Codonopsis pilosula is an important Chinese herbal medicine. However, fresh C. pilosula is prone to decay during storage due to microorganism infections, seriously affecting the medicinal value and even causing mycotoxin accumulation. Therefore, it is necessary to study the pathogens present and develop efficient control strategies to mitigate their detrimental effects on the herbs during storage. In this study, fresh C. pilosula was collected from Min County in Gansu Province, China. The natural disease symptoms were observed during different storage stages, and the pathogens causing C. pilosula postharvest decay were isolated from the infected fresh C. pilosula. Morphological and molecular identification were performed, and pathogenicity was tested using Koch’s postulates. In addition, the control of ozone was examined against the isolates and mycotoxin accumulation. The results indicated that the naturally occurring symptom increased progressively with the extension of storage time. The mucor rot caused by Mucor was first observed on day 7, followed by root rot caused by Fusarium on day 14. Blue mold disease caused by Penicillum expansum was detected as the most serious postharvest disease on day 28. Pink rot disease caused by Trichothecium roseum was observed on day 56. Moreover, ozone treatment significantly decreased the development of postharvest disease and inhibited the accumulations of patulin, deoxynivalenol, 15-Acetyl-deoxynivalenol, and HT-2 toxin.

H₂S plays an important role in various inflammatory diseases. However, the role of H₂S and synthetic enzymes in Holstein cows with CM is unknown. The aim of this study was to identify DEPs associated with sulfide metabolism and further investigate their roles in dairy cows with CM. From 3739 DEPs generated by data-independent acquisition proteomics, we identified a total of 17 DEPs included in 44 GO terms and five KEGG pathways related to sulfide metabolism, including CTH and cystathionine-β-synthase (CBS). Immunohistochemical and immunofluorescence staining results showed that CTH and CBS proteins were present mainly in the cytoplasm of mammary epithelial cells. Endogenous H₂S production in the serum of the CM group was significantly lower than that of the healthy Holstein cows. CTH and CBS mRNA and protein levels in the mammary glands of the CM group were significantly downregulated compared to those of the healthy group. These results indicate that CTH and H₂S were correlated with the occurrence and development of CM in Holstein cows, which provides important insights into the function and regulatory mechanism of CTH/H₂S in Holstein cows.

Nuclear hormone receptors NR1D1 and NR4A2 play important roles in the synthesis and metabolism of hormones that are thought to be strictly regulated by the hypothalamus-pituitary-gonad axis (HPG) tissues via gene expression. However, in the yak, the function and regulatory mechanisms of NR1D1 and NR4A2 are not clearly understood. The current study is aimed to investigate the expression patterns, distribution and functions of these two receptors in HPG tissues in male Tianzhu white yaks. Immunohistochemical staining showed NR1D1 and NR4A2 proteins were present in all yak HPG tissues with differential expression patterns and degrees of staining, particularly in Leydig cells that were strongly positive in accordance with the immunofluorescence results. qRT-PCR and Western blot results suggested that the highest expression levels of NR1D1 and NR4A2 mRNA were present in the hypothalamus, while the expression levels of NR1D1 and NR4A2 proteins were higher in the testis and epididymis than in the hypothalamus or pituitary gland. In addition, expression levels of NR1D1 and NR4A2 mRNA and protein in testicular tissues differed by age. Expression levels were significantly higher at 6 years of age. Gene ontology (GO) and pathway analysis enrichment revealed that NR1D1 may directly regulate the synthesis and metabolism of steroid hormones via interaction with different targets, while NR4A2 may indirectly regulate the synthesis and metabolism of steroid hormones. These results showed that NR1D1 and NR4A2, as important mediators, are involved in the regulation of male yak reproduction, and especially of steroid hormones and androgen metabolism. These results will be helpful for the further understanding of the regulatory mechanisms of NR1D1 and NR4A2 in yak reproduction.

Increased poll gland secretion is a major characteristic and indicator of estrus in male Bactrian camels; however, research on these poll glands and their secretion is extremely rare. In this study, we determine the chemical composition of poll gland secretions and identify the key functional substances that regulate seasonal estrus in male camels. A GC/LC-MS dual platform was used to analyze ventral hair (control) and neck mane samples containing poll gland secretions from male Bactrian camels during estrus. Multidimensional and single-dimensional analyses were used to screen differentially expressed metabolites (DEMs) between groups. Functional prediction of enriched metabolites was performed using a Human Metabolome Database comparison and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, which were then compared with a behavioral analysis of male Bactrian camels in estrus. A total of 1172 DEMs and 34 differential metabolic pathways were identified. One metabolite group was found to relate to steroid synthesis and metabolism, and another metabolite group was associated with neural metabolism. Therefore, we speculate that steroids and neurochemicals jointly regulate estrous behavior in male Bactrian camels, thus providing theoretical insights into the development and function of poll glands in Bactrian camels.

Radix Astragali (RA) is the root of Astragalus membranaceus var. mongholicus (Bunge) P.K. Hsiao. In order to determine targeted and effective methods for treating mold infections in fresh RA, pathogenic fungi were isolated and identified using morphology and molecular biology techniques, and factors affecting the growth of the pathogenic fungi, such as temperature, pH, light, and carbon and nitrogen sources, were analyzed. Changes in the main active ingredients of RA, such as carbohydrates, flavonoids, and saponins, were detected before and after infection with pathogenic fungi. The results showed that Penicillium polonicum, Trichothecium roseum, and Fusarium equiseti were the main pathogens causing postharvest disease in fresh RA. The optimum growth temperature for the three pathogens was 25 °C; P. polonicum was able to grow at an alkaline pH of 8–9, while T. roseum and F. equiseti grew better and had higher levels of sporulation under acidic conditions with pH values of 5–6. Different carbon and nitrogen sources had different effects on mycelium growth and sporulation. Darkness was favorable for the growth and sporulation of the three pathogens. Compared with healthy RA, the total carbohydrate, flavonoid, and saponin contents of the RA samples infected with the three fungi decreased. This study provides a theoretical basis for the scientific and precise control of RA postharvest disease during storage by targeting different pathogens and their growth conditions.

The type II Na/Pi co-transporter (NaPi2b), encoded by the solute carrier (SLC) transporter 34A2 (SLC34A2), is responsible for calcium (Ca) and phosphorus (P) homeostasis. Unbalanced Ca/P metabolism induces mastitis in dairy cows. However, the specific role of SLC34A2 in regulating this imbalance in Holstein cows with clinical mastitis (CM) remains unclear. The aim of this study was to investigate the role of SLC34A2 and identify differentially expressed proteins (DEPs) that interact with SLC34A2 and are associated with Ca/P metabolism in dairy cows with CM. Immunohistochemical and immunofluorescence staining results showed that SLC34A2 was located primarily in the mammary epithelial cells of the mammary alveoli in both the control (healthy cows, Con/C) and CM groups. Compared to the Con/C group, the relative expression of the SLC34A2 gene and protein were significantly downregulated in the CM group. We identified 12 important DEPs included in 11 GO terms and two pathways interacting with SLC34A2 using data-independent acquisition proteomics. The PPI (protein-and-protein interaction) network results suggested that these DEPs were associated with ion metabolism and homeostasis, especially SLC34A2. These results demonstrate that SLC34A2 downregulation is negatively correlated with the occurrence and development of CM in Holstein cows, providing a basis for exploring the function and regulatory mechanism of SLC34A2 in Ca/P metabolism and homeostasis in Holstein cows with CM.