Isolation and Characterization of Yeasts from Rumen Fluids for Potential Use as Additives in Ruminant Feeding
<i>Saccharomyces cerevisiae</i> is a yeast strain often used to improve the feed quality of ruminants. However, <i>S. cerevisiae</i> has limited capacity to provide biomass when inoculated with carbon sources and a low ability to produce cellulase enzymes. Here, we hypothesiz...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-03-01
|
| Series: | Veterinary Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2306-7381/8/3/52 |
| _version_ | 1797540693647818752 |
|---|---|
| author | Chanon Suntara Anusorn Cherdthong Metha Wanapat Suthipong Uriyapongson Vichai Leelavatcharamas Jutaporn Sawaengkaew Pin Chanjula Suban Foiklang |
| author_facet | Chanon Suntara Anusorn Cherdthong Metha Wanapat Suthipong Uriyapongson Vichai Leelavatcharamas Jutaporn Sawaengkaew Pin Chanjula Suban Foiklang |
| author_sort | Chanon Suntara |
| collection | DOAJ |
| description | <i>Saccharomyces cerevisiae</i> is a yeast strain often used to improve the feed quality of ruminants. However, <i>S. cerevisiae</i> has limited capacity to provide biomass when inoculated with carbon sources and a low ability to produce cellulase enzymes. Here, we hypothesized that yeast in the rumen produces a large amount of biomass and could release cellulase enzymes to break down fiber content. Therefore, the aim of this study was to screen, isolate and identify yeast from the rumen fluids of Holstein Friesian steers and measure the efficiency of biomass production and cellulase activity. A fermentation medium containing sugarcane molasses as a carbon source and urea as a nitrogen source was optimized. Two fistulated–crossbred Holstein Friesian steers averaging 350 ± 20 kg body weight were used to screen and isolate the ruminal yeast. Two experiments were designed: First, a 12 × 3 × 3 factorial was used in a completely randomized design to determine biomass and carboxymethyl cellulase activity. Factor A was the isolated yeast and <i>S. cerevisiae</i>. Factor B was sugarcane molasses (M) concentration. Factor C was urea (U) concentration. In the second experiment, potential yeasts were selected, identified, and analyzed for 7 × 4 factorial use in a completely randomized design. Factor A was the incubation times. Factor B was the isolated yeast strains, including codes H-Khon Kaen University (KKU) 20 (as <i>P. kudriavzevii</i>-KKU20), I-KKU20 (<i>C. tropicalis</i>-KKU20), and C-KKU20 (as <i>Galactomyces</i> sp.-KKU20). Isolation was imposed under aerobic conditions, resulting in a total of 11 different colonies. Two appearances of colonies including asymmetric colonies of isolated yeast (indicated as A, B, C, E, and J) and ovoid colonies (coded as D, F, G, H, I, and K) were noted. Isolated yeast from the rumen capable of providing a high amount of biomass when inoculant consisted of the molasses 15% + urea 3% (M15 + U3), molasses 25% + urea 1% (M25 + U1), molasses 25% + urea 3% (M25 + U3), and molasses 25% + urea 5% (M25 + U5) when compared to the other media solution (<i>p</i> < 0.01). In addition, 11 isolated biomass-producing yeasts were found in the media solution of M25 + U1. There were 4 isolates cellulase producing yeasts discovered in the media solution of M25 + U1 and M25 + U5 whereas molasses 5% + urea 1% (M5 + U1), molasses 5% + urea 3% (M5 + U3), molasses 5% + urea 5% (M5 + U5), molasses 15% + urea 1% (M15 + U1), molasses 15% + urea 3% (M5 + U3), and M25 + U3 were found with 2, 3, 1, 2, 1, and 2 isolates, respectively. Ruminal yeast strains H-KKU20, I-KKU20, and C-KKU20 were selected for their ability to produce biomass. Identification of isolates H-KKU20 and I-KKU20 revealed that those isolates belonged to <i>Pichia kudriavzevii</i>-KKU20 and <i>Candida tropicalis</i>-KKU20 while C-KKU20 was identified as <i>Galactomyces</i> sp.-KKU20. Two strains provided maximum cell growth: <i>P. kudriavzevii</i>-KKU20 (9.78 and 10.02 Log cell/mL) and <i>C. tropicalis</i>-KKU20 (9.53 and 9.6 Log cells/mL) at 60 and 72 h of incubation time, respectively. The highest ethanol production was observed in <i>S. cerevisiae</i> at 76.4, 77.8, 78.5, and 78.6 g/L at 36, 48, 60, and 72 h of incubation time, respectively (<i>p</i> < 0.01). The <i>P. kudriavzevii</i>-KKU20 yielded the least reducing sugar at about 30.6 and 29.8 g/L at 60 and 72 h of incubation time, respectively. The screening and isolation of yeasts from rumen fluids resulted in 11 different yeasts being obtained. The potential yeasts discovered in the rumen fluid of cattle were <i>Pichia kudriavzevii</i>-KKU20, <i>Candida tropicalis</i>-KKU20, and <i>Galactomyces</i> sp.-KKU20. <i>P. kudriavzevii</i>-KKU20 had higher results than the other yeasts in terms of biomass production, cellulase enzyme activity, and cell number. |
| first_indexed | 2024-03-10T13:04:49Z |
| format | Article |
| id | doaj.art-66978afa8ff94e17b2e76508bd7fc9cf |
| institution | Directory Open Access Journal |
| issn | 2306-7381 |
| language | English |
| last_indexed | 2024-03-10T13:04:49Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj.art-66978afa8ff94e17b2e76508bd7fc9cf2023-11-21T11:11:46ZengMDPI AGVeterinary Sciences2306-73812021-03-01835210.3390/vetsci8030052Isolation and Characterization of Yeasts from Rumen Fluids for Potential Use as Additives in Ruminant FeedingChanon Suntara0Anusorn Cherdthong1Metha Wanapat2Suthipong Uriyapongson3Vichai Leelavatcharamas4Jutaporn Sawaengkaew5Pin Chanjula6Suban Foiklang7Tropical Feed Resources Research and Development Center (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen 40002, ThailandTropical Feed Resources Research and Development Center (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen 40002, ThailandTropical Feed Resources Research and Development Center (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen 40002, ThailandTropical Feed Resources Research and Development Center (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen 40002, ThailandFermentation Research Center for Value Added Agricultural Products (FerVAAP), Department of Biotechnology, Faculty of Technology, Khon Kaen University, Khon Kaen 40002, ThailandDepartment of Microbiology, Faculty of Science, Khon Kaen University, Khon Kaen 40002, ThailandAnimal Production Innovation and Management Division, Faculty of Natural Resources, Hat Yai Campus, Prince of Songkla University, Songkhla 90112, ThailandFaculty of Animal Science and Technology, Maejo University, Chiangmai 50290, Thailand<i>Saccharomyces cerevisiae</i> is a yeast strain often used to improve the feed quality of ruminants. However, <i>S. cerevisiae</i> has limited capacity to provide biomass when inoculated with carbon sources and a low ability to produce cellulase enzymes. Here, we hypothesized that yeast in the rumen produces a large amount of biomass and could release cellulase enzymes to break down fiber content. Therefore, the aim of this study was to screen, isolate and identify yeast from the rumen fluids of Holstein Friesian steers and measure the efficiency of biomass production and cellulase activity. A fermentation medium containing sugarcane molasses as a carbon source and urea as a nitrogen source was optimized. Two fistulated–crossbred Holstein Friesian steers averaging 350 ± 20 kg body weight were used to screen and isolate the ruminal yeast. Two experiments were designed: First, a 12 × 3 × 3 factorial was used in a completely randomized design to determine biomass and carboxymethyl cellulase activity. Factor A was the isolated yeast and <i>S. cerevisiae</i>. Factor B was sugarcane molasses (M) concentration. Factor C was urea (U) concentration. In the second experiment, potential yeasts were selected, identified, and analyzed for 7 × 4 factorial use in a completely randomized design. Factor A was the incubation times. Factor B was the isolated yeast strains, including codes H-Khon Kaen University (KKU) 20 (as <i>P. kudriavzevii</i>-KKU20), I-KKU20 (<i>C. tropicalis</i>-KKU20), and C-KKU20 (as <i>Galactomyces</i> sp.-KKU20). Isolation was imposed under aerobic conditions, resulting in a total of 11 different colonies. Two appearances of colonies including asymmetric colonies of isolated yeast (indicated as A, B, C, E, and J) and ovoid colonies (coded as D, F, G, H, I, and K) were noted. Isolated yeast from the rumen capable of providing a high amount of biomass when inoculant consisted of the molasses 15% + urea 3% (M15 + U3), molasses 25% + urea 1% (M25 + U1), molasses 25% + urea 3% (M25 + U3), and molasses 25% + urea 5% (M25 + U5) when compared to the other media solution (<i>p</i> < 0.01). In addition, 11 isolated biomass-producing yeasts were found in the media solution of M25 + U1. There were 4 isolates cellulase producing yeasts discovered in the media solution of M25 + U1 and M25 + U5 whereas molasses 5% + urea 1% (M5 + U1), molasses 5% + urea 3% (M5 + U3), molasses 5% + urea 5% (M5 + U5), molasses 15% + urea 1% (M15 + U1), molasses 15% + urea 3% (M5 + U3), and M25 + U3 were found with 2, 3, 1, 2, 1, and 2 isolates, respectively. Ruminal yeast strains H-KKU20, I-KKU20, and C-KKU20 were selected for their ability to produce biomass. Identification of isolates H-KKU20 and I-KKU20 revealed that those isolates belonged to <i>Pichia kudriavzevii</i>-KKU20 and <i>Candida tropicalis</i>-KKU20 while C-KKU20 was identified as <i>Galactomyces</i> sp.-KKU20. Two strains provided maximum cell growth: <i>P. kudriavzevii</i>-KKU20 (9.78 and 10.02 Log cell/mL) and <i>C. tropicalis</i>-KKU20 (9.53 and 9.6 Log cells/mL) at 60 and 72 h of incubation time, respectively. The highest ethanol production was observed in <i>S. cerevisiae</i> at 76.4, 77.8, 78.5, and 78.6 g/L at 36, 48, 60, and 72 h of incubation time, respectively (<i>p</i> < 0.01). The <i>P. kudriavzevii</i>-KKU20 yielded the least reducing sugar at about 30.6 and 29.8 g/L at 60 and 72 h of incubation time, respectively. The screening and isolation of yeasts from rumen fluids resulted in 11 different yeasts being obtained. The potential yeasts discovered in the rumen fluid of cattle were <i>Pichia kudriavzevii</i>-KKU20, <i>Candida tropicalis</i>-KKU20, and <i>Galactomyces</i> sp.-KKU20. <i>P. kudriavzevii</i>-KKU20 had higher results than the other yeasts in terms of biomass production, cellulase enzyme activity, and cell number.https://www.mdpi.com/2306-7381/8/3/52ruminal yeastscreeningisolationbiomass of yeastcellulase enzyme |
| spellingShingle | Chanon Suntara Anusorn Cherdthong Metha Wanapat Suthipong Uriyapongson Vichai Leelavatcharamas Jutaporn Sawaengkaew Pin Chanjula Suban Foiklang Isolation and Characterization of Yeasts from Rumen Fluids for Potential Use as Additives in Ruminant Feeding Veterinary Sciences ruminal yeast screening isolation biomass of yeast cellulase enzyme |
| title | Isolation and Characterization of Yeasts from Rumen Fluids for Potential Use as Additives in Ruminant Feeding |
| title_full | Isolation and Characterization of Yeasts from Rumen Fluids for Potential Use as Additives in Ruminant Feeding |
| title_fullStr | Isolation and Characterization of Yeasts from Rumen Fluids for Potential Use as Additives in Ruminant Feeding |
| title_full_unstemmed | Isolation and Characterization of Yeasts from Rumen Fluids for Potential Use as Additives in Ruminant Feeding |
| title_short | Isolation and Characterization of Yeasts from Rumen Fluids for Potential Use as Additives in Ruminant Feeding |
| title_sort | isolation and characterization of yeasts from rumen fluids for potential use as additives in ruminant feeding |
| topic | ruminal yeast screening isolation biomass of yeast cellulase enzyme |
| url | https://www.mdpi.com/2306-7381/8/3/52 |
| work_keys_str_mv | AT chanonsuntara isolationandcharacterizationofyeastsfromrumenfluidsforpotentialuseasadditivesinruminantfeeding AT anusorncherdthong isolationandcharacterizationofyeastsfromrumenfluidsforpotentialuseasadditivesinruminantfeeding AT methawanapat isolationandcharacterizationofyeastsfromrumenfluidsforpotentialuseasadditivesinruminantfeeding AT suthiponguriyapongson isolationandcharacterizationofyeastsfromrumenfluidsforpotentialuseasadditivesinruminantfeeding AT vichaileelavatcharamas isolationandcharacterizationofyeastsfromrumenfluidsforpotentialuseasadditivesinruminantfeeding AT jutapornsawaengkaew isolationandcharacterizationofyeastsfromrumenfluidsforpotentialuseasadditivesinruminantfeeding AT pinchanjula isolationandcharacterizationofyeastsfromrumenfluidsforpotentialuseasadditivesinruminantfeeding AT subanfoiklang isolationandcharacterizationofyeastsfromrumenfluidsforpotentialuseasadditivesinruminantfeeding |