieves (Eriksson et al, 2004), sackcloth (Martinele et al, 2002), fiberglass (Nheta et al, 2005) and mousseline (Rezaeian et al, 2004;Naik et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Quantitative and differential analysis of ciliate protozoa in rumen content samples filtered before and after fixation

2013

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-The objective of this study was to assess whether the straining of rumen content samples influences the estimation of protozoal density. Ninety rumen samples were obtained from 30 cattle (three samples per animal). The samples were subjected to one of three treatments at the moment of collection: 1) fixation in formalin without straining (control treatment), 2) straining before fixation in formalin, or 3) straining after fixation in formalin. To test the hypothesis of the variation in the protozoa composition in the samples, multivariate analyses with non-metric multidimensional scaling (NMDS) were carried out. The diversity and density of rumen protozoa were negatively affected by straining before fixation. In the pre-filtered sample, the number of ciliates from the genus Entodinium was reduced, and no individuals from the Diploplastron, Elytroplastron and Eudiplodinium genera were detected; these effects were not observed in the other two treatments. Straining after fixation did not interfere with the diversity of the ruminal community, but the abundance of protozoa was greater than in the control treatment and significantly greater than in the samples filtered before fixation. These factors suggest that postfixation straining is the recommended technique to analyze rumen protozoa.

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Methods for the isolation, culture and assessment of the status of anaerobic rumen chytrids in both in vitro and in vivo systems

Cited by 23 publications

References 40 publications

Enzymatic characteristics of crude feruloyl and acetyl esterases of rumen fungus Neocallimastix sp. YAK11 isolated from yak (Bos grunniens)

Enzymatic characteristics of crude feruloyl and acetyl esterases of rumen fungus Neocallimastix sp. YAK11 isolated from yak (Bos grunniens)

Application of Rumen Microorganisms for Enhancing Biogas Production of Corn Straw and Livestock Manure in a Pilot-Scale Anaerobic Digestion System: Performance and Microbial Community Analysis

Quantitative and differential analysis of ciliate protozoa in rumen content samples filtered before and after fixation

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