Conserving purple prairie clover (Dalea purpurea Vent.) as hay and silage had little effect on the efficacy of condensed tannins in modulating ruminal fermentation in vitro.

BACKGROUND: The objective of this study was to determine the effects of forage conservation method and condensed tannins (CT) in conserved forage on rumen fermentation. Purple prairie clover (PPC; Dalea purpurea Vent.) containing 84.5 g kg-1 dry matter (DM) of CT was harvested at full flower and preserved as freeze-dried green chop (F), hay (H) or silage (S). Batch culture incubations were conducted using conserved forages as a substrate and treatments were arranged as 3 × 2 factorial design of forage type (F, H and S) with or without polyethylene glycol (PEG). PEG was used to isolate the effects of CT on fermentation. Incubation was repeated twice with quadruplicate vials for each treatment in each incubation. 15 N-labelled ammonium sulfate was used as microbial N marker and headspace gas was sampled to determine methane production. RESULTS: Concentrations of neutral detergent fiber (NDF) and acid detergent fiber were lower (P < 0.01) in F than in H or S. Ensiling decreased (P < 0.001) total phenolics and extractable CT, but had no effect on total CT, whereas none of these phenolic fractions were altered in H. Hay and silage had lower (P < 0.01) true DM disappearance (TDMD) and NDF disappearance (NDFD) than F. Inclusion of PEG did not affect TDMD or NDFD after 8, 24 or 72 h of incubation. Productions of total gas, methane and total volatile fatty acid and the rate of gas production (c) were also similar, but ammonia was higher (P < 0.05) and microbial N was lower (P < 0.05) with than without PEG after 72 h of incubation. After 72 h, a lower (P < 0.001) proportion of acetate but higher (P < 0.05) proportion of propionate was noted with S, resulting in a lower (P < 0.001) acetate:propionate ratio as compared to F or H after 8 h of incubation. CONCLUSION: Condensed tannins in PPC decreased protein degradation in vitro, but had minimal effects on overall rumen fermentation, and conservation of PPC as hay or silage had little effect on the efficacy of CT in modulating rumen fermentation. © 2020 Her Majesty the Queen in Right of Canada. Journal of The Science of Food and Agriculture © 2020 Society of Chemical Industry.

Condensed Tannins Affect Bacterial and Fungal Microbiomes and Mycotoxin Production during Ensiling and upon Aerobic Exposure.

Purple prairie clover (PPC; Dalea purpurea Vent.) containing 84.5 g/kg dry matter (DM) of condensed tannin (CT) was ensiled without (control) or with polyethylene glycol (PEG) for 76 days, followed by 14 days of aerobic exposure. Changes in fermentation characteristics were determined, and the composition of bacterial and fungal communities were assessed using metagenomic sequencing. The addition of polyethylene glycol (PEG) that deactivated CT at ensiling increased (P < 0.05 to ∼0.001) soluble N, nonprotein N, lactic acid, total volatile fatty acids, ammonia N, deoxynivalenol (DON), and ochratoxin A (OTA) but decreased (P < 0.001) pH and water-soluble carbohydrates. The concentrations of DON and OTA increased (P < 0.001) for both silages, with the extent of increase being greater for control than for PEG-treated silage during aerobic exposure. The PEG-treated silage exhibited higher (P < 0.01 to ∼0.001) copy numbers of total bacteria, Lactobacillus, yeasts, and fungi than the control. The addition of PEG decreased (P < 0.01) bacterial diversity during both ensiling and aerobic exposure, whereas it increased (P < 0.05) fungal diversity during aerobic exposure. The addition of PEG at ensiling increased (P < 0.05) the abundances of Lactobacillus and Pediococcus species but decreased (P < 0.01) the abundances of Lactococcus and Leuconostoc species. Filamentous fungi were found in the microbiome at ensiling and after aerobic exposure, whereas Bacillus spp. were the dominate bacteria after aerobic exposure. In conclusion, CT decreased protein degradation and improved the aerobic stability of silage. These desirable outcomes likely reflect the ability of PPC CT to inhibit those microorganisms involved in lowering silage quality and in the production of mycotoxins.IMPORTANCE The present study reports the effects of condensed tannins on the complex microbial communities involved in ensiling and aerobic exposure of purple prairie clover. This study documents the ability of condensed tannins to lower mycotoxin production and the associated microbiome. Taxonomic bacterial community profiles were dominated by Lactobacillales after fermentation, with a notable increase in Bacillus spp. as a result of aerobic exposure. It is interesting to observe that condensed tannins decreased bacterial diversity during both ensiling and aerobic exposure but increased fungal diversity during aerobic exposure only. The present study indicates that the effects of condensed tannins on microbial communities lead to reduced lactic acid and total volatile fatty acid production, proteolysis, and mycotoxin concentration in the terminal silage and improved aerobic stability. Condensed tannins could be used as an additive to control unfavorable microbial development and maybe enhanced feed safety.

Molecular improvement of alfalfa for enhanced productivity and adaptability in a changing environment.

Due to an expanding world population and increased buying power, the demand for ruminant products such as meat and milk is expected to grow substantially in coming years, and high levels of forage crop production will therefore be a necessity. Unfortunately, urbanization of agricultural land, intensive agricultural practices, and climate change are all predicted to limit crop production in the future, which means that the development of forage cultivars with improved productivity and adaptability will be essential. Because alfalfa (Medicago sativa L.) is one of the most widely cultivated perennial forage crops, it has been the target of much research in this field. In this review, we discuss progress that has been made towards the improvement of productivity, abiotic stress tolerance, and nutrient-use efficiency, as well as disease and pest resistance, in alfalfa using biotechnological techniques. Furthermore, we consider possible future priorities and avenues for attaining further enhancements in this crop as a means of contributing to the realization of food security in a changing environment.

Fecal microbiota of lambs fed purple prairie clover (Dalea purpurea Vent.) and alfalfa (Medicago sativa).

The present study assessed the effect of purple prairie clover (PPC) and PPC condensed tannins (CT) on the fecal microbiota of lambs using high-throughput 16S rRNA gene pyrosequencing. A total of 18 individual lambs were randomly divided into three groups and fed either green chop alfalfa (Alf), a 40:60 (DM basis; Mix) mixture of Alf and PPC, or Mix supplemented with polyethylene glycol (Mix-P) for 18 days. Fecal samples were collected on days 13 through 18 using digital rectal retrieval. The DNA of fecal samples was extracted and the microbial 16S rRNA gene amplicons were sequenced using 454 pyrosequencing. Regardless of diet, the bacterial community was dominated by Firmicutes and Bacteroidetes with many sequences unclassified at the genus level. Forage type and CT had no effect on the fecal microbial composition at the phylum level or on α-diversity. Compared to the Alf diet, the Mix diet reduced the relative abundance of Akkermansia (P = 0.03) and Asteroleplasma (P = 0.05). Fecal microbial populations in Alf and Mix-P clustered separately from each other when assessed using unweighted UniFrac (P < 0.05). These results indicate that PPC CT up to 36 g/kg DM in the diet had no major effect on fecal microbial flora at the phyla level and exerted only minor effects on the genera composition of fecal microbiota in lambs.

Characterization of Condensed Tannins from Purple Prairie Clover (Dalea purpurea Vent.) Conserved as either Freeze-Dried Forage, Sun-Cured Hay or Silage.

Conservation methods have been shown to affect forage nutrient composition and value, but little information is available about the effect of forage conservation on plant condensed tannins (CT). The objective of this study was to assess the effects of conservation method on the concentration, chemical composition and biological activity of CT. Whole-plant purple prairie clover (PPC, Dalea purpurea Vent.) was harvested at full flower and conserved as freeze-dried forage (FD), hay (HAY) or silage (SIL). Concentration of CT in conserved PPC was determined by the butanol-HCl-acetone method. Structural composition, protein-precipitation capacity and anti-bacterial activity of CT isolated from conserved forage were determined by in situ thiolytic degradation followed by HPLC-MS analysis, a protein precipitation assay using bovine serum albumin and ribulose 1,5-disphosphate carboxylase as model proteins and by an Escherichia coli (E. coli) growth test, respectively. Conservation method had no effect on concentration of total CT, but ensiling decreased (p < 0.001) extractable CT and increased (p < 0.001) protein- and fiber-bound CT. In contrast, hay-making only increased (p < 0.01) protein-bound CT. Regardless of conservation method, epigallocatechin (EGC), catechin (C) and epicatechin (EC) were the major flavan-3-ol units, and gallocatechin (GC) was absent from both terminal and extension units of PPC CT. The SIL CT had the lowest (p < 0.001) EGC, but the highest (p < 0.01) EC in the extension units. Similarly, SIL CT exhibited a lower (p < 0.001) mean degree of polymerization (mDP), but higher (p < 0.001) procyanidins (PC) than FD or HAY CT. The protein-precipitating capacity of CT in conserved PPC ranked (p < 0.001) as FD > HAY > SIL. E. coli growth n M9 medium was inhibited by 25-100 µg/mL of CT isolated from FD, HAY and SIL (p < 0.05), but preservation method had no effect on the ability of CT to inhibit bacterial growth. The results demonstrated that ensiling decreased the extractability and protein-precipitating capacity of CT by increasing the proportions of PC. Purple prairie clover conserved as hay retained more biologically active CT than if it was conserved as silage.

Culture-negative endocarditis with multifocal spread and flail mitral valve leaflet: a case report.

Introduction: Blood culture-negative infective endocarditis is the condition in which a causative organism cannot be identified after inoculation of at least three samples using standard blood-culture systems for 7 days. It has a low reported incidence of about 2.5-31%. Causes may be infectious or non-infectious; use of prior antibiotic therapy is usually the leading factor. Case presentation: The authors present a case of true culture-negative endocarditis involving the mitral valve, with multiple foci of spread including brain, spleen, liver, and Intervertebral disc, which remained persistent despite treatment with intravenous broad-spectrum antibiotics on an inpatient and outpatient basis but eventually improved after upgrading alternative broad-spectrum antibiotic for an extended duration. The patient had complications in the form of a flail mitral valve with persistent mitral regurgitation, requiring mitra-clip placement. Discussion: Positive blood culture is one of the major diagnostic criteria to establish infective endocarditis. Patients may have persistent negative cultures due to previous antibiotic use, the presence of fastidious organisms, or the use of inappropriate techniques or media. Involvement of a multidisciplinary team, use of multimodal investigations, and appropriate antibiotic stewardship are crucial. Extended duration of treatment and upgrading antibiotics can be helpful next steps in highly suspicious cases. With multifocal spread as in our case, it further becomes challenging to control and treat the infection as it is frequently connected with higher morbidity and mortality. Conclusion: Blood culture-negative endocarditis is an entity that can present with early complications. It is diagnostically and therapeutically challenging to treat such patients. Multimodal approaches for early diagnosis and appropriate treatment are crucial owing to its high morbidity and mortality.

Liver injury: the therapeutic dilemma of homeopathy - a case report from Bangladesh.

Despite the lack of scientific evidence supporting its effectiveness, homeopathic treatment is increasingly being used as a form of alternative medicine, with many people taking homeopathic remedies instead of drug therapies. It is based on the principle of 'like cures like', meaning that a remedy similar to the illness can be used to treat it. However, there have been several reports suggesting the risks of homeopathic remedies, among which homeopathy-induced liver injury is widely discussed. Here, we report a case of a 35-year-old well-oriented male patient with a typical clinical presentation of liver injury as presented by yellowish discoloration of sclera and skin along with generalized body itching following the use of homeopathic medicine for musculoskeletal pain. Laboratory reports of increased liver markers along with bilirubin were also suggestive. Excluding other differentials like viral hepatitis, alcoholic hepatitis, hemochromatosis, Wilson disease, and standard drug and toxin-induced hepatitis, the recent use of homeopathic remedies was a contributing factor in leading to the diagnosis of homeopathy-induced liver injury. He was then treated with the discontinuation of homeopathic medicine and supportive care. This case highlights the need for public awareness of the possible complications such as headache, tiredness, skin eruption, dizziness, bowel dysfunction, allergic reactions to acute pancreatitis, renal failure, neurological dysfunction, possible liver injury, and even mortality in those patients who pursue homeopathic treatments and health care professionals should take this into account when making a differential diagnosis in patients with liver injury.

Diesel siphoning induced chemical pneumonitis: a case report.

Diesel fuel aspiration can occasionally result in an uncommon but potentially deadly condition called chemical pneumonitis. Case presentation: In this case study, a 16-year-old boy gave a history of diesel fuel siphoning from a motor vehicle tank, which brought him to our emergency room. He complained of coughing, breathing difficulties, and chest discomfort upon admission to the hospital. Patchy bilateral parenchymal lung opacities consistent with acute chemical pneumonitis were seen in radiological imaging tests. Treatment included supportive care, oxygen supplementation, and intravenous antibiotics. The patient's symptoms improved gradually throughout his hospitalization, and he was eventually discharged home with a good prognosis. Clinical discussion: Siphoning is a common practice in developing countries like Bangladesh. Workers at automobile transfer hydrocarbon products from one vehicle to other. However, its aspiration can cause a pneumonia like features and may wrongly misdiagnose. Diagnosis is made mainly on history taking. Conclusion: Physicians ought to know that patients exposed to diesel fuel may develop chemical pneumonitis, and they should consider this for an early diagnosis and effective treatment that can lead to favourable outcomes.

Elucidation of Physiological, Transcriptomic and Metabolomic Salinity Response Mechanisms in Medicago sativa.

Alfalfa (Medicago sativa L.) is a widely grown perennial leguminous forage crop with a number of positive attributes. However, despite its moderate ability to tolerate saline soils, which are increasing in prevalence worldwide, it suffers considerable yield declines under these growth conditions. While a general framework of the cascade of events involved in plant salinity response has been unraveled in recent years, many gaps remain in our understanding of the precise molecular mechanisms involved in this process, particularly in non-model yet economically important species such as alfalfa. Therefore, as a means of further elucidating salinity response mechanisms in this species, we carried out in-depth physiological assessments of M. sativa cv. Beaver, as well as transcriptomic and untargeted metabolomic evaluations of leaf tissues, following extended exposure to salinity (grown for 3-4 weeks under saline treatment) and control conditions. In addition to the substantial growth and photosynthetic reductions observed under salinity treatment, we identified 1233 significant differentially expressed genes between growth conditions, as well as 60 annotated differentially accumulated metabolites. Taken together, our results suggest that changes to cell membranes and walls, cuticular and/or epicuticular waxes, osmoprotectant levels, antioxidant-related metabolic pathways, and the expression of genes encoding ion transporters, protective proteins, and transcription factors are likely involved in alfalfa's salinity response process. Although some of these alterations may contribute to alfalfa's modest salinity resilience, it is feasible that several may be disadvantageous in this context and could therefore provide valuable targets for the further improvement of tolerance to this stress in the future.

Iodine deficiency and thyroid dysfunction: Current scenario in Nepal.

Iodine deficiency is a major cause of thyroid disorders worldwide. Nepal lies in the endemic area of iodine deficiency which was previously referred to as Himalayan goiter belt, with high prevalence of iodine deficiency disorders including goiter, cretinism and hypothyroidism. With effective implementation of universal salt iodization program, Nepal has a successful public health story to share having drastically reduced the iodine deficiency disorders. Moreover, challenge has appeared with rising number of excess iodine states. Thyroid dysfunction is growing higher and the increasing proportion of hyperthyroidism is particularly concerning. Time has come for us to suitably review the standards of salt iodization and control the increasing number of cases with thyroid dysfunction by the coordinated efforts of all stakeholders, along with sustaining the optimal level of iodine.

High-altitude illness: Menace in Himalayas of Nepal.

Nepal is a country of Himalayas including Mt Everest, the tallest mountain in the world, where a lot of people travel to high altitudes. Significant number of people develop high-altitude illness which includes acute mountain sickness (AMS), high altitude pulmonary edema (HAPE) and high-altitude cerebral edema (HACE) leading even to death. Addressing this issue demands more efforts on both the preventive and treatment aspects to decrease the sufferings of people in this modern time of advanced healthcare and technology.

In vitro ruminal fermentation of fenugreek (Trigonella foenum-graecum L.) produced less methane than that of alfalfa (Medicago sativa).

OBJECTIVE: The objective of this study was to compare fenugreek (FG) with alfalfa (Alf) in ruminal fermentation and methane (CH4) production in vitro. METHODS: Whole-plant FG harvested at 11- and 15-wk and Alf harvested at early and mid-bloom maturities, alone or as 50:50 mixture of FG and Alf at the respective maturity, were assessed in a series of 48-h in vitro batch culture incubations. Total fermentation gas and methane gas production, dry matter (DM) disappearance, volatile fatty acids, microbial protein and 16S RNA gene copy numbers of total bacteria and methanogens were determined. RESULTS: Compared to early bloom Alf, FG harvested at 11-wk exhibited higher (p<0.05) in vitro DM and neutral detergent fibre disappearance, but this difference was not observed between the mid-bloom Alf and 15-wk FG. Regardless plant maturity, in vitro ruminal fermentation of FG produced less (p<0.001) CH4 either on DM incubated or on DM disappeared basis than that of Alf during 48-h incubation. In vitro ruminal fermentation of FG yielded similar amount of total volatile fatty acids with higher (p<0.05) propionate percentage as compared to fermentation of Alf irrespective of plant maturity. Microbial protein synthesis was greater (p<0.001) with 11-wk FG than early bloom Alf as substrate and 16S RNA gene copies of total bacteria was higher (p<0.01) with 15-wk FG than mid-bloom Alf as substrate. Compared to mid-bloom Alf, 15-wk FG had lower (p<0.05 to 0.001) amount of 16S RNA methanogen gene copies in the whole culture during 48-h incubation. CONCLUSION: In comparison to Alf, FG emerges as a high quality forage that can not only improve rumen fermentation in vitro, but can also remarkably mitigate CH4 emissions likely due to being rich in saponins.

Effects of 3-nitrooxypropanol manure fertilizer on soil health and hydraulic properties.

Supplementing beef cattle with 3-nitrooxypropanol (3-NOP) decreases enteric methane production, but it is unknown if fertilizing soil with 3-NOP manure influences soil health. We measured soil health indicators 2 yr after manure application to a bromegrass (Bromus L.) and alfalfa (Medicago sativa L.) mixed crop. Treatments were: composted conventional manure (without supplements); stockpiled conventional manure; composted manure from cattle supplemented with 3-NOP; stockpiled 3-NOP manure; composted manure from cattle supplemented with 3-NOP and monensin (3-NOP+Mon), a supplement that improves digestion; stockpiled 3-NOP+Mon manure; inorganic fertilizer (150 kg N ha-1 and 50 kg P ha-1 ); and an unamended control. Select chemical (K+ , Mg2+ , Mn+ , Zn+ , pH, and Olsen-P), biological (soil organic matter, active C, respiration, and extractable protein), physical (wet aggregate stability, bulk density, total porosity, and macro-, meso-, and micro-porosity), and hydraulic (saturation, field capacity, wilting point, water holding capacity, and hydraulic conductivity) variables were measured. The inclusion of monensin decreased soil Zn+ concentrations by 70% in stockpiled 3-NOP+Mon compared with stockpiled conventional manure. Active C and protein in composted conventional manure were 37 and 92% higher compared with stockpiled manure, respectively, but did not vary between 3-NOP treatments. 3-Nitrooxypropanol did not significantly alter other soil health indicators. Our results suggest that composted and stockpiled 3-NOP manure can be used as a nutrient source for forage crops without requiring changes to current manure management because it has minimal influence on soil health.

Composition and Protein Precipitation Capacity of Condensed Tannins in Purple Prairie Clover (Dalea purpurea Vent.).

This study aimed to determine the concentration and composition of condensed tannins (CT) in different tissues of purple prairie clover (PPC; Dalea purpurea Vent.) at different maturities and to determine their protein-precipitating capacity. The compositions of CT were elucidated after thiolysis with benzyl mercaptan followed by high-performance liquid-chromatography (HPLC) and 1H-13C heteronuclear single quantum coherence (HSQC) NMR spectroscopy. The results indicated that PPC flowering heads contained the highest CT concentration. Purple prairie clover CT consisted mainly of epicatechin (EC) and epigallocatechin (EGC) subunits. CT in the leaves were composed of more EC and less EGC than CT in stems and flowering heads at both the early flowering (EF) and late flowering (LF) head stages. The mean degree of polymerization was the highest for CT in stems and increased with maturity. CT isolated from PPC leaves at the early flowering head stage exhibited the greatest biological activity in terms of protein precipitation. Overall, the CT in PPC were predominantly procyanidins and the concentration and composition varied among the plant tissues and with maturity.

The CRISPR/Cas9-Mediated Modulation of SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE 8 in Alfalfa Leads to Distinct Phenotypic Outcomes.

Alfalfa (Medicago sativa L.) is the most widely grown perennial leguminous forage and is an essential component of the livestock industry. Previously, the RNAi-mediated down-regulation of alfalfa SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE 8 (MsSPL8) was found to lead to increased branching, regrowth and biomass, as well as enhanced drought tolerance. In this study, we aimed to further characterize the function of MsSPL8 in alfalfa using CRISPR/Cas9-induced mutations in this gene. We successfully generated alfalfa genotypes with small insertions/deletions (indels) at the target site in up to three of four MsSPL8 alleles in the first generation. The efficiency of editing appeared to be tightly linked to the particular gRNA used. The resulting genotypes displayed consistent morphological alterations, even with the presence of up to two wild-type MsSPL8 alleles, including reduced leaf size and early flowering. Other phenotypic effects appeared to be dependent upon mutational dosage, with those plants with the highest number of mutated MsSPL8 alleles also exhibiting significant decreases in internode length, plant height, shoot and root biomass, and root length. Furthermore, MsSPL8 mutants displayed improvements in their ability to withstand water-deficit compared to empty vector control genotypes. Taken together, our findings suggest that allelic mutational dosage can elicit phenotypic gradients in alfalfa, and discrepancies may exist in terms of MsSPL8 function between alfalfa genotypes, growth conditions, or specific alleles. In addition, our results provide the foundation for further research exploring drought tolerance mechanisms in a forage crop.

Identification of Differential Drought Response Mechanisms in Medicago sativa subsp. sativa and falcata through Comparative Assessments at the Physiological, Biochemical, and Transcriptional Levels.

Alfalfa (Medicago sativa L.) is an extensively grown perennial forage legume, and although it is relatively drought tolerant, it consumes high amounts of water and depends upon irrigation in many regions. Given the progressive decline in water available for irrigation, as well as an escalation in climate change-related droughts, there is a critical need to develop alfalfa cultivars with improved drought resilience. M. sativa subsp. falcata is a close relative of the predominantly cultivated M. sativa subsp. sativa, and certain accessions have been demonstrated to exhibit superior performance under drought. As such, we endeavoured to carry out comparative physiological, biochemical, and transcriptomic evaluations of an as of yet unstudied drought-tolerant M. sativa subsp. falcata accession (PI 641381) and a relatively drought-susceptible M. sativa subsp. sativa cultivar (Beaver) to increase our understanding of the molecular mechanisms behind the enhanced ability of falcata to withstand water deficiency. Our findings indicate that unlike the small number of falcata genotypes assessed previously, falcata PI 641381 may exploit smaller, thicker leaves, as well as an increase in the baseline transcriptional levels of genes encoding particular transcription factors, protective proteins, and enzymes involved in the biosynthesis of stress-related compounds. These findings imply that different falcata accessions/genotypes may employ distinct drought response mechanisms, and the study provides a suite of candidate genes to facilitate the breeding of alfalfa with enhanced drought resilience in the future.

Potential effects of a high CO2 future on leguminous species.

Legumes provide an important source of food and feed due to their high protein levels and many health benefits, and also impart environmental and agronomic advantages as a consequence of their ability to fix nitrogen through their symbiotic relationship with rhizobia. As a result of our growing population, the demand for products derived from legumes will likely expand considerably in coming years. Since there is little scope for increasing production area, improving the productivity of such crops in the face of climate change will be essential. While a growing number of studies have assessed the effects of climate change on legume yield, there is a paucity of information regarding the direct impact of elevated CO2 concentration (e[CO2]) itself, which is a main driver of climate change and has a substantial physiological effect on plants. In this review, we discuss current knowledge regarding the influence of e[CO2] on the photosynthetic process, as well as biomass production, seed yield, quality, and stress tolerance in legumes, and examine how these responses differ from those observed in non-nodulating plants. Although these relationships are proving to be extremely complex, mounting evidence suggests that under limiting conditions, overall declines in many of these parameters could ensue. While further research will be required to unravel precise mechanisms underlying e[CO2] responses of legumes, it is clear that integrating such knowledge into legume breeding programs will be indispensable for achieving yield gains by harnessing the potential positive effects, and minimizing the detrimental impacts, of CO2 in the future.

Interrelationships of Fiber-Associated Anaerobic Fungi and Bacterial Communities in the Rumen of Bloated Cattle Grazing Alfalfa.

Frothy bloat is major digestive disorder of cattle grazing alfalfa pastures. Among the many factors identified to contribute to the development of frothy bloat, the disruption of rumen microbiota appears to be of central importance. Anaerobic rumen fungi (ARF) play an important role in sequential breakdown and fermentation of plant polysaccharides and promote the physical disruption of plant cell walls. In the present study, we investigated the dynamics of ARF during the development of alfalfa-induced frothy bloat and in response to bloat preventive treatments. By sequencing the internal transcribed spacer （ITS1）region of metagenomic DNA from the solid fraction of rumen contents, we were able to identify eight distinct genera of ARF, including Neocallimastix, Caecomyces, Orpinomyces, Piromyces, Cyllamyces, Anaeromyces, Buwchfawromyces, and unclassified Neocallimastigaceae. Overall, transition of steers from a baseline hay diet to alfalfa pastures was associated with drastic changes in the composition of the fungal community, but the overall composition of ARF did not differ (p > 0.05) among bloated and non-bloated steers. A correlation network analysis of the proportion of ARF and ruminal bacterial communities identified hub fungal species that were negatively correlated with several bacterial species, suggesting the presence of inter-kingdom competition among these rumen microorganisms. Interestingly, the number of negative correlations among ARF and bacteria decreased with frothy bloat, indicating a potential disruption of normal microbial profiles within a bloated rumen ecosystem. A better understanding of fungal-bacterial interactions that differ among bloated and non-bloated rumen ecosystem could advance our understanding of the etiology of frothy bloat.

Variation in diosgenin levels among 10 accessions of fenugreek seeds produced in western Canada.

A collection of 10 accessions of fenugreek (Trigonella foenum-graecum L.), an annual legume, was grown during two summers at three plot locations in western Canada to assess whether genetic (accession) and environmental factors (site and year of production) influenced levels of diosgenin, a steroidal sapogenin. The 60 harvested seed samples, each analyzed by single determinations on three subsamples of defatted and dried seed material, were hydrolyzed by a microscale procedure in water containing 2-propanol (70%) and sulfuric acid (1 M). The extracts were analyzed by capillary gas chromatography with 6-methyldiosgenin as internal standard. Diosgenin levels from mature seeds ranged from 0.28 to 0.92% (28-92 microg/10 mg). Analysis of variance on combined diosgenin levels from the three sites and two years revealed that accession, accession x year, and site x year effects were significant for diosgenin content, whereas site, year, and site x accession effects were not. Four accessions, CN 19062, CN 19067, CN 19070, and CN 19071, were identified with high levels of diosgenin on the basis of the 2-year data set. In these accessions, mean levels of diosgenin plus yamogenin from seven site years were estimated at 0.70, 0.98, 0.84, and 0.87%, respectively.

Biochemical monitoring in fenugreek to develop functional food and medicinal plant variants.

Many plants used as functional foods or for medicinal purposes have been criticized for their inconsistent physiological effects. Variation in genotype and environmental conditions under which plants are produced can contribute to this inconsistency in biochemical composition. Fenugreek (Trigonella foenum-graecum L.) is a medicinal plant that not only can lower blood glucose and cholesterol levels in animals, but also can be used as a forage crop for livestock feed. Seed content for the bioactive compounds diosgenin, galactomannan and 4-hydroxyisoleucine was characterized for ten fenugreek genotypes under rainfed and irrigated conditions. High and low seed yielding genotype/environment combinations were identified that possessed distinct biochemical and seed production traits. In general high seed yielding genotype/environment combinations exhibited a more stable biochemical composition and consisted largely of irrigated fenugreek. This research indicates that comprehensive biochemical analysis of plant products would facilitate the development of more reliable produce for use by the functional food/medicinal plant industry.