Exploring the Rumen Microbiota and Serum Metabolite Profile of Hainan Black Goats with Different Body Weights before Weaning.

The critical role of the rumen microbiota in the growth performance of livestock is recognized, yet its significance in determining the body weight of goat kids before weaning remains less understood. To bridge this gap, our study delved into the rumen microbiota, serum metabolome, rumen fermentation, and rumen development in goat kids with contrasting body weights before weaning. We selected 10 goat kids from a cohort of 100, categorized into low body weight (LBW, 5.56 ± 0.98 kg) and high body weight (HBW, 9.51 ± 1.01 kg) groups. The study involved sampling rumen contents, tissues, and serum from these animals. Our findings showed that the HBW goat kids showed significant enrichment of VFA-producing bacteria, particularly microbiota taxa within the Prevotellaceae genera (UCG-001, UCG-003, and UCG-004) and the Prevotella genus. This enrichment correlated with elevated acetate and butyrate levels, positively influencing rumen papillae development. Additionally, it was associated with elevated serum levels of glucose, total cholesterol, and triglycerides. The serum metabonomic analysis revealed marked differences in fatty acid metabolism between the LBW and HBW groups, particularly in encompassing oleic acid and both long-chain saturated and polyunsaturated fatty acids. Further correlational analysis underscored a significant positive association between Prevotellaceae_UCG-001 and specific lipids, such as phosphatidylcholine (PC) (22:5/18:3) and PC (20:3/20:1) (r > 0.60, p < 0.05). In summary, this study underscores the pivotal role of the rumen microbiota in goat kids' weight and its correlation with specific serum metabolites. These insights could pave the way for innovative strategies aimed at improving animal body weight through targeted modulation of the rumen microbiota.

Comprehensive profiling of the metabolome in corn silage inoculated with or without Lactiplantibacillus plantarum using different untargeted metabolomics analyses.

Silage fermentation is a complicated biochemical process involving interactions between microbes and metabolites. However, the overall metabolome feature of ensiled forage and its response to lactic acid bacteria inoculation is poorly understood. Hence, in this study metabolome profiles of whole-plant corn silage inoculated with or without Lactiplantibacillus plantarum were characterised via solid-phase microextraction/gas chromatography/mass spectrometry (SPME-GC-MS), gas chromatography/time-of-flight mass spectrometry (GC-TOF-MS), and Liquid chromatography/Q Exactive HFX mass spectrometry (LC-QE-MS/MS) analysis. There were 2087 identified metabolites including 1143 reliably identified metabolites in fresh and ensiled whole-plant corn. After ensiling, the increased metabolites in whole-plant corn were mainly composed of organic acids, volatile organic compounds (VOC), benzene and substituted derivatives, carboxylic acids and derivatives, fatty acyls, flavonoids, indoles and derivatives, organooxygen compounds (including amines and amides), phenols, pyridines and derivatives, and steroids and steroid derivatives, which includes neurotransmitters and metabolites with aromatic, antioxidant, anti-inflammatory, and antimicrobial activities. Phenylacetaldehyde was the most abundant aromatic metabolite after ensiling. L-isoleucine and oxoproline were the major free amino acids in silage. Ensiling markedly increased the relative abundances of 3-phenyllactic acid, chrysoeriol, 6-O-acetylaustroinulin, acetylcholine, γ-aminobutyric acid, pyridoxine, and alpha-linoleic acid. Inoculation with L. plantarum remarkably changed silage VOC composition, and essential amino acids, 3-phenyllactic acid, and cinnamaldehyde compared with untreated silage. The present study does not only provide a deeper insight into metabolites of the ensiled whole-plant corn but also reveals metabolites with specific biological functions that could be much helpful in screening novel lactic acid bacteria to well ensile forages. Inoculation with L. plantarum significantly affects the metabolome in ensiled whole-plant corn.

Effects of nisin on bacterial community and fermentation profiles, in vitro rumen fermentation, microbiota, and methane emission of alfalfa silage.

BACKGROUND: Alfalfa (Medicago sativa L.) has been used widely in preparing silage. However, forage legumes are prone to contamination by spoilage bacteria during fermentation. Nisin has broad-spectrum antibacterial properties and has been applied as an inhibitor of rumen methane emissions. However, little research has been carried out on the application of nisin in silage. This study therefore aimed to investigate the impacts of different nisin concentrations on the bacterial community and fermentation dynamics, in vitro ruminal fermentation characteristics, microbiota, and methane emissions of alfalfa silage. RESULTS: The detection limits of organic acid in nisin-treated silages were not reached in 0.09 g kg-1 nisin (0.09 level) from days 1 to 7 of ensiling. With increasing nisin concentrations, the silage dry matter increased linearly (P < 0.05), and dry matter loss decreased linearly (P < 0.05). Moreover, both the 0.06 g kg-1 nisin (0.06 level) and 0.09 level treatments increased the relative abundance of Pediococcus acidilactici during ensiling. Concurrently, as the nisin concentrations increased, ruminal methane production decreased linearly (P < 0.05), while the relative abundances of ruminal Succinivibrio, Fibrobacter succinogenes and Ruminobacter amylophilus increased linearly (P < 0.05). The populations of ruminal total bacteria, methanogens, protozoa, and fungi decreased linearly with increasing nisin concentrations (P < 0.05). CONCLUSION: The addition of nisin delayed the fermentation process, preserved more nutrients in alfalfa silage, and promoted fermentation dominated by P. acidilactici in the late phase of ensiling. Moreover, nisin reduced in vitro rumen methane emissions without adverse effects on dry matter digestibility. © 2023 Society of Chemical Industry.

Lignocellulose conversion of ensiled Caragana korshinskii Kom. facilitated by Pediococcus acidilactici and cellulases.

To explore the biofuel production potential of Caragana korshinskii Kom., Pediococcus acidilactici and an exogenous fibrolytic enzyme were employed to investigate the fermentation profile, structural carbohydrates degradation, enzymatic saccharification and the dynamics of bacterial community of C. korshinskii silage. After 60 d of ensiling, all additives increased the fermentation quality. The highest lactic and acetic acids and lowest non-protein nitrogen (NPN) and ammonia nitrogen (NH3 -N) were observed in P. acidilactici and Acremonium cellulase (PA + AC) treated silage. Additionally, all additives significantly increased the ferulic acid content and fibre degradability with the highest values obtained from PA + AC silage. The bacterial community in all silages was dominated by P. acidilactici throughout the entire fermentation process. The bacterial community was also modified by the silage additives exhibiting a relatively simple network of bacterial interaction characterized by a lower bacterial diversity in P. acidilactici (PA) treated silage. The highest 6-phospho-beta-glucosidase abundance was observed in PA-treated silage at the mid-later stage of ensiling. PA treatment exhibited lower structural carbohydrates degradation but performed better in lignocellulose conversion during enzymatic saccharification. These results indicated that pretreating C. korshinskii improved its silage quality and potential use as a lignocellulosic feedstock for the production of bio-product and biofuel.

Storage Temperature Is More Effective Than Lactic Acid Bacteria Inoculations in Manipulating Fermentation and Bacterial Community Diversity, Co-Occurrence and Functionality of the Whole-Plant Corn Silage.

The objective of this study was to investigate effects of different lactic acid bacteria (LAB) on the fermentation process of whole-plant corn silage stored at different temperatures based on bacterial community successions, interaction networks, and predicted functions. Before ensiling, whole-plant corn was inoculated with L. plantarum (LP) or L. buchneri (LB) and the silage bags were stored at 20 or 30°C, and sampled after 0.5, 1, 3, 7, 14, and 60 d of ensiling. The higher abundances of Leuconostoc, Pedicoccus and Weissella were observed in silage stored at 30°C after 12 h of ensiling, thereby rapidly decreased pH to about 4.5. According to meta-network analysis, the bacterial communities were more sensitive to storage temperature than LAB inoculants during whole-plant corn ensiling. Species of Lactobacillus and Weissella were sensitive to 30°C, while Leuconostoc species were sensitive to 20°C in whole-plant corn silage. The storage temperature of 30°C decreased bacterial diversity and network complexity of whole-plant corn silage compared with 20°C. Additionally, LP inoculation changed the bacterial community successions during the early and middle ensiling periods, while LB inoculation affected bacterial community successions in the later stage of ensiling. The metabolic pathways of bacterial community were totally different in LB-inoculated silage from that in control and LP-inoculated silage. As the bacterial compositions became simple along with the ensiling process, the functional structure of bacterial community became simplified as well. In general, the storage temperature had a greater impact on the fermentation characteristics, bacterial community and predicted function of whole-plant corn silage compared with LAB inoculations. IMPORTANCE Increased understanding of effects of regulation measures on whole-plant corn silage is important from bacterial community succession, interaction network and predicted functions. According to alpha diversity and meta co-occurrence network, the bacterial communities were more sensitive to storage temperature than LAB inoculants during whole-plant corn ensiling. The storage temperature of 30°C decreased bacterial diversity and network complexity of whole-plant corn silage compared with 20°C. In addition, 30°C promoted the initiation of LP and LB inoculants, and 20°C was conducive to the long-term growth of LP and LB inoculants. According to the changes of bacterial community and predicated functions, it was further confirmed that the effect of LB inoculation was more obvious on whole-plant corn silage.

Microbial mechanisms of using feruloyl esterase-producing Lactobacillus plantarum A1 and grape pomace to improve fermentation quality and mitigate ruminal methane emission of ensiled alfalfa for cleaner animal production.

This study was conducted to investigate the influence of feruloyl esterase-producing Lactobacillus plantarum A1 (Lp A1) and grape pomace (GP) alone, or in combination (LG) on ensiling characteristics and bacterial community, in vitro ruminal fermentation, methane (CH4) emission, and the microbiota of ensiled alfalfa. Alfalfa at 42% dry matter (DM) was treated in a 2 × 2 factorial design: with the application of Lp A1 at 0 (control) or 1 × 106 cfu/g of fresh forage, and GP at 0 or 5% of fresh forage. After 60 d of ensiling, a decrease in nonprotein nitrogen (NPN) was observed in GP treated silage. Lp A1 inoculated silage had a lower fiber content than silages without Lp A1. The lowest NPN was found in silage treated with LG, and an obvious increase in the relative abundance of Lactobacillus paracasei was detected in silages treated with Lp A1 and LG, respectively. In vitro ruminal experiments indicated that, although the application of GP deceased ruminal total gas, CH4 production, nitrogen degradation and the number of methanogenic archaea in alfalfa silage, it also reduced silage DM digestibility. In contrast, inoculation with Lp A1 not only increased DM digestibility and populations of ruminal Ruminococcus flavefaciens and fungi, but also improved ruminal total gas and CH4 production. As expected, LG treatment decreased alfalfa silage ruminal total gas and CH4 production relative to Lp A1 treatment alone, and increased silage DM digestibility compared with GP treated silage. In conclusion, the application of LG before ensiling alfalfa, balanced silage proteolysis, feed digestibility, and CH4 emission, and could be a promising strategy for using food industry by-products to produce a nutritional and environmentally-friendly legume silage that will mitigate N and greenhouse gas emissions from ruminants.

Effect of Bacillus amyloliquefaciens and Bacillus subtilis on fermentation, dynamics of bacterial community and their functional shifts of whole-plant corn silage.

BACKGROUND: Bacillus amyloliquefaciens (BA) and Bacillus subtilis (BS) are usually used as feed supplements directly or bacterial inoculants in biological feeds for animals. However, few research have reported the effects of BA and BS on fermentation characteristics and bacterial community successions of whole-plant corn silage during ensiling. If the BA and BS inoculants have positive effects on silages, then they could not only improve fermentation characteristics, but also deliver BA or BS viable cells to ruminants, which would play its probiotic effect. Therefore, the objectives of this study were to investigate the effects of BA and BS on the fermentation, chemical characteristics, bacterial community and their metabolic pathway of whole-plant corn silage. RESULTS: Freshly chopped whole-plant corn was inoculated without or with BA and BS, respectively, and ensiled for 1, 3, 7, 14 and 60 d. Results showed that BA and BS inoculations increased lactic acid concentrations of whole-plant corn silages compared with control, and BA inoculation decreased acetic acid concentrations, whereas BS inoculation decreased fiber contents and increased crude protein (CP) content. Higher water-soluble carbohydrate contents and lower starch contents were observed in BA- and BS-inoculated silages compared with that in control. The decreased CP content and increased non-protein nitrogen content were observed in BA-inoculated silage, which was consistent with the higher amino acid metabolism abundances observed in BA-inoculated silage. In addition, it was noteworthy that BA and BS inoculations increased the metabolism of cofactors and vitamins, and decreased the relative abundances of drug resistance: antimicrobial pathways. We also found that the bacterial metabolism pathways were clearly separated into three clusters based on the ensiling times of whole-plant corn silage in the present study. There were no significant differences in bacterial community compositions among the three groups during ensiling. However, BA and BS inoculations decreased the relative abundances of undesirable bacteria such as Acetobacter and Acinetobacter. CONCLUSION: Our findings suggested that the BS strain was more suitable as silage inoculants than the BA strain in whole-plant corn silage in this study.

Erlotinib combined with bevacizumab and chemotherapy in first line osimertinib-resistant NSCLC patient with leptomeningeal metastasis: A case report.

RATIONALE: Leptomeningeal metastasis (LM) is a fatal complication of advanced non-small cell lung cancer (NSCLC) with a poor prognosis. Osimertinib is a promising option for NSCLC with LM harboring epidermal growth factor receptor (EGFR) mutation. However, therapeutic approaches remain a challenge for osimertinib resistant NSCLCs with LM. Although studies have reported that the first/second-generation EGFR-tyrosine kinase inhibitors were active against osimertinib-resistant NSCLC with EGFR C797S and sensitive mutation (SM), the resistance inevitably occurred due to the development of the EGFR SM/C797S/T790M triple mutations. PATIENT CONCERNS: A 48-year-old woman was diagnosed with stage IV lung adenocarcinoma harboring the EGFR mutation in the combination of chest computed tomography, biopsy and amplification refractory mutation system-polymerase chain. One year and a half after oral administration of osimertinib, the patient progressed to extensive LM. DIAGNOSES: Magnetic resonance images of the brain showed extensive LM. Exfoliated tumor cells from cerebrospinal fluid (CSF) were positive detected by lumbar puncture and the cytology examination. EGFR mutations (exon19 E746_T751delinsI and exon20 C797S) in CSF circulating tumor DNA were detected by next-generation sequencing (NGS). INTERVENTIONS: Pemetrexed (800 mg day 1), cis-platinum (40 mg day 1-3) combined with bevacizumab (400 mg day 1) every 3 weeks were administered to the patient. After 1 cycle, due to optic nerve invasion, erlotinib was applied 150 mg/d combined with previous regimen. The patient continued erlotinib monotherapy after 6 cycles. OUTCOMES: After LM, erlotinib combined with pemetrexed, cis-platinum and bevacizumab were administered to the patient for 4.25 months based on the CSF NGS. Then, the patient continued erlotinib monotherapy and appeared disease progression after 10 months. The overall survival is 35 months. LESSONS: LM is a fatal complication of advanced NSCLC with a poor prognosis. NGS profiling of CSF circulating tumor DNA is important in NSCLC patients with LM and erotinib plus bevacizumab and chemotherapy is a promising option for patients with LM harboring EGFR C797S/SM.

Effects of Replacing Ensiled-Alfalfa with Fresh-Alfalfa on Dynamic Fermentation Characteristics, Chemical Compositions, and Protein Fractions in Fermented Total Mixed Ration with Different Additives.

Alfalfa (Medicago sativa) is one of the high protein ingredients of fermented total mixed ration (FTMR). Additionally, FTMR is widely used to satisfy the nutrition requirements of animals. This study was conducted to confirm the fermentation characteristics, chemical compositions and protein fractions changes when replacing ensiled-alfalfa with fresh-alfalfa in FTMR with additives. Three additives were separately applied to fresh-alfalfa total mixed ration (TMR) and ensiled-alfalfa TMR, including molasses (MOL), Lactobacillus plantarum (LP) and MOL plus LP (MOL+LP). The same volume of distilled water was sprayed onto the prepared TMR as performed for the control (CK). Each treatment included 18 repetitions and opened 3 repetitions at each fermenting day (1, 3, 7, 15, 30 and 60 d). The results showed that fresh-alfalfa FTMR (F-FTMR) exhibited slight changes in the fermentation characteristics during the first 7 d and showed similar trends in terms of the pH and organic acids content to ensiled-alfalfa FTMR (E-FTMR). The lactic acid contents of F-FTMR were significantly lower than those of E-FTMR at 60 d fermentation and the ammonia nitrogen contents were lower than E-FTMR during the entire fermenting period. The crude protein of the F-FTMR was enhanced after 60 d of fermenting. F-FTMR supplemented with MOL+LP exhibited a lower nonprotein nitrogen content, variable to slow protein and indigestible protein contents, and higher fast degradable protein and true protein degraded intermediately contents at 60 d fermenting, indicating that it effectively inhibited protein degradation.

A case report of a patient with first phenotype of papillary thyroid carcinoma and heterochronous multiprimary tumor harboring germline MUTYH Arg19*/Gly286Glu mutations.

MUTYH-associated polyposis (MAP) is an autosomal recessively inherited disease with multiple system tumors mainly in alimentary system. Tumor occurrence of MAP patients is highly heterogeneous in space and time. MAP is associated with germline biallelic mutations in MUTYH. The targeted next­generation sequencing technology and Sanger sequencing are the important methods to screen MUTYH mutations now. Herein, we identified a patient with heterochronous multiprimary tumor carring MUYTH Arg19*/Gly286Glu compound heterozygous mutations. The patient in this case had a first phenotype of thyroid cancer at age 44, which earlier 2 years than the alimentary system cancers. In conclusion, our case report creases the in-depth understanding of the MAP heterogeneous phenotype and further reminds recommendations for improvement of health management and genetic counseling, special treatment plans.

Effects of the Application of Lactobacillus plantarum Inoculant and Potassium Sorbate on the Fermentation Quality, In Vitro Digestibility and Aerobic Stability of Total Mixed Ration Silage Based on Alfalfa Silage.

This study aimed to evaluate the effect of the application of an inoculant and a preservative on the fermentation quality, in vitro digestibility, and aerobic stability of alfalfa silage-based fermented total mixed ration (TMR). The TMR was ensiled with (1) no additives (control), (2) Lactobacillus plantarum (LP), or (3) potassium sorbate (PS). The V-scores of all silages were higher than 80 points during the 30 days of ensiling. The addition of LP and PS had no effects on the in vitro parameters, such as in vitro digestibility and in vitro gas production (p > 0.05). LP-treated silage showed similar fermentation quality and comparable aerobic stability to the control (110 h). The LP only decreased the ammonia nitrogen (NH3-N) content (p < 0.05) during ensiling. The PS significantly increased the pH of TMR silages (p < 0.05). Meanwhile, the addition of PS improved the aerobic stability (>162 h) of TMR silage, indicated by the higher water-soluble carbohydrate content and lower NH3-N content in comparison with those in the control after aerobic exposure (p < 0.05). The improvement in fermentation quality is extremely small in terms of applying LP in TMR silage based on a large percentage of other silage ingredients. The PS is effective in conserving unpacked TMR silage and showed the potential to reduce the risk of ruminal acidosis in livestock.

Effects of antibacterial peptide-producing Bacillus subtilis and Lactobacillus buchneri on fermentation, aerobic stability, and microbial community of alfalfa silage.

This study assessed the effects of antibacterial peptide-producing Bacillus subtilis (BS), Lactobacillus buchneri (LB), or their combination on fermentation, proteolysis, aerobic stability, and microbial communities during ensiling and aerobic exposure phases of alfalfa silage. The results showed that the BS-treated silage displayed a higher lactic acid concentration, less proteolysis, and higher aerobic stability than those in the control silage. Both LB and BS treatments increased Lactobacillus and Ascochyta abundance, and decreased Enterococcus and Sporormiacea abundance after 60 d of fermentation. LB and BS also inhibited the growth of Enterococcus after 3 d of aerobic exposure but similar to the control silage, the fungal community of BS silage was dominated by Candida and Pichia after 9 d of aerobic exposure. Therefore, inoculation of BS improved silage fermentation quality, aerobic stability and bacterial community during ensiling and after 3 d of aerobic exposure.

Effect of inoculants and storage temperature on the microbial, chemical and mycotoxin composition of corn silage.

OBJECTIVE: To evaluate the effect of lactic acid bacteria and storage temperature on the microbial, chemical and mycotoxin composition of corn silage. METHODS: Corn was harvested at 32.8% dry matter, and chopped to 1 to 2 cm. The chopped material was subjected to three treatments: i) control (distilled water); ii) 1×106 colony forming units (cfu)/g of Lactobacillus plantarum; iii) 1×106 cfu/g of Pediococcus pentosaceus. Treatments in triplicate were ensiled for 55 d at 20°C, 28°C, and 37°C in 1-L polythene jars following packing to a density of approximately 800 kg/m3 of fresh matter, respectively. At silo opening, microbial populations, fermentation characteristics, nutritive value and mycotoxins of corn silage were determined. RESULTS: L. plantarum significantly increased yeast number, water soluble carbohydrates, nitrate and deoxynivalenol content, and significantly decreased the ammonia N value in corn silage compared with the control (p<0.05). P. pentosaceus significantly increased lactic acid bacteria and yeast number and content of deoxynivalenol, nivalenol, T-2 toxin and zearalenone, while decreasing mold population and content of nitrate and 3-acetyl-deoxynivalneol in corn silage when stored at 20°C compared to the control (p<0.05). Storage temperature had a significant effect on deoxynivalenol, nivalenol, ochratoxin A, and zearalenone level in corn silage (p<0.05). CONCLUSION: Lactobacillus plantarum and Pediococcus pentosaceus did not decrease the contents of mycotoxins or nitrate in corn silage stored at three temperatures.