Postnatal feeding with high-fat combined with high-glucose diet induces precocious puberty in Sprague‒Dawley rat pups.

With economic development and overnutrition, including high-fat diets (HFD) and high-glucose diets (HGD), the incidence of obesity in children is increasing, and thus, the incidence of precocious puberty is increasing. Therefore, it is of great importance to construct a suitable animal model of overnutrition-induced precocious puberty for further in-depth study. Here, we fed a HFD, HGD, or HFD combined with a HGD to pups after P-21 weaning, while weaned pups fed a normal diet served as the control group. The results showed that HFD combined with a HGD increased the body weight (BW) of weaned rat pups. In addition, a HFD, HGD, and HFD combined with a HGD lowered the age at which vaginal opening occurred and accelerated the vaginal cell cycle. Furthermore, a HFD combined with a HGD increased the weight of the uterus and ovaries of weaned rat pups. Additionally, a HFD combined with a HGD promoted the development of reproductive organs in weaned female rat pups. Ultimately, a HFD combined with a HGD was found to elevate the serum levels of gonadotropin-releasing hormone (GnRH), luteinizing hormone (LH), follicle stimulating hormone (FSH), leptin, adiponectin, and oestradiol (E2) and increase hypothalamic GnRH, Kiss-1, and GPR54 expression levels in weaned female rat pups. The current study found that overnutrition, such as that through a HFD combined with HGD, could induce precocious puberty in weaned female rat pups. In addition, a rat model of overnutrition-induced precocious puberty was established.

First report of leaf spot on passion fruit in China, caused by Alternaria alternata.

Passion fruit (Passiflora edulis Sims), which is native to South America, is an important fruit crop in tropical and subtropical countries. Passion fruit growing areas have increased rapidly in southern China. In 2018 to 2019, circular spots on passion fruit were observed in Shangsi, Guangxi, China (21°15'N, 107°98'E). The disease occurred from June to April of the following year. The disease incidence was generally between 10% to 30%, but could reach up to 50% in purple passion fruit 'Tainong No.1'. The initial lesions on the fruits were small, with a brown center and a greasy margin, and then became sunken and lighter brown with a diameter of about 1 cm in later stages. The spots on the leaves were often surrounded by a yellow halo and turned into larger lesions after coalescence.. Five typical symptomatic fruit and leaves were collected from Shangsi county for the presumed pathogen isolation. Section of the samples were surface sterilized to isolate the fungus on potato dextrose agar (PDA) at 28°C. Five fungal isolates with similar morphology on PDA were obtained by single spore isolation. Colonies at the age of 7 days accompany with flourishing aerial hyphae, showed surface color varying from white to grey. Conidia were ovate or elliptic, light brown to brown, with 2 to 5 diaphragms, 0 to 4 longitudinal-oblique diaphragms, and mostly 8.2 to 36.7 µm × 5.4 to 15.8 µm. The morphology of the fungus resembled Alternaria alternata (Fr.) Keissl (Simmons, 2007). Each of the five isolates (SF-001, SF-002, SF-003, SF-004 and SF-005) was molecularly identified using genomic regions of 18S nrDNA (SSU), 28S nrDNA (LSU), RNA polymerase second largest subunit (RPB2), internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and translation elongation factor 1-alpha (TEF1) (Jayawardena et al., 2019). Nucleotide sequences of SSU (MZ275254, ON055696, ON055697, ON055698 and ON055699), LSU (MZ275253, ON062947, ON062948, ON062949, ON062950), RPB2 (MZ275251, ON055377, ON055378, ON055379 and ON055380), ITS (MW866522, MW866523, ON053451, ON053452 and ON053453), GAPDH (MZ286628, ON055381, ON055382, ON055383 and ON055384) and TEF1 (MZ275255, ON055373, ON055374, ON055375 and ON055376) were deposited in GenBank database. The LSU, GAPDH and TEF1 sequences showed 100% identity with A. alternata in NCBI (KX609773, MK683852 and MK637432, respectively). The SSU, RPB2 and ITS sequences showed 99% identity to A. alternata (U05194, MK605898 and MN856409, respectively). In pathogenicity test (Zhang et al., 2020), 3-month-old grafted 'Tainong No.1' seedlings and mature fruit were used. Five-mm-diameter mycelial plugs taken from 7-day-old PDA colonies of each of 5 isolates were placed on the leaves and fruit that were wounded with a sterilized needle to form 3 pinpricks. Sterile PDA plugs were used as control. Three plants and three fruits were used in each treatment, and the test was repeated twice. The inoculated plants and fruit were kept in plastic bags and grown in a chamber at 28℃. Typical lesions were observed on inoculated plants and fruit after 3 days, but the controls remained healthy. A. alternata was consistently reisolated from these typical lesions. Previously, leaf spot on passion fruit caused by A. alternata has only been recorded in New Zealand (Rheinländer, 2010). To our knowledge, this is the first report of A. alternata (Fr.) Keissl. causing leaf spot on passion fruit in China. The identification of the pathogen may help to take effective management strategies of controlling this disease.

Effect of alfalfa (Medicago sativa L.) saponins on meat color and myoglobin reduction status in the longissimus thoracis muscle of growing lambs.

The effect of alfalfa saponins (AS) supplementation on the meat quality especially the color for growing lamb was investigated. Fifty Hu male lambs with body weights (BW, 19.21 ± 0.45 kg) were divided into five groups and supplemented AS with 0, 500, 1,000, 2,000, and 4,000 mg/kg of dietary dry matter intake. After 90 days, all lambs were slaughtered. The longissimus thoracis muscle in lamb displayed significant changes in the content of intramuscular fat, especially n-3 polyunsaturated fatty acids, and drip loss within AS treatment (p < .05) between control and treatments groups. Redness (a*) significantly improved in both 0-day and 7-day storage with the AS supplementation coupled with the percentage of met-myoglobin reduction (p < .05). The redness (a*) change may result from improved met-myoglobin reducing activity, antioxidant enzymes, lactate dehydrogenase, and succinate dehydrogenase (p < .05) by AS supplementation in muscle. These enzymes may help to protect mitochondria function and reduce met-myoglobin, which bring a bright and red meat color.

The Effect of Dietary Lycopene Supplementation on Drip Loss during Storage of Lamb Meat by iTRAQ Analysis.

This study was designed to investigate the impact of dietary lycopene (antioxidant extracted from tomato) supplementation on postmortem antioxidant capacity, drip loss and protein expression profiles of lamb meat during storage. Thirty male Hu lambs were randomly divided into three treatment groups and housed in individual pens and received 0, 200 or 400 mg·kg-1 lycopene in their diet, respectively. All lambs were slaughtered after 3 months of fattening, and the longissimus thoracis (LT) muscle was collected for analyses. The results indicated that drip loss of LT muscle increased with storage days (P < 0.05). After storage for 7 days, significantly lower drip loss of meat was found in fed the lycopene-supplemented diet (P < 0.05). Dietary lycopene supplementation increased the activity of antioxidant enzymes (total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT)) (P < 0.05) and decreased the thiobarbituric acid reactive substance (TBARS) and carbonyl contents (P < 0.05). During the storage period (days 0, 5 and 7), a number of differentially abundant proteins (DAPs), including oxidases, metabolic enzymes, calcium channels and structural proteins, were identified based on iTRAQ data, with roles predominantly in carbon metabolism, oxidative phosphorylation, cardiac muscle contraction and proteasome pathways, and which contribute to decreased drip loss of lamb meat during storage. It can be concluded that dietary lycopene supplementation increased antioxidant capacity after slaughter, and the decreased drip loss during postmortem storage might occur by changing the expression of proteins related to enzyme activity and cellular structure in lamb muscle.

Nitroethanol in Comparison with Monensin Exhibits Greater Feed Efficiency Through Inhibiting Rumen Methanogenesis More Efficiently and Persistently in Feedlotting Lambs.

This study was conducted to determine the dietary supplemental effects of nitroethanol (NEOH) in comparison with monensin on growth performance and estimated methane (CH4) production in feedlotting lambs. Sixty male, small-tailed Chinese Han lambs were arranged at random into three dietary treatment groups: (1) a basal control diet (CTR), (2) the basal diet added with 40 mg/kg monensin (MON), (3) the basal diet added with 277 mg/kg nitroethanol (NEOH). During the 32-day lamb feeding, monensin and nitroethanol were added in period 1 (day 0-16) and then withdrawn in the subsequent period 2 (day 17-32) to determine their withdrawal effects. The average daily gain (ADG) and feed conversion rate in the whole period ranked: NEOH > MON > CTR (p < 0.01), suggesting that the dietary addition of NEOH in comparison with monensin presented a more lasting beneficial effect on feed efficiency. Methane emission was estimated with rumen VFA production and gross energy intake. Both monensin and NEOH addition in comparison with the control remarkably decreased CH4 emission estimate (24.0% vs. 26.4% decrease; p < 0.01) as well as CH4 emission per kg ADG (8.7% vs. 14.0% decrease; p < 0.01), but the NEOH group presented obvious lasting methanogenesis inhibition when they were withdrawn in period 2. Moreover, the in vitro methanogenic activity of rumen fluids was also decreased with monensin or NEOH addition (12.7% vs. 30.5% decrease; p < 0.01). In summary, the dietary addition of NEOH in comparison with monensin presented a greater promoting effect on growth performance in feedlotting lambs by inhibiting rumen methanogenesis more efficiently and persistently.

Identification of candidate genes in regulation of spermatogenesis in sheep testis following dietary vitamin E supplementation.

Dietary vitamin E supplementation is beneficial to semen quality in different sheep and goat breeds. The aim of this research was to further investigate the effect of vitamin E in sheep on spermatogenesis and its regulatory mechanisms using RNA-seq. Thirty male Hu lambs were randomly divided into three groups. The animals received 0, 200 or 2000 IU/day vitamin E dietary supplementation for 105 days, and its effects were subsequently evaluated. The results indicate vitamin E supplementation increased the number of germ cells in the testes and epididymides. The positive effects were reduced, however, in animals that received 2000 IU/d vitamin E. Using the RNA-seq procedure, there was detection of a number of differentially expressed genes such as NDRG1, FSCN3 and CYP26B1 with these genes being mainly related to the regulation of spermatogenesis. Supplementation with 2000 IU/d vitamin E supplementation resulted in a lesser abundance of skeleton-related transcripts such as TUBB, VIM and different subtypes of collagen, and there was also an effect on the ECM-receptor interaction pathway. These changes appear to be responsible for the lesser beneficial effect of the greater vitamin E concentrations. The results provide a novel insight into the regulation of spermatogenesis by vitamin E at the molecular level, however, for a precise understanding of functions of the affected genes there needs to be further study.

Screening of α-Tocopherol Transfer Protein Sensitive Genes in Human Hepatoma Cells (HepG2).

α-Tocopherol transfer protein (α-TTP) is a ~32 kDa protein expressed mainly in hepatocytes. The major function of the protein is to bind specifically to α-tocopherol and, together, the complex transfers from late lysosomes to the cell membrane. A previous study indicated that some factors might be required in the transferring process. However, there is little information available about the potential transferring factors. In addition, there remains much to learn about other physiological processes which α-TTP might participate in. Thus, in this study a human α-TTP eukaryotic expression vector was successfully constructed and expressed in human hepatoma cells (HepG2). The sensitive genes related to α-TTP were then screened by microarray technology. Results showed that expression of the vector in HepG2 cells led to the identification of 323 genes showing differential expression. The differentially expressed transcripts were divided into four main categories, including (1) cell inflammation; (2) cell cycle and cell apoptosis; (3) cell signaling and gene regulation; and (4) cellular movement. A few cellular movement related transcripts were selected and verified by quantitative real-time PCR. Expressions of some were significantly increased in α-TTP-expressed group, which indicated that these factors were likely to play a role in the transferring process.

Regulation of sheep α-TTP by dietary vitamin E and preparation of monoclonal antibody for sheep α-TTP.

α-Tocopherol transfer protein (α-TTP) is a cytosolic protein that plays an important role in regulating concentrations of plasma α-tocopherol (the most bio-active form of vitamin E). Despite the central roles that α-TTP plays in maintaining vitamin E adequacy, we have only recently proved the existence of the α-TTP gene in sheep and, for the first time, cloned its full-length cDNA. However, the study of sheep α-TTP is still in its infancy. In the present study, thirty-five local male lambs of Tan sheep with similar initial body weight were randomly divided into five groups and fed with diets supplemented with 0 (control group), 20, 100, 200, 2000IU·sheep(-1)·d(-1) vitamin E for 120 days. At the end of the experiment, the plasma and liver vitamin E contents were analyzed first and then α-TTP mRNA and protein expression levels were determined by quantitative real-time PCR (qRT-PCR) and Western-blot analysis, respectively. In addition, as no sheep α-TTP antibody was available, a specific monoclonal antibody (McAb) against the ovine α-TTP protein was prepared. The effect of vitamin E supplementation was confirmed by the significant changes in the concentrations of vitamin E in the plasma and liver. As shown by qRT-PCR and Western-blot analysis, dietary vitamin E does not affect sheep α-TTP gene expression, except for high levels of vitamin E supplementation, which significantly increased expression at the protein level. Importantly, the specific sheep anti-α-TTP McAb we generated could provide optimal recognition in ELISA, Western-blot and immunohistochemistry assays, which will be a powerful tool in future studies of the biological functions of sheep α-TTP.

Dietary vitamin E affects α-TTP mRNA levels in different tissues of the Tan sheep.

The α-tocopherol transfer protein (α-TTP) is a ~32kDa cytosolic protein that plays an important role in the efficient circulation of plasma α-tocopherol in the body, a factor with great relevance in reproduction. The α-TTP gene has been studied in a number of tissues; however, its expression and function in some ovine tissues remain unclear. A previous study from our laboratory has demonstrated α-TTP expression in sheep liver. In the present study we determined whether α-TTP is expressed in non-liver tissues and investigated the effects of dietary vitamin E on the α-TTP mRNA levels. Thirty-five male Tan sheep with similar body weight were randomly allocated into five groups and supplemented 0, 20, 100, 200 and 2000IUsheep(-1)day(-1) vitamin E, for four months, respectively. At the end of the study, the animals were slaughtered and tissue samples from the heart, spleen, lung, kidney, longissimus dorsi muscle and gluteus muscle were immediately collected. We found that the α-TTP gene is expressed in sheep tissues other than the liver. Moreover, dietary vitamin E levels had influenced the expression levels of α-TTP gene in these tissues in a tissue-specific way. The technique of immunohistochemistry was used to detect α-TTP in tissues of the heart, spleen, lung, and kidney and we found that α-TTP was mainly located in the cytoplasm while no α-TTP immunoreactivity was detected in the cytoplasm of longissimus dorsi and gluteus muscle samples. Importantly, our findings lay the foundation for additional experiments focusing on the absorption and metabolism of vitamin E in tissues other than the liver.

Molecular cloning and characterization of the sheep α-TTP gene and its expression in response to different vitamin E status.

The α-tocopherol transfer protein (α-TTP) is a ~32 kDa protein that exhibits a marked ligand specificity and selectively recognizes of α-tocopherol, which is the most active form of vitamin E. The α-TTP gene has been cloned and its physiological functions have been studied in numbers of species, however, the understanding of sheep α-TTP is still in his infancy. In this study, the full-length cDNA of sheep α-TTP gene was cloned from sheep liver by using of rapid amplification of complementary DNA ends (RACE). As a result, the sheep α-TTP gene was 1098 bp in nucleotide which contained 23 bp 5'-untranslated region (UTR), 226 bp 3'-UTR and 849 bp open reading frame (ORF) that encoded a basic protein of 282 amino acids. Further bioinformatic analysis indicated that the sheep α-TTP gene had a high homologous of both nucleotide and amino acid sequences compared with that of other species and had a Sec14p-like lipid-binding domain which called the CRAL-TRIO domain. Moreover, the expression of sheep α-TTP mRNA and protein in response to different vitamin E supplemented levels were observed according to quantitative real-time PCR (qRT-PCR) and Western blotting analysis. The results showed that dietary vitamin E levels did not affect α-TTP mRNA expression significantly while the low vitamin E supplemented level groups of sheep had significantly higher α-TTP protein compared to high-vitamin E groups.

[Effects of different years of planting Pennisetum sp. on the plant- and insect diversity in Pennisetum sp. communities].

This paper studied the effects of 1-, 2- and 3 years of planting Pennisetum sp. on the plant- and insect diversity in the Pennisetum sp. communities, taking the barren mountain land without planting Pennisetum sp. as the control (CK). Compared with CK, the plant species richness in Pennisetum sp. communities with different years of planting was lower, but the coverage was higher. The coverage in the Pennisetum sp. community having been planted for 3 years was the highest, up to 91.6%, and 75.8% higher than the CK. The insect species richness in the Pennisetum sp. communities having been planted for 1, 2 and 3 years was 3.6, 5.3 and 5.6 times of the CK, respectively. The plant- and insect diversity indices, including Simpson index, Shannon index, evenness, Brillouin index, and McIntosh index for the Pennisetum sp. communities with different years of planting were significantly higher than the CK, which indicated that the growth of Pennisetum sp. could affect the plant- and insect diversity. With the increasing year of planting, the plant- and insect diversity in Pennisetum sp. communities tended to be stable.