Chinese Horse Research Articles

Characterizing flavor development in low-salt Chinese horse bean-chili paste through integrated metabolomics and metagenomics

This study utilized metabolomics and metagenomics to investigate the microbial composition and functions in low- and high-salt Chinese horse bean-chili pastes (CHCPs). The results showed that 25 key metabolites were identified to distinguish the flavor attributes between the two samples. Leuconostoc was identified as the dominant microbiota in low-salt CHCP, while Pantoea prevailed in the high-salt CHCP. Compared to traditional high-salt fermentation, low-salt and inoculated fermentation promoted the increase in the relative abundances of Companionlactobacillus, Levilactobacillus, Tetragenococcus, Zygosaccharomyces and Wickerhamiella as well as the enrichment of carbohydrate and amino acid metabolic pathways, which contributed to the enhancement of characteristic flavor compounds. Further metabolic pathway reconstruction elucidated 21 potential microbial genera associated with the formation of key metabolites, such as Leuconostoc, Levilactobacillus, Pantoea, and Pectobacterium. This study may provide insights for optimizing the fermentation process and improving the flavor quality of low-salt CHCP and similar fermentation products. KeywordsLow-salt fermentationHight-salt fermentationChinese horse-bean chili pasteFlavor formationMetabolomicsMetagenomics

Microbial community variation and flavor metabolism in low-salt Chinese horse bean-chili-paste: Insights from bioturbation fermentation

Chinese horse bean-chili-paste (CHCP) is a traditional and famous high-salt fermented condiment in southwest China, and salt reduction strategy in this industry is imperative. In this study, we investigated the effect of inoculation with two endogenous strains (Tetragenococcus halophilus and Wickerhamiella versatilis) on the products quality at different salinity. The results showed that salt reduction combined with microbial inoculation showed prominent impact on the volatile profile. A total of 15 compounds was identified as key flavors, and bioturbation fermentation could significantly promote the concentrations of many pleasant aroma volatiles. Quantitative microbial profile analysis identified 29 key genera that might benefit microbial stability. In addition, bioturbation fermentation stimulated more microbial communities to contribute to the formation of key flavors. Moreover, the metabolic pathways of key flavor compounds were further investigated and preliminary results suggest that bioturbation under low-salt conditions promotes high expression of enzymes involved in the formation of these compounds, thereby increasing the abundance of key flavor compounds. These results revealed the variations of microbial communities in low-salt CHCP after biofortification fermentation and their influences on flavor metabolism, which could provide references for the manufacture of low-salt fermented foods.

Unveiling the aromatic differences of low-salt Chinese horse bean-chili-paste using metabolomics and sensomics approaches

To achieve salt reduction while ensuring flavor quality of Chinese horse bean-chili-paste (CHCP), we comprehensively explored the effect of indigenous strains Tetragenococcus halophilus and Candida versatilis on the aroma profiles of low-salt CHCP by metabolomics and sensomics analysis. A total of 129 volatiles and 34 aroma compounds were identified by GC × GC-MS and GC-O-MS, among which 29 and 20 volatiles were identified as significant difference compounds and aroma-active compounds, respectively. Inoculation with the two indigenous strains could effectively relieve the undesired acidic and irritative flavor brought by acetic acid and some aldehydes in salt-reduction samples. Meanwhile, inoculated fermentation provided more complex and richer volatiles in low-salt batches, especially for the accumulation of 3-methylbutanol, 1-octen-3-ol, benzeneacetaldehyde, phenylethyl alcohol, and 4-ethyl-phenol etc., which were confirmed as essential aroma compounds of CHCP by recombination and omission tests. The research elucidated the feasibility of bioturbation strategy to achieve salt-reducing fermentation of fermented foods.

Abortion storm of Yili horses is associated with Equus caballus papillomavirus 2 variant infection.

Nine different species of Equus caballus papillomavirus (EcPV) and three bovine papillomaviruses (BPVs) have been reported to infect horses. However, there are few descriptions of such infections in China. In our pioneer study on Chinese horses, we identified EcPV-2 in the nasal swabs (4/230, 1.7%) of Yili horses, and the semen (3/18, 16.7%) of thoroughbred horses. This indicated that EcPV is indeed hosted by horses in China, and that EcPV-2 might be transmitted though breeding. Further detection of EcPVs in the lung tissues of aborted fetuses of Yili horses, which were originally negative for equid herpes viruses, demonstrated EcPV-2 positivity in 19 of 50 samples, thereby indicating that EcPV-2 may be a new pathogen responsible for causing abortion. Thereafter, sequence analyses of the L1 genes of 26 EcPV-2 in China were performed, indicating that EcPV-2, which primarily infects horses in China, shared 98.3-99.9% nt identity with the published sequences for EcPV-2. These observations indicated that EcPV-2 identified in the current study were highly similar variants of the previously identified strains of EcPV-2. Phylogenetic analysis based on L1 gene sequences from GenBank showed that the EcPV-2 found in Chinese horses was closely related to and clustered together with an already known EcPV-2a lineage. Our study provides the first evidence related to EcPV-2 infection in Chinese horses, which can serve as a causative agent for Yili horse abortions, and may thus lay the foundation for a systematic and detailed epidemiological study of this infection in Chinese horses.

CKM intron: an appropriate marker for the determination of the genetic relationships among horse populations and breeds

To date, the origins, domestication, and genetic structure of Chinese Mongolian horses (CMH) are poorly understood. Furthermore, there have been sparse reports on the genetic differences between CMH and Thoroughbred. In order to determine their genetic structure, understand their genetic relationships, and explore their domestication processes, we performed an extensive survey of creatine kinase (muscle isoenzyme; CKM) variations among six populations of indigenous CMH, cultivated Sanhe horses, and imported Thoroughbred. Twenty-three single-nucleotide polymorphisms were found among the 343 horse sequences. From these, 40 haplotypes were inferred. Haplotype diversity (H) values differed from 0.6424 to 0.7881 and nucleotide diversity (π) values ranged from 0.00150 to 0.00211. The differences between Thoroughbred population and other Chinese horse populations were large, but only small differences were observed among Chinese horse populations with respect to CKM intron sequences suggesting that the domestication history, breeding measures, and origins of these horse populations are completely different. Results suggest that Sanhe and CMH are very closely related and the introgression (interbreeding) between them is serious. Our results suggest that Sanhe and Wushen require prompt and powerful protection. Overall, CKM intron was an appropriate marker for the determination of genetic relationships among horse populations and breeds.

Comparison of muscle metabolomics between two Chinese horse breeds.

With their enormous muscle mass and athletic ability, horses are well-positioned as model organisms for understanding muscle metabolism. There are two different types of horse breeds-Guanzhong (GZ) horses, an athletic breed with a larger body height (~148.7 cm), and the Ningqiang pony (NQ) horses, a lower height breed generally used for ornamental purposes-both inhabited in the same region of China with obvious differences in muscle content. The main objective of this study was to evaluate the breed-specific mechanisms controlling muscle metabolism. In this study, we observed muscle glycogen, enzyme activities, and LC-MS/MS untargeted metabolomics in the gluteus medius muscle of six, each of GZ and NQ horses, to explore differentiated metabolites that are related to the development of two muscles. As expected, the glycogen content, citrate synthase, and hexokinase activity of muscle were significantly higher in GZ horses. To alleviate the false positive rate, we used both MS1 and MS2 ions for metabolite classification and differential analysis. As a result, a total of 51,535 MS1 and 541 MS2 metabolites were identified, and these metabolites can separate these two groups from each other. Notably, 40% of these metabolites were clustered into lipids and lipid-like molecules. Furthermore, 13 significant metabolites were differentially detected between GZ and NQ horses (fold change [FC] value ≥ 2, variable important in projection value ≥1, and Q value ≤ 0.05). They are primarily clustered into glutathione metabolism (GSH, p = 0.01), taurine, and hypotaurine metabolism (p < 0.05) pathways. Seven of the 13 metabolites were also found in thoroughbred racing horses, suggesting that metabolites related to antioxidants, amino acids, and lipids played a key role in the development of skeleton muscle in horses. Those metabolites related to muscle development shed a light on racing horses' routine maintenance and improvement of athletic performance.

Genetic characterization of three recently discovered parvoviruses circulating in equines in China.

The family Parvoviridae comprises many major viral pathogens that can infect humans and multiple other species, causing severe diseases. However, knowledge of parvoviruses that infect equids is limited. In the present study, we found that three equine parvoviruses (EqPVs), namely, equine parvovirus-hepatitis (EqPV-H), equine parvovirus-cerebrospinal fluid (EqPV-CSF) and equine copivirus (EqCoPV) cocirculated among horses in China. We examined the prevalence of these three EqPVs in 225 horse serum samples in China and found EqPV-H, EqPV-CSF and EqCoPV viremia in 7.6% (17/225), 2.7% (6/225) and 2.2% of samples (5/225), respectively. We also obtained the complete genomes of one EqPV-H strain, six EqPV-CSF strains and one EqCoPV strain. After phylogenetic analysis of the EqPVs, we found that EqPV-CSF and EqCoPV may have evolved from the same ancestor. The EqPV-CSF strains (E111 and A27) and EqCoPV strain (F124) were genetically similar to foreign strains, but the EqPV-CSF strains (B48, E96, C61 and F146) comprised unique clades. This study determined the prevalence of three EqPVs in Chinese horses and analyzed the genetic characteristics of EqPVs prevalent strains in Chinese horse herds. Our data provide a theoretical basis for follow-up research on the prevention and control of EqPVs.

Whole-genome identification of transposable elements reveals the equine repetitive element insertion polymorphism in Chinese horses.

Transposable elements (TEs) are diverse, abundant, and complicated in genomes. They not only can drive the genome evolution process but can also act as special resources for adaptation. However, little is known about the evolutionary processes that shaped horses. In this work, 126 horse assemblages involved in most horse breeds in China were used to investigate the patterns of TE variation for the first time. By using RepeatMasker and melt software, we found that the horse-specific short interspersed repetitive elements family, equine repetitive elements (ERE1), exhibited polymorphisms in horse genomes. Phylogenetic analysis based on these ERE1 loci (minor allele frequency ≥0.05) revealed three major horse groups, namely, those in northern China, southern China, and Qinghai-Tibetan, which mirrors the result determined by SNPs to some extent. The present ERE1 family emerged ~0.26 to 1.77 Mya ago, with an activity peak at ~0.49 Mya, which matches the early stage of the horse lineage and decreases after the divergence of Equus caballus and Equus ferus przewalskii. To detect the functional ERE1(s) associated with adaptation, locus-specific branch length, genome-wide association study, and absolute allele frequency difference analyses were conducted and resulted in two common protein-coding genes annotated by candidate ERE1s. They were clustered into the vascular smooth muscle contraction (p=0.01, EDNRA) and apelin signalling pathways (p = 0.02, NRF1). Notably, ERE1 insertion into the EDNRA gene showed a higher association with adaptation among southern China horses and other horses in 15 populations and 451 individuals (p = 4.55 e-8). Our results provide a comprehensive understanding of TE variations to analyse the phylogenetic relationships and traits relevant to adaptive evolution in horses.

Salt tolerance and ester production mechanisms of Candida etchellsii in Chinese horse bean-chili-paste

Candida etchellsii is a predominant strain in Chinese horse bean-chili-paste, a famous and traditional condiment with high salt content. In this study, physiological properties, ester production capacity and gene expression of C. etchellsii under salt stress were investigated, and the results showed that the strain evoked miscellaneous regulations to resist high salt stress. Under salt stress, C. etchellsii would accumulate intracellular glycerol, Na+ and K+ to balance osmotic pressure. Interestingly, the strain showed much stronger capacity to yield ethyl hexanoate and hexyl acetate after 5% and 20% (w/v) NaCl treatment whether in esterification or alcoholysis system. Furthermore, transcriptome data indicated that the genes associated with carbohydrate metabolism and amino acid metabolism had significantly different expression, and their regulations were consistent with the results of physiological analysis. The results obtained in this study could contribute to understand the salt tolerance mechanism of C. etchellsii and provide a theoretical support for its application in traditional food fermentation.

The Development of Equestrian Policies in China between 2015 and 2020.

Simple SummaryOne of the most important industries in China’s history has been the horse industry, and the modern horse industry is still in the process of being transformed. The purpose of this paper is to introduce relevant policies through an analysis of information related to the horse industry in order to promote the economic development of the horse industry and gradually create a chain for the horse industry by selecting for this analysis the fastest-growing period in China’s modern horse industry. This can support the economic development of the horse industry and gradually create an integrated horse industry with Chinese characteristics. Equestrian education in China should be supported so that it gradually develops Chinese characteristics, and, thus, horse breeding and welfare will also improve in the future.China competed in equestrian sports for the first time at the 2008 Beijing Olympics. China’s modern equine business has developed significantly over the past decade, and the lessons from the expansion between 2015 and 2020 are important considerations as China implements further legislation to aid in the development of its modern equestrian sports. Equestrian sports can propel the Chinese horse industry forward, and the horse industry is a unique business in that it integrates one, two, and three industries, with much growth potential. This paper assesses the development of equestrianism in China from four perspectives: the general sports economic environment; the development of equestrian activities; the economic industries driven by equestrian activities; and relevant equestrian policies. Equestrian sports within China are currently facing problems, and recommendations are given. This paper is a single case study. The research utilized a qualitative approach, collecting data from official and semi-official documents. Through understanding the data collated and its analysis, equestrian sports can improve the speed and quality of their development under the influence of better-informed policy-making and a relevant economic model. It is expected that the wider related horse industry in China will also undergo more significant development.

Genome-Wide Detection of Copy Number Variants in Chinese Indigenous Horse Breeds and Verification of CNV-Overlapped Genes Related to Heat Adaptation of the Jinjiang Horse.

In the present study, genome-wide CNVs were detected in a total of 301 samples from 10 Chinese indigenous horse breeds using the Illumina Equine SNP70 Bead Array, and the candidate genes related to adaptability to high temperature and humidity in Jinjiang horses were identified and validated. We determined a total of 577 CNVs ranging in size from 1.06 Kb to 2023.07 Kb on the 31 pairs of autosomes. By aggregating the overlapping CNVs for each breed, a total of 495 CNVRs were detected in the 10 Chinese horse breeds. As many as 211 breed-specific CNVRs were determined, of which 64 were found in the Jinjiang horse population. By removing repetitive CNV regions between breeds, a total of 239 CNVRs were identified in the Chinese indigenous horse breeds including 102 losses, 133 gains and 4 of both events (losses and gains in the same region), in which 131 CNVRs were novel and only detected in the present study compared with previous studies. The total detected CNVR length was 41.74 Mb, accounting for 1.83% of the total length of equine autosomal chromosomes. The coverage of CNVRs on each chromosome varied from 0.47% to 15.68%, with the highest coverage on ECA 12, but the highest number of CNVRs was detected on ECA1 and ECA24. A total of 229 genes overlapping with CNVRs were detected in the Jinjiang horse population, which is an indigenous horse breed unique to the southeastern coast of China exhibiting adaptability to high temperature and humidity. The functional annotation of these genes showed significant relation to cellular heat acclimation and immunity. The expression levels of the candidate genes were validated by heat shock treatment of various durations on fibroblasts of horses. The results show that the expression levels of HSPA1A were significantly increased among the different heat shock durations. The expression level of NFKBIA and SOCS4 declined from the beginning of heat shock to 2 h after heat shock and then showed a gradual increase until it reached the highest value at 6 h and 10 h of heat shock, respectively. Breed-specific CNVRs of Chinese indigenous horse breeds were revealed in the present study, and the results facilitate mapping CNVs on the whole genome and also provide valuable insights into the molecular mechanisms of adaptation to high temperature and humidity in the Jinjiang horse.

Unraveling the Glucosylation of Astringency Compounds of Horse Chestnut via Integrative Sensory Evaluation, Flavonoid Metabolism, Differential Transcriptome, and Phylogenetic Analysis.

The seeds of Chinese horse chestnut are used as a source of starch and escin, whereas the potential use of whole plant has been ignored. The astringency and bitterness of tea produced from the leaves and flowers were found to be significantly better than those of green tea, suggesting that the enriched flavonoids maybe sensory determinates. During 47 flavonoids identified in leaves and flowers, seven flavonol glycosides in the top 10 including astragalin and isoquercitrin were significantly higher content in flowers than in leaves. The crude proteins of flowers could catalyze flavonol glucosides' formation, in which three glycosyltransferases contributed to the flavonol glucosylation were screened out by multi-dimensional integration of transcriptome, evolutionary analyses, recombinant enzymatic analysis and molecular docking. The deep exploration for flavonol profile and glycosylation provides theoretical and experimental basis for utilization of flowers and leaves of Aesculus chinensis as additives and dietary supplements.

A single-nucleotide mutation within the TBX3 enhancer increased body size in Chinese horses

SummaryChinese ponies are endemic to the mountainous areas of southwestern China and were first reported in the archaeological record at the Royal Tomb of Zhongshan King, Mancheng, dated to approximately ∼2,100 YBP.1 Previous work has started uncovering the genetic basis of size variation in western ponies and horses, revealing a limited number of loci, including HMGA2,2LCORL/NCAPG,3ZFAT, and LASP1.4,5 Whether the same genetic pathways also drive the small body size of Chinese ponies, which show striking anatomical differences to Shetland ponies,6 remains unclear.2,7 To test this, we combined whole-genome sequences of 187 horses across China. Statistical analyses revealed top association between genetic variation at the T-box transcription factor 3 (TBX3) and the body size. Fine-scale analysis across an extended population of 189 ponies and 574 horses narrowed down the association to one A/G SNP at an enhancer region upstream of the TBX3 (ECA8:20,644,555, p = 2.34e−39). Luciferase assays confirmed the single-nucleotide G mutation upregulating TBX3 expression, and enhancer-knockout mice exhibited shorter limbs than wild-type littermates (p < 0.01). Re-analysis of ancient DNA data showed that the G allele, which is most frequent in modern horses, first occurred some ∼2,300 years ago and rose in frequency since. This supports selection for larger size in Asia from approximately the beginning of the Chinese Empire. Overall, this study characterized the causal regulatory mutation underlying small body size in Chinese ponies and revealed size as one of the main selection targets of past Chinese breeders.

Bio-augmented effect of Bacillus amyloliquefaciens and Candida versatilis on microbial community and flavor metabolites during Chinese horse bean-chili-paste fermentation

Chinese horse bean-chili-paste (CHCP), a fermented condiment in China, is traditionally manufactured through naturally spontaneous semi-solid fermentation procedures without intentionally inoculated microorganisms. The aim of this study was to investigate the effect on microbiota and quality variations during CHCP fermentation by inoculation of selected autochthonous microorganisms Bacillus amyloliquefaciens and Candida versatilis. The results showed that relative abundance of Bacillus in the samples inoculated with B. amyloliquefaciens were increased from about 0.6% to almost 25%, and the batches bio-augmented with C. versatilis exhibited clearly 0.7 Lg copies/g higher biomass than that of the other samples. By bio-augmentation, six enzyme activities, namely acid protease, leucine aminopeptidase, α-amylase, cellulose, β-glucosidase and esterase, were considerably enhanced. As a result, inoculation of these two strains exhibited significant effect on the volatile profiles of CHCP. B. amyloliquefaciens herein was found to contribute mainly to the accumulation of acids, sulfur-containing compounds and pyrazines, whereas C. versatilis was considerably associated with the formation of alcohols, esters and phenols. This study proved that combination of B. amyloliquefaciens and C. versatilis could obtain more extensive aroma profiles, especially for the enrichment of miso-like and fruity flavors, which could provide a guideline for the tailored control of CHCP fermentation process.

Ester synthesis mechanism and activity by Bacillus licheniformis, Candida etchellsii, and Zygosaccharomyces rouxii isolated from Chinese horse bean chili paste.

Esters are indispensable aroma compounds and contribute significantly to the fruity aromas in fermented condiments. The ester synthesis activity and pathways of Bacillus licheniformis, Candida etchellsii, and Zygosaccharomyces rouxii, isolated from Chinese horse bean chili-paste (CHCP), were investigated. Chemical buffer models containing esterification and alcoholysis systems inoculated with extracellular extracts of these three strains were established. The ester synthesis activity of C. etchellsii was stronger than that of the other two strains. Zygosaccharomyces rouxii could synthesize acetate esters via esterification, whereas the biosynthesis pathways of B. licheniformis and C. etchellsii were esterification and alcoholysis. Esterification exhibited relatively high activity at pH4, whereas alcoholysis activity improved with an increase in the pH from 4 to 8. Candida etchellsii could synthesize C6 -C8 of acetate esters, and its activity improved with the number of alcohol carbon atoms. These three strains could synthesize C10 -C18 of ethyl esters. Their ethyl ester synthesis activity decreased with the aliphatic acid carbon number. Candida etchellsii has the potential to be used in CHCP fermentation to accumulate esters and improve flavor compared with the other two strains. This research is helpful in explaining the mechanism of ester synthesis in fermented condiments. © 2021 Society of Chemical Industry.

Comparative population genomic analysis uncovers novel genomic footprints and genes associated with small body size in Chinese pony

BackgroundBody size is considered as one of the most fundamental properties of an organism. Due to intensive breeding and artificial selection throughout the domestication history, horses exhibit striking variations for heights at withers and body sizes. Debao pony (DBP), a famous Chinese horse, is known for its small body size and lives in Guangxi mountains of southern China. In this study, we employed comparative population genomics to study the genetic basis underlying the small body size of DBP breed based on the whole genome sequencing data. To detect genomic signatures of positive selection, we applied three methods based on population comparison, fixation index (FST), cross population composite likelihood ratio (XP-CLR) and nucleotide diversity (θπ), and further analyzed the results to find genomic regions under selection for body size-related traits.ResultsA number of protein-coding genes in windows with the top 1% values of FST (367 genes), XP-CLR (681 genes), and log2 (θπ ratio) (332 genes) were identified. The most significant signal of positive selection was mapped to the NELL1 gene, probably underlies the body size and development traits, and may also have been selected for short stature in the DBP population. In addition, some other loci on different chromosomes were identified to be potentially involved in the development of body size.ConclusionsResults of our study identified some positively selected genes across the horse genome, which are possibly involved in body size traits. These novel candidate genes may be useful targets for clarifying our understanding of the molecular basis of body size and as such they should be of great interest for future research into the genetic architecture of relevant traits in horse breeding program.

Diversity and specificity of the bacterial community in Chinese horse milk cheese.

The nutrition and flavor of cheese are generated by the microbial community. Thus, horse milk cheese with unique nutrition and flavor, an increasingly popular local cheese of the Xinjiang Uygur Autonomous Region of China, is considered to have diverse and specific bacterial community. To verify this hypothesis, horse, cow, and goat milk cheese samples produced under the same environmental conditions and manufacturing process were collected, and the 16S rRNA gene was targeted to determine the bacterial population size and community composition by real‐time quantitative PCR and high‐throughput sequencing. The bacterial community of horse milk cheese had a significantly larger bacterial population size, greater species richness, and a more diverse composition than those of cow and goat milk cheeses. Unlike the absolute dominance of Lactococcus and Streptococcus in cow and goat milk cheeses, Lactobacillus and Streptococcus dominated the bacterial community as the starter lactic acid bacteria in horse milk cheese. Additionally, horse milk cheese also contains a higher abundance of unclassified secondary bacteria and specific secondary bacteria (e.g., Psychrobacter, Sulfurisoma, Halomonas, and Brevibacterium) than cow and goat milk cheeses. These abundant, diverse, and specific starter lactic acid bacteria and secondary bacteria may generate unique nutrition and flavor of horse milk cheese.

Ancient Patrilineal Lines and Relatively High ECAY Diversity Preserved in Indigenous Horses Revealed With Novel Y-Chromosome Markers.

Extremely low nucleotide diversity of modern horse Y-chromosome has been reported, and only poor phylogenetic resolution could be resulted from limited Y-chromosome markers. In this study, three types of horse Y-chromosome markers, including Single-nucleotide polymorphisms (SNPs), copy number variants (CNVs), and allele-specific CNVs, were developed by screening more than 300 male horses from 23 indigenous Chinese horse populations and 4 imported horse breeds. Fourteen segregating sites including a novel SNP in the AMELY gene were found in approximately 53 kb of male-specific Y-chromosome sequences. CNVs were detected at 11 of 14 sites, while allele-specific CNVs at 6 polymorphic sites in repeated fragments were also determined. The phylogenetic analyses with the SNPs identified in this study and previously published 51 SNPs obtained mainly from European horses showed that indigenous Chinese horses exhibit much deeper divergence than European and Middle Eastern horses, while individuals of Chinese horses with the C allele of the AMELY gene constituted the most ancient group. Via SNPs, CNVs, and allele-specific CNVs, much higher diversity of paternal lines can be detected than those identified with merely SNPs. Our results indicated that there are ancient paternal horse lines preserved in southwestern China, which sheds new light on the domestication and immigration of horses, and suggest that the priorities of the conservation should be given to the ancient and rare paternal lines. These three marker types provided finer phylogenetic resolution of horse patrilineal lines, and the strategies used in the present study also provide valuable reference for the genetic studies of other mammalian patrilineages.

Flavor volatiles evolution of Chinese horse bean-chili-paste during ripening, accessed by GC×GC-TOF/MS and GC-MS-olfactometry

ABSTRACT The evolution of volatile compounds during the ripening of Chinese horse bean-chili-paste (CHCP) was investigated in this study. GC×GC-TOF/MS allowed an identification of 748 volatiles in three CHCP samples, respectively, ripened for 3 months (M3), 12 months (M12) and 24 months (M24), and it showed that a longer ripening resulted in a richer diversity of the volatiles. In addition, M3 contained more aldehydes and organic acids in content and M12 had more alcohols, ketones, and esters, while M24 was abundant in heterocyclics. A total of 69 among the 748 volatiles were discriminated as the key flavor compounds of CHCP by method of Fisher ratio together with odor activity value and flavor dilution, including 12 alcohols, 9 aldehydes, 4 ketones, 5 organic acids, 16 esters, 4 phenols, 6 sulfur-containing compounds, and 13 heterocyclics. Further, principal component analysis suggested that some key aldehydes contributed the flavor of M3, while some key alcohols, esters and heterocyclics existed in M12, and some key phenols and heterocyclics were in M24. Varying of the key flavor volatiles during CHCP ripening gave it different fragrance configurations. Results presented details for the volatile profiles of CHCP during the natural ripening period, beneficial to shorten fermentation time by functional microorganisms, and could enrich the desire flavors.

Physicochemical properties and microbial community dynamics during Chinese horse bean-chili-paste fermentation, revealed by culture-dependent and culture-independent approaches

The production of Chinese horse bean-chili-paste (CHCP) involves three fermentation phases: chili-to-moromi fermentation (CF) phase, horse bean-to-meju fermentation (HF) phase and moromi-meju mixed fermentation (MF) phase. To understand the microbial dynamics among these three phases and the potential roles of viable microbes for fermentation, microbial community dynamics was investigated by using culture-dependent and culture-independent methods. Furthermore, the capacities of enzyme-producing of the isolates were determined. During the CF phase, reducing sugar content increased from 3.1% to 3.49%, while pH declined from 4.85 to 4.5. The protein content in the HF phase and MF phase reduced sharply from 22.23% to 10.29% and 4.39%-1.19%, respectively. Bacillus sp., Staphylococcus sp., Oceanobacillus sp., Candida sp., Zygosaccharomyces sp. and Aspergillus sp. dominated the CF phase, while Bacillus sp., Candida sp. and Zygosaccharomyces sp. were the dominant microorganisms in both the HF and MF phases. B. amyloliquefaciens, B. methylotrophicus, B. subtilis, B. licheniformis and A. oryzae possessed strong capacities of producing enzymes, i.e. α-amylase, cellulase and xylanase, acid protease and leucine aminopeptidase, and could make a great contribution to CHCP fermentation.