To investigate the impact of total flavonoids of Epimedium (TFE) on the development of follicles of laying hens, 3 types of follicles including primary, prehierarchical, and preovulatory follicles were selected to obtain the follicular granulosa cells cultured in vitro. First, extraction of TFE was conducted by alcohol-soluble and ultrasonic methods. The effects of TFE on activity and proliferation of follicular granulosa cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and measuring the expression of proliferating cell nuclear antigen mRNA through real-time quantitative polymerase chain reaction, and the expression of the follicle-stimulating hormone receptor, luteinizing hormone receptor, steroidogenic acute regulatory protein, and cytochrome P450 family 11 subfamily A member 1 mRNA was detected to study the functions of TFE affecting the differentiation and hormone secretion by granulosa cells. The results showed that TFE significantly improved the proliferation of 3 types of granulosa cells and promoted the differentiation of granulosa cells and accelerated the conversion of primary follicles to prehierarchical follicles. Total flavonoids of Epimedium played an important role in promoting progesterone secretion by prehierarchical and preovulatory granulosa cells. The results indicated that TFE could promote proliferation and differentiation of follicular granulosa cells and improve hormone secretion and follicle development, which provided reference data for TFE used as a feed additive or safe Chinese veterinary medicine to promote the laying rate.

In the present study, for the purpose of investigating the effects of the total flavonoids of Epimedium (TFE) in regard to preventing the development of atrophied oviducts and follicles induced by forced molting, 300-day-old Hy-Line Brown layer hens were divided into 3 study groups as follows: the control (CON) group was the normal group, without forced molting and TFE treatments; the TFE1 group was treated by adding a 1‰ TFE treatment after forced molting; and the TFE0 group was not treated by TFE after forced molting. During this study's experimental process, the egg production rates were recorded each day. In addition, the hens were randomly chosen to be weighed every 4 D and also randomly selected to be sacrificed every 7 D. Then, sample tissues of albumen-secreting part and uterus from the fallopian tube of the layer hens were collected for PCR and hematoxylin-eosin staining tests. The results showed that the body weights, number of follicles, and weights and sizes of the fallopian tube for the TFE1 and TFE0 groups were significantly reduced when compared with those of the control group on the 15th D of the experiment. Furthermore, at the end of study, it was found that the egg production rates, weights of the fallopian tube, and ovarian follicles of TFE1 had recovered to normal levels. At the same time, the serum estrogen and the expressions of the progesterone receptor and estrogen receptor mRNA in fallopian tube were higher than those observed for the TFE0 group. The results of this study provided valuable evidence that TFE could improve the development of atrophied oviducts and increase the egg laying rates, thereby making it a potential multicomponent natural drug for egg production in the future.

Few data have suggested how norepinephrine (NE) and acetylcholine (Ach) regulate the development of Leydig cells in mice at prepuberty, except for data indicating endocrine effects. The present study aims to elucidate the roles of NE and Ach on the differentiation and proliferation of Leydig cells. Firstly, the expression of adrenergic receptors and muscarinic acetylcholine receptors in Leydig cells was investigated. It was found that adrenergic receptors (β1AR,β2AR, andα1D) and muscarinic acetylcholine receptors (M1 and M3) mRNA are expressed in adult Leydig cells. Then, the effects of NE and Ach on the differentiation and proliferation of Leydig cells were analyzed. The results showed that NE and Ach at 10 μM significantly increased the number of 3β-hydroxysteroid-dehydrogenase- (3β-HSD-) positive Leydig cells and improved the expression of proliferating cell nuclear antigen (PCNA) in Leydig cells on postnatal day (PD) 15 (P<0.05). NE and Ach at 10 μM had no impact on the expression of PCNA mRNA (P>0.05), but reduced the expression of 3β-HSD mRNA in adult Leydig cells and a murine Leydig tumor cell line (MLTC-1) (P<0.05). Therefore, a conclusion may be reached that NE and Ach participated in stimulating the development of Leydig cells in mice from prepuberty to adult stage.

Enteric viruses, as a potential pathogen, have been found to be vital causes of economic losses in poultry industry worldwide. The enteric viruses widely studied to date mainly include avian nephritis virus (ANV), avian reovirus (ARe), chicken astrovirus (CAstV), chicken parvovirus (ChPV), fowl adenovirus group I (FAdV-1), infectious bronchitis virus (IBV), and avian rotavirus (ARoV). This paper aimed to identify single and multiple infections of the seven enteric viruses using the data obtained from positive 145 enteric virus samples in poultry flocks from different areas in Hebei Province, throughout the period from 2019 to 2021. Next, the correlation between bird age and clinical signs was investigated using PCR and RT-PCR techniques. Furthermore, the whole genomes of seven parvovirus strains and open reading frame 2 (ORF2) of six CAstV strains and eight ANV strains were sequenced for phylogenetic analysis and recombination analysis, to characterize the viruses and evaluate species correlation and geographic patterns. A total of 11 profiles of virus combinations were detected; 191 viruses were detected in 145 samples; 106 single infections were reported in 73.1% of the samples; and multiple infections were detected in the remaining 26.9%. For viruses, 69% of ChPV was correlated with single infection, while ANV (61.4%) and CAstV (56.1%) were correlated with multiple infections. However, IBV and ARe were not detected in any of the samples. Recombination events were reported in parvovirus, and all CAstV sequences investigated in this paper were included within genotype Bii. The eight ANV strains pertained to different subtypes with significant differences. The above results revealed for the first time the complexity of enteric viruses over the past several years, thus contributing to disease prevention and control in the future.

Previously, a fungus was isolated from a diseased pigeon group clinically suspected of being infected with Candida. The fungus was subsequently identified as Candida glabrata using morphology, physiology, biochemistry, and molecular biology testing methods. In the present study, to determine the controlling effects of Chinese herbal medicine for C. glabrata, the bacteriostatic effects of the ethanol extracts Acorus gramineus, Sophora flavescens, Polygonum hydropiper, Cassia obtusifolia, Pulsatilla chinensis, Dandelion, and Cortex phellodendri on C. glabrata in vitro were analyzed. The results showed that the minimum inhibitory concentrations (MIC80) of Cortex phellodendri was 0.25 μg/μL. Meanwhile, that of S. flavescens was 32 μg/μL; C. obtusifolia was 56 μg/μL; A. gramineus and Polygonum hydropiper was 64 μg/μL; and P. chinensis was 112 μg/μL. However, MIC80 for Dandelion was undetectable. In addition, improved drug sensitivity tests revealed that colonies had grown after 24 h in the blank group, as well as the Polygonum hydropiper, P. chinensis, Dandelion, and ethanol groups. The colonies first appeared at the 48-hour point in the other drug-sensitive medium of Chinese herbal medicine. However, no colony growth was found in Cortex phellodendri medium, and the formation of the maximum colony diameter in that group was later than the blank group (e.g., 96 h in the blank group and 120 h in the Chinese herbal medicine group). It was observed that only 17 colony-forming units had grown in 125 μg/μL of the S. flavescens medium, which was significantly different from other groups. Also, the final colony diameter was significantly smaller than that of the other experimental groups. Therefore, it was determined that the A. gramineus, S. flavescens, Polygonum hydropiper, Cassia obtusifolia, P. chinensis, and Cortex phellodendri had certain inhibitory effects on the growth of the C. glabrata. Among those, it was observed that the Cortex phellodendri had the strongest inhibitory effects, followed by the S. flavescens. In the future, these Chinese herbal medicines are expected to be used to treat the fungal infections related to C. glabrata in poultry to improve production performance.

Aspergillus flavus and Aspergillus fumigatus were derived and identified from the ducks infected with fungi. In order to investigate the effectiveness of Chinese herbal medicines against Aspergillus flavus and Aspergillus fumigatus, in vitro antibacterial test and animal infection control test were conducted to study the antibacterial activity of the Chinese medicine mixture which was compatible with Acorus gramineus, Phellodendron chinensis and Cassia obtusifolia. According to the results, the liver of chickens infected with Aspergillus flavus and Aspergillus fumigatus displayed granulomatous lesions, indicating that the isolation of pathogen from the lungs of sick ducks is also pathogenic to chickens. As suggested by the results of in vitro drug sensitivity test, the mixture 1 MIC₈₀ was the minimum, the MIC₈₀ of Aspergillus flavus was 16 μg/μL, and the MIC₈₀ of Aspergillus fumigatus was 4 μg/μL. In a petri dish of the same concentration, the colony diameter of Aspergillus flavus and Aspergillus fumigatus in Mixture 1 was the minimum. Besides, Aspergillus flavus colonies grew when the concentration was 64 μg/μL, and Aspergillus fumigatus colonies grew when the concentration was 4 μg/μL, which suggests the more significant inhibitory effect of Mixture 1 on Aspergillus flavus and Aspergillus fumigatus. According to the results of animal experiments, there was a significantly lower activity level of Glutamic oxaloacetic transaminase (GOT) and Glutamate pyruvic transaminase (GPT) in the protection group and the treatment group than in the bacterial infection group. As indicated by the blood smear results, there were more neutrophils in the infected group than in the prevention group and the treatment group. Thus, it can be seen from that the Mixture 1 produced preventive and therapeutic effects on the chickens infected with Aspergillus flavus and Aspergillus fumigatus.