A method of identifying microorganisms with reduced adaptability to a particular environment, wherein (1) each microorganism is independent by insertional inactivation of a gene with a nucleic acid comprising a unique marker sequence. Mutating to provide a plurality of microorganisms, or clones of said microorganisms, such that each mutation has a different label sequence; (2) providing storage samples of each mutation produced by step (1), respectively, and providing storage nucleic acids each comprising a unique label sequence from each mutation; (3) introducing a plurality of mutations produced by step (1) into the specific environment such that the microorganisms capable of growing in the specific environment grow in the above environment; (4) recovering the microorganisms or selected portions thereof from the environment and separating nucleic acids from the recovered microorganisms; (5) comparing all label sequences in the nucleic acid isolated in step (4) with the unique label sequences of each of the stored mutations as in step (2); and (6) any label isolated in step (4). Screening for individual mutations that also do not have a sequence.

Identification of genes

Probiotics are being considered as valuable microorganisms related to human health. Hu sheep is referred as one of the important sheep breeds in China. Goat milk produced by Hu sheep is characterized with high nutritional value and hypoallergenic in nature. Particularly, this milk contains plenty of milk prebiotic and probiotic bacteria. This study was aimed to scrutinize more bacterial strains from Hu sheep milk with potential probiotic activity. Based on 16S rRNA sequence analysis, pool of forty bacterial strains were identified and evaluated their antimicrobial activities against Staphylococcus aureus, enterohemorrhagic Escherichia coli (EHEC), Salmonella typhimurium, Listeria monocytogenes enterotoxigenic E. coli (ETEC) and Aeromonas caviae. Four out of these isolated strains demonstrated their efficient bacteriostatic ability and potential healthy properties. We also examined the safety aspects of these bacterial candidates including three Lactococcus lactis strains (named as HSM-1, HSM-10, and HSM-18) and one Leuconostoc lactis strain (HSM-14), and were further evaluated via in vitro tests, including antimicrobial activity, cell surface characteristics (hydrophobicity, co-aggregation, and self-aggregation), heat treatment, antibiotic susceptibility, simulated transport tolerance in the gastrointestinal tract, and acid/bile tolerance. The obtained results revealed that HSM-1, HSM-10, HSM-14, and HSM-18 showed high survival rate at different conditions for example low pH, presence of bovine bile and demonstrated high hydrophobicity. Moreover, HSM-14 had an advantage over other strains in terms of gastrointestinal tract tolerance, antimicrobial activities against pathogens, and these results were significantly better than other bacterial candidates. Hu sheep milk as a source of exploration of potential lactic acid bacteria (LAB) probiotics open the new horizon of probiotics usage from unconventional milk sources. The selected LAB strains are excellent probiotic candidates which can be used for animal husbandry in the future. Rationale of the study was to utilize Hu sheep milk as a source of potential probiotic LABs. The study has contributed to the establishment of a complete bacterial resource pool by exploring the Hu sheep milk microflora.

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Functional probiotics of lactic acid bacteria from Hu sheep milk

Intramuscular fat (IMF) contributes importantly to various aspects of meat quality, and genetic regulation is an effective pathway to improve IMF deposition in sheep. In this study, we systematically explored the effect of IMF content on meat quality in Tan sheep and investigated the regulatory mechanism of flavor precursors metabolism and IMF deposition. The results revealed that IMF significantly affected meat color, total muscle fiber numbers, and muscle fiber types in Tan sheep. Widely-targeted metabolomic analysis showed that amino acids were the main differential flavor precursors between lambs with different IMF content. Importantly, the comparison of fatty acid profiles revealed that saturated fatty acids and monounsaturated fatty acids are beneficial for IMF deposition. Furthermore, integrated analysis between metabolome and transcriptome indicated that MME is a key gene resulting in the reduction of amino acids in lambs with high IMF content; and the joint analysis between fatty acid profiles and transcript profiles showed that ADIPOQ, FABP4, PLIN1, PPARGC1A, SLC2A1 accelerated IMF deposition through positive regulation of saturated fatty acids and monounsaturated fatty acids metabolism. These results revealed key changes in meat quality affected by IMF content and the corresponding genetic mechanism, which may provide a new insight for understanding the IMF differential deposition and for improving meat quality in Tan sheep.

https://www.frontiersin.org/articles/10.3389/fnut.2022.908355/pdf

Effects of intramuscular fat on meat quality and its regulation mechanism in Tan sheep

Ovine ELOVL5 and FASN genes polymorphisms and their correlations with sheep tail fat deposition

For further understanding the genetic control mechanisms of growth and development in Tan sheep, and culturing good traits on meat performance, which is very important to both in developing local species and improving economic efficaciously. In our study, we recruited a total of 250 Tan sheep and 174 healthy Hu sheep to detect 32 SNPs in GH, GHR, NPY, Leptin, H-FABP, MSTN, and CAST by using direct sequencing techniques, in order to explore genetic marking loci which were an association with growth characters. From the results, we found different SNPs with an obvious difference for the growth traits. In the different genetic model analysis, we found SNP12, SNP29, SNP41, SNP8, SNP34, SNP35, SNP9, SNP10, SNP36, SNP45, and SNP39 were a significantly negative association with the two kinds of sheep. And SNP46, SNP42, and SNP69 with the positive association between the different trait in sheep were analyzed. From the LD and haplotype analysis, we found three blocks with the positive association in growth traits between Tan sheep and Hu sheep. The block of SNP29, SNP32, SNP34, SNP35, SNP36, SNP39, SNP41, SNP42, SNP45, and SNP46 with the genotype "AATCTACTTA" is the most significantly association with the traits. In summary, the study initially explored the genes for growth and reproduction between Tan sheep and Hu sheep and found some statistically significant results which demonstrate that there are genetic differences. These differential molecular markers may provide a scientific theoretical basis for the preferred species of Tan sheep which with good meat performance and better utilization of species resources.

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Genetic polymorphism association analysis of SNPs on the species conservation genes of Tan sheep and Hu sheep.

The present invention relates to lick bricks with interaction effects of combining nutrition and improving immunity for cattle and sheep and a technology thereof. The lick bricks contain the following components: lycium barbarum polysaccharides, astragalus polysaccharides, methionine zinc, yeast selenium, sodium bicarbonate, magnesium oxide, corn bran, edible salt and molasses. The lycium barbarum polysaccharides, astragalus polysaccharides, methionine zinc, yeast selenium, sodium bicarbonate, magnesium oxide and corn bran are mixed, the mixture is stirred, and the stirred mixture is superfinely crushed to obtain a first mixture; in the first mixture, the edible salt is added and the materials are stirred to obtain a second mixture; the molasses at 70-150 parts by weight are heated at a temperature of 80-100 DEG C to prepare syrup; and the second mixture and the syrup are uniformly mixed, the mixture is pressed and shaped to obtain the lick bricks. The lick bricks are used for improving immunity and production performance of animals, particularly the immunity and production performance of the cattle or sheep.

Lick bricks with interaction effects of combining nutrition and improving immunity for cattle and sheep and technology thereof

The invention discloses a traditional Chinese medicine composition for preventing and treating postpartum abdominal pain of dairy cows. The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 35 to 45 parts of pollen typhae, 35-45 parts of trogopterus dung, 45-65 parts of angelica sinensis, 30-50 parts of ligusticum wallichii, 20-30 parts of rhizoma cyperi, 35-45 parts of mangnolia officinalis, 50-70 parts of fennel, 60-80 parts of herba leonuri, 30-40 parts of herba lycopi, 40-60 parts of radix cyathulae, 40-60 parts of codonopsis, 50-70 parts of astragalus membranaceus, 10-15 parts of amethyst and 40-60 parts of lily. The invention also provides a preparation method thereof and an application thereof. The traditional Chinesemedicine composition can be used for effectively treating postpartum pain of dairy cows; the animal welfare of modern animal husbandry is improved, the milk yield of the tested dairy cow is increased,the elimination rate, the hysteritis occurrence rate and the mastitis occurrence rate of the dairy cow within 30 days after delivery are reduced, the postpartum health management level of pasture cows is promoted, the problem of lack of clinical treatment drugs is solved, and the medication requirements of green and healthy breeding in the modern milk industry are met.

Yue Caijuan

Papers

Genetic polymorphism association analysis of SNPs on the species conservation genes of Tan sheep and Hu sheep.

Lick bricks with interaction effects of combining nutrition and improving immunity for cattle and sheep and technology thereof

Traditional Chinese medicine composition for preventing and treating postpartum abdominal pain of dairy cows and preparation method thereof