Home Fermentation parameters and nutritional value of silages from fodder mallow (Malva verticillata L.), white sweet clover (Melilotus albus Medik.), and their mixtures
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Fermentation parameters and nutritional value of silages from fodder mallow (Malva verticillata L.), white sweet clover (Melilotus albus Medik.), and their mixtures

  • Antonín Kintl , Igor Huňady , Jiří Třináctý , Michal Richter , Julie Sobotková , Tereza Hammerschmiedt , Oldřich Látal , Martin Brtnický and Jakub Elbl EMAIL logo
Published/Copyright: May 10, 2025
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Open Agriculture
From the journal Open Agriculture Volume 10 Issue 1

Abstract

The goal of this experiment was to compare fermentation parameters and the nutritional value of silages made of fodder mallow (Malva verticillata L.), white sweet clover (Melilotus albus Medik.), and their mixtures. Chopped plants of Malva verticillata L., Melilotus albus Medik., and Malva verticillata L./Melilotus albus Medik. were used to prepare silages in the ratios of 1:0, 3:7, 1:1, 7:3, and 0:1, respectively. The crops were harvested at BBCH 51–61 vegetation stage (formation of flowers → flowering) with the stand height at harvest being 75 cm; a stubble height after the harvest was 8 cm. Fermentation parameters of the silage process, composition of carbohydrate fibre complex, concentration of nutrients, metabolizable energy (ME3×), net energy for lactation (NEL3×), protein fractions, and rumen non-degraded proteins (RUP5) were evaluated. Due to the low dry matter content and higher degree of proteolysis in the Malva verticillata L. silage (P < 0.001), the concentration of ammonia (NH3) was significantly higher than in the Melilotus albus Medik. variant. Differences in the concentrations of ME3× and NEL3× between silages from solitary Malva verticillata L. and Melilotus albus Medik. were significant, respectively, but low. The nutritional value of silages from both evaluated alternative crops and their mixtures represented by NEL3× and RUP5 concentrations is comparable to the nutritional value of silages from the conventional forages mentioned in feed databases.

Keywords: metabolizable energy; net energy for lactation; protein fractions; rumen non-degraded protein

Abbreviations

AA

acetic acid

ADFom

ash-free acid detergent fibre

ADL

acid detergent lignin

aNDFom

neutral detergent fibre with α-amylase and sodium sulphite corrected for ash

BA

butyric acid

BFN

biological fixation of nitrogen

CF

crude fibre

CP

crude protein

DM

dry matter

LA

lactic acid

LAB

lactic acid bacteria

ME3×

metabolizable energy

NEL3×

net energy for lactation

RUP5

rumen non-degraded proteins

1 Introduction

The cultivation of maize for silage production increased in the Czech Republic in recent decades. In the last 6 years, the area of land used for growing maize for feed production in the Czech Republic ranged from 231,000 ha in 2018 [1] to 213,500 ha in 2024 [2]. Maize silage is a major component of cattle diet [3]; however, its production is negatively influenced by periods of drought and extreme rainfall as a result of global climate changes [4].

The extreme conditions simultaneously increase the risk of water erosion due to the low vegetative soil cover during maize growing [5]. For these reasons, research teams are looking for alternative fodder plants that would be drought-tolerant, contribute to soil sustainability, and exhibit nutritional values comparable to conventional ones [6]. Fodder mallow (Malva verticillata L.) and white sweet clover (Melilotus albus Medik.) are plants that are close to the above-mentioned properties [7,8]. Both crops can be successfully grown under harsh conditions in which the production of conventional fodder is impossible. There are, however, no studies that would be focused on details of cultivating these crops in the mixed cropping system used to produce fodder. In this respect, this study differs from the previous study [5,6,7,8], which dealt mainly with the use of individual crops. The potential of growing the two crops is considerably high.

M. verticillata originated in the warm regions of eastern Asia. A characteristic feature of its root system is a taproot, which is strongly branched in each soil layer and can reach a depth of 1 m. Shoots of this plant are plentifully covered with round leaves, develop numerous branches, and may reach up to 2 m [9]. M. verticillata (Malvaceae species), as a promising forage, contains, on average, 20% crude protein (CP), 3% raw fats, and an optimal content of crude cellulose of about 29–32% [10,11]. Its inclusion as a silage in the diet of lactating cows increases milk production [12]. One problematic feature of this crop is the high moisture content in the harvested matter [10], which can negatively affect the ensiling process [13].

M. albus is a species from the Fabaceae family with typical properties such as taproots penetrating up to greater depths and a nitrogen fixation rate that is superior to those of other legumes, making it beneficial for crop rotations [14]. Because of its Eurasian origin, it has been adapted to extreme environments, such as drought and cold. However, Melilotus species plants are specific in their high content of coumarin, and their concentrations reach up to 5% of dry matter (DM) depending on the cultivar [15]. Silages prepared from the plants of this family reach 13–19% CP, 6.8–11% ash, 44–56% neutral detergent fibre (NDF), 12–17% hemicellulose, 24–33% acid detergent fibre (ADF), 12–14% lignin, and 26–32% cellulose. Although the silages made of Melilotus species can feature interesting feeding parameters from the nutritional point of view, there is also a risk of coumarin occurrence and higher content of less biodegradable substances as compared with the Malvaceae species [15,16]. In mouldy silages (excess moisture or aerobic conditions), coumarin can be converted to toxic dicoumarol, which is a potent anticoagulant causing anaemia, haematoma formation, uncontrollable bleeding, and abortion [17].

This may be a problem in silages prepared of forage legumes that contain natural concentrations of coumarin, for example, Trifolium pratense L., Trifolium repens L., Melilotus spp., and others [18]. The issue of dicoumarol development has been studied for a long time [17,18,19,20]. Causal relationships between mouldy sweet clover and mysterious bleeding diseases of farm animals were first described in 1923 [21]. Since then, efficient methods are sought how to prevent the development of excessively high concentrations of dicoumarol and other mycotoxins in silages prepared from forage legumes [18,19,20,22]. One of the most important factors is to follow the correct technological procedure of ensiling [20], such as optimum moisture content of harvested plant biomass, sufficient compaction of plant biomass, and addition of conservatives (bacterial inoculants) [23,24]. Optimum moisture content is particularly important when harvesting forage legumes and should be thoroughly monitored. Bacterial inoculants are mostly represented by lactic acid bacteria (LAB) [24,25]. LAB can support the conversion of water-soluble carbohydrates to lactic acid (LA), which results in decreased pH. Acidification of the environment then results in the decreased activity of microorganisms that could contribute to the production of mycotoxins [26]. Fan et al. [24] demonstrated a positive influence of applied LAB inoculants on the reduced concentration of aflatoxin and deoxynivalenol in alfalfa (Medicago sativa L.) silage. Other authors [27] maintain that the production of dicoumarol can also be reduced if LAB is present in the silage.

Another possibility is to influence the representation of individual plant species in silage so that the biomass of plants with a high content of coumarin does not dominate. Undesirable properties of the above-mentioned crops (high moisture, coumarin content) can be compensated by their mixing [28]. In the future, an option opens up to breed new varieties of legumes, which would naturally contain lower coumarin concentrations than the currently available ones [29].

In our experiment, we evaluated ensiled mixtures with different ratios of M. verticillata and M. albus as to their influence on the process of ensiling and nutritional value (energy content, protein quality). Forage legumes are advantageous for the protein supply to dairy cattle because of their high CP content [30]. In the nutrition of dairy cows, legumes represent a domestic source of CP (N-substances) in ensiled bulky feeds and one of the ways how to reduce their share in purchased core feeds [31]. However, CP is subjected to extensive degradation in the rumen. Part of CP escapes ruminal degradation (in the sense of rumen non-degraded crude protein assuming ruminal passage rate, RUP5) and may become available for digestion and amino acid absorption in the small intestine. That is why RUP5 is an indicator of feed CP quality. Ruminal degradation characteristics of CP for RUP5 determination are usually obtained using the in situ method [32]. However, this method is laborious and cost-intensive because of its dependence on ruminally cannulated animals. Shannak et al. [33] and Kirchhof et al. [30] developed regression equations to estimate in situ RUP5 values from the chemical fractionation of CP, according to Licitra et al. [34]. A regression equation developed for forage by Kirchhof et al. [30] was used in our experiment for the RUP5 calculation.

The goal of this study was to compare the fermentation parameters and the nutrient composition of silages from M. verticillata, M. albus, and their mixtures. The results of this comparison will allow us to select a mixture with optimal properties. A partial objective of the study was to decide whether the mixture of M. verticillata and M. albus legumes can represent a domestic source of CP (N-substances) in ensiled bulky feeds used for the nutrition of dairy cows (cattle). Based on the setup research goal, two hypotheses were tested: null hypothesis (H0) and alternative hypothesis (H1). H0 = the growing of fodder mallow (Malva verticillata L.) in the mixed cropping system with white sweet clover (Melilotus albus Medik.) has no demonstrable positive influence on the quality of resulting silage from the viewpoint of its nutritional value for feeding farm animals. H1 = the legume (white sweet clover) inclusion into the mixed cropping system with fodder mallow will bring demonstrable changes in nutritional parameters in the resulting silage, particularly the increased content of N-substances in the silage.

2 Materials and methods

2.1 Field experiment site

The experimental plot is situated not far from the Troubsko station of Agricultural Research, Ltd. (49.1709261N, 16.4916056E). The experimental crop was grown in 2019 on plots with a pre-crop of winter wheat (Triticum aestivum L.). Geological subsoil in the area is loess and loess loam of the Bohemian Massif, soil type is Haplic Luvisol [35]. In terms of agro-ecological classification [36], the experimental site is situated in the mildly warm and mildly dry climatic zone with an average altitude of 290 m a. s. l., long-term (1981–2010) mean annual temperature of 8.95°C, and annual precipitation amount of 525.6 mm.

Prior to sowing the experimental crops, a mixed NPK fertilizer (10:26:26) was applied by HOKR, Ltd. (Czech Republic) at a dose of 300 kg/ha. The crops were sown using the zero-waste seeder model Oyord. The experimental stands were then treated with the post-emergent graminicide TARGA SUPER 5 EC manufactured by NISSAN Chemical Industries Ltd. (Tokyo, Japan) at a dose of 0.7 l/ha. The stand was not irrigated.

2.2 Plant biomass production

Biomass for the experiment was obtained as follows:

  1. Monoculture: curled cheese weed (M. verticillata var. crispa L., breed Palina Figure 1) and fodder mallow (M. verticillata). The sowing rate was determined according to Zielewicz and Wróbel [9] and used in Kintl et al. [35]. The total number of individuals per hectare was 9.20 million individuals (23.0 kg).

  2. Mixed culture: M. verticillata + M. albus (white sweet clover; Figure 1). The M. verticillata and M. albus combination was sown at a ratio of 50:50: 4.60 million individuals (11.5 kg) of M. verticillate per hectare and 2.93 million individuals (6 kg) of M. albus per hectare according to Kintl et al. [35].

  3. Monoculture: white sweet clover (M. albus, var. Meba; Figure 1). The sowing rate was determined according to Rigal et al. [37] identically as in the previous variant and used, for example, in Kintl et al. [35]. The total number of individuals per hectare was 5.85 mil. (12.0 kg).

Figure 1 Experimental crops: (a) monoculture of M. verticillata, (b) monoculture of M. albus, and (c) mixed culture of M. verticillata + M. albus (authors’ archives).
Figure 1

Experimental crops: (a) monoculture of M. verticillata, (b) monoculture of M. albus, and (c) mixed culture of M. verticillata + M. albus (authors’ archives).

The system of growing crops used to evaluate the feed parameters of prepared model silages is described in detail in the study of Kintl et al. [35].

2.3 Production of mixed culture silage – preparation of model silage

The crops were harvested at BBCH 51–61 phenological stage (formation of flowers; 17 July 2019) using the Deutz – Fahr MH 650 S chopper (Deutz – Fahr, Lauingen, Germany) pursuant to the procedure described in the study of Mlejnková et al. [38] (Table 1). The average stand height before the harvest was 75 cm, and the height of stubble left on the plot after the harvest was not more than 8 cm. Chopped plants of M. verticillata and M. albus monocultures and mixed cropping of M. verticillata/M. albus – 50/50 were homogenized and used for silage 1:0 (M. verticillata monoculture), 1:1 (mixed cropping of M. verticillata + M. albus), and 0:1 (M. albus monoculture). The above-mentioned variants and a detailed description of the harvest were published in the study of Kintl et al. [35].

Table 1

Basic information about the harvest of grown crops

Harvest date of both crops BBCH 51–61 (formation of flowers → flowering)
Size of shreddings 15–20 mm
Type of chopper Deutz – Fahr MH 650 S chopper (Deutz – Fahr, Lauingen, Germany)

Only the 1:1 mixed culture variant was harvested directly in the field. Further silages in ratios 3:7 and 7:3 were prepared by mixing the shreddings of M. verticillata and M. albus grown in monocultures (i.e. after harvest). The prepared shreddings served to produce model micro-silages characterized in detail in the study of Kintl et al. [35,39]. The procedure was the same for all variants.

The prepared shreddings were placed in airtight tubes (mini silo containers) made of unplasticized polyvinyl chloride with a diameter of 150 mm and a height of 1,000 mm. The tubes were provided with a plug on both sides and with a system for the discharge of gas generated during the fermentation process. Each mini silo container was filled with 8 kg of plant biomass shreddings. Each variant (silo) contained an addition of inoculant with the following bacteria: Lactococcus lactis, Lactobacillus plantarum, Lactococcus lactis, Lactobacillus plantarum, and Enterococcus faecium. The type of inoculant was Silo Solve EF(Chr. Hansen Holding Ltd., Starovice, Czech Republic), and the dose of the inoculant was 5 g + 3.5 H2O/t. The final concentration of microorganisms was 250 CFU × 103/g in fresh chop at the above dose (5 g + 3.5 l H2O/t). An overview of prepared silages is presented in Table 2.

Table 2

Overview of prepared model silages

No. Treatment M. verticillate in silage† (%) M. albus in silage (%) Average density of model silage in DM (kg/m3)
1 M. verticillata* 100 0 112.66
2 M. albus* 0 100 164.66
3 M. verticillata + M. albus* 50 50 150.23
4 M. verticillata + M. albus** 70 30 157.33
5 M. verticillata + M. albus** 30 70 129.92

†Percentage w/w.; *published in the study of Kintl et al. [35]; **prepared model silages not yet evaluated. There were five variants of experimental silage, each in triplicate.

2.4 Analytical procedures

Fermentation parameters characterizing the ensiling process and silage quality were determined from the aqueous silage extract prepared according to the following procedure: a 50 g sample of silage was blended in 450 ml of deionized water for 4 min and filtered through Whatman 54 filter paper.

The value of pH in the water extract was determined by a glass electrode using the bench-top pH meter (Radelkis OP 211, Radelkis Electrochemical Instruments, Budapest, Hungary). Water silage extract acidity (free acidity) was determined after titration of 0.1 M KOH to pH 8.5 and measured again using the bench-top pH meter according to Bernardes et al. [40] and Křížová et al. [41]. Ammonia (NH3) content was determined according to the Australian Fodder Industry Association [42] by NH3 gas sensing ISE (Elektrochemické detektory, s. r. o., Czech Republic) using Radelkis OP 211 digital pH meters (Radelkis Electrochemical Instruments, Budapest, Hungary).

LA and volatile fatty acids, including acetic acid (AA) and butyric acid (BA) in the silage extract, were determined by using the capillary isotachophoresis method [43] with Ionosep 2003 (RECMAN – laboratory equipment, Czech Republic) and alcohols using the Varian CP-3380 gas chromatograph (Varian, USA) according to Porter and Murray [44].

Silage samples for the determination of nutritional value parameters were dried at 55°C for 48 h and ground to pass through a 1 mm sieve of a knife mill (Brabender Technologie GmbH & Co. KG, Duisburg, Germany). In the ground samples, the following parameters were estimated according to European Union standards for sampling and analysis [45]: DM (L 54:12), CP (L 54:15), ash (L 54:50), crude fibre (CF) (L 54:40), and ether extract (L 54:37). Neutral detergent fibre with α-amylase and sodium sulphite corrected for ash (aNDFom) was estimated according to International Standardization Organization [46], ash-free acid detergent fibre (ADFom), and acid detergent lignin (ADL) according to International Standardization Organization [47]. Soluble protein, neutral detergent insoluble crude protein, and acid detergent insoluble crude protein (acid detergent insoluble crude protein [ADICP]) were estimated using the procedure reported by Licitra et al. [34] and modified by Higgs et al. [48].

2.5 Calculation

The net energy for lactation at three times the maintenance level of intake (NEL3×) was calculated using the following equations [49]:

tdNFC ( % ) = 0.98 × ( 100 – ( aNDFom + CP + EE + Ash ) ) ,

tdCP ( % ) = CP × e ( − 1.2 × ( ADICP / CP ) ) ,

tdFA ( % ) = FA,

tdNDF ( % ) = 0.75 × ( aNDFom – ADL ) × ( 1 – ( ADL / aNDFom ) 0.667 ) ,

DE1 × ( Mcal / kg ) = ( tdNFC / 100 ) × 4.2 + ( tdNDF / 100 ) × 4.2 + ( tdCP / 100 ) × 5.6 + ( FA / 100 ) × 9.4 – 0.3 ,

Discount = ( TDN 1 × – ( 0.18 × TDN1 × – 103 ) × Intake ) / TDN1 × ,

DE3 × ( Mcal / kg ) = Discount × DE1 × ,

ME3 × ( Mcal / kg ) = ( 1.01 × DE3 × ) – 0.45 ,

NEL3 × ( Mcal / kg ) = ( 0.703 × ME3 × ) – 0.19 ,

NEL3 × ( MJ / kg ) = 4.184 × NEL3 × ( Mcal / kg ) ,

where tdNFC denotes truly digestible non-fibre carbohydrates, tdCP denotes truly digestible CP, tdFA denotes truly digestible fatty acids (FA (%) = EE – 1), tdNDF denotes truly digestible aNDFom, DE1× (Mcal/kg) denotes digestible energy, Discount denotes the coefficient for the conversion of digestible energy at maintenance to that at production level of intake, TDN1× denotes total digestible nutrients for the entire diet (in our case TDN1× = 74%, i.e. average value assumed from NRC [49], Intake denotes incremental intake above maintenance (in our case for a cow consuming 3× maintenance Intake = 2).

The content of rumen non-degraded crude protein assuming ruminal passage rate Kp = 5%/h (RUP5) was calculated on the basis of protein and carbohydrate fractions using a regression equation developed by Kirchhof et al. [30]:

RUP5 ( % CP ) = 0.1 × ( 321.9023 + 0.1676 × ADFom – 0.0022 × ( CP × ( A 1 + A 2 ) ) + 0.0001 × ( CP × C2 ) ) ,

where parameters ADFom and CP are expressed in g/kg DM and fractions A1, A2, and C are expressed in g/kg CP.

2.6 Statistical analysis

All measured values from silages obtained in the experiment were analysed using the program Statistica 7.0 (StatSoft, Inc., USA). At first, the entire set of data was analysed using Exploratory data analysis (EDA) to detect possible extreme values and errors in the data set. Part of the input analysis was the calculation of normality tests, basic statistical parameters (min, max, range, median, average, etc.), and the building of histograms. None of the partial EDA confirmed systematic errors in the set of measured data that were then analysed using one-way ANOVA according to the following model:

Y i j = m + M i + ε i j ,

where m is the general mean, M i is the effect of the mixture (i = 5), ɛ ij is the residual error of the ith treatment, and jth observation (j = 3).

The ANOVA was subsequently complemented with the Tukey test to compare the means when significant differences were observed. All statistical analyses were performed at a significance level of P < 0.001.

3 Results and discussion

The presented results represent the first and unique characterization of the parameters (fermentation parameters, carbohydrate fibre complex, chemical composition, and energy value) featured by the mixed culture of M. verticillate and M. albus from the viewpoint of its potential use for feeding farm animals. Kintl et al. [35,39] primarily focused on the use of these crops for the anaerobic fermentation process in biogas plants; in other words, for the production of biogas. The data described below are about the potential use of singular mixed culture, which has not yet been described in terms of feeding parameters. However, their interpretation should take into account the limits of the experiment relating to the number of tested crops.

3.1 Fermentation parameters

The basic parameters of fermentation measured are summarized in Table 3. The data indicate that the measured values could have been affected by the compaction of shreddings. The compaction of shreddings in mini-silos ranged from 112.7 kgTS/m3 in Variant 1 M. verticillata * 100% up to 164.7 kgTS/m3 in Variant 2 M. albus 100% (Table 2). Harrison and Fransen [50] mentioned a significant influence of shredding type on final compaction. It is assumed that the preparation of plant biomass for ensiling (size of shreddings) has the same influence on the resulting silage quality as the course of the ensiling process. The fact affected, for example, the compaction of shreddings. According to Hutnik and Kobielak [51], the resulting density of silage is affected by the length of shreddings and by DM related to crop ripeness or the use of wilting technology. Johnson et al. [52] confirmed that the use of LAB-producing LA improves the aerobic stability of maize silage more than ripeness and mechanical treatment. Production of high-quality silage has to take into account the above-mentioned factors. The measured values are presented in Table 3. The greatest difference in the DM content (over 12%) was found between the silages from solitary (monoculture) M. verticillata and M. albus. Except for the silage from the solitary M. albus (32.35%), DM content in the remaining mixtures was lower than 30%. In this case, an important role is played by the so-called critical pH value [53], which provides anaerobic stability of silages. According to the author, critical pH values for DM contents of 20 and 35% are 4.20 and 4.60, respectively, and the pH values observed in the present experiment are close to this range (4.20). The closest variant to this interface was 0:1 (4.20); the other variants were below this value (Table 3). The DM content determines a further use of harvested biomass in the silage, particularly in legumes where inappropriate biomass moisture content may lead to the degradation of the final product – fodder [54]. DM values in model plants correspond with the findings of other authors who measured 11.46–21.33% in the case of M. verticillata [12] and, on average, 30% in the case of M. albus [55]. In leguminous plants, DM is generally affected mainly by meteorological conditions [54] and also by breeding, which affects the level of their yield [56]. Thus, it is possible to state that changes in DM contents and fermentation parameters of silages depend on the M. verticillata/M. albus ratio in the mixture. The increasing share of M. albus in silages resulted in the increasing values of DM, pH, and other changes in the values of fermentation parameters (see below). This situation could have been due to the chemical composition of the biomass of Melilotus species dominated by poorly degradable substances (cellulose, hemicellulose, etc.) as compared with the Malva species [11,55].

Table 3

DM and fermentation parameters of silages from fodder mallow, white sweet clover, and their mixtures

Parameter Unit Mixtures (M. verticillata + M. albus) P SEM
1:0 7:3 1:1 3:7 0:1
DM % 20.75e 24.47d 27.79c 29.38b 32.35a <0.001 0.12
pH 4.15c 4.15c 4.16bc 4.18ab 4.20a <0.001 0.01
NH3 % CP 10.55ab 14.27a 3.73c 6.19bc 3.56c <0.001 0.98
LA % DM 12.62a 10.45ab 8.73ab 8.56b 8.69ab 0.031 0.86
AA % DM 3.93a 2.97a 4.2a 3.2a 2.4a 0.366 0.66
BA % DM 0.03 0 0 0 0 0.452 0.01
LA:AA 3.49 3.52 2.45 2.86 3.73 0.468 0.55
Alc. % DM 0.7 0.35 0.83 1.09 0.79 0.074 0.15

DM = dry matter, LA = lactic acid, AA = acetic acid, BA = butyric acid, Alc. = alcohols, and SEM = standard error of the mean. Lowercase letters indicate significant differences (P < 0.05) between individual variants within the selected parameter.

Typical concentrations of LA in legume and grass silages (DM contents between 30 and 55%) range from 2 to 10% DM but this one can be considerably higher in silages with DM contents lower than 30% [13]. In spite of the low DM contents, the concentrations of LA mentioned in Table 3 are within the reported range. Only a slightly higher value of LA concentration (12.62% DM) was recorded in the silage with the lowest DM content (20.75%) from solitary M. verticillata. This LA concentration significantly differed only from the value for the silage of M. verticillata + M. albus mixed in the ratio of 3:7. The pH values and corresponding LA concentrations indicated a standard fermentation process during ensiling. As a marker of the activity of clostridial organisms is the concentration of BA higher than 0.5% DM [13]. Trace concentration of BA (0.03% DM) was observed only in the silage from the solitary M. verticillata with the lowest DM content. If we further focus only on LA, similar values for Melilotus officinalis L. Lam. were measured by Kara [57]: LA concentration of 6.30% during the vegetative stage and BA content of 0.02% during the vegetative stage. The LA content was apparently very similar to the data determined by us; however, there are differences in the concentration of BA. It is very interesting to note that Kara [57] observed that the amount of LA decreased linearly with increasing vegetative stage (plant maturation) while the amount of BA fluctuated irregularly.

The measured data indicate that the LA values decreased with increasing share of M. albus in the tested silages. Similarly, as in the case of fermentation parameters, this development was closely related to the composition of the respective silages. The addition of M. albus into the silage resulted in a small increase of pH (mixtures from 1:1 to 0:1). This slight increase of pH could have negatively affected the activity of clostridial organisms [13] and hence decreased the concentration of BA, most likely showing the decreased concentration of LA. These results reject the H0 hypothesis while confirming the alternative H1 hypothesis, claiming that the addition of M. albus brings a demonstrable change in silage quality.

Next, the two fermentation parameters such as AA concentration and LA: AA ratio, correspond to the values reported by Kung et al. [13] for wet silages (range from 3 to 4% DM for AA concentration; 2.5 to 3.0 and greater in the case of LA:AA ratio). High concentrations of ethanol usually indicate excessive metabolism of yeasts. Usual concentrations of ethanol in silages range from 0.5 to 1.5% DM [13]. Our concentrations of alcohol are within the mentioned interval. A similar concentration of AA (0.6–0.7%) depending on the vegetative stage was found in M. officinalis Lam. silages and also in other legumes determined for feeding farm animals but of the same genus (Melilotus) [57]. Thus, we can state that the measured values of AA, BA, and LA:AA ratio were within the expected range [13,56]. No demonstrable differences were observed between the silages made of pure cultures (M. verticillate and M. albus) and the silages made of mixed crops (M. verticillata + M. albus).

Concentrations of NH3 higher than 12–15% CP result from excessive protein breakdown caused by the slow drop in pH or clostridial action [13]. In general, wet silages have higher concentrations of NH3 than drier silages because of the overall more robust fermentation, which is in agreement with the results of our experiment, such as a significant (P < 0.001) difference in NH3 concentration between M. verticillata (10.55% CP) and M. albus (3.56% CP). This difference is quite interesting as it was expected in the context of natural properties of legumes (high content of N substances in the plant biomass) that the presence of M. albus in silages would increase the content of proteins (N-substances). Such a situation did not occur, and the explanation can be potentially the date of harvest in the flowering period of crops. M. albus reached neither full inflorescence nor fruits (beans), in which the plants of the Fabaceae family store (synthesize) most N-substances [55,58]. On the other hand, this conclusion can be used only partly in the case of M. verticillata + M. albus mixtures. Mixtures 7:3 exhibited that the share of CP increased to 14.27% as compared to the other variants, but the other mixtures (1:1, 3:7) showed a significant decrease when compared with the silage made of pure M. verticillata. This fluctuation within one variant could be due to, for example, the deviation in the process of ensiling, which could have resulted in a part of CP being transformed into simpler organic substances. However, this assumption cannot be significantly confirmed by the measured and below presented values.

In the context of feed nutritional value, a higher degree of proteolysis impairs the quality of the original protein. Monitoring of NH3 concentration in the silage of legumes is immensely important for their use in feeding farm animals, especially with regard to milk production [59,60]. The increasing concentration of NH3 is reducing DM intake, which has a negative impact on the amount and quality of milk [60]. According to Kung and Shaver [61], the legume silage (DM 30–40%) reaches, on average, an NH3 concentration of 10–15% of CP. These concentrations correspond to values measured in the silage of M. verticillate, while M. albus exhibited lower values. For comparison, maize silage (DM 30–40%) contains, on average, 5–7% NH3 [61]. By evaluating the fermentation parameters in general, we can state that the null hypothesis H0 has been disproved only partly. The creation of mixed silages from M. verticillata + M. albus resulted only in a partial change of fermentation parameters. In the case of CP, the change conflicted with expectations (assumed increase of N-substances in the silage in the context of using Fabaceae family crops).

3.2 Carbohydrate fibre complex

The carbohydrate fibre complex represented by aNDFom includes cellulose, hemicelluloses, and lignin, which are calculated from the determined parameters ADFom and ADL. A digestible portion of aNDFom is a major source of energy in forages. The digestibility of fibre is largely determined by the ratio of major components (cellulose, hemicelluloses, and lignin) in aNDFom [62]. Comparisons of all mentioned parameters determined in the silages from M. verticillata, M. albus, and their mixtures are presented in Table 4. Differences in the concentration of aNDFom (P < 0.001), ADFom (P < 0.001), ADL (P < 0.001), hemicellulose (P < 0.05), and cellulose (P < 0.001) between the silages from solitary M. verticillata and M. albus were high, and concentrations of all mentioned parameters in the individual mixtures corresponded to the ratio of M. verticillata and M. albus.

Table 4

Composition of carbohydrate fibre complex in silages from fodder mallow, white sweet clover, and their mixtures

Parameter Unit Mixtures (M. verticillata + M. albus) P SEM
1:0 7:3 1:1 3:7 0:1
aNDFom % DM 29.06d 34.82c 39.12b 38.58b 43.43a <0.001 0.24
ADFom % DM 24.78d 30.60c 34.09b 33.16b 37.17a <0.001 0.40
ADL % DM 3.59c 4.34bc 5.46ab 5.32ab 6.05a <0.001 0.25
% aNDFom 12.34 12.46 13.96 13.78 13.95 0.212 0.62
Hemicell. % DM 4.28b 4.22b 5.03ab 5.42ab 6.26a 0.014 0.36
% aNDFom 14.74 12.13 12.86 14.04 14.40 0.350 0.98
Cell. % DM 21.19d 26.26c 28.63b 27.8bc 31.1a <0.001 0.36
% aNDFom 72.92 75.41 73.18 72.19 71.65 0.068 0.82

aNDFom = neutral detergent fibre with α-amylase and sodium sulphite corrected for ash, ADFom = ash-free acid detergent fibre, ADL = acid detergent lignin, Hemicell. = hemicelluloses, Cell. = cellulose, DM = dry matter, SEM = standard error of the mean.

In the case of silage from the solitary M. verticillata, the concentration of aNDFom in our experiment was lower (29.06% DM) than the values reported for the M. verticillata green matter by Zielewicz and Wróbel [9] (from 47.7 to 53.6% DM) and of Malva crispa L. by Ţîţei and Teleuță [11] (70.1% DM). A lower value (24.78% DM) as compared to the mentioned authors (36.6–40.7 and 47.1% DM, respectively) was also observed for ADFom. Our ADL concentration (3.59% DM) was comparable to values reported by Zielewicz and Wróbel [9] (3.51–3.98% DM) but lower than those reported by Ţîţei and Teleuță [11] (6.9% DM).

In the case of silage from the solitary M. albus, our concentration of aNDFom (43.43% DM) was lower than in the M. officinalis Lam. hay (53.0% DM) compared to that of Howard et al. [63] but higher than in the M. albus green matter (33.6% DM) [64] and approximate to the value reported by Kintl et al. [39] for the M. albus silage (48.7% DM). ADFom concentration (37.17% DM) determined in the M. albus silage in our study was considerably higher as compared to 24.2% DM stated by Bozhanska et al. [64] and approximate values of 40.3% DM [63] and 31.6% DM [39]. Our ADL concentration (6.0% DM) was in agreement with the value of 6.24% DM [63], which is two times higher than 3.05% DM [64] and half in comparison to 14.3% DM, as reported by Kintl et al. [39] in the M. albus silage. There are relatively only a few data in the scientific literature for the evaluation of nutritional substances contained in the M. albus silage that could be used for comparison with values measured by us since the topic has not been paid enough attention so far, and available data are often incomplete. Nevertheless, the hitherto available sources [55,57,58] indicate that the composition of plant biomass in M. albus significantly differs in the period at the beginning of flowering and in the period at the peak of flowering, possibly in the period of fruit formation, namely as to the contents of proteins, ash, cellulose, hemicellulose, etc. According to the above-mentioned studies, the plants of M. albus harvested at the stage of butonization exhibited a higher content of total protein, crude fat, and ash but a lower content of CF. In our experiment, the biomass was harvested in the period after the beginning of flowering and it is therefore well possible that the date affected the composition of M. albus biomass in terms of its greater amount of less accessible substances at the cost of, e.g. N-substances.

As reported by Jančík et al. [62], the degree of correlation between NDF digestibility and the concentration of individual major components of carbohydrate fibre complex expressed as a coefficient of linear correlation (R) decreases in the following order: hemicelluloses (R = −0.41), cellulose (R = −0.71), and ADL (R = −0.87). For the evaluation of carbohydrate fibre complex as to its digestibility, there is a more representative expression of the above-mentioned components in the percent of NDF. Particularly, the degree of lignification expressed by the ADL:aNDFom ratio strongly correlated with NDF digestibility and serves as a component in the equation for the calculation of tdNDF (see Section 2.5) according to NRC [49]. As shown in Table 4, significant differences between individual mixtures observed by ADL, hemicelluloses, and cellulose expressed in % of DM were eliminated after their expression in % of aNDFom. Based on this finding, we can anticipate minimal differences in aNDFom digestibility between the evaluated mixtures.

For fermentation parameters, the null hypothesis (H0) was not rejected in full. For the carbohydrate fibre complex, it can be stated that its composition in mixed silages was significantly affected by the addition of M. albus and that H0 was rejected, and the alternative H1 hypothesis was confirmed.

3.3 Chemical composition and energy value

Concentrations of basic nutrients, ME3× and NEL3×, in the evaluated mixtures are shown in Table 5. Silage from the solitary M. verticillata shows a relatively high concentration of ash (13.71% DM) in comparison to that from M. albus (7.76% DM, P < 0.001). This information is very interesting as the available sources indicate [39,55,58,64] that M. albus contains less degradable substances (hemicellulose, lignin, etc.) when compared with M. verticillata [11]. The observed increased content of ash in M. verticillata could be due to, for example, the harvesting method when shreddings of this crop could have been partly contaminated with soil particles [65] in spite of the fact that the harvesting methodology eliminated the risk as much as possible. Comparable values were reported for the green matter of M. verticillata (10.5–27.7% DM) by Bonnemaire et al. [10] and of M. crispa (12.0% DM) by Ţîţei and Teleuță [11]. In the case of silage from M. albus, the ash concentration in our experiment agreed with the value for M. officinalis Lam. hay (8.25% DM) reported by Howard et al. [63] and was slightly higher as compared with the value for green matter of M. albus (5.28% DM) reported by Bozhanska et al. [64].

Table 5

Chemical composition, content of metabolizable (ME3×), and net energy for lactation (NEL3×) at 3 times maintenance level of intake of silages from fodder mallow, white sweet clover, and their mixtures

Parameter Unit Mixtures (M. verticillata + M. albus) P SEM
1:0 7:3 1:1 3:7 0:1
Ash % DM 13.71a 11.35b 9.55c 9.08d 7.76e <0.001 0.09
CF % DM 20.56d 25.43c 29.52b 28.63b 31.81a <0.001 0.42
CP % DM 16.23a 15.96ab 15.19bc 16.02ab 14.48c <0.001 0.18
EE % DM 2.62 2.87 2.83 2.91 2.88 0.658 0.15
NFC % DM 38.38a 34.99b 33.31c 33.41bc 31.45d <0.001 0.35
ME3× MJ/kg 9.31a 9.24a 8.98b 9.18ab 8.89b 0.004 0.06
NEL3× MJ/kg 5.75a 5.70ab 5.52bc 5.66abc 5.45c 0.004 0.04

CF = crude fibre, CP = crude protein, EE = ether extract, NFC = non-fibre carbohydrates, ME3× = metabolizable energy at 3 times the maintenance level of intake, NEL3× = net energy for lactation at 3 times the maintenance level of intake, DM = dry matter, and SEM = standard error of the mean.

CF is a constituent of the Weende analysis system. Namely, in some French references, it is known as crude cellulose (cellulose brutte). The values of CF concentrations in Table 5 are very near to the cellulose concentration in Table 4, with identical differences (P < 0.001) between the evaluated mixtures. Changes in the content of CF were obvious with the addition of M. albus, which clearly resulted in the increased content of CF in silage. This situation was expected, as the biomass of M. albus is characterized by a higher content of less degradable organic substances. Our value of CF concentrations in the M. verticillata silage (20.56% DM) was within the range (19.5–27.8% DM) reported by Bonnemaire et al. [10] and lower than the value (32.0% DM) stated by Ţîţei and Teleuță [11].

The CP concentration in the M. albus silage (14.48% DM) was lower in comparison with M. verticillata (15.96% DM). The values were rather surprising as M. albus belongs to the family of Fabaceae, whose representatives are characterized by the capability of biological fixation of atmospheric N and its storage in biomass [66,67]. Therefore, the measured values of CP are very low in M. albus when compared with values reported by Bozhanska et al. [64] (32.6% DM) and by Kintl et al. [39] (28.2% DM). In the case of CP concentration, the difference between silages from the solitary M. verticillata and M. albus is significant (P < 0.001). Its value in both evaluated crops is relatively high and comparable with values exhibited by conventional forages such as legumes. The addition of M. albus had no significant influence on the increase or decrease of CP content in the mixtures of M. verticillata and M. albus. The addition of legume to the silage was assumed to increase the content of CP (N-substances in general); however, this was not observed. This phenomenon is not easy to explain. Based on Phillips and Williams [67] and Oso and Ashafa [68], we assume that during the growth of M. albus, N-substances obtained through BFN were gradually stored in the plant biomass with the highest intensity of this storage taking place during the formation of fruits. As the plants of M. albus were harvested for the purpose of ensiling prior to the formation of fruits (beans), the resulting content of CP in plant biomass could have been significantly affected. Our value of CP concentrations in the M. verticillata silage (16.23% DM) was within the range (13.0–24.0% DM) reported by Bonnemaire et al. [10], lower than the value of 19.1% DM stated by Ţîţei and Teleuță [11], and within the range (16.8–19.0% DM) reported by Zielewicz and Wróbel [9]. CP concentration in the M. albus silage (14.48% DM) was in agreement with the value of 14.1% DM reported by Howard et al. [63] and higher than the values of 11.8% DM reported by Bozhanska et al. [64], and 11.6% DM by Kintl et al. [39].

The concentration of EE was not influenced (P > 0.05) by the ratio of evaluated crops in the mixtures, and its contribution to energy value was only minor. The NFC parameter, also known as nitrogen-free extract, represents starch, water-soluble carbohydrates, pectins, and organic acids such as LA in the case of silages. NFC concentration in silages from the solitary M. verticillata (38.38% DM) and M. albus (31.45% DM) was significantly different (P < 0.001). This parameter was reported only for M. crispa (34.7% DM) by Ţîţei and Teleuță [11]. Differences in the concentrations of ME3× and NEL3× between silages from the solitary M. verticillata and M. albus were significant (P < 0.01; P < 0.001, respectively) but low. The main reasons for this low difference can be similar nutrient compositions (Table 5) and fibre digestibility (Table 4) of both forage crops, as discussed above in Section 3.2. The concentration of NEL3× in M. verticillata silage (5.75 MJ/kg) ranged from 4.4 to 5.8 MJ/kg after conversion of values for the green matter of M. verticillate, as reported by Bonnemaire et al. [10]; our ME3× concentration (9.31 MJ/kg) was slightly higher than that for M. crispa (8.55 MJ/kg) reported by Ţîţei and Teleuță [11]. In the case of M. albus silage, the concentration of NEL3× in our study (5.45 MJ/kg) was higher than 4.91 MJ/kg after conversion of the value reported by Bozhanska et al. [64]. After comparing the NEL3× concentrations from both evaluated forage crops and their mixtures with the conventional forages, we found out that our values were in the upper limit of the interval from 4.6 to 5.6 MJ/kg for legumes of different ripeness with various maturity listed in NRC [49] feed database.

3.4 Protein fractions

Concentrations of protein fractions and RUP5 of silages from M. verticillata, M. albus, and their mixtures are presented in Table 6. The measured values are very interesting since the expected increase of protein fraction in the silage mixtures (7:3; 1:1 and 3:7) did not occur. The phenomenon might have been related to the date of harvest, as mentioned above.

Table 6

Concentrations of protein fractions and rumen non-degraded protein of silages from fodder mallow, white sweet clover, and their mixtures

Parameter Unit Mixtures (M. verticillata + M. albus) P SEM
1:0 7:3 1:1 3:7 0:1
A1 % CP 10.55ab 14.27a 3.73c 6.19bc 3.56c <0.001 0.98
A2 % CP 46.17c 43.34c 56.37ab 53.22b 59.68a <0.001 1.3
B1 % CP 34.73a 32.85a 29.90ab 31.48a 26.02b 0.004 1.16
B2 % CP 2.69 3.17 2.9 2.34 3.49 0.697 0.59
C % CP 5.86 6.37 7.11 6.76 7.25 0.178 0.4
RUP5 % CP 16.15 17.15 17.9 16.86 18.36 0.086 0.52

A1 = NH3, A2 = soluble true protein, B1 = insoluble true protein, B2 = fibre-bound protein, C = indigestible protein, RUP5 = rumen non-degraded crude protein assuming ruminal passage rate Kp = 5%/h, and SEM = standard error of the mean.

Fraction A1 represents the concentration of NH3 expressed in % CP and is identical to the parameter of NH3 in Table 3. From the nutritional point of view, the A1 fraction is soluble and immediately utilized by rumen bacteria. Its significantly (P < 0.001) higher value (10.55% CP) in the M. verticillata silage as compared with that of M. albus (3.56% CP) is discussed in Section 3.1.

Compared with fraction A1, the soluble fraction A2 as a true protein source (free amino acids, soluble peptides) is used slowly in the rumen. Only in this fraction protein, silages with the addition of M. albus (1:1, 3:7) exhibited increased protein contents when compared with the mono-silage of M. verticillata. Its lower concentration (P < 0.001) in the M. verticillata silage (46.17% CP) in comparison to that of M. albus (59.68% CP) was probably caused by its part shifting to fraction A1 due to the above-mentioned higher degree of proteolysis in the M. verticillata silage with higher moisture content. Another possible explanation is the potentially higher content of CP in the plants of the Fabaceae family [13,16]. It is interesting, however, that this assumption was not confirmed in silage from the solitary M. albus.

Fraction B1, with an intermediate rate of degradation in the rumen, represents a protein with long chains, including the content of cells soluble in water but not in neutral detergent. Its concentration is higher (P < 0.01) in the M. verticillata silage (34.73% CP) than in the M. albus silage (26.02% CP). Fractions B2 (slowly degraded protein associated with the cell wall) and C (assumed to be non-degradable and non-digestible) do not show any significant differences (P > 0.05) between M. verticillata and M. albus. No results with protein fractionation of evaluated forage crops have been reported to date. Regarding the fact that M. albus is a leguminous species, the determined concentration of protein fractions can be compared with values reported for conventional legumes [69]. Nevertheless, it should be realized here that there is a certain limit in the present study, which is the number of experimental plants as the typical conventional plant for the production of feed silage (Zea mays L.) was not tested. According to Kintl et al. [70], Zea mays combined with legume could help to reach increased content of N-substances (proteins) in the silage, which would then favourably reflect in the nutritional value of the feed. Concentrations of fraction A1 reported by Stojanović et al. [69] for alfalfa and red clover silage (8.38; 7.26% CP, respectively) were higher; concentrations of fraction A2 (53.9; 48.8% CP, respectively) were lower; concentrations of fraction B1 (22.2; 21.9% CP, respectively) were also lower; concentrations of fraction B2 (11.5; 16.7% CP, respectively) were higher and concentrations of fraction C (3.99; 5.32% CP, respectively) were lower than our values. Based on the compositions of the mentioned protein fractions, it is possible to approximately predict the nutritional properties of CP; however, the simple final parameter RUP5 is better comparable. In our study, only the tendency (P < 0.1) to higher value of this parameter in the M. albus silage (18.36% CP) was observed as compared with the M. verticillata silage (16.15% CP).

The final concentrations of RUP5 obtained in our experiment were compared with conventional forages listed in the NRC [49] feed database. We confirm that our values were within the interval from 16.5 to 26.7% CP for legumes of different ripeness and DM intake by cattle. Regarding the fact that RUP5 values measured after the end of the ensiling process correspond to the values of conventional feed silages, we can state that the method of preparing the shreddings and the ensiling process are likely to have had no impact on the silage quality. Also, the mentioned values do not indicate any potentially inappropriate agronomic measures (early or poorly executed harvest).

4 Conclusion

The presented study confirms that new alternative crops (M. verticillata and M. albus) can represent alternative sources of biomass for feeding farm animals. However, the measured values should be interpreted with caution as it is just a comparison of pure crops of M. verticillata and M. albus and their mixed culture with no direct comparison with Zea mays. Each of them has a specific biomass composition, and together in the mixed culture, they are a good prerequisite for the production of high-quality silage to be used in the nutrition of farm animals. For example, the silage prepared only from M. albus exhibited a higher content of CP than the silage prepared only from M. verticillata. On the other hand, it should be pointed out that future studies should focus on a direct comparison with other crops, for example, Zea mays monoculture and some legumes. In general, it is possible to state that the addition of M. albus biomass to the silage made of M. verticillata affected the silage qualitative parameters. Thus, the H0 hypothesis was rejected, while the alternative H1 hypothesis was confirmed. The measured data, however, do not indicate that the addition would always have a positive effect, for example, from the viewpoint of increasing the content of proteins in silage. The nutritional value of silages from the two evaluated alternative crops and their mixtures, as represented by the concentrations of NEL3× and RUP5, is comparable to silages from conventional forages (alfalfa, clover). Based on the acquired data, it is possible to state that a mixture of M. verticillata and M. albus can represent a domestic source of CP (N-substances) ensiled in bulky feeds used in the nutrition of dairy cows (cattle), similarly as it is, for example, in alfalfa. It could be supposed that these types of silages must be tested in trials with ruminants where DM intake will be measured. Determination of an optimum ratio of M. verticillata and M. albus biomass in silage is not simple and should be a subject of further research. Until now, the 1:1 ratio appears to be the most balanced compared to nutritional parameters. In such silage, it is, however, necessary to control the content of coumarin, impairing feed silage digestibility. Thus, from a practical point of view, it paves the way for the use of new crops in producing silages to feed farm animals. Moreover, these crops are expected to be better adapted to climate change in the conditions of central Europe than the hitherto used crops such as Zea mays.

  1. Funding information: The results were obtained within the framework of institutional support MZE-RO1725.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and consented to its submission to the journal. All authors have read and agreed to the published version of the manuscript. Conceptualization, A.K. I.H., J.T., M.R., and J.E.; methodology, A.K., I.H., J.T., and J.E.; validation, A.K., I.H., J.S., T.H., O.L., and M.B.; formal analysis, A.K., I.H., J.S., and J.E.; investigation, A.K. I.H., J.T., M.R., and J.E; resources, A.K.; data curation, A.K., I.H., and J.T.; writing – original draft preparation, A.K., I.H., J.T., and J.E.; writing – review and editing, A.K. and J.E.; visualization, A.K., J.S., and J.E.; supervision, A.K.; project administration, A.K.; and funding acquisition, A.K.

  3. Conflict of interest: Authors state no conflict of interest.

  4. Data availability statement: The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Received: 2024-08-20
Revised: 2025-03-05
Accepted: 2025-03-26
Published Online: 2025-05-10

© 2025 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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https://doi.org/10.1515/opag-2025-0435
Keywords for this article
metabolizable energy; net energy for lactation; protein fractions; rumen non-degraded protein
Creative Commons
BY 4.0

Articles in the same Issue

  1. Research Articles
  2. Optimization of sustainable corn–cattle integration in Gorontalo Province using goal programming
  3. Competitiveness of Indonesia’s nutmeg in global market
  4. Toward sustainable bioproducts from lignocellulosic biomass: Influence of chemical pretreatments on liquefied walnut shells
  5. Efficacy of Betaproteobacteria-based insecticides for managing whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae), on cucumber plants
  6. Assessment of nutrition status of pineapple plants during ratoon season using diagnosis and recommendation integrated system
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  10. Antioxidant responses of black glutinous rice to drought and salinity stresses at different growth stages
  11. Differential efficacy of salicylic acid-induced resistance against bacterial blight caused by Xanthomonas oryzae pv. oryzae in rice genotypes
  12. Yield and vegetation index of different maize varieties and nitrogen doses under normal irrigation
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  14. Organizational-economic efficiency of raspberry farming – case study of Kosovo
  15. Application of nitrogen-fixing purple non-sulfur bacteria in improving nitrogen uptake, growth, and yield of rice grown on extremely saline soil under greenhouse conditions
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  21. Metagenomic and metabolomic analyses of bacterial communities in short mackerel (Rastrelliger brachysoma) under storage conditions and inoculation of the histamine-producing bacterium
  22. Fostering women’s engagement in good agricultural practices within oil palm smallholdings: Evaluating the role of partnerships
  23. Increasing nitrogen use efficiency by reducing ammonia and nitrate losses from tomato production in Kabul, Afghanistan
  24. Physiological activities and yield of yacon potato are affected by soil water availability
  25. Vulnerability context due to COVID-19 and El Nino: Case study of poultry farming in South Sulawesi, Indonesia
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