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10.1017/anr.2024.28
Supplementation of VLT and marine-derived probiotic BA-9
promotes the growth performance and antioxidant capacity at early
life of ruminants
Jia Kang1, Jiangjiang Zhu2, Kerui Li2, Junwei Wang2, Kai Zhang1, Yu Chen3, Tao
Luo1 and Hengbo Shi1, 4*
1 Institute of Dairy Science, College of Animal Sciences, Zhejiang University,
Hangzhou 310015, P. R. China
2Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization Key
Laboratory of Sichuan Province, Southwest Minzu University, Chengdu, China,
3 Institute of Nanjiang Yellow Goat Sciences, Bazhong, Sichuan, China
4 Zhejiang Key Laboratory of Cow Genetic Improvement & Milk Quality Research,
Zhejiang University, Hangzhou 310015, P. R. China
*Correspondence: shihengbo@zju.edu.cn;
This is an Open Access article, distributed under the terms of the Creative Commons
Attribution licence (http://creativecommons.org/licenses/by/4.0), which permits
unrestricted re-use, distribution and reproduction, provided the original article is
properly cited.
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ABSTRACT
This study aims to investigates the effects of the vine of Lonicera japonica Thunb
(VLT) and marine-derived Bacillus amyloliquefaciens-9 (BA-9) supplementation on
the growth performance, antioxidant capacity, and gut microbiota of goat kids. A total
of 32 four-week-old kids were randomly assigned into four groups: a control group
(CON), a group supplemented with 0.3% BA-9 (BA-9), a group supplemented with 2%
VLT (VLT), and a group supplemented with both 0.3% BA-9 and 2% VLT (MIX).
The results indicated that VLT supplementation significantly increased both average
daily (P < 0.001) and total weight gain (P < 0.001), while BA-9 alone had no
significant effect (P > 0.05) on the average daily and total weight gain. Biomarker
analysis of oxidative stress revealed that supplementation of VLT or BA-9 alone
enhanced antioxidant capacity. The MIX group showing a higher total antioxidant
capacity (T-AOC) compared with the CON, VLT, and BA-9 groups (P < 0.05).
Plasma albumin levels were significantly increased in the both VLT and BA-9 groups.
Microbiota analysis revealed significant differences in α-diversity and β-diversity
between the MIX and CON groups, with specific genera such as
Prevotellaceae_UCG.004 and Rikenellaceae_RC9_gut_group negatively correlated
with ADG, while Alistipes was positively correlated with T-AOC. These findings
suggest that the combined supplementation of VLT and BA-9 can significantly
enhance growth performance and antioxidant capacity in goat kids by modulating the
composition of gut microbiota and reducing oxidative stress.
KEYWORDS: antioxidant capacity; growth performance; goat kids; probiotic;
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INTRODUCTION
The young animals are born with immature immune and digestive systems,
which makes them highly susceptible to oxidative stress [1]. The incidence of
oxidative stress often leads to decreased growth performance, feed efficiency, and
survival rates in livestock [2, 3]. A healthy and balanced microbiome is particularly
important for the growth, development, and metabolism of ruminants. Specifically,
there has been increasing interest in the early development of the gut or rumen
microbiome in farm livestock as a means of maintaining health [4, 5]. However,
feeding microorganisms alone presents challenges, such as low colonization rates or
weak efficiency [6, 7]. Therefore, it is essential to develop feeding additives that can
improve the health of young animals, maximizing their overall growth performance.
Marine-derived bioactive compounds and probiotics provide a wide range of
health benefits, including antioxidant, immunomodulatory, and anti-inflammatory
effects [8–10]. These compounds have significant potential for applications in health
supplements. Bacillus amyloliquefaciens-9 (BA-9), which is isolated from the
intestinal tract of the white-spotted bamboo shark (Chiloscyllium plagiosum), can
secrete potential antibacterial materials, such as β-1,3-1,4-glucanase and antimicrobial
peptides [11]. Animal feeding experiments have demonstrated that this probiotic
reduces the occurrence of diarrhea in goat kids and decreases oxidative stress in the
mammary gland by altering the diversity of the intestinal microbial community [12,
13]. Combining probiotics with Chinese herbal polysaccharides has been found to
improve the growth performance of lambs and the diversity of rumen bacteria [14].
Recent data combining probiotics with traditional Chinese medicine suggests that this
combination has an enhanced effect in inhibiting intestinal inflammatory responses
and reducing disease recurrence compared to using probiotics or traditional Chinese
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medicine alone [15]. Combined with plant-derived bioactive compounds provides an
ideal strategy to expand the usage of BA-9 in animal feeding.
Lonicera japonica Thunb (LT) is a widely used traditional Chinese herb with
medicinal value attributed to the entire plant. This plant is rich in primary bioactive
components including chlorogenic acid and luteolin glucoside. These components
have antioxidant and immunomodulatory properties [16–18]. Supplementation of LT
extract significantly decreases the respiratory rate in heat-stressed dairy cows and
promote their antioxidant and immune functions [19]. However, the use of LT extract
is challenged by the high costs of extraction, which can increase production expenses
in large-scale applications [20]. Studies suggest that the vine of Lonicera japonica
Thunb (VLT) also exhibits biological activity and can serve as a cost-effective
alternative [21].
BA-9 and LT have demonstrated significant antioxidant and health-promoting
effects when used individually [22–24]. However, the mechanisms underlying their
combined effects remain unclear. BA-9, as a probiotic, may synergize with the
bioactive compounds in VLT, such as flavonoids and phenolic acids, to enhance
antioxidant capacity, modulate gut microbiota, and reduce oxidative stress more
effectively than when used alone [22–24].
This study aims to investigate the dietary addition of VLT and the BA-9 and their
interaction on the growth performance and health status of young ruminants using the
Nanjiang Yellow goats as a model.
2 MATERIALS AND METHODS
2.1 Material Preparation
Bacillus amyloliquefaciens-9 (China General Microbiological Culture Collection
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Center number: 13337, accession number: CP021011) was isolated from the intestinal
tract of the white spotted bamboo shark (C. plagiosum) [25]. The fermentation of
Bacillus-9 was prepared following the reported procedure [26]. The Bacillus-9
powder was prepared using a spray dryer (L-217, 1.0 mm nozzle, Lai Heng, Beijing,
China). The inlet air temperature, aspirator, liquid flow, and compressed spray airflow
were set at 55℃, 2 L/h, and 50 L/h, respectively. Corn starch was used as an adhesion
agent at a ratio of 100 g/L. The colony-forming units in the powder were more than 2
× 109 /g. The production of BA-9 is gifted from Professor Zhengbing Lv in Zhejiang
Sci-tech University.
The dry VLT (produced in Nanjiang County, Sichuan province, China) is ground
using a grinder to obtain fine VLT powder. The powder was then sifted through a
mesh sieve to remove larger particles, ensuring uniform and fine consistency. The
prepared VLT powder was stored in dry, sealed containers.
2.2 Animal Management and Experimental Design
This study was approved by the Experimental Animal Management Committee
of Zhejiang University. A total of 32 healthy suckling goat kids (30 ± 3 days old)
with similar body weights (8.68 ± 0.69 kg) were selected from the Nanjiang Yellow
Goat Breeding Farm in Nanjiang County, Sichuan Province, China. Prior to the
experiment, the health status of all goat kids was confirmed by veterinary examination.
All goat kids were sourced from the same farm to minimize genetic and
environmental variation and were uniformly managed during the pre-experimental
and experimental periods. Weights were recorded before morning feeding.
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During the trial, all goat kids remained in the suckling phase and were not
weaned. Natural nursing by their dams was maintained throughout the study, allowing
each goat kid to consume milk freely according to its needs without any artificial
restrictions. To support growth and gastrointestinal development, a gradually
increasing amount of starter feed (pelleted) was introduced. Starter feed was provided
twice daily at 8:00 AM and 3:00 PM. The specific feeding regimen was as follows:
100 g/day per kid during weeks 1 to 3, increased to 300 g/day per kid during weeks 4
to 5, and further increased to 500 g/day per kid during weeks 6 to 8. Pellets were
evenly distributed at each feeding, and leftover feed was recorded after each meal to
monitor feed intake and assess feeding behavior. Efforts were made to ensure
adequate feed intake while minimizing wastage.
Using a randomized block design, the goat kids were randomly assigned to one
of four groups (n = 8 per group). The groups were as follows: the control group (CON)
fed only the basal diet (pellets, provided by Advanced Feed Co., Ltd., Chengdu,
China); the BA-9 group, fed the basal diet supplemented with 0.3% BA-9; the VLT
group, fed the basal diet supplemented with 2% VLT; and the MIX group, fed the
basal diet supplemented with both 0.3% BA-9 and 2% VLT. To ensure consistency in
nutrient intake, the feed for each group was thoroughly mixed with the respective
supplements and processed into pellets. Water was provided ad libitum, and neck
collars were used during feeding to reduce competition for feed.
The nutrient composition of the basal diet is presented in Supplementary Table
S1. The experiment consisted of a 1-week adaptation period followed by a 7-week
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formal feeding period. Initial body weights were recorded at the start of the trial, and
subsequent body weight measurements were conducted every two weeks in the
morning before feeding to ensure accuracy and consistency.
Health monitoring was conducted daily throughout the study. Any signs of illness,
such as diarrhea, lethargy, or reduced feed intake, were promptly recorded and treated
as necessary. No major health issues were observed during the trial. These
management practices ensured consistent experimental conditions and reliable
outcomes.
2.3 Sample Collection and Analysis
Blood samples were collected from all kids on the last day. Blood was collected
from the posterior jugular vein of each kid from 9:00 a.m. to 10:00 a.m. The plasma
was separated by centrifugation. The plasma levels of malondialdehyde (MDA),
Glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC),
immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M (IgM),
interleukin-2 (IL-2), interleukin-4 (IL-4) and interleukin-6 (IL-6) were measured
using specific commercial kits (Nanjing Jiancheng Biotech, Nanjing, Jiangsu, China)
[26]. Fecal samples were collected at last two days at the end of the experiment. The
homogenized samples were snap-frozen in liquid nitrogen and stored at −80℃ for
subsequent DNA analysis.
2.4 16S rRNA Gene Sequencing
Total genomic DNA was extracted from the fecal samples using a commercial kit
(Tiangen Biotech, Beijing, China). The V3–V4 regions of the bacterial 16S rRNA
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genes were amplified and then paired-end sequenced (2×300 bp) on the Illumina
MiSeq platform following standard protocols (Novogene Technology Co., LTD,
Tianjing, China).
2.5 Sequencing Data Analysis
Raw reads from different samples were demultiplexed and quality-filtered
according to established methods [27]. Bioinformatics analysis was performed using
QIIME 2. Shannon and Chao1 indices were calculated to estimate bacterial richness
and community diversity [28]. Principal Coordinate Analysis (PCoA) and Linear
Discriminant Analysis Effect Size (LEfSe) were conducted using the Novomagic
platform (https://magic.novogene.com). Correlation heatmaps were generated using
the pheatmap package in R studio. Weighted Gene Co-expression Network Analysis
(WGCNA) was performed using the WGCNA package in R studio, and the resulting
network was visualized using Cytoscape (Version 3.8.0).
2.6 Statistical Analysis
The data of growth performance was analyzed using a one-way ANOVA with the
SPSS software (SPSS v.19, SPSS Inc., Chicago, IL, USA). The statistical analyses of
various factors in plasma and microbiol diversity were performed by a one-way
ANOVA. Data are presented as means plus SEM. P < 0.05 was considered
statistically significant. Correlation networks were generated using Spearman’s rank
correlation coefficients and visualized using the Cytoscape. The significant
correlation between bacterial genus and the immune globulins and cytokines was
considered when |R| > 0.2 and P < 0.05).
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3 RESULT
3.1 Supplementation of VLT and BA-9 significantly promotes the daily weight
gain of goat kids
To investigate the effects of VLT and BA-9 on the growth performance of kids,
we first assessed their impact on weight gain. As shown in Table 1, the initial body
weight (IBW) of the kids did not differ significantly among the four groups, but there
was a trend towards differences in final body weight (FBW) (P = 0.094). Compared
with the CON group, feeding goat kids with VLT alone significantly increased their
average daily gain (ADG) (P < 0.001) and total weight gain (TWG) (P < 0.001). No
significant changes were observed in the BA-9 group. Interestingly, the combined
VLT powder and BA-9 (MIX group) outperformed the individual additions (P <
0.001). Compared to the CON group, feeding BA-9 alone did not significantly
increase TWG or ADG. However, goat kids in the VLT group showed a significant
increase in ADG (P = 0.007), which was further enhanced in the MIX group.
Additionally, goat kids in the MIX group had higher average daily weight gain
compared to both the CON group (P < 0.001) and BA-9 group (P = 0.014).
Furthermore, there was an increasing trend in ADG compared to the VLT group (P =
0.081).
3.2 VLT and BA-9 alter biomarkers in oxidative status and nutrition metabolism
To further investigate the physiological and metabolic mechanisms responsible
for the increased daily weight gain in goat kids with the supplementation of VLT,
BA-9, or their combination, we examined biomarkers associated with oxidative status
and nutrition metabolism. As shown in Table 2, MDA (P = 0.504) and GSH-Px (P =
0.353) activities did not differ significantly among the different treatment groups.
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However, the combined supplementation significantly increased the T-AOC of the
kids (P = 0.039). The single supplementation of VLT (P = 0.083) or BA-9 (P = 0.084)
had minor effects on T-AOC. The MIX group exhibited a significantly higher T-AOC
to the CON group (P = 0.021), the BA-9 group (P = 0.022), and the VLT group (P =
0.012). There were no significant changes in triglyceride (P = 0.387) and glucose (P =
0.661) among the different treatment groups.
3.3 VLT and BA-9 alter biomarkers in immune response
Compared with the CON group, BA-9 (P = 0.004) and VLT (P = 0.035)
significantly increased the plasma albumin (ALB) level. There was an observed
increasing trend in the MIX group compared with the CON group (P = 0.081). There
were no significant changes in IgA (P = 0.115), IgG (P = 0.387), IgM (P = 0.257),
IL2 (P = 0.950), IL4 (P = 0.446), and IL6 (P = 0.880).
3.4 Supplementation of VLT and BA-9 decreases the diversity indices of fecal
microbiota
To investigate the combined effects of VLT powder and BA-9 in goat kids, the
fecal microbiota was assessed using 16S rRNA sequencing. The amplicon sequencing
data were assessed using rarefaction curves. These curves reached a plateau as the
sample size increased, indicating that the species distribution within the samples was
even and that we had sufficient sequencing depth to cover the major species (Figure
S1). This outcome provides a reliable basis for our data and ensures the accuracy of
subsequent analyses. We classified all fecal microbiota into 6214 ASVs through
amplicon sequencing of the 16S rRNA gene. The Venn diagram illustrates that there
were 552 unique ASVs in the CON group, 384 in the BA-9 group, 1072 in the VLT
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group, and 1071 in the MIX group, with 1173 ASVs shared among all four groups
(Figure 1A). We used the Chao1, Simpson, and Shannon indices to examine the
α-diversity of the fecal microbiota (Figure 1B-D). Compared with the CON group, the
MIX group showed significantly decreased Chao1 (P < 0.001), Simpson (P < 0.001),
and Shannon (P < 0.001) indices, indicating a lower diversity of microbial species
(Figure 1B-D). We also found significant differences in α-diversity indices between
the MIX group and the BA-9 and VLT groups for these indices (P < 0.001). There
were no significant differences in Chao1, Simpson and Shannon indices when the diet
was supplemented with BA-9 or VLT alone compared to the CON group. PCoA based
on unweighted UniFrac distances showed distinct clustering of samples based on the
different treatments (PCoA1 = 28.92%, PCoA2 = 9.93%) (Figure 1E).
3.5 Comparison of fecal microbial profiling between CON and MIX groups
To further explore the combined effects of combination VLT and BA-9 on kids,
the microbiota profiling between the MIX and CON groups was compared. At the
phylum level, the MIX group showed higher abundances of Verrucomicrobiota (P =
0.007) and Patescibacteria (P = 0.037) compared to the CON group (Figure 2A).
Additionally, the MIX group had lower abundances of Campylobacterota (P = 0.029)
and Desulfobacterota compared to the CON group (P = 0.008). At the genus level, the
abundances of Akkermansia (P = 0.006), Candidatus_Stoquefichus (P = 0.008),
Faecalibacterium (P = 0.048), Candidatus_Saccharimonas (P = 0.038) and UBA1819
(P = 0.004) were significantly increased in the MIX group. Conversely, the
abundances of Rikenellaceae_RC9_gut_group (P = 0.001), Campylobacter (P =
0.035), Turicibacter (P = 0.009), Desulfovibrio (P = 0.005), NK4A214 group (P =
0.019), Clostridium_sensu_stricto_1 (P = 0.008), Ruminococcus (P = 0.009),
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Romboutsia (P = 0.011), and Defluviitaleaceae_UCG_011 (P = 0.030) were
significantly decreased in MIX group compared to the CON group (Figure 2B). At the
species level, the MIX group had higher abundances in Bacteroides_massiliensis (P =
0.007), Akkermansia_muciniphila (P = 0.022), Alistipes_finegoldii (P = 0.011),
Parabacteroides_merdae (P = 0.029) and Bacteroides_caecigallinarum (P = 0.010)
but a lower level of Romboutsia_sp_DR1 (P = 0.011) compared to the CON group
(Figure 2C). Furthermore, the LEfSe further identified differential abundances in
bacterial taxa between the MIX and CON groups. The MIX group was enriched with
o_Pseudomonadales, f_Moraxellaceae, g_Acinetobacter, g_Bacteroides,
f_Bacteroidaceae, and f_Muribaculaceae, while the CON group was enriched with
f_Barnesiellaceae, g_Prevotellaceae_UCG_004, g_Rikenellaceae_RC9_gut_group,
and f_Rikenellaceae in When LDA was greater than 4. (Figure 2D-E).
3.6 Hub microbiota correlated with growth performance and antioxidant
capacity
Spearman correlation analysis was conducted to determine the correlation
between fecal microbiota and ADG, T-AOC and ALB at the genus level (Figure 3A).
The results showed that three genera in feces including Prevotellaceae_UCG_004 (P
< 0.01, R = -0.77), UCG_005 (P < 0.01, R = -0.53) and
Rikenellaceae_RC9_gut_group (P < 0.01, R = -0.58) were negatively correlated with
ADG. Alistipes was positively correlated with T-AOC (P < 0.01, R = 0.76), while
Blautia was negatively correlated with T-AOC (P = 0.03, R = -0.44).
The correlation between ALB, T-AOC or ADG and the microbial abundance at
the genus level was further investigated using WGCNA. A total of six microbiota
modules were identified (Figure 3B). The MEblue was significantly associated with
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ADG (P = 0.035, R = -0.399), the MEyellow was significantly associated with ADG
(P = 0.02, R = -0.437) and MEyellow was significantly associated with T-AOC (P =
0.043, R = -0.385). The network derived from the MEblue module indicated that
o_Bacteroidales_RF16_group, g_Clostridium_sensu_stricto_1,
f_Peptostreptococcaceae and g_Romboutsia were the hub microbiota within the
MEblue module (Figure 3C). The network derived from the MEyellow module
indicated that g_Desulfovibrio, Spirochaetes_bacterium_GWE2_31_10, and
Lachnospiraceae_NK4B4_group were the hub microbiota within the MEyellow
module (Figure 3C).
4 DISUSSION
Young livestock are highly susceptible to various diseases because of their
immature antioxidant and immune systems, which leads to a low rate of growth
performance [29–31]. This study aims to optimize rearing strategies for goat kids by
promoting the status of their antioxidant and immune systems through the addition of
BA-9 and VLT. Our data found that VLT and BA-9 exhibited significant synergistic
effects in promoting the growth performance of goat kids by enhancing the total
antioxidant capacity. Furthermore, the association analysis demonstrated that
combined administration of VLT and BA-9 modulates the fecal microbial community
of kids, thereby optimizing their physiological functions and health status.
Young kids, due to their immature digestive and immune systems, are susceptible
to environmental changes, often resulting in a low average daily weight gain [12]. The
increase in average daily weight gain in the VLT group is consistent with previous
data in Saanen Kids [31]. It is worth noting that when both VLT and BA-9 were added
to the kids’ feed, the weight gain effect was superior to that of adding either alone.
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This indicates a synergistic effect in promoting the growth of kids. One possible
reason for this is that chlorogenic acid in Lonicera japonica Thunb increases
transepithelial electrical resistance and reducing horseradish peroxidase flux, which
shows potential in repairing the intestinal barrier and maintaining intestinal health
[32]. Moreover, BA-9 derived from marine organisms contains various antimicrobial
peptides [13]. These peptides can enhance the bioavailability of Lonicera japonica
Thunb by altering the metabolic functions of the gut microbiota or directly improving
the overall function of the intestine in young goats. This idea of a synergistic
interaction between VLT and BA-9 is further supported by the effects of VLT in
promoting the growth of kids. Our data indicate that adding both VLT and BA-9
provides an ideal strategy for promoting the health of young goats.
Our study found that adding VLT and BA-9 to the diet did not impact total
protein, triglycerides or blood glucose levels, indicating that these additives do not
influence kids’ nutritional metabolism. However, when VLT and BA-9 were added
simultaneously, there was a significant increase in plasma ALB levels in goat kids.
This suggests that there is a synergistic effect between these two additives, which may
involve complex interactions between intestinal immune regulatory cells and immune
signaling molecules. This provides new insights into the regulation of immune
function in goat kids [32].
During nutritional metabolism, an amount of reactive oxygen species (ROS) are
produced, which can damage cells and tissues and cause oxidative status [34]. T-AOC
(total antioxidant capacity) and GSH-Px activity are key markers used to assess the
antioxidant capacity [34]. The current study showed that adding VLT or BA-9 alone
did not affect the antioxidant capacity of goat kids. However, when VLT and BA-9
were fed together, there was a significant increase in total antioxidant capacity of kids.
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This suggests that this combination has an antioxidant activity. VLT and BA-9 may
interact to enhance the absorption and utilization of antioxidant components in
Lonicera japonica Thunb, thereby further enhancing its antioxidant effects.
The observation of a positive correlation between Alistipes and T-AOC agrees
with the fact that Alistipes improves gut health and enhances the host's antioxidant
capacity through its metabolic products, such as short-chain fatty acids [35]. WGCNA
revealed microbiome modules significantly associated with ADW and T-AOC,
particularly the MEblue and MEyellow modules. The central species within these
modules, such as Clostridium_sensu_stricto_1 and Desulfovibrio, may play crucial
roles in influencing the growth performance and antioxidant status of kids by
modulating the gut microenvironment and affecting host nutrient absorption and
energy metabolism. The activity of Desulfovibrio, related to sulfate reduction, impacts
the redox state of the gut, thereby influencing the host's antioxidant capacity [36]. The
correlation between the microbiota and antioxidants provides an insight into the
interactions between microbiota and growth performance as well as antioxidant status
in kids. Future research should further explore the specific mechanisms of action of
VLT and BA-9, particularly how they modulate the composition and metabolic
activities of the gut microbiota. These data may provide evidence for
microbiome-based interventions to optimize livestock growth performance during the
early stage of life.
5 CONCLUSION
Developing additives that improve the health of young animals can maximize
their overall growth performance and productivity throughout their lives. In the
current study, our data firstly underscore the significant improvement in growth
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performance and antioxidant capacity of goat kids through the combined
supplementation of VLT and marine-derived BA-9. This synergistic effect is likely
attributed to the modulation of the gut microbiota, as evidenced by significant
differences in microbial diversity and the presence of specific bacterial genera that are
correlated with growth and antioxidant indices. These findings highlight the potential
of using these novel feed additives to enhance the health and productivity of young
ruminants, providing a promising strategy for improving livestock rearing practices.
Further research should explore the underlying mechanisms of this synergistic
interaction to optimize its application in animal husbandry.
Data availability statement
The datasets presented in this study were deposited in the NCBI Sequence Read
Archive (SRA) under the accession PRJNA1143899.
Author contributions
JK: data curation, and writing—original draft. JZ, KL, JW, TL, YC, TL and BM:
data curation. HS, JL and JZ: funding acquisition and supervision, and editing draft.
All authors contributed to the article and approved the submitted version.
Funding
This study was jointly supported by Key R&D program of Zhejiang Province
(2022C04017 and 2021C02068-6).
Acknowledgments
The authors thank the owners and staff of Nanjiang Yellow Goat Original
Breeding Farm (Bazhong, China) for allowing the use of their goats in this experiment
and their kind help for the sample collection.
Conflict of interest
The authors declare there are no conflicts of interest.
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Table 1. Effects of BA-9 and VLT on the growth performance of Nanjiang Yellow Goat kids
Items2
Groups
SEM
p Value
CON3
BA-93
VLT3
MIX3
IBW, kg
8.26±1.46
8.80±1.94
7.85±5.37
9.79±5.67
0.005
0.311
FBW, kg
10.29±2.19
11.34±4.16
10.9±5.12
13.28±1.94
0.009
0.094
TWG, kg/hd
2.03±4.68c
2.54±5.02bc
3.05±3.89ab
3.49±5.93a
0.007
<0.001
ADG, g
65.32±2.64c
81.86±2.90bc
98.39±4.84ab
112.50±2.42a
0.004
<0.001
Notes：
1 The data are expressed as the mean±SEM, n = 8 per group .
a,b,c Means without a common superscript differ significantly between the two groups at the same time point (P < 0.05).
2 IBW, initial body weight (weight of a seven-day-old lamb); FBW, final body weight (weight of a fifty-day-old lamb);
TWG, total weight gain (the difference between FBW and IBW); ADG, average daily gain.
3 CON, control group; BA-9, BA-9 probiotic supplementation group; VLT, VLT powder supplementation group; MIX,
combined supplementation group of both VLT powder and BA-9 probiotic.
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Table 2. Effects of BA-9 and VLT on indicators of oxidative status and biochemical indices in
Nanjiang Yellow Goat kids
Items2
Groups
SEM
p
Value
CON4
BA-94
VLT4
MIX4
Oxidative Stress2
GSH-Px, U/ml
384.49±3.82
420.61±5.42
363.06±1.95
382.65±2.36
11.99
0.353
MDA, nmol/ml
2.99±2.71
3.20±1.83
3.48±5.02
3.12±6.84
0.21
0.504
T-AOC, mmol/L
2.20±5.75b
2.20±2.76b
2.18±1.74b
2.40±3.35a
0.05
0.039
Nutrition Metabolism3
TG, mmol/L
0.93±2.88
0.77±0.73
0.86±1.36
0.75±0.32
0.04
0.387
GLU, mmol/L
5.02±1.41
5.21±1.93
4.93±2.21
5.09±2.52
0.06
0.661
TP, g/L
69.70±2.05
71.46±1.11
67.77±2.60
76.51±5.81
1.87
0.084
Notes:
1 The data are expressed as the mean±SEM, n = 8 per group.
a,b,c Means without a common superscript differ significantly between the two groups at the same time point (P < 0.05).
2 GSH-Px, glutathione peroxidase; MDA, malondialdehyde; T-AOC, total antioxidant capacity.
3 TG, triglyceride; GLU, glucose; TP, totol protein.
4 CON, control group; BA-9, BA-9 probiotic supplementation group; VLT, VLT powder supplementation group; MIX,
combined supplementation group of both VLT powder and BA-9 probiotic.
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Table 3. Effects of BA-9 and VLT on immune indices of Nanjiang Yellow Goat kids
Items2
Groups
SEM
p
Value
CON3
BA-93
VLT3
MIX3
IgA, mg/mL
2.35±0.19
2.38±5.33
2.25±0.45
1.91±2.66
0.11
0.115
IgG, mg/mL
10.89±7.50
10.55±4.41
9.49±1.53
9.90±3.30
0.32
0.387
IgM, mg/mL
3.00±1.14
2.82±1.08
2.69±4.93
2.45±2.31
0.12
0.257
IL2, pg/mL
47.07±3.27
48.30±0.05
49.87±3.52
49.67±3.26
0.67
0.950
IL4, pg/mL
79.02±1.88
68.81±0.52
75.35±3.23
71.96±0.89
2.20
0.446
IL6, pg/mL
104.21±6.92
102.59±4.91
98.67±3.48
98.85±3.76
1.38
0.880
ALB, g/L
25.34±4.57c
28.56±5.78a
27.63±5.50ab
26.14±4.23b
0.72
0.019
Notes:
1 The data are expressed as the mean±SEM, n = 8 per group.
a,b,c Means without a common superscript differ significantly between the two groups at the same time point (P < 0.05).
2 ALB, Albumin.
3 CON, control group; BA-9, BA-9 probiotic supplementation group; VLT, VLT powder supplementation group; MIX,
combined supplementation group of both VLT powder and BA-9 probiotic.
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Figure 1
Figure 1. Diversity analysis of fecal microbiota in Nanjiang Yellow Goat kids. (A)
Venn diagram of OTUs. (B) Chao 1 index comparing α-diversity among CON, BA-9,
VLT and MIX treatment groups. (C-D) Simpson index and Shannon index further
confirm significant differences in α-diversity among the groups. (E) PCoA analysis
demonstrating intergroup differences in β-diversity. * P < 0.05, ** P < 0.01, *** P <
0.001, **** P < 0.0001.
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Figure 2
Figure 2 Analysis of the microbial community structure in the CON and MIX groups.
(A) Displays the differential bacteria at the phylum level. (B) Displays the differential
bacteria at the genus level. (C) Displays the differential bacteria at the species level.
(D) Shows the key differential bacteria through Linear Discriminant Analysis (LDA)
score plot. (E) The partial phylogenetic tree (Cladogram) reveals the phylogenetic
relationships of various microbial categories and their changes in different treatment
groups.
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Figure 3
Figure 3 (A) Correlation between environmental factors and fecal bacteria at the
genus level, with * P < 0.05 and ** P < 0.01. Weighted Gene Co-Expression Network
Analysis (WGCNA) was used to analyze the related modules. Correlation networks
were generated using Spearman's rank correlation coefficient. (B) Heatmap of the
WGCNA modules. (C) Interaction network and hub microbes in the MEblue and
MEyellow modules.
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