Elucidation of potential relationship between endogenous proteases and key flavor substances in dry-cured pork coppa

Dry-cured meat products are considerably popular around the world due to unique flavor.Proteolysis is one of the enzymatic reactions from which flavor substances are derived,which is affected by endogenous proteases.The purpose aimed to reveal the potential relationship between endogenous proteases and key flavor substances in dry-cured pork coppa in this paper.The dynamic changes of endogenous proteases activity,free amino acids,and volatiles during dry-cured pork coppa processing were characterized.The results showed that 5 kinds of free amino acids,Glu,Lys,Val,Ala,and Leu,were identified as significant contributors to taste.Meanwhile,key volatiles,such as hexanal,nonanal,octanal,benzaldehyde,3-methyl butanoic acid,2-methyl propanoic acid,and ethyl octanoate,greatly contributed to the flavor characteristics of dry-cured pork coppa.Further partial correlation analysis was performed to better elucidate the relationship among parameters.The results revealed that close relationship between endogenous proteases and key substances.RAP not only significantly affected the accumulation of key active-amino acids,but also affected the accumulation of ethyl octanoate,2,3-pentanedione,and 2,3-octanedione by regulating the accumulation of octanoic acid and Leu.In addition,cathepsin B and D,DPP II,DPP IV and RAP notably affected accumulation of hexanal.

Effect of the Inclusion of Chestnut in the Finishing Diet on Volatile Compounds of Dry-Cured Ham from Celta Pig Breed

The effect of the inclusion of chestnut in pigs ifnishing diet on volatile compounds of dry-cured Celta ham was studied. Twelve hams of each type （from three different pigs ifnishing diets：concentrate （CO）, mixed （MI） and chestnut （CH）） were used. Volatiles were extracted using a purge-and-trap method and analyzed by gas chromatography/mass spectrometry （GC/MS）. Thirty-nine volatile compounds were identified in dry-cured Celta ham samples. Most abundant volatiles in ham samples were aldehydes, which represented respectively, 53% （CO）, 51% （MI） and 46% （CH） of the total volatile composition. With the exception of 2-butenal, 2-methyl, all aldehydes were affected by feeding system. On the other hand, hydrocarbons n-alkanes were the second major group in the volatile proifle of dry-cured Celta hams and represented 28.9, 35.7 and 32.4%of the total volatile composition for CO, MI and CH groups, respectively. Ham samples from chestnut group showed a higher content of alcohols and this result could be related with the inclusion of chestnut in the ifnishing diet of pigs. Principal component analysis showed a good separation among groups. The discriminant analysis selected eight variables （butanoic acid, hexanal, octanal, nonenal （E）, decenal （E）, tetradecane, decane trimethyl and pyridine 2-methyl） and calculated two discriminating functions to predict if chestnut has been included in the ifnishing diet. Thus, it was possible to discriminate between groups fed with ifnishing diets containing chestnuts in their composition （mixed and chestnut group）.

Study Progress on Pests in Dry-cured Ham

Dry-cured ham can become infested with pests during the curing and ag-ing process, which not only damages the ham quality, but also causes economic loss. The advance of study on the effects of ham pests on the ham quality, the main species and distributions of ham pests in process of dry-cured ham were intro-duced in this article.

Evaluation and selection of yeasts as potential aroma enhancers for the production of dry-cured ham

Yeasts play a critical role in the flavor formation of dry-cured ham.In this study,41 yeast isolates from the dry-cured ham at different processing stages were evaluated for their technological properties.Debaryomyces hansenii was the most dominant yeast and has been detected at each phase of dry-cured ham,followed by Candida zeylanoides which was mainly detected in salting phase.Yarrowia bubula and Yarrowia alimentaria were found at the first two-phase of dry-cured ham.All isolates of yeast showed enzymatic activities against milk protein and tributyrin,while only 4 strains displayed proteolytic activity on meat protein.Yeast strains were grown in a meat model medium and volatile compounds were identified.The result showed that inoculated yeast strains could promote the production of volatiles and there were significant differences among strains.D.hansenii S25 showed the highest production of volatile compounds,followed by the strain C.zeylanoides C4.D.hansenii S25 was the highest producer of alcohols showing the highest production of benzeneethanol and 3-(methylthio)-1-propanol.Based on OAV and PLS analysis,D.hansenii S25 was strongly correlated with overall flavor and key volatile compounds of dry-cured ham,which could be selected as potential starter cultures.

Reducing the Levels of Sodium, Saturated Animal Fat, and Nitrite in Dry-Cured Pork Meat Products: A Major Challenge

The curing of meat is a conservation technique widely used since ancient times to prolong shelf-life. It consists in exposing meat to a mixture of sodium chloride and nitrate/nitrite. Sodium chloride affects the flavor, texture and shelf-life of meat products. Animal fat mainly affects the flavor and texture, and nitrate and nitrite affect the color and flavor, and give cured meat products their typical aroma. Excessive intake of sodium has been linked to arterial hypertension and increased risk of cardiovascular diseases. Excessive intakes of saturated fatty acids in pork fat, and also of nitrite, have been identified as factors promoting some cancers. There is consequently an increasing consumer demand to reduce these ingredients in processed meat and so develop healthier cured meat products. This paper reviews how and to what extent sodium, animal fat rich in saturated fatty acids, and nitrite contents can be reduced in the production of dry-cured hams and dry-fermented sausages.

Flavor release from traditional dry-cured pork during oral processing

The rate of flavor release in the mouth from traditional dry-cured pork was evaluated in this study.It was hypothesized that a slow steady release of flavor would occur on chewing and hydration.To test this,high salt dry-cured pork was chewed and held in oral cavity without swallowing for 4 different chewing stages(B,NC_(30),C_(30),EC).Saliva and food bolus from 16 healthy subjects were collected during oral processing,and analyzed for saliva flow rate,total saliva protein content,saliva pH,conductivity,saliva and sodium,and taste components.Results show that oral processing behavior and salivary release have important effects on flavor release of dry-cured pork.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)analysis showed that oral processing has a certain effect on the protein composition and concentration in saliva.This study suggests that the interaction between food and saliva that accompanies chewing can lead to significant changes in sensory perception during oral processing.

Pangenome and multi-tissue gene atlas provide new insights into the domestication and highland adaptation of yaks

Background The genetic diversity of yak,a key domestic animal on the Qinghai-Tibetan Plateau(QTP),is a vital resource for domestication and breeding efforts.This study presents the first yak pangenome obtained through the de novo assembly of 16 yak genomes.Results We discovered 290 Mb of nonreference sequences and 504 new genes.Our pangenome-wide presence and absence variation(PAV)analysis revealed 5,120 PAV-related genes,highlighting a wide range of variety-specific genes and genes with varying frequencies across yak populations.Principal component analysis(PCA)based on binary gene PAV data classified yaks into three new groups:wild,domestic,and Jinchuan.Moreover,we pro-posed a‘two-haplotype genomic hybridization model'for understanding the hybridization patterns among breeds by integrating gene frequency,heterozygosity,and gene PAV data.A gene PAV-GWAS identified a novel gene(Bos-Gru3G009179)that may be associated with the multirib trait in Jinchuan yaks.Furthermore,an integrated transcrip-tome and pangenome analysis highlighted the significant differences in the expression of core genes and the muta-tional burden of differentially expressed genes between yaks from high and low altitudes.Transcriptome analysis across multiple species revealed that yaks have the most unique differentially expressed m RNAs and lnc RNAs(between high-and low-altitude regions),especially in the heart and lungs,when comparing high-and low-altitude adaptations.Conclusions The yak pangenome offers a comprehensive resource and new insights for functional genomic studies,supporting future biological research and breeding strategies.

Composition Analysis on Intermuscular Fatty Acid of Qinghai Datong Yak

[ Objective] The study aimed to provide a theoretical basis for rational utilization of Datong yak resource. [ Method] By qualitative and quantitative determination with gas chromatograph, the composition and content of intermuscular fatty acid of Datong yak were compared with those of Datong yellow cattle. [ Result] Comparing with Datong yellow cattle, Datong yak had lower saturated fatty acid （SFA） content and higher polyunsaturated fatty acid （PUFA） content at extremely significant level （P 〈0.01 ）, while the difference of monounsaturated fatty acid （MUFA） contents between them was not significant （P〉0.05）. The PUFA/SFA and n-6/n-3 PUFA of Datong yak were in accordance with the recommended proportions. [ Conclusion] The intermuscular fatty acid composition of Datong yak is superior to that of local yellow cattle, which indicates that Datong yak meat has high nutritive value and wide development prospect.

Cloning and Bioinformatics Analysis of Lfcin Gene of Datong Yak

[Objective] This study was to clone Lfcin gene from Datong yak, so as to provide reference for applying this gene in feed industry and breeding industry. [Method] Using PCR technology, the lactoferricin(Lfcin)-encoding gene was obtained from genome of Datong yak; then it was cloned into pGEM-T easy vector, and then sequenced; the sequencing results were subsequently aligned with the sequences of dairy cow accessed in GenBank. Moreover, amino acid sequences of Lfcin gene from various species including yak, dairy cow, human and mouse were used for sequence alignment and phylogenesis analysis. [Result] The second exon of lactoferrin(LF) from Datong yak, which is 778 bp in length, was obtained, within which the coding region of Lfcin gene is 75 bp (25 amino acid residues); sequence analysis showed that there is discrepancy of eleven bases between Datong yak and dairy cow; Lfcin proteins from various species shared high homeology, of which that from Datong yak and dairy cow were completely identical; phylogenesis analysis showed that cladogram based on Lfcin was consistent with species evolutionary law. [Conclusion] This study laid a foundation for the prokaryotic or eukaryotic expression of Lfcin gene and further understanding the activity of Lfcin protein.

Study on Carcass Traits of Changtai Yak

For studying the germplasm charactedsUcs and carcass traits of Changtai yak, 8 adult yaks （5 male, 3 female） were selected and tested. The results showed that the pre-slaughter weight of the adult male and female Changtai yak was 364.32 and 266.83 kg, respectively, which presented extremely significant difference （P〈0.01）. The carcass weight, net bone weight net meat weight, and the ratio of bone to meat also showed extremely significant difference （P〈O.01）, which were 186.60 and 125.67 kg, 39.74 and 25.00 kg, 147.84 and 100.83 kg, 1：3.73 and 1： 4.03, respectively. The dressing percentage had significant difference which was 51.15% and 49.34%, respectively （P〈0.05）. The neat meat percentage and carcass meat production rate showed no significant difference which were 40.54% and 37.66%, 79.29% and 80.24%, respectively （P〉0.05）. It indicated that Changtai yak has better growth performance and meat performance potential, and worth of further studying and breeding.

Study on the Correlation between Blood Enzyme Activity and Production Performance in Tibetan Yaks

Yak ( Bos grunniens ) is classified as Bovine genus,they live in Qingzang Plateau with elevation more than 3500m and it's neighbor highland,where the weather is high chilly,short of oxygen and large difference of temperature between day and night.But yak have adapted that circumstance after thousands of years' artificial and natural selection and has great anti adversity capacity.Yak is an important sustenance and means of production to Tibetan.China is a country with the largest number of yak,whose amount of livestock on hand accounts for 95% of the world and Tibet region takes the second place next to Qinghai in the number of yak.Tibetan yak has formed some groups for Tibetan's large area,complicated geological ecosystem conditions,geological isolation.The typical better groups are Yadong yak,Jiali yak and Sibu yak. Study about yak fell behind other animals due to the restriction of inconvenient transportation,difficult sampling and poor experimental technology.Study about the relationship between yak blood enzyme activities and milk production was little,only Jing (1992) studied correlation of milk production with serum amylase,esterase in Gansu Jiulong yak,Maiwa yak and no report was found in other groups.This study discussed six Jiali yak blood enzyme activities lactate dehydrogenase (LDH),alkaline phosphatases (AKP),acid phosphatase (ACP),catalase (CAT),amylase (Amy) and superoxide dismutase (SOD),and explored their relationship with milk production in Jiali yak,so as to supply with theoretical evidence by using blood enzyme activity as biochemical assisted selection marker in the breeding of Tibetan yak milk yield. This experiment investigated activity of six blood enzymes from 28 Tibetan yaks and explored its relationship to production performance.The results showed a extremely positive significant difference ( P <0.01) between activity of LDH and milk yield,CAT activity and length of belly hair respectively,while a positive significant difference ( P <0.05) was observed between AKP activity and body weight,Amy activity and body weight and a negative significant difference ( P <0.05) for SOD activity and body weight.Stepwise Regression Analysis showed that activity of LDH could be used for the prediction of milk yield,AKP,CAT,Amy for body weight,CAT,Amy for length of belly hair.So it is expected to use activity of LDH,AKP,CAT,Amy as biochemical genetic marker in the selection of milk yield,body weight and length of belly hair.

Determination and Analysis on Nutritional Components in Placenta of Plateau Yak

Yaks live in Qinghai-Tibet Plateau （4 km above sea level）, and the nutrient components of their placenta were determined, including 17 kinds of amino acids and 11 kinds of trace elements. The results showed that there are abundant amino acids and trace elements in the placenta of yak, equal to that of human and deer, indicating some medical and nutritional value of yak placenta.

Hair and Villus Production Performance of Tianzhu White Yak

[ Objective ] To analyze the hair and villus production performance of Tianzhu white yak and thus provide conditions for the genetic breed ing and product development. [Method] The hair and villus yield as well as villus rate of male and female Tianzhu white yaks from one year to six years old was detected and the villus yield was also determined. And these indexes of hair and villus were analyzed to evaluate the production per formance. [ Result] In Tianzhu white yaks from one year to two years old, the hair and villus yield was lower, but the villus rate and yield were high er. The hair and villus yield, villus rate, and villus yield were all higher in Tianzhu white yaks from three years to six years old but significantly de creased in 7yearold Tianzhu white yaks. In addition, the hair and villus yield was significantly （ P 〈 0.05） or very significantly （ P 〈 0.01 ） higher in male yaks than in female yaks. The villus rate was very significantly （ P 〈0.01 ） higher than in female yaks from five years to seven years old than in male yaks at the same age. [ Conclusion] Tianzhu white yaks from one year to two years old have the best hair and villus production perform ance; Tianzhu white yaks from three years to six years old are in the main phase of hair and villus production; Tianzhu white yaks older than seven years old should not be used for hair and villus production.

Study on the Datong Yak's Skin Development

[Objective] This study was to explore the skin development law of Datong Yak.[Method]Taking Datong yaks at four various growth stages（1 day old,30 days old,180 days old and adult）as experimental materials,we studied histological structure of Datong yak skin by employing the histological methods.[Results]The skins of Datong yak at four various growth stages all showed the characteristics of thinner epidermis and thicker hypodermis,and of less capillary vessels in hypodermis and more pigment cells in epidermis.Datong yak at four various growth stages all showed a fluctuating trend in entire skin thickness,in detail thickening from 1 day old to 30 days old,then thinning to 180 days old,again thickening to adult.[Conclusion]The results provided theoretical basis for further understanding the structural characteristics of Datong yak to adapt to the abominable plateau environment.

Comparative Study on Hydrolase Activities in Tianzhu White Yak Milk and Gannan Yak Milk of Gansu

This study was conducted to compare the hydrolase activities in the milk of Tianzhu white yak and Gannan yak in Gansu. The results showed that the ACP activity in Tianzhu yak milk was significantly higher than that in Gannan yak milk （P〈0.05）; the AKP activity in Tianzhu yak milk was higher than that in Gannan yak milk, but not significant; and the AMC activity in Tianzhu yak milk was significantly lower than that in Gannan yak milk （P〈0.05）. There were no remarkable changes in the activities of the 3 hydrolases with the change of parity, i.e., the effects of parity on enzyme activities in milk were not significant. The comparative study on hydrolase activities in the milk of Tianzhu while yak and Gannan yak helps to understand the characteristics of the milk of the 2 yak breeds, and further provides certain reference basis for the development of yak milk.

Optimization of Parameters of Exogenous Gene Mediated by Liposome to Transfect Yak Mammary Epithelial Cells in Vitro

[Objective] The aim of this study was to optimize conditions of exogenous gene mediated by liposome to transfect yak mammary epithelial cells in Vitro.[Method] Yak mammary epithelial cells were isolated and cultivated in Vitro by the methods of collagenase digestion and tissue adhesion.The expression of cytokeratin in yak mammary epithelial cell was detected by immunocytochemistry technique.With green fluorescence protein as the report gene,yak mammary epithelial cells were transfected with exogenous gene m...

Potential antibacterial activity of berberine against multi drug resistant enterovirulent Escherichia coli isolated from yaks(Poephagus grunniens) with haemorrhagic diarrhoea

Objective:To evaluate the antimicrobial efficacy of berberine,a plant alkaloid.Methods:Five multi-drug resistant（MDR） STEC/EPEC and five MDR ETEC isolates from yaks with haemorrhagic diarrhoea were selected for the study.Antibacterial activity of berberine was evaluated by broth dilution and disc diffusion methods.The binding kinetics of berberine to DNA and protein was also enumerated.Results:For both categories of enterovirulent Escherichia coli（E.roli） isolates, berberine displayed the antibaclerial effect in a dose dependent manner.The MIC50 of berberine chloride for STEC/EPEC isolates varied from 2.07μM to 3.6μM with a mean of（2.95±0.33）μM where as for ETEC strains it varied from 1.75 to 1.96μM with a mean of（1.87±0.03）μM. Berberine bind more tightly with double helix DNA with Bmax and Kd of（24.68±2.62） and（357.8±57.8）,respectively.Berberine reacted with protein in comparatively loose manner with Bmax and Kd of（18.9±3.83） and 【286.2±113.6),respectively.Conclusions:The results indicate clearly that berberine may serve as a good antibacterial against multi drug resistant E.coli.

Comparison of rumen archaeal diversity in adult and elderly yaks(Bos grunniens)using 16S rRNA gene high-throughput sequencing

This study was conducted to investigate the phylogenetic diversity of archaea in the rumen of adult and elderly yaks. Six domesticated female yaks, 3 adult yaks （（5.3±0.6） years old）, and 3 elderly yaks （（10.7±0.6） years old）, were used for the rumen contents collection. Illumina MiSeq high-throughput sequencing technology was applied to examine the archaeal composition of rumen contents. A total of 92 901 high-quality archaeal sequences were analyzed, and these were assigned to 2 033 operational taxonomic units （OTUs）. Among these, 974 OTUs were unique to adult yaks while 846 OTUs were unique to elderly yaks; 213 OTUs were shared by both groups. At the phylum level, more than 99% of the obtained OTUs belonged to the Euryarchaeota phylum. At the genus level, the archaea could be divided into 7 archaeal genera. The 7 genera （i.e., Methanobrevibacter, Methanobacterium, Methanosphaera, Thermogymnomonas, Methanomicrobiu, Meth- animicrococcus and the unclassified genus） were shared by all yaks, and their total abundance accounted for 99% of the rumen archaea. The most abundant archaea in elderly and adult yaks were Methanobrevibacterand Thermogymnomonas, respectively. The abundance of Methanobacteria （class）, Methanobacteriales （order）, Methanobacteriaceae （family）, and Methanobrevibacter （genus） in elderly yaks was significantly higher than in adult yaks. In contrast, the abundance of Ther-mogymnomonas in elderly yaks was 34% lower than in adult yaks, though the difference was not statistically significant. The difference in abundance of other archaea was not significant between the two groups. These results suggested that the structure of archaea in the rumen of yaks changed with age. This is the first study to compare the phytogenetic differences of rumen archaeal structure and composition using the yak model.

Analysis of genetic diversity and population structure of Chinese yak breeds (Bos grunniens) using microsatellite markers

Nine Chinese yak breeds （Maiwa, Tianzhu White, Qinghai Plateau, Sibu, Zhongdian, Pali, Tibetan High Mountain, Jiulong, and Xinjiang） and Gayal were analyzed by means of 16 microsatellite markers to determine the level of genetic variation within populations, genetic relationship between populations, and population structure for each breed. A total of 206 microsatellite alleles were observed. Mean F-statistics （0.056） for 9 yak breeds indicated that 94.4% of the genetic variation was observed within yak breeds and 5.6% of the genetic variation existed amongst breeds. The Neighbor-Joining phylogenetic tree was constructed based on Nei＇s standard genetic distances and two clusters were obtained. The Gayal separated from the yaks far away and formed one cluster and 9 yak breeds were grouped together. The analysis of population structure for 9 yak breeds and the Gayal showed that they resulted in four clusters; one cluster includes yaks from Tibet Autonomous Region and Qinghai Province, one cluster combines Zhongdian, Maiwa, and Tianzhu White, and Jiulong and Xinjiang come into the third cluster. Pali was mainly in the first cluster （90%）, Jiulong was mainly in the second cluster （87.1%）, Zhongdian was primarily in the third cluster （83%）, and the other yak breeds were distributed in two to three clusters. The Gayal was positively left in the fourth cluster （99.3%）.

Novel SNP of EPAS1 gene associated with higher hemoglobin concentration revealed the hypoxia adaptation of yak(Bos grunniens)

Endothelial PAS domain protein 1 gene （EPAS1） is a key transcription factor that activates the expression of oxygen-regu- lated genes. In this study, in order to better understand the effects of EPAS1 gene on hematologic parameters in yak, we firstly quantified the tissue expression patterns for EPASl mRNA of yak, identified polymorphism in this gene and evaluated its association with hematologic parameters. Expression of EPAS1 mRNA was detected in all eight tissues （heart, liver, lung, spleen, pancreas, kidney, muscles and ovary）. The expressions of EPAS1 in lung and pancreas were extremely higher than other tissues examined. Three novel single nucleotide polymorphisms （SNPs） （g.83052 C〉T, g.83065 G〉A and g.83067 C〉A） within the EPAS1 were identified and genotyped in Pali （PL）, Gannan （GN） and Tianzhu White （TZW） yak breeds. Significant higher frequencies of the AA and GA genotypes and A allele of the g.83065 G〉A were observed in the PL and GN breeds than that in the TZW breed （P〈0.01）. Association analysis of the PL breed indicated that the g.83065 G〉A polymorphism was significantly associated with hemoglobin （HGB） concentration in yaks （P〈0.05）. Individuals with genotype AA had significantly higher HGB concentration （P〈0.05） than those with genotype GA and GG. All these results will help our further understanding of biological functional of yak EPAS1 gene in responding to hypoxia and also indicate EPAS1 might contribute to the hypoxia adaptation of the yak.