Tannin-degrading Bacteria Research Articles

Isolation and Identification of Tannin-Degrading Bacteria From Goat Feces, Ruminal Fluid, and Rumen Gut

Tannins are toxic polyphenols present in various plants, contributing to microbial attacks and plant protection due to their astringence and bitter taste. However, high tannin inclusion in poultry diets will result in dyspepsia, hampering nutrient absorption and digestion. Interestingly, several bacteria occupying the rumen and gastrointestinal tract (GIT) of animals may tolerate tannins and degrade them by wielding tannase enzymes. The study aims to isolate and characterize potential tannin-degrading bacteria (TDB) from several ruminant specimens. The TDBs were isolated based on their tannin hydrolyzing ability on a minimal salt medium (MSM) agar complemented with 0.2% tannic acid as the sole source of carbon and energy. The maximum tannin tolerance of the isolates was characterized using increased tannin concentrations on the MSM agar plates. Furthermore, the tannase activity was also evaluated over a five-day incubation. A total of 42 tannin degraders were isolated, and 10 TDBs were chosen for further characterization based on the hydrolyzed zone produced. Molecular identification revealed the presence of Bacillus cereus (TDB536), Lysinibacillus macroides (TDB17), Acinetobacter nosocomialis (TDB18, 20, 23, 24, 30, 35), and Staphylococcus saprophyticus (TDB40). TDB17, TDB18, and TDB24 showed the highest tannic acid tolerance at 1.0%, while TDB36 and TDB40 exhibited the lowest tolerance at 0.4%. Each TDB displayed varying tannase activities, ranging from 11.56 to 42.08 U/mL over a five-day incubation period. TDB5 and TDB35 demonstrated significantly higher tannase activity on day 2 (p<0.05). Meanwhile, TDB23 and TDB24 showed the highest tannase on day 4 (p<0.05). Among the isolates, A. nosocomialis strain AE6 (TDB24) from feces exhibited the highest tannase activity (42.08 U/mL) and represented the best TDB. The isolated strains demonstrate their capabilities in reducing tannin's antinutritional effects in poultry feed.

Isolation and characterization of tanninolytic bacteria from sheep rumen contents: Assessment of tannin degrading, fibrolytic and feed digestibility potential

To counteract the negative effects of phytotannins, ruminants host a diverse community of bacteria that break down tannins. These bacteria not only possess tanninolytic activity but also have other functional relevance which has not been properly examined. Therefore, the present study was conducted to isolate and characterize tannin degrading bacteria (TDB) with possible fibrolytic activity from rumen of sheep fed tannin-rich diet. Based on the development of a clear zone on tannin-enriched agar plates, a total of 24 isolates were screened as tannin hydrolyzing bacteria. Majority of the isolates were gram positive cocci. The isolates could tolerate phenolic monomers such as ferulic acid, gallic acid, vanillic acid up to 30 mM and pyrogallol up to 10 mM however, the isolates failed to grow in presence of syringic acid, p-coumaric acid and p-hydroxybenzoic acid. Highest tannase activity was observed in isolate TDB23 while, lowest in isolates TDB2 and TDB5. In terms of fibrolytic activity, maximum endoglucanase, exoglucanase, and FPase activity was observed in isolate TDB9. In vitro digestibility of Prosopis cineraria leaves was considerably enhanced by inoculation of isolate TDB23 followed by TDB9. Therefore, the screened isolates demonstrated promising tannin and fibre degrading potential which can be further explored as direct-fed microbial in ruminants for effective utilization of tannin-rich fibrous feeds.

Isolation and characterization of tannin-degrading bacteria from the rumen of wild Hokkaido sika deer (Cervus nippon yezoensis).

We isolated tannin-degrading bacteria from the rumen of wild Hokkaido sika deer and characterized their phylogeny and tannase activity in relation to sample sources. The condensed tannin level was higher in all deer rumen samples (n = 20) than in forage-fed cattle rumen samples (n = 6), whereas no hydrolyzable tannins were detected in any of the rumen samples. Rumen bacteria were enumerated on nonselective brain heart infusion (BHI) agar medium and then transferred onto tannic acid-containing BHI agar plates to screen for bacteria only showing growth (tannin-resistant bacteria) and those showing both growth and a clear zone (tannin-degrading bacteria). Summer samples provided only tannin-resistant bacteria, none of which showed tannin-degrading activity. Although winter samples also provided tannin-resistant bacteria, most isolates exhibited tannin-degrading activity. A total of 70 isolates exhibiting tannin-degrading activity were classified as Streptococcus bovis group based on 16S rRNA gene sequencing and further classified into two groups, either group A or group B. Group A consisted of isolates showing weak tannase activity, whereas group B included a majority of the isolates exhibiting high tannase activity. These results suggest that wild Hokkaido sika deer develop tannin-degrading Streptococcus in the rumen during winter, which allows access to woody food materials rich in tannins.

Use of tannase-producing bacteria isolated from the rumen to improve the nutritional value of pomegranate peel for fattening lambs.

The use of plants and by-products, which are containing a high amount of secondary and anti-nutritional compounds such as tannins, in animal feed is limited. The methods that can reduce these compounds make facilitate their use in animal feed. The aim of this study was to reduce the adverse effects of pomegranate peel (PP) tannin for fattening lambs using the tannase-producing bacteria. Twenty-one Arabi male lambs (averagely 35±3.8kg weight and 8±1.0 months age) were used in a completely randomized design with three treatments and seven replications in the present experiment. The experimental treatments included 1 - control diet (CNT, no PP), 2 - diet containing untreated PP (raw PP, UTPP) and 3 - diet containing PP treated with tannase-producing bacteria (bacteria treating PP, BTPP). Using UTPP decreased nutrient intake compared to the control and treatment with tannase-producing bacteria again significantly increased nutrient intake compared to the UTPP (p<0.05). The digestibilities of organic matter, neutral detergent fibre and acid detergent fibre in the control treatment were significantly higher than UTPP and BTPP and in the BTPP were significantly higher than the UTPP (p<0.05). The use of UTPP in the diet significantly decreased the pH, ammonia nitrogen concentration and the total protozoa population of the rumen compared to the control (p<0.05), and treatment with bacteria increased them again. The lowest total protozoa population was observed in UTPP treatments (p<0.05). The highest concentration of blood glucose was observed in UTPP; however, the highest concentrations of blood urea nitrogen, cholesterol, triglyceride, high-density lipoprotein (non-significant) and low-density lipoprotein were in the control treatment. The effect of experimental treatments on the dry matter consumption of the whole period was significant; however, there was no significant effect on average daily gain, feed conversion ratio, feed efficiency and longissimus muscle colorimetric systems. Therefore, considering the positive effects of treatment PP with tannin-degrading bacteria relative to raw PP, using these bacteria is a proper way to reduce tannin, thus improving the nutritional value of PP for ruminants.

Tannin Degrading Bacteria as Biotherapeutic and Energy Recovering Additives of Agricultural Waste's Silage

Tannin Degrading Bacteria as Biotherapeutic and Energy Recovering Additives of Agricultural Waste's Silage

Effect of treated Conocarpus erectus L. leaves with Klebsiella pneumoniae and Acinetobacter as tannin-degrading bacteria on digestion activity of rumen microorganisms

The current research focuses on the effect of Conocarpus erectus L. leaves (CL) processed with two tannin-degrading bacteria Klebsiella pneumoniae (K. p) and Acinetobacter (A. b) on digestion activity and fermentation parameters of rumen bacteria and fungi in Arabian sheep. These isolates capable of utilizing tannic acid as sole carbon and energy source. Eight species of Klebsiella pneumoniae (A1, A2, A3, A4, A5, A7, A8, A9) follow by an Acinobacter sp. were used for biological treatment of CL for ten days. Then, digestion activity and some rumen fermentation parameters of ruminal bacteria and fungi in specific culture medium (SCM) were determined in Arabian sheep. Treated CL by bacterial isolates, reduced total tannin (TT) compared with unprocessed groups (P &lt; 0.05). Dry matter disappearance (DMD) was affected by biological treatment for both rumen bacteria and fungi in different incubation times, except for 3 and 6 h of incubation in SCM of mixed rumen fungi, which indicated same pattern with untreated CON group (P &gt; 0.05). The NDF disappearance (NDFD) and CP disappearance (CPD) in SCM of mixed rumen bacteria and fungi were increased during all incubation times (P &lt; 0.05) due to biological prosses of CL compared to CON treatment. Biologically treated CL increased gas production from the fermentable fraction (b) pattern in both SCM of mixed rumen bacteria and fungi during all incubation times (P &lt; 0.05), but, the gas production rate constant (c) was increased only at SCM of mixed rumen fungi (P &lt; 0.05). Overall, the data indicated that bacterial inoculation of K. p and A. b. sp. could improve digestion activity and some fermentation parameters of rumen bacteria and fungi in Arabian sheep.

Effects of tannins on population dynamics of sympatric seed-eating rodents: the potential role of gut tannin-degrading bacteria.

Chemical compounds in seeds exert negative and even lethal effects on seed-consuming animals. Tannin-degrading bacteria in the guts of small mammals have been associated with the ability to digest seeds high in tannins. At the population level, it is not known if tannins influence rodent species differently according to the composition of their gut microbiota. Here, we test the hypothesis that sympatric tree species with different tannins exert contrasting effects on population fluctuations of seed-eating rodents. We collected a 10-year dataset of seed crops and rodent population sizes and sequenced 16S rRNA of gut microbes. The abundance of Apodemus peninsulae was not correlated with seed crop of either high-tannin Quercus mongolica or low-tannin Corylus mandshurica, but positively correlated with their total seed crops. Abundance of Tamias sibiricus was negatively correlated with seed crop of Q. mongolica but positively correlated with C. mandshurica. Body masses of A. peninsulae and T. sibiricus decreased when given high-tannin food; however, only the survival of T. sibiricus was reduced. The abundance of microbial genus Lactobacillus exhibiting potential tannin-degrading activity was significantly higher in A. peninsulae than in T. sibiricus. Our results suggest that masting tree species with different tannin concentrations may differentially influence population fluctuations of seed predators hosting different gut microbial communities. Although the conclusion is based on just correlational analysis of a short time-series, seeds with different chemical composition may influence rodent populations differently. Future work should examine these questions further to understand the complex interactions among seeds, gut microbes, and animal populations.

Effect of feeding tannin-degrading bacteria Streptococcus gallolyticus strain TDGB 406 on meat quality of goats fed with Quercus semicarpifolia leaves.

The effect of feeding tannin-degrading bacteria (Streptococcus gallolyticus strain TDGB 406) on carcass characteristics of goats fed with oak (Quercus semicarpifolia) leaves was studied on 18 male goats (4months old, average body weight 9.50 ± 1.50kg), distributed into three groups of six animals each. The animals of group 1 served as control, while the animals of groups 2 and 3 were given (at 5ml/kg live weight) autoclaved and live culture of isolate TDGB 406 (10(6)cells/ml), respectively. The animals were fed with oak leaves as a basal roughage source and maize hay along with fixed quantity of concentrate mixture. After 4months of feeding, the animals were slaughtered for carcass studies. The feeding of live culture of isolate TDGB 406 did not cause any effect (P > 0.05) on pre-slaughter weight, empty body weight, carcass weight, dressing percent, and yield of wholesale cuts (neck, rack, shoulder, breast, shank, loin, leg, and flank) of the goat meat. The chemical composition of longissimus dorsi muscle was comparable (P > 0.05) among the groups. The organoleptic evaluation of pressure-cooked meat in terms of tenderness and overall palatability was increased significantly (P < 0.05) in the meat of group 3 where live culture was supplemented. The other attributes were similar among the groups. It was concluded that supplementation of tannin-degrading bacteria S. gallolyticus strain TDGB 406 to goats fed with oak leaves did not affect the carcass characteristics and meat quality.

Tannin-degrading bacteria with cellulase activity isolated from the cecum of the Qinghai-Tibet plateau zokor (Myospalax baileyi)

Tannins, which are polyphenols present in various plants, have anti-nutritional activity; however, their negative effects are mitigated by the presence of tannin-degrading microorganisms in the gastrointestinal tract of animals. This has never been investigated in the plateau zokor (Myospalax baileyi) – the predominant small herbivore in the alpine meadow ecosystem of Qinghai Province, China – which consumes tannin-rich herbaceous plants. Tannase activity in the feces of the plateau zokor increased from June to August corresponding to the increase in hydrolyzable tannin concentrations in plants during this period, and three tannin-degrading facultative anaerobic strains (designated as E1, E2, and E3) were isolated from the cecum of these animals. Sequencing of the 16S rDNA gene identified isolates of strain E1 as belonging to the genusEnterococcus, and E2 and E3 to the genusBacillus. All of the bacteria had cellulose-degrading capacity. This study provides the first evidence of symbiotic bacterial strains that degrade tannic acid and cellulose in the cecum of plateau zokor.

Inoculation of tannin-degrading bacteria into novel hosts increases performance on tannin-rich diets.

It has been hypothesized that herbivores host tannin-degrading bacteria (TDB) to overcome the toxic challenges posed by plant tannins. While TDB have been isolated from the guts of numerous mammals, their functional significance to their hosts has never been explicitly tested. We introduced TDB into lab rats, which do not host TDB, and measured host performance on tannin-rich diets. We first isolated three species of TDB, Escherichia coli, Bacillus subtilis and Enterococcus faecalis, from the guts of the desert woodrat (Neotoma lepida), which regularly feeds on tannin-rich plants. Then, we inoculated isolated TDB, as well as full woodrat microbial communities into laboratory rats. A control group was inoculated with sterilized woodrat faeces. Recipient lab rats were fed increasing concentrations of tannic acid, and we monitored tannic acid intake, body mass and liver damage as measured by serum alanine aminotransferase activity. Lab rats given TDB as isolates or full communities exhibited increased tannic acid intake, higher maintenance of body mass and lower indicators of liver damage compared with control animals. These differences were maintained when the trial was repeated after 6 weeks of feeding on tannin-free diets. Our results are the first to demonstrate that TDB significantly increase host performance on tannin-rich diets.

Isolation and Characterization of Tannin-Degrading Bacteria from the Rumen of Goats Fed Oak (Quercus semicarpifolia) Leaves

Tannin-degrading bacteria (TDGB) were isolated from the rumen of goats fed oak leaves containing diet. The isolates were screened for their ability to increase in vitro digestibility of oak leaves (Quercus semicarpifolia), and fifteen best isolates were selected for further characterisation. The isolates TDGB 19, 20, 406, 415, 417, 420, 425, 437, 446, and 450 were Gram-positive cocci with medium- and long-chains, and isolates TDGB 409, 428, 430, and 433 were Gram-positive cocco-bacilli of short chain length. All the isolates exhibited tannase activity with the maximum in TDGB 417 (72.0 units; nmol methyl gallate degraded/min/ml) and minimum in TDGB 433 (28.92 units). All the fifteen isolates utilized fructose, galactose, lactose, starch, maltose, glycerol, and mannose as carbon source. The isolates were tested for their tolerance to phenolic monomers like ferulic acid, vanillic acid, coumaric acid, pyrogallol acid, and gallic acid. All the isolates could tolerate the presence of pyrogallol and gallic acid up to 20 mM, but ferulic was toxic at 20 mM level for all the isolates. The isolates were able to degrade tannic acid to pyrogallol except isolates TDGB 409, 428, 430, and 433, which could degrade tannic acid only up to gallic acid after 24 h of incubation as assessed by thin-layer chromatography. None of the isolates degraded tannic acid to resorcinol even after 96 h of incubation. The phylogenetic analysis of the isolates using sequences of PCR amplicons generated by the three sets of primers, two primers sets targeting 16S rRNA gene and one targeting sodA gene for superoxide dismutase enzyme, revealed that the isolates TDGB 7, 19, 20, 406, 417, 446, and 450 formed a close cluster with Streptococcus gallolyticus with 99 to 100 % similarity in all the three phylogenetic trees. There was a maximum of 11 % improvement in vitro true degradability of tannin-rich feed in the presence of live culture of TDGB 406 and can be exploited as a probiotic to improve nutrient utilization of tannin-containing feed given to the livestock.

Effects of phenolic compounds in three oak species on in vitro gas production using inoculums of two breeds of indigenous Iranian goats

The capabilities of rumen fluid (RF) from two indigenous Iranian goats (i.e., Markhoz, Alamout) to ferment tanniniferous (i.e., oak leaves) and non-tanniniferous (i.e., a mixture of lucerne hay and barley grain) feeds were studied. Five air dried samples from each of three oak species (i.e., Quercus persica, Quercus infectoria, and Quercus libani) were used to determine their chemical composition, phenolic compounds and protein precipitable phenolics (PPP). In three runs of in vitro gas production, the influence of non-tannin containing substrate incubated with either RF (i.e., Markhoz or Alamout) was assessed. Gas production (GP) was recorded from 1 to 96h of incubation. Effects of tannins on in vitro gas production, in vitro organic matter disappearance (IVOMD) and estimated metabolisable energy (ME) were assessed by a polyethylene glycol (PEG) tannin bioassay. There were no differences in gas production and IVOMD for a non-tannin substrate incubated with RF from either breed of goat. However, RF from Markhoz goats produced consistently more gas than RF from Alamout goats among oak species. The PEG treatment improved (P<0.01) the IVOMD in RF and estimated ME from both breeds of goats for all oak species. The RF from Markhoz goats fermented oak leaves more effectively than RF from Alamout goats, possibly due to the presence of tannin-degrading bacteria.

Two-phase olive mill waste composting: Community dynamics and functional role of the resident microbiota

In this study, physico-chemical modifications and community dynamics and functional role of the resident microbiota during composting of humid husk from a two-phase extraction system (TPOMW) were investigated. High mineralization and humification of carbon, low loss of nitrogen and complete degradation of polyphenols led to the waste biotransformation into a high-quality compost. Viable cell counts and denaturing gradient gel electrophoresis (DGGE) profiling of the 16S rRNA genes showed that the thermophilic phase was characterized by the strongest variations of cell number, the highest biodiversity and the most variable community profiles. The isolation of tannin-degrading bacteria (e.g. Lysinibacillus fusiformis, Kocuria palustris, Tetrathiobacter kashmirensis and Rhodococcus rhodochrous) suggested a role of this enzymatic activity during the process. Taken together, the results indicated that the composting process, particularly the thermophilic phase, was characterized by a rapid succession of specialized bacterial populations with key roles in the organic matter biotransformation.

Phylogenetic and functional characterization of tannin-degrading bacteria isolated during composting of olive mill waste

Phylogenetic and functional characterization of tannin-degrading bacteria isolated during composting of olive mill waste

Isolation of tannin-degrading bacteria isolated from feces of the Japanese large wood mouse, Apodemus speciosus, feeding on tannin-rich acorns

Bacteria with tannase activity were isolated from the feces of the Japanese large wood mouse, Apodemus speciosus. They were largely classified into two groups: Gram-positive cocci and Gram-positive bacilli. Genotypic analysis using a species-specific PCR assay as well as biochemical tests identified all cocci as Streptococcus gallolyticus. A PCR assay targeting a genus-specific sequence in the 16S/23S rDNA spacer region and additional 16S rDNA sequencing indicated that the bacilli belong to the genus Lactobacillus, with L. animalis and L. murinus being closely related taxa. Subsequent estimation of guanine-plus-cytosine content, amplified ribosomal DNA restriction analysis, and DNA/DNA hybridization assay confirmed that the bacilli are homologous to each other but different from L. animalis or L. murinus. Consequently, a novel species of the genus Lactobacillus may be proposed. To date, this study is the first to report on the isolation of tannase-positive bacteria from the feces of a rodent species. These bacteria may play an essential role for the host organism in digesting tannin-rich acorns available in their natural habitats, thereby endowing them with a greater ecological advantage.

Isolation and characterization of tannin-degrading bacteria from faecal samples of some wild ruminants in Ethiopia

The aim of this study was to isolate and characterize bacteria capable of hydrolysing tannins from the faeces of Ethiopian ruminants adapted to feed on tannin-rich leaves. Faecal samples were collected from dikdik ( Madoqua guentheri), camel ( Camelus dromedaries), Grant's gazelle ( Gazella granti), zebra ( Equus quagga) and hartebeest ( Alcelaphus buselaphus). Mixed cultures of the samples were screened for their tannin-hydrolysing capacities by thin layer chromatography (TLC). Twelve bacterial isolates with the ability to hydrolyse tannin were characterized morphologically, physiologically and by restriction fragment length polymorphism (RFLP) of the 16S rRNA genes of the isolates. Isolate Eden Ephraim 9 (EE9) completely degraded tannic acid, gallic acid and chestnut tannin. The other isolates could hydrolyse tannin to pyrogallol. Characterization of EE9 shows that the isolate is different from previously isolated tannin-tolerant bacteria. The bacteria isolated from some Ethiopian ruminants could partially or completely hydrolyse tannins and EE9 is possibly a new isolate, which may warrant further taxonomic studies. This study has shown the presence of bacteria in faecal materials of ruminants that can completely degrade tannin and which may be used to alleviate tannin toxicity problems in ruminants feeding on tannin-rich but nutritious fodder tree leaves.