Researchers prepared five isocaloric and isonitrogenous total mixed rations for an in vitro gas-production assay using rumen fluid collected from three slaughtered Holstein cows. Each diet contained 50% clover hay and 50% concentrate, with yellow corn progressively replaced by fungi-degraded date pits at 0%, 25%, 50%, 75% or 100%. Six analytical bottles were prepared per treatment: three for fermentation and protozoal measurements and three for estimating truly degraded dry and organic matter.

The statistical analysis used three inoculum replicates, and the experiment was conducted in a single run. To produce the experimental ingredient, the date pits were washed, dried at 80 °C for eight hours and ground to pass a 0.6 mm sieve. Approximately 25 kg of the powder was then inoculated with A. parasiticus during an aerobic solid-state fermentation carried out in autoclavable bags for two weeks.

The fermentation medium was adjusted to pH 5.5 and supplemented with lactose, with continuous aeration, agitation and cooling during incubation. After fermentation, the colonized material was autoclaved twice and exposed to ultraviolet radiation for one hour before the bags were opened, a biosafety measure intended to reduce viable fungal spores and limit environmental dispersal during sample handling. Safety was assessed by measuring four aflatoxins—AFB1, AFB2, AFG1 and AFG2—with an enzyme-linked immunosorbent assay.

The researchers reported a total aflatoxin concentration of 0.72 ppm, below the maximum limit they cited for animal feed and indicating no apparent safety concern in the tested material. That result applies to the particular fermented sample analyzed; it does not establish that every batch produced through the same process would have identical contamination levels. During the 24-hour incubation at 39 °C, cumulative gas production declined from 177.24 mL per gram of dry matter in the all-corn control to 153.09 mL per gram when fungi-degraded date pits completely replaced corn.

Truly degraded dry matter fell from 760.10 to 695.00 g/kg, while truly degraded organic matter decreased from 769.98 to 703.17 g/kg. Statistical tests identified both linear and quadratic responses for these three measures, each with p = 0.001, indicating systematic changes across the replacement series rather than isolated differences between adjacent diets. Despite the lower overall gas production, methane production did not differ significantly among the diets: values ranged from 18.92 to 19.84 mL per gram of degraded organic matter.

Rumen pH moved in the opposite direction, rising from 6.32 in the control to 6.49 in the diet containing only fungi-degraded date pits. The increase was significant for both the linear trend, with p = 0.012, and the quadratic trend, with p = 0.032. The authors associated this response with the date-pit diets’ greater fiber content and potentially lower fermentable-carbon availability relative to corn.

They also noted that sugars can be incorporated into microbial glycogen, which may limit the production of fermentation acids. By contrast, the partitioning factor, total protozoa and ammonia nitrogen did not differ significantly among diets. The partitioning factor, expressed as milligrams of truly degraded organic matter per milliliter of gas production, ranged from 3.70 to 4.02 mg/mL.

The main short-chain fatty acids also remained stable. Acetic acid accounted for 42.92% to 43.82% of the measured molar proportions across the diets, propionic acid for 24.26% to 24.91%, and n-butyric acid for 19.45% to 20.06%. None of these major fatty acids showed significant treatment effects.

Heptanoic acid was the exception: its molar proportion increased linearly as fungi-degraded date pits replaced corn, with p = 0.029, while the quadratic tendency did not meet the conventional significance threshold. Thus, the largest observed differences in rumen chemistry were concentrated in the extent of substrate degradation and gas generation rather than in the balance of the principal volatile fatty acids used to characterize fermentation. Molecular analysis offered only a limited view of the methane-producing community.

The researchers extracted DNA from rumen samples, used nested polymerase chain reaction with methanogen-specific primers and sequenced products approximately 179 base pairs long. Phylogenetic analysis placed 11 operational taxonomic units in a major cluster associated with Methanobrevibacter, including sequences from several date-pit replacement levels. One subcluster contained nine units with bootstrap support ranging from 39% to 67%, while another included two units supported at 58%.

The authors explicitly cautioned that these results described phylogenetic affiliations among detected sequences rather than a comprehensive characterization of the entire rumen methanogenic community. The modest bootstrap values also limit the strength of the inferred evolutionary clustering. There was no notable association between the phylogenetic grouping and methane production across the diets.

The researchers noted that this agrees with the absence of a clear relationship between methanogen abundance and methane emissions reported previously, but the current experiment neither established such a relationship nor ruled one out. More broadly, the findings show that fungi-degraded date pits can replace yellow corn in a short-term laboratory rumen system without detectable disruption of several major fermentation indicators. They do not show that the ingredient improves ruminant productivity, lowers greenhouse-gas emissions at farm level or provides an economic advantage.

Feed costs, palatability, intake, growth, milk yield, carcass traits and animal health were outside the scope of the study. For the authors, the results support further investigation of fungi-degraded date pits as a locally available energy source that could reduce reliance on conventional concentrate in ruminant diets. The ingredient offers a route to adding value to a date-processing by-product, and the absence of significant changes in methane output and principal fermentation measures provides a rationale for in vivo testing.

Nevertheless, the reduced gas production and nutrient degradability at higher replacement levels remain important constraints.

Subject of Research: In vitro assessment of fungi-degraded date pits as a corn replacement for ruminant diets

Article Title: Efficacy of fungi-degraded date pits as a potential alternative dietary energy source in farm animal diets

Article References: Sallam, S. M. A., Attia, M. F. A., El-Nile, A. E., El-Azab, M., Shehab El-Deen, M. A., Aladhadh, M., Ghareeb, A. Z., Hafez, E. E., Salama, A. M. M., & Ghareeb, D. A. (2026). Efficacy of fungi-degraded date pits as a potential alternative dietary energy source in farm animal diets. Frontiers in Veterinary Science, 13, Article 1892086. https://doi.org/10.3389/fvets.2026.1892086

Image Credits: AI Generated

DOI: 10.3389/fvets.2026.1892086

Keywords: ruminant nutrition, date pits, corn replacement, rumen fermentation, methanogens, fungal fermentation, short-chain fatty acids, livestock feed, agricultural by-products, in vitro study, Efficacy, fungi-degraded

The core problem the review identifies is what researchers call the breeder’s dilemma. Conventional forage breeding has historically prioritized yield, stress tolerance, and pest resistance, while traits that matter enormously to ruminant nutrition, such as digestibility, crude protein content, and the reduction of anti-nutritional compounds, have received far less attention. The result is a generation of improved cultivars that perform beautifully in agronomic trials yet fail to deliver meaningful gains in animal productivity. Improved cereal and forage varieties often produce more biomass but retain high lignin or phytate levels that limit how much energy and protein livestock can actually extract from them.

The consequences of this mismatch are most visible during the dry season. Studies from Tanzania and Ethiopia cited in the review show that crude protein and metabolizable energy levels in available forages can drop by 20 to 28 percent when the rains fail, driving milk production down by as much as 40 percent. In Nigeria, ruminant systems face acute forage scarcity during dry months, forcing farmers to rely on low-nutrient crop residues or expensive commercial supplements. These seasonal feed gaps are compounded by drought-induced pasture loss, livestock mortality, and declining soil fertility, undermining household food security and the long-term viability of mixed farms across the tropics.

Forages are not a minor input in these systems. Grasses and legumes supply more than 90 percent of the feed energy consumed by herbivorous livestock worldwide, effectively converting non-human-edible biomass into high-quality animal protein. The review catalogs the workhorses of tropical pasture science: Brachiaria grasses such as the Marandu, Piata, and Xaraes cultivars, prized for drought tolerance and persistence; Guinea grass, which offers high biomass but demands fertile soils; and Napier grass, a staple of cut-and-carry systems despite its vulnerability to stunt and smut diseases. On the legume side, species like Stylosanthes, Leucaena leucocephala, Desmodium, and Lablab purpureus boost crude protein in forage mixtures and fix nitrogen biologically, enriching the soil in the process.

The nutritional chemistry of these plants tells a striking story. Many tropical grasses, including Urochloa brizantha and the Urochloa hybrid Cayman, deliver crude protein concentrations of just 68 to 77 grams per kilogram of dry matter, below the threshold needed for efficient rumen microbial function. Legumes such as Leucaena diversifolia and Canavalia brasiliensis reach 195 to 256 grams per kilogram, while also contributing more metabolizable energy and lowering neutral detergent fiber and acid detergent lignin, both of which improve digestibility. Mineral deficits in calcium, phosphorus, and magnesium are common in dry-season tropical grasses and must be corrected through supplementation or by integrating mineral-rich legumes into pastures.

But legumes come with their own chemical complications. Leucaena diversifolia can carry condensed tannin concentrations of up to 47 grams per kilogram of dry matter, along with phenolics that bind proteins and reduce digestibility. The review notes that moderate tannin levels can actually be beneficial, slowing ruminal protein degradation and reducing nitrogen losses, but excessive amounts depress intake and nutrient absorption. Palatability adds another layer of complexity: birdsfoot trefoil, for example, is often less palatable than lucerne or white clover, and rapidly degradable legume proteins can cause ruminal ammonia accumulation and environmental nitrogen losses if not balanced with sufficient dietary energy.

On the resilience side, the review details the physiological traits that separate drought survivors from casualties. Deep and dense root systems allow plants like cocksfoot and tall fescue to tap subsoil moisture; deep-rooting tall fescue cultivars such as Flecha and Centurion demonstrate improved survival by accessing deeper water reserves. Osmotic adjustment, the accumulation of compatible solutes like proline, glycine betaine, and trehalose, maintains cell turgor and stabilizes membranes and proteins during water stress. Transpiration efficiency matters too, and C4 grasses including several Brachiaria cultivars hold an inherent advantage thanks to their photosynthetic pathway and reduced stomatal conductance. The cocksfoot cultivar Kasbah takes an even more dramatic approach, entering complete summer dormancy to survive prolonged drought, though at the cost of relatively low water-use efficiency during active growth.

The breeding toolkit itself is evolving, though unevenly. Conventional methods remain the cornerstone: mass selection, recurrent selection, and hybridization have produced landmark cultivars such as Marandu in Brazil, which combined high biomass yield with adaptation to acidic soils. Yet these approaches are slow, typically requiring 10 to 15 years to release a new cultivar, and apomictic grasses that propagate clonally through seeds complicate genetic recombination. Modern molecular tools promise acceleration but face real barriers. Marker-assisted selection has underperformed in forages because traits like digestibility and drought tolerance are polygenic and expensive to screen for. Genomic selection and genome-wide association studies have transformed maize, wheat, and dairy cattle breeding but lag in forage crops, where large training populations are required and commercial investment is thin. CRISPR gene editing offers tantalizing prospects for tailoring lignin and tannin levels, but regulatory restrictions and public acceptance remain significant hurdles.

Perhaps the most compelling section of the review addresses what happens when the two disciplines genuinely collaborate. Australia’s EverCrop initiative provides a working model, integrating crop improvement with animal performance trials from the design stage of breeding programs. Feed evaluation trials, including in vitro digestibility assays and animal model studies measuring intake, growth, reproduction, and feed efficiency, should become integral to selection pipelines rather than afterthoughts. Case examples show the payoff: forage sorghum delivers both biomass and energy density for dry-season feeding, while Desmodium intortum grown with Napier grass and Brachiaria improves crude protein, metabolizable energy, feed intake, and animal productivity while simultaneously enriching soil through nitrogen fixation. Trials with different ryegrass cultivars similarly demonstrate that evaluating plant and animal outcomes together identifies genuinely superior forages.

The review is candid about the obstacles that go beyond the laboratory. Adoption of improved forages in sub-Saharan Africa has been hampered by weak seed markets, high establishment costs, and limited extension services, in sharp contrast to Brazil, where strong seed systems and government-private sector partnerships enabled large-scale uptake of Brachiaria cultivars and measurable gains in beef productivity. Genetic trade-offs also persist: breeding for drought tolerance often raises lignin content or delays maturity, both of which reduce digestibility, while selecting for high protein or reduced fiber can compromise persistence and pest resistance. The authors call for holistic breeding programs with embedded nutritional testing, integrated forage innovation hubs connecting research institutions, seed companies, and extension networks, and policies that fund integrative research and guarantee seed availability. They also acknowledge the limitations of their own evidence base, noting that most cultivar evaluations remain confined to controlled experiments and that socioeconomic factors such as labor availability, gender roles, and market incentives are poorly integrated into breeding priorities. The message is clear: resilient, nutritious forages exist on the drawing board, but turning them into farm-level transformation will require sustained investment, participatory research, and delivery systems that finally connect plant science to the animals and farmers who depend on it.

Subject of Research: Integrating plant breeding and animal nutrition to develop drought-tolerant, high-nutrient forage grasses and legumes for sustainable ruminant livestock production

Article Title: Synergizing plant breeding and animal nutrition: A review on the development of drought-tolerant and high-nutrient forage grasses and legumes

Article References: Adepoju, S. A., & Faniyi, T. O. (2025). Synergizing plant breeding and animal nutrition: A review on the development of drought-tolerant and high-nutrient forage grasses and legumes. Innovations in Agriculture, 8, 1-10. https://doi.org/10.3897/ia.2025.163461

Image Credits: AI Generated

DOI: 10.3897/ia.2025.163461

Keywords: forage breeding, drought tolerance, ruminant nutrition, forage legumes, forage grasses, crude protein, digestibility, sub-Saharan Africa, livestock productivity, seed systems, genomic selection, climate resilience

Pica is far more than an odd behavioral quirk. In sheep production systems it manifests as the persistent ingestion of non-feed substances: geophagia, trichophagia, urine drinking, fence licking, and even plastic chewing. Beyond disrupting normal feeding, it raises the risk of gastrointestinal injury, impaired nutrient absorption, parasitic infection, and lost productivity. Although the full etiology remains unresolved, researchers have long considered pica a multifactorial disorder rooted in nutritional imbalance, metabolic disturbance, and environmental stress, with mineral deficiency or imbalance widely recognized as a major driver. Shortages of elements such as phosphorus, copper, iron, zinc, and selenium can undermine antioxidant defenses and metabolic homeostasis, potentially pushing animals into the abnormal oral behaviors that define the syndrome.

Simple mineral supplementation has shown promise before—earlier work on wool-eating lambs found that dosing with iron, copper, and zinc sulfate salts raised blood mineral levels and produced complete recovery within eight weeks. But the picture is complicated by the fact that pica may also involve oxidative stress, endocrine dysregulation, and metabolic dysfunction. Moreover, whether supplemented minerals actually reach the bloodstream depends on their chemical form, antagonistic interactions among minerals, the rumen environment, and the intricacies of gastrointestinal absorption. This is where the new study departs from convention: instead of relying on minerals alone, the researchers combined them with Shariga-5, a traditional Mongolian multi-herb formula derived from local ethnoveterinary practice and rich in bioactive flavonoids and phenolic acids.

The Mongolian herbal–mineral supplement, or MHMS, was manufactured into solid 5-kilogram lick blocks containing target concentrations of calcium, phosphorus, copper, iron, manganese, zinc, selenium, and cobalt, along with vitamins D3 and E. Embedded within the mineral matrix were five medicinal herbs of the Mongolian Plateau: turmeric rhizome (Shariga), forsythia fruit (Saolong-Jimusi), clove flower buds (Gaoletu-Baori), chebulic myrobalan fruit (Arula), and gardenia fruit (Zhurula). Widely targeted metabolomic profiling of the individual herbs identified 1,427 metabolite annotation records, with phenolic acids accounting for nearly 39 percent and flavonoids another 32 percent—two chemical classes renowned for free-radical scavenging, metal-ion chelation, and anti-inflammatory activity. The design logic was synergy: trace elements supply the cofactors for antioxidant enzymes, while plant secondary metabolites provide complementary redox support.

To test the formula, the team screened East Friesian rams over seven consecutive days, enrolling only animals that showed five to ten instances of pica behaviors per observation day. Twenty rams, averaging about 1.5 years of age and 42.6 kilograms, were randomly split into two groups of ten, each housed in its own pen. The control group received only a basal diet of corn silage, oat hay, alfalfa, and Leymus chinensis hay; the treatment group received the same diet plus continuous free-choice access to an MHMS lick block. After a 14-day adaptation and a 60-day experimental period, blood was drawn before morning feeding, and serum from six animals per group underwent mineral, biochemical, antioxidant, hormonal, and untargeted metabolomic analyses.

The mineral results were strikingly selective. Serum copper roughly doubled, rising from 4.86 to 9.55 micromoles per liter, while zinc climbed from 10.62 to 15.29 and selenium from 0.43 to 0.59 micromoles per liter. Yet calcium, phosphorus, magnesium, sodium, potassium, iron, and manganese remained unchanged. This selectivity, the authors argue, suggests the supplement primarily restored the trace elements most closely tied to redox defense, metalloenzyme activity, immune regulation, and metabolic homeostasis. Copper underpins mitochondrial respiration and iron metabolism, zinc is a structural and catalytic factor for countless enzymes and supports epithelial barriers, and selenium is an essential component of glutathione peroxidase, a key peroxide-removing selenoprotein. The pattern also echoes previous work showing that mineral responses in ruminants are shaped not just by dose but by body reserves, absorption efficiency, and antagonistic interactions among elements.

The downstream physiological shifts extended well beyond minerals. Total antioxidant capacity in serum rose significantly, even though the individual enzyme activities of superoxide dismutase, glutathione peroxidase, and catalase did not change—an indication that the improvement came from the composite, largely non-enzymatic antioxidant pool, plausibly driven by the herbal polyphenols. Biochemical markers told a similar story of de-escalation: serum glucose fell from 4.44 millimoles per liter, sitting at the upper edge of the ovine reference range, to 3.60, a healthier mid-range value, and aspartate aminotransferase activity dropped from 160.70 to 125.35 units per liter, returning from above the normal range to within it. Because AST reflects hepatic metabolic burden and tissue injury—and pica-affected calves have previously shown elevated transaminases linked to copper deficiency—this normalization suggests the supplement eased tissue metabolic stress in the treated animals.

Perhaps the most dramatic changes appeared in the endocrine panel. Serum triiodothyronine, thyroxine, insulin-like growth factor 1, and growth hormone all rose significantly, signaling enhanced metabolic rate and activated anabolic growth signaling, while cortisol—the primary stress hormone of the hypothalamic–pituitary–adrenal axis—fell markedly, from 943 to 746 picograms per milliliter. The authors note that the supplement’s selenium, copper, zinc, and vitamin E all carry antioxidant functions that could plausibly dampen the HPA axis burden imposed by chronic nutritional stress and oxidative pressure. Previous studies lend support to the mechanism: curcumin supplementation has raised thyroid hormones and lowered cortisol in goats, and vitamin E and selenium combinations have reduced cortisol in transportation-stressed dairy heifers.

The untargeted metabolomics added an exploratory molecular layer. Out of 1,062 putatively annotated serum metabolites, 112 differed significantly between groups—51 higher and 61 lower in the supplemented rams. Pathway analysis ranked porphyrin metabolism, the biochemical route for heme synthesis, as the top candidate pathway, with glycine abundance more than doubled and porphobilinogen reduced to less than half of control levels. Because glycine is the starting substrate for heme biosynthesis and also a precursor for glutathione, the body’s master intracellular antioxidant, its rise fits a scenario of improved antioxidant substrate supply. The drop in porphobilinogen, an early heme precursor that can accumulate abnormally when the pathway falters, correlated positively with serum AST, hinting that reduced accumulation of this intermediate accompanied the easing of tissue metabolic pressure. The authors are careful to flag that this pathway enrichment did not survive multiple-testing correction and the metabolites lack validation against authentic standards, so these findings are hypothesis-generating rather than definitive.

The study also candidly acknowledges its limits: individual lick-block intake was not recorded, the composite design cannot separate the effects of minerals from herbs or their synergy, the sample was modest, and rumen fluid and microbiota were not examined. Even so, the overall picture is compelling—a traditional Mongolian herbal formula fused with trace-element nutrition appears to shift pica-affected rams from a stress-laden, oxidatively burdened metabolic state toward physiological balance, with elevated thyroid and growth hormones, subdued cortisol, stronger antioxidant capacity, and a metabolome inching back toward order. For a condition as old as animal husbandry itself, the remedy may lie partly in knowledge herders on the Mongolian Plateau have carried for generations, now given rigorous, molecule-by-molecule confirmation.

Subject of Research: Effects of a Mongolian herbal–mineral supplement on serum minerals, antioxidants, hormones, and metabolomics in rams with pica

Article Title: Free-choice Mongolian herbal-mineral supplementation improves serum mineral, antioxidant, endocrine, and metabolic indices in rams with pica

Article References: Man, D., Chen, L., Ao, X., Zhang, W., & Na, R. (2026). Free-choice Mongolian herbal-mineral supplementation improves serum mineral, antioxidant, endocrine, and metabolic indices in rams with pica. Frontiers in Veterinary Science, 13, Article 1946016. https://doi.org/10.3389/fvets.2026.1946016

Image Credits: AI Generated

DOI: 10.3389/fvets.2026.1946016

Keywords: pica, sheep, herbal-mineral supplement, trace minerals, antioxidant capacity, cortisol, thyroid hormones, serum metabolomics, porphyrin metabolism, Mongolian ethnoveterinary medicine, ruminant nutrition, oxidative stress

The research, led by Pawan Kumar Dash and supervised by Ganesan Kalipatty Nalliappan, focused on four chemical traits that animal nutritionists consider the cornerstones of fodder quality: crude protein, crude fiber, acid detergent fiber, and neutral detergent fiber. Crude protein is straightforwardly desirable, since ruminants need dietary protein for milk production and muscle growth. The fiber fractions are more nuanced. Neutral detergent fiber, often abbreviated as NDF, represents the total cell wall material of the plant, including hemicellulose, cellulose, and lignin, and it essentially governs how much feed an animal can physically consume; higher NDF generally means lower intake. Acid detergent fiber, or ADF, captures the less digestible components, chiefly cellulose and lignin, and is inversely related to how well the animal can digest the feed. Crude fiber, an older but still widely used measure, reflects the structural bulk of the plant tissue. In short, breeders want more protein and less of the indigestible fiber fractions, a combination that translates directly into more productive animals per hectare of fodder.

To find the genes behind these traits, the team employed one of the classic workhorses of quantitative genetics: a mapping population. They crossed two fodder maize lines with contrasting characteristics, DM 94 and African Tall, and then advanced the offspring to produce F2:3 families, generations in which each family carries a unique random mosaic of DNA segments inherited from the two parents. By measuring the quality traits in these families and simultaneously tracking genetic markers scattered across the maize genome, the researchers could detect chromosomal regions where marker inheritance consistently tracked with trait performance. These regions are the QTLs, stretches of DNA that harbor genes influencing the measured characteristic. Because traits like fiber content and protein content are controlled by many genes each contributing a modest effect, rather than by a single gene, this statistical approach is the only practical way to dissect their genetic architecture.

For the analysis itself, the team used Inclusive Composite Interval Mapping, or ICIM, a modern QTL mapping method implemented in software designed for quantitative genetics research. ICIM refines earlier mapping strategies by more effectively separating the effect of the QTL being tested from the background effects of other regions, which reduces the risk of false positives and improves the precision of the estimated positions. The method worked well: the study identified a total of nine QTLs across the four traits. One QTL was associated with crude protein, two with crude fiber, one with acid detergent fiber, and five with neutral detergent fiber. The abundance of QTLs for NDF is consistent with the biology of that trait, since neutral detergent fiber aggregates many cell wall components and is therefore likely to be influenced by a larger number of underlying genes.

What makes these QTLs particularly valuable to breeders is their effect size. All nine of the identified loci explained more than ten percent of the phenotypic variation in their respective traits, with the proportion of variation explained ranging from 14.9 percent to 19.7 percent, the latter recorded for a neutral detergent fiber QTL. In the world of quantitative genetics, where individual QTLs for complex traits often explain only a few percent of variation, loci contributing fifteen to twenty percent are considered major-effect regions. Such substantial effects mean that these DNA markers can serve as reliable signposts in marker-assisted selection, a breeding technique in which scientists screen seedlings for specific genetic markers rather than waiting years to observe the mature plant’s traits. A breeder can test a young plant’s DNA, know whether it carries the favorable allele at a major QTL, and make selection decisions long before the plant ever reaches the field or the feeding trough.

One of the most intriguing findings of the study was the co-localization of QTLs for crude fiber and neutral detergent fiber, meaning that loci influencing both traits were detected in the same chromosomal regions. This overlap was accompanied by a strong positive correlation between the two traits, which makes biological sense: crude fiber and neutral detergent fiber both measure components of the plant’s structural tissue, so genes that build more cell wall material naturally raise both measurements together. Co-localized QTLs of this kind are double-edged for breeders. On the one hand, a single marker in such a region could be used to select for or against both traits simultaneously, making breeding programs more efficient. On the other hand, the tight linkage means that selecting to reduce one fiber component will tend to reduce the other as well, and disentangling their effects may require breaking up the linked region through recombination or identifying additional independent loci.

The new QTLs are described by the authors as novel, adding to a growing but still incomplete map of the maize forage quality genome. Previous efforts by other research groups, and by some of the same authors in an earlier study published in Euphytica, have mapped QTLs for fiber and digestibility traits in various maize populations, including recombinant inbred lines derived from elite European and Chinese germplasm. Meta-analyses of silage quality QTLs have also compared mapped regions with the positions of candidate genes involved in cell wall biosynthesis and lignification. Each new population and each new cross adds pieces to this puzzle, because QTL detection depends on whether the two parents happen to carry different alleles at the relevant genes. The DM 94 by African Tall cross evidently uncovered variation that earlier populations did not, which is precisely why the authors highlight the novelty of their findings.

The practical implications extend well beyond the laboratory. Livestock productivity is a pressing concern in countries like India, where national statistics on animal husbandry document the enormous demand for quality fodder and where research institutes such as the Indian Grassland and Fodder Research Institute track persistent gaps between fodder supply and demand. Forage maize is among the most nutritious and widely cultivated non-legume forage crops globally, prized for its high biomass yield and energy content. Yet the digestibility of that biomass is limited by its cell wall composition, and improving it through conventional phenotypic selection is slow and expensive, since measuring fiber fractions requires laboratory analysis of harvested material. Marker-assisted breeding using the newly identified QTLs offers a faster and cheaper alternative: breeders can genotype plants at the seedling stage and assemble combinations of favorable alleles for higher protein and optimized fiber content without destructive sampling.

The research also connects to broader scientific questions about lignin and cell wall biology. Lignin, the polymer that gives plant tissues their rigidity, is a major determinant of forage digestibility, and it has been the target of intensive study both in agriculture and in biofuel research, where reducing lignin makes plant biomass easier to convert into fermentable sugars. Understanding which genomic regions control fiber deposition in maize stalks and leaves contributes to this wider effort, and the QTLs identified here may overlap with genes involved in monolignol biosynthesis and cell wall assembly. The authors acknowledge the support of the Department of Forage Crops and the Center of Excellence in Molecular Breeding at Tamil Nadu Agricultural University, facilities that enabled the combination of field phenotyping and molecular marker work the study required.

For now, the nine QTLs represent a toolkit rather than a finished product. The next steps for the research group and for breeders elsewhere will be to validate these loci in additional genetic backgrounds, develop flanking markers suitable for high-throughput screening, and test whether stacking favorable alleles across the QTLs produces measurable improvements in animal-relevant outcomes such as digestibility and milk yield. The authors note that the analysed data are included within the article and that raw datasets are available from the corresponding author on reasonable request, an openness that should accelerate follow-up work. If the promise of these major-effect loci holds up, the humble maize plant, already a global staple, may become an even more powerful engine of livestock production, with benefits flowing from a chromosome map to the milk pail and the meat supply.

Subject of Research: Mapping of quantitative trait loci for fodder quality traits in forage maize

Article Title: Identification of QTLs Responsible for Fodder Quality Traits in Fodder Maize (Zea mays L.)

Article References: Dash, P. K., Subramani, P., Nalliappan, G. K., Narayana, M., Sampathrajan, V., & Natesan, S. (2026). Identification of QTLs Responsible for Fodder Quality Traits in Fodder Maize (Zea mays L.). Indian Journal of Genetics and Plant Breeding, 86(1), 11-16. https://doi.org/10.1007/s44489-026-00010-7

Image Credits: AI Generated

DOI: 10.1007/s44489-026-00010-7

Keywords: fodder maize, QTL mapping, forage quality, crude protein, neutral detergent fiber, acid detergent fiber, crude fiber, marker-assisted selection, plant breeding, ruminant nutrition, quantitative genetics, Zea mays

The research team, led by Setiasih Setiasih and colleagues at Indonesia’s National Research and Innovation Agency (BRIN) and partner institutions, published their findings in Results in Chemistry. Their strategy centered on a single deceptively simple question: what exactly is inside rose myrtle leaves, and how does the answer change depending on how you extract it? To find out, they harvested leaves from an experimental garden in Riau Province, dried them at 40 degrees Celsius for three days, and ground them into a flour. That flour was then subjected to maceration with three solvents spanning the polarity spectrum: 96 percent ethanol as a polar solvent, ethyl acetate as a semipolar solvent, and n-hexane as a nonpolar solvent. Each extract was subsequently analyzed by liquid chromatography coupled to high-resolution mass spectrometry, or LC-HRMS, at BRIN’s Advanced Characterization Laboratories.

The choice of solvent turned out to matter enormously. LC-HRMS profiling tentatively identified 191 distinct compounds in the ethanol extract, 226 in the ethyl acetate extract, and 206 in the n-hexane extract. Ethanol preferentially pulled out polar molecules, yielding a profile dominated by flavonoid glycosides, phenolic acids and related derivatives, including myricitrin, myricetin, quinic acid derivatives and multiple dihydroxybenzoic acids. These hydroxyl-rich compounds dissolve readily in polar solvents through hydrogen bonding and are classic free-radical scavengers, which helps explain the high antioxidant activity long associated with the plant. The ethyl acetate extract displayed the greatest metabolite diversity of all, dominated by pentacyclic triterpenoids such as ursolic acid, asiatic acid, oleanolic acid, arjungenin and glycyrrhetinic acid derivatives, alongside phenolics including ellagic acid, gallic acid, myricetin and taxifolin. The n-hexane extract, by contrast, was rich in lipophilic sesquiterpenes such as caryophyllene oxide, alpha-farnesene and curcumene, together with assorted triterpenoid derivatives.

The analytical power of the method lies in its ability to go beyond a simple mass readout. Using higher-energy collisional dissociation fragmentation, the researchers could watch molecules break apart in characteristic ways, dramatically raising confidence in their identifications. Myricitrin, for example, produced a precursor ion at mass-to-charge ratio 463.0881 that fragmented to give a product ion at 316.0224, corresponding to the loss of its rhamnose sugar and formation of the myricetin aglycone. Oleanolic acid showed a precursor at 455.3534 with a diagnostic fragment at 411.3472 reflecting loss of carbon dioxide, while caryophyllene oxide in the n-hexane extract yielded sequential ring-cleavage fragments at 161.0962 and 109.1014. The authors are careful to stress that this was qualitative screening rather than quantitative metabolomics, conducted without biological replicates, so peak areas reflect relative signal intensity within a run rather than absolute concentrations. Even so, the picture that emerges is of a plant whose chemistry shifts meaningfully with every solvent a chemist chooses.

With the chemical inventory in hand, the team turned to biological testing. Using the agar disc diffusion method, they challenged four bacterial strains with each extract at concentrations of 25, 50 and 80 percent: Escherichia coli ATCC 25922 and Salmonella typhimurium ATCC 14028 as Gram-negative representatives, and Staphylococcus aureus ATCC 25923 and Listeria monocytogenes ATCC 19115 as Gram-positive representatives. The result was a clean split. Every extract, regardless of solvent, inhibited both Gram-positive species and failed entirely to inhibit either Gram-negative one. Chloramphenicol, the reference antibiotic, produced a 14.82 millimeter zone against S. aureus, larger than the 7.11 to 7.70 millimeter zones produced by the extracts, but the n-hexane extract at 80 percent actually outperformed chloramphenicol against Listeria, generating a 7.17 millimeter zone compared with the antibiotic’s 5.23 millimeters.

The selectivity is rooted in bacterial architecture. Gram-negative bacteria wrap themselves in an outer membrane studded with lipopolysaccharides, an exceptionally effective permeability barrier that excludes hydrophobic molecules and is reinforced by selective porins and efflux pumps that expel antimicrobials before they reach lethal intracellular concentrations. Gram-positive bacteria, lacking that outer membrane, rely on a thick but porous peptidoglycan layer some 20 to 80 nanometers deep, through which small plant metabolites can diffuse relatively freely toward the cytoplasmic membrane. The researchers also observed that solvent type shaped potency: ethanol extracts, rich in phenolics and flavonoids, were most effective against S. aureus, while n-hexane extracts, loaded with membrane-disrupting terpenoids, were the champions against Listeria, a reminder that even within the Gram-positive category, membrane composition and defensive machinery differ enough to matter. The findings align with earlier reports that rose myrtle extracts and the pure compound rhodomyrtone, an antibiotic candidate with activity comparable to vancomycin, are most effective against Gram-positive organisms.

The second half of the study addresses climate. Ruminant livestock account for roughly a quarter of anthropogenic greenhouse gas emissions measured by methane’s contribution, with enteric fermentation in the digestive tract responsible for 80 to 89.5 percent of that methane and manure contributing the rest. Methanogenic archaea in the rumen generate the gas, and feed additives that suppress them are among the most actively pursued mitigation strategies. Because flavonoids, tannins and saponins have documented anti-methanogenic effects, the team tested the ethyl acetate extract, the richest in bioactive triterpenoids and phenolics, in an in vitro rumen fermentation system using Brachiaria ruziziensis grass as the basal feed, at inclusion levels of 1 to 4 percent, with methane measured by gas chromatography over 48 hours.

The outcome was striking in one particular way. Total gas production was unaffected across all treatment levels, meaning fermentation efficiency was preserved, but methane output fell steadily as the extract dose increased. The methane percentage of total gas dropped from 7.72 percent in the untreated control to 3.77 percent at the 4 percent inclusion level, a statistically significant reduction, and methane volume declined as well with a P value of 0.056, right at the threshold of significance. The chemical fingerprint offers a plausible mechanism in several complementary parts. Triterpenoids such as ursolic acid are membrane-active against rumen protozoa, which maintain physical symbioses with methanogenic archaea and supply them with hydrogen; suppressing protozoa often drags methane down with it. Phenolic compounds can selectively inhibit methanogens while redirecting metabolic hydrogen toward propionate synthesis, an energetically favorable route for the animal. Terpenoids such as caryophyllene oxide may interfere with membrane integrity and ion gradients in anaerobic microbes. The authors also cite prior molecular docking work showing that rose myrtle compounds bind strongly to methyl coenzyme M reductase, the enzyme at the heart of methanogenesis, with rhodomyrtial A achieving a predicted binding affinity of minus 10.4 kilocalories per mole.

The researchers are appropriately measured about what these results do and do not prove. The antibacterial assay was a preliminary screen, and they recommend follow-up with minimum inhibitory and bactericidal concentration determinations, plus in vivo testing, to establish therapeutic potential; earlier work suggests possible veterinary applications including mastitis treatment in dairy cows. Similarly, they emphasize that the methane findings provide a chemical rationale rather than a demonstrated field benefit, and that in vivo feeding trials will be required to confirm efficacy and safety. Nevertheless, the study is the first LC-HRMS-based phytochemical characterization of rose myrtle in the context of ruminant nutrition, and it reveals an intertwined network of triterpenoids, phenolics, flavonoids and lipophilic terpenoids whose combined effects may prove more stable than any single isolated compound. In an era when agriculture is under intense pressure to cut emissions without compromising animal productivity, a shrub that grows wild across tropical Asia, fights Gram-positive pathogens in the laboratory, and measurably dampens methane in simulated rumen fluid is exactly the kind of natural lead worth chasing.

Subject of Research: Phytochemical characterization of Rhodomyrtus tomentosa leaves and their antibacterial and methane-mitigating bioactivities

Article Title: LC–HRMS-based phytochemical characterization and biological activities of Rhodomyrtus tomentosa leaves extracts: Antibacterial and methane mitigation potentials

Article References: Setiasih, S., Anggraeny, Y. N., Puspito, S., Widodo, S., Wardi, W., Prihandini, P. W., Antonius, A., Shiddieqy, M. I., Sasongko, W. T., Istiqomah, N., Haryanto, B., Kurniawati, A., Prihartini, I., Wulansari, W. I., & Ernawati, A. (2026). LC–HRMS-based phytochemical characterization and biological activities of Rhodomyrtus tomentosa leaves extracts: Antibacterial and methane mitigation potentials. Results in Chemistry, 30, Article 103812. https://doi.org/10.1016/j.rechem.2026.103812

Image Credits: AI Generated

DOI: 10.1016/j.rechem.2026.103812

Keywords: Rhodomyrtus tomentosa, rose myrtle, LC-HRMS, phytochemical profiling, antibacterial activity, triterpenoids, flavonoids, enteric methane, ruminant nutrition, feed additives, methanogens, Results in Chemistry