Lignin represents one of the most abundant plant-derived polymers. It is mostly present in the cell wall, and its primary role is to provide mechanical support to the plant. Chemical processes during wood-pulping yield diverse technical lignins with distinct characteristics. Due to their complex and variable nature, technical lignins are often undervalued and are mainly used as burning fuel in mills. However, various technical lignins have been shown to possess antimicrobial properties. Consequently, there is an increasing interest in understanding the properties and conditions that underlie their antimicrobial characteristics and how we can utilize them for practical applications. This review, for the first time, comprehensively summarized the antimicrobial activities of technical lignins and their potential antimicrobial applications.

Our objectives were to evaluate technical lignins for their antifungal properties against 3 molds and 1 yeast causing hay spoilage, and their ability to preserve ground high-moisture alfalfa hay nutritive value in vitro. In experiment 1, 8 technical lignins and propionic acid (PRP; positive control) were tested at a dose of 40 mg/mL. The experiment had a randomized complete block design (RCBD, 4 runs) and a factorial arrangement of 3 molds × 10 additives (ADV). The effects of the ADV on yeast were evaluated separately with a RCBD. Sodium lignosulfonate (NaL) and PRP were the only treatments with 100 ± 2.8% inhibition of fungi. In experiment 2, the minimum inhibitory concentration (MIC) for selected lignins and PRP were determined. At pH 4, NaL had the lowest MIC across the molds (20-33.3 mg/mL) and magnesium lignosulfonate (MgL) for the yeast (26.7) among the lignins. However, PRP had MIC values that were several-fold lower across all fungi (1.25-3.33). In experiment 3, a RCBD (5 blocks) with a 3 (ADV; NaL, MgL, and PRP) × 4 (doses: 0, 0.5, 1, and 3% wt/wt fresh basis) factorial arrangement of treatments was used to evaluate the preservative effects of ADV in ground high-moisture alfalfa hay inoculated with a mixture of the fungi previously tested and incubated under aerobic conditions in vitro. After 15 d, relative to untreated hay (14.9), dry matter (DM) losses were lessened by doses as low as 1% for NaL (3.39) and 0.5% for PRP (0.81 ± 0.77%). The mold count was reduced in both NaL at 3% (3.92) and PRP as low as 0.5% (3.94) relative to untreated hay (7.76 ± 0.55 log cfu/fresh g). Consequently, sugars were best preserved by NaL at 3% (10.1) and PRP as low as 0.5% (10.5) versus untreated (7.99 ± 0.283% DM), while keeping neutral detergent fiber values lower in NaL (45.9) and PRP-treated (45.1) hays at the same doses, respectively, relative to untreated (49.7 ± 0.66% DM). Hay DM digestibility was increased by doses as low as 3% for NaL (67.5), 1% MgL (67.0), and 0.5% PRP (68.5) versus untreated hay (61.8 ± 0.77%). The lowest doses increasing neutral detergent fiber digestibility relative to untreated hay (23.3) were 0.5% for MgL and PRP (30.5 and 30.1, respectively) and 1% for NaL (30.7 ± 1.09% DM). Across technical lignins, NaL showed the most promise as a potential hay preservative. However, its effects were limited compared with PRP at equivalent doses. Despite not having an effect on preservation, MgL improved DM digestibility by stimulating neutral detergent fiber digestibility. This study warrants further development of NaL under field conditions.

A meta-analysis was performed to evaluate the effects of chemical (50 articles) and microbial (21 articles) additives on hay preservation during storage. Multilevel linear mixed-effects models were fit with response variables calculated as predicted differences (Δ) between treated and untreated samples. Chemical preservatives were classified into five groups such as propionic acid (PropA), buffered organic acids (BOA), other organic acids (OOA), urea, and anhydrous ammonia (AA). Moderators of the models included preservative class (PC), forage type (FT; grass, legumes, and mixed hay), moisture concentration (MC), and application rate (AR). Dry matter (DM) loss during storage was affected by PC × FT (P = 0.045), PC × AR (P < 0.001), and PC × MC (P = 0.009), relative to the overall effect of preservatives (−0.37%). DM loss in PropA-treated hay was numerically reduced to a greater extent in grasses (−16.2), followed by mixed hay (−1.76), but it increased (+2.2%) in legume hay. Increasing AR of PropA resulted in decrease in DM loss (slope = −1.34). Application of BOA, OOA, PropA, and AA decreased visual relative moldiness by −22.1, −29.4, −45.5, and −12.2 percentage points, respectively (PC; P < 0.001). Sugars were higher in treated grass hay (+1.9) and lower in treated legume hay (−0.8% of DM) relative to their untreated counterparts (P < 0.001). The application of all preservatives resulted in higher crude protein (CP) than untreated hay, particularly urea (+7.92) and AA (+5.66% of DM), but PropA, OOA, and BOA also increased CP by 2.37, 2.04, and 0.73 percentage points, respectively. Additionally, preservative application overall resulted in higher in vitro DM digestibility (+1.9% of DM) relative to the untreated hay (⁠ $x¯=$58.3%), which increased with higher AR (slope = 1.64) and decreased with higher MC (slope = −0.27). Microbial inoculants had small effects on hay spoilage because the overall DM loss effect size was −0.21%. Relative to untreated (⁠ $x¯=$4.63% DM), grass hay preserved more sugars (+1.47) than legumes (+0.33) when an inoculant was applied. In conclusion, organic acid-based preservatives prevent spoilage of hay during storage, but their effectiveness is affected by FT, MC, and AR. Microbial inoculants had minor effects on preservation that were impaired by increased MC. Moreover, legume hay was less responsive to the effects of preservatives than grass hay. Storing hay that has not been properly dried to a moisture concentration (MC) below 20% can lead to the growth of undesirable microbes, such as molds, that decrease the nutritive value of the hay and compromise animal performance and health. Hay preservatives such as propionic acid (PropA), buffered organic acids (BOA), other organic acids (OOA), urea, anhydrous ammonia (AA), and microbial inoculants have been commonly applied to high moisture (>20%) hay to reduce microbial growth during hay storage. The present study compiled the results of 62 published articles on hay preservatives and performed a quantitative analysis (meta-analysis) to determine the effect of preservative treatment on dry matter (DM) loss, moldiness, bale heating, nutritional composition, and DM digestibility, and their interactions with forage type, application rate, and bale moisture. Organic acid-based preservatives (PropA, BOA, and OOA) were effective at reducing DM loss, moldiness, bale maximum temperature, and insoluble nitrogen, and at preserving sugars and DM digestibility during storage to different extents. Microbial inoculants had small effects on the prevention of hay spoilage, and were negatively affected by the increase of hay MC. Legume hay was less responsive to the effects of preservatives than grass hay during storage.

Objective: Examine the effects of supplementing bahiagrass hay (BG) with potentially anthelmintic quantities of hays of perennial peanut (PEA) or sericea lespedeza (LES) or seeds of velvet bean (Mucuna pruriens L.; MUC) or papaya (PAP) on the intake and nutritive value (Experiment 1), and the performance and parasite burden (Experiment 2) of goats. Methods: In Experiment 1, 38 male goats (27.4 +/- 5.7 kg body weight) were randomly assigned to each of 5 treatments: i) BG alone and BG plus; ii) PEA; iii) LES; iv) MUC; and v) PAP. Goats were fed for ad libitum consumption and adapted to the diets for 14 d followed by 7 d of measurement. The PEA, LES, MUC (50%, 50%, and 10% of the diet dry matter [DM], respectively), and PAP (forced-fed at 10 g/d) were fed at rates that would elicit anthelmintic effects. In Experiment 2, goats remained in the same treatments but were allocated to 15 pens (3 pens per treatment) from d 22 to 63. All goats were infected with parasites by grazing an infected bahiagrass pasture from 0800 to 1500 h daily and then returned to the pens. Results: Dry matter intake tended to be greater in goats fed PEA and LES than those fed BG (757 and 745 vs 612 g/d, respectively). Digestibility of DM (59.5% vs 54.9%) and organic matter (60.8% vs 56.0%) were greater in goats fed MUC vs BG, respectively. In Experiment 2, feeding PAP, LES, and PEA to goats reduced nematode fecal egg counts by 72%, 52%, and 32%, reduced abomasal adult worm counts by 78%, 52%, and 42%, and decreased plasma haptoglobin concentrations by 42%, 40%, and 45% relative to feeding BG alone, respectively. Conclusion: Supplementation with PEA, LES, and PAP decreased the parasite burden of goats but did not increase their performance. PAP was the most effective anthelmintic supplement.

This study evaluated the effects of chemical and biological preservatives and ensiling stage on spoilage, ruminal in vitro fermentation, and methane production of wet brewer’s grain (WBG) silage. Treatments (TRT) were sodium lignosulfonate at 10 g/kg fresh WBG (NaL1) and 20 g/kg (NaL2), propionic acid at 5 g/kg fresh WBG (PRP, 99%), a combination inoculant (INO; Lactococcus lactis and Lactobacillus buchneri each at 4.9 log cfu/fresh WBG g), and untreated WBG (CON). Fresh WBG was treated and then ensiled for 60 d, after which mini silos were opened and aerobically exposed (AES) for 10 d. Data were analyzed as a RCBD (5 blocks) with a 5 TRT × 3 stages (STG; Fresh, Ensiled, and AES) factorial arrangement. Results showed that Ensiled PRP-treated WBG markedly preserved more water-soluble carbohydrates and starch than all other Ensiled TRT (P<0.001). Dry matter losses of Ensiled PRP-treated WBG were 48% lower than all other Ensiled TRT (P=0.009) but were not different than CON in AES (P=0.350). Due to its greater concentration of digestible nutrients, PRP-treated AES was less aerobically stable than CON (P=0.03). Preservation was not improved by INO, NaL1 or NaL2 but the latter prevented the increase of neutral detergent fiber across STG (P=0.392). Apparent in vitro DM digestibility (IVDMD) decreased only in Ensiled CON, INO and NaL1 relative to Fresh WBG and AES NaL2 had greater IVDMD than all other AES TRT (P≤0.032). In vitro ruminal fermentation of Fresh WBG resulted in a greater methane concentration and yield than the other STG (P<0.033). In conclusion, PRP was the most effective at preserving WBG during ensiling but failed to improve aerobic stability under the conditions tested.

We evaluated the effects of applying a combination inoculant to four corn hybrids harvested at high moisture on their nutritive value and microbial populations. The treatment design was the factorial combination of corn hybrids ensiled with (INO) and without (CON) inoculant. The hybrids were TMF2R737 (MCN), F2F817 (MBR), P2089YHR (PCN), and PI144XR (PBR), ensiled at dry matter (DM) concentrations of 30.5%, 26.3%, 31.1%, and 31.5%, respectively; MBR and PBR were brown midrib mutants (BMR). The inoculant contained Lactobacillus buchneri and Pediococcus pentosaceus (4 × 10⁵ and 1 × 10⁵ cfu/g of fresh corn). The experiment had a complete randomized design with treatments replicated six times. Corn was treated or not with inoculant, packed into 7.6 L bucket silos, and stored for 100 d. At d 0, the relative abundance (RA, %) of Enterobacteriaceae was lower in PBR vs. the other hybrids [51.3 vs. $x¯$ = (average of) 58.4] and in the case of fungi, incertae sedis (i.s.) Tremellales and Mucoraceae were more and less abundant, respectively, in conventional hybrids vs. BMRs (⁠ $x¯$= 25.8 vs. $x¯$ = 13.9 and $x¯$ = 3.64 vs. $x¯$ = 7.52; P < 0.04). After ensiling, INO had higher LAB (9.3 vs. 7.1 log cfu/g of fresh corn) and acetic acid (3.44% vs. 1.32% of DM) and lower yeast (3.1 vs. 4.6) and molds (1.5 vs. 3.0), and also extended the aerobic stability (582 vs. 111 h) but decreased DM recovery (95.6% vs. 97.4%) vs. CON (P < 0.02). Inoculation reduced bacterial phylogenetic diversity (6.75 vs. 14.4) but increased fungal observed taxonomical units (46 vs. 20) vs. CON (P < 0.01). Also, a higher relative abundance (RA) for Lactobacillaceae (99.2% vs. 75.7%) and lower for Enterobacteriaceae (0.28 vs. 9.93) was observed due to inoculation (P < 0.001). For fungi, INO had a lower RA compared to CON for Monascaceae (12.6 vs. 44.7) and increased i.s. Tremellales (8.0 vs. 1.2) and i.s. Saccharomycetales (6.4% vs. 0.3%; P < 0.006). Inoculation changed the diverse bacterial community found in the phyllosphere across hybrids to a taxonomically uneven one dominated by Lactobacillaceae. In the case of fungi, INO application increased the fungal diversity at d 100 mainly by reducing the dominance of Monascaceae vs. CON. In conclusion, the INO treatment overwhelmed the disparate microbial populations found across BMR and conventional hybrids ensiled at low DM concentrations and ensured a significant concentration of acetic acid that modified fungal populations and in turn extended the aerobic stability of all hybrids.