A New Method to Detect Buffalo Mastitis Using Udder Ultrasonography Based on Deep Learning Network.

Mastitis is one of the most predominant diseases with a negative impact on ranch products worldwide. It reduces milk production, damages milk quality, increases treatment costs, and even leads to the premature elimination of animals. In addition, failure to take effective measures in time will lead to widespread disease. The key to reducing the losses caused by mastitis lies in the early detection of the disease. The application of deep learning with powerful feature extraction capability in the medical field is receiving increasing attention. The main purpose of this study was to establish a deep learning network for buffalo quarter-level mastitis detection based on 3054 ultrasound images of udders from 271 buffaloes. Two data sets were generated with thresholds of somatic cell count (SCC) set as 2 × 105 cells/mL and 4 × 105 cells/mL, respectively. The udders with SCCs less than the threshold value were defined as healthy udders, and otherwise as mastitis-stricken udders. A total of 3054 udder ultrasound images were randomly divided into a training set (70%), a validation set (15%), and a test set (15%). We used the EfficientNet_b3 model with powerful learning capabilities in combination with the convolutional block attention module (CBAM) to train the mastitis detection model. To solve the problem of sample category imbalance, the PolyLoss module was used as the loss function. The training set and validation set were used to develop the mastitis detection model, and the test set was used to evaluate the network's performance. The results showed that, when the SCC threshold was 2 × 105 cells/mL, our established network exhibited an accuracy of 70.02%, a specificity of 77.93%, a sensitivity of 63.11%, and an area under the receiver operating characteristics curve (AUC) of 0.77 on the test set. The classification effect of the model was better when the SCC threshold was 4 × 105 cells/mL than when the SCC threshold was 2 × 105 cells/mL. Therefore, when SCC ≥ 4 × 105 cells/mL was defined as mastitis, our established deep neural network was determined as the most suitable model for farm on-site mastitis detection, and this network model exhibited an accuracy of 75.93%, a specificity of 80.23%, a sensitivity of 70.35%, and AUC 0.83 on the test set. This study established a 1/4 level mastitis detection model which provides a theoretical basis for mastitis detection in buffaloes mostly raised by small farmers lacking mastitis diagnostic conditions in developing countries.

Integrating mid-infrared spectroscopy, machine learning, and graphical bias correction for fatty acid prediction in water buffalo milk.

BACKGROUND: Buffalo milk, constituting 15% of global production, has higher fatty acids content than Holstein milk. Fourier-transform mid-infrared (FT-MIR) spectroscopy is widely used for dairy analysis, but its application to buffalo milk, with larger fat globules, remains understudied. The ultimate goal of this study is to develop machine learning models based on FT-MIR spectroscopy for predicting fatty acids in buffalo milk and to assess the accuracy of commercial milk analyzers. This research provides a convenient, fast, and environmentally friendly method for detecting the fatty acid composition in buffalo milk. RESULTS: We employed six machine learning algorithms to establish a detection model for 34 fatty acids in buffalo milk. The predictive models demonstrated robust capabilities for high-content fatty acids [C14:0, C15:0, C16:0, C17:0, C18:0, C18:1, saturated fatty acid (SFA), monounsaturated fatty acid (MUFA)], with errors within a 15% range. Traditional FT6000 detection methods exhibited limitations in measuring SFAs and polyunsaturated fatty acids (PUFA). Implementing a mean difference correction of 0.21 for MUFAs and applying regression equations (SFA × 1.0639 + 0.0705; PUFA × 0.5472 + 0.0047) significantly improved measurement accuracy. CONCLUSION: This study successfully developed a predictive model for fatty acids in Mediterranean buffalo milk based on FT-MIR spectroscopy. Additionally, a correction was applied to the existing measurement device, FT6000, enabling more accurate measurements of fatty acids in buffalo milk. The findings have practical implications for the food industry, offering a faster and more reliable approach to assess and monitor fatty acid composition in buffalo milk, potentially influencing product development and quality control processes. © 2024 Society of Chemical Industry.

Identification of Milk Adulteration in Camel Milk Using FT-Mid-Infrared Spectroscopy and Machine Learning Models.

Camel milk, esteemed for its high nutritional value, has long been a subject of interest. However, the adulteration of camel milk with cow milk poses a significant threat to food quality and safety. Fourier-transform infrared spectroscopy (FT-MIR) has emerged as a rapid method for the detection and quantification of cow milk adulteration. Nevertheless, its effectiveness in conveniently detecting adulteration in camel milk remains to be determined. Camel milk samples were collected from Alxa League, Inner Mongolia, China, and were supplemented with varying concentrations of cow milk samples. Spectra were acquired using the FOSS FT6000 spectrometer, and a diverse set of machine learning models was employed to detect cow milk adulteration in camel milk. Our results demonstrate that the Linear Discriminant Analysis (LDA) model effectively distinguishes pure camel milk from adulterated samples, maintaining a 100% detection rate even at cow milk addition levels of 10 g/100 g. The neural network quantitative model for cow milk adulteration in camel milk exhibited a detection limit of 3.27 g/100 g and a quantification limit of 10.90 g/100 g. The quantitative model demonstrated excellent precision and accuracy within the range of 10-90 g/100 g of adulteration. This study highlights the potential of FT-MIR spectroscopy in conjunction with machine learning techniques for ensuring the authenticity and quality of camel milk, thus addressing concerns related to food integrity and consumer safety.

Relationships between body- and udder-related type traits with somatic cell counts and potential use for an early selection method for water buffaloes (Bubalus bubalis).

Water buffalo milk is a reliable source of high-quality nutrients; however, the susceptibility of mastitis in buffaloes must be taken into consideration. An animal with somatic cell count (SCC) of greater than 250,000 cells/mL is reported to be likely to have mastitis which has serious adverse effects on animal health, reproduction, milk yield, and milk quality. Type traits (TTs) of water buffalo can affect SCC in animal milk to some extent, but few reports on the correlation between SCC and TTs are available. In this study, a total of 1908 records collected from 678 water buffaloes were investigated. The general linear model was used to identify factors associated with phenotypic variation of the somatic cell score (SCS) trait, including parity, lactation length, calving year, and calving season as fixed effects. Using PROC CORR analysis method, taking calving year and lactation length as covariates, the correlation co-efficient between TT and SCS was obtained. Our results showed that correlation co-efficients between the 45 TTs with SCS ranged from 0.003 to 0.443 (degree of correlation). The correlation between udder traits and SCS was greater than that between body structure traits and SCS. Among udder traits, distance between teats (including front and rear teat distance [r = 0.308], front teat distance [r = 0.211], and teat crossing distance [r = 0.412]) and teat circumference (r = 0.443) had the highest correlation with SCS, followed by the leg traits including rear leg height (r = -0.354) and hock bend angle (r = -0.170). Animal with high rear legs (>48 cm) and short teat crossing distance (<17 cm), and narrow teat circumference (<11 cm) exhibited low SCS. Using four nonlinear models (Von Bertalanffy, Brody, Logistic, and Gompertz), the optimal growth curves of the TTs highly correlated with the SCS (rear leg height and teat crossing distance) were fitted, and the correction co-efficients of these two TTs rear leg height and teat crossing distance of animal from young age (2 mo old) to first lactation (35 mo old) were attained for establishment of early selection method for water buffaloes with low SCS. This study provides theoretical support for early selection of low-SCS water buffaloes and lays a foundation for improving milk quality and promoting healthy development of water buffalo's dairy industry.

Some type traits (TTs) have been reported to affect somatic cell count (SCC) in bovine milk, which result in mastitis to some extent, but the correlation between SCC and TTs of water buffalo has been poorly understood. Here, a total of 1908 records from 678 buffaloes were investigated. The correlation between 45 TTs and somatic cell score (SCS) was analysed, and the optimal growth curves of TTs highly correlated with SCS were fitted. Our result showed that high rear legs (>48 cm) and short teat crossing distance (<17 cm), and narrow teat circumference (<11 cm) were correlated with low SCS. We obtained correction co-efficients for two TTs highly correlated with SCS of water buffalos from young age (2 mo old) to first lactation (35 mo) by fitting the optimal growth curve for rear leg height and teat crossing distance. This study provides theoretical support for early selection for water buffaloes that are less susceptible to mastitis, and lays a foundation for improving milk quality and promoting healthy development of water buffalo dairy industry.

Establishment and Validation of Fourier Transform Infrared Spectroscopy (FT-MIR) Methodology for the Detection of Linoleic Acid in Buffalo Milk.

Buffalo milk is a dairy product that is considered to have a higher nutritional value compared to cow's milk. Linoleic acid (LA) is an essential fatty acid that is important for human health. This study aimed to investigate and validate the use of Fourier transform mid-infrared spectroscopy (FT-MIR) for the quantification of the linoleic acid in buffalo milk. Three machine learning models were used to predict linoleic acid content, and random forest was employed to select the most important subset of spectra for improved model performance. The validity of the FT-MIR methods was evaluated in accordance with ICH Q2 (R1) guidelines using the accuracy profile method, and the precision, the accuracy, and the limit of quantification were determined. The results showed that Fourier transform infrared spectroscopy is a suitable technique for the analysis of linoleic acid, with a lower limit of quantification of 0.15 mg/mL milk. Our results showed that FT-MIR spectroscopy is a viable method for LA concentration analysis.

Evaluation of Parity Effect on Characteristics and Minerals in Buffalo (Bubalus Bubalis) Colostrum and Mature Milk.

Colostrum is a vital performance for buffaloes and potentially functional foods in the future. Therefore, this study aimed to evaluate the difference between the parity of buffalo colostrum and mature milk. Twenty pregnant buffaloes (primiparous = 10; multiparous = 10) were assigned to the same diet prepartum and milking routine postpartum. Calves were separated from the dams immediately after birth and colostrum was harvested within 2 h, whilst mature milk was harvested at 7 days postpartum. The colostrum was analyzed for immunoglobulin G and milk composition as the mature milk. The results showed that there was a higher level of protein, solid not fat, and milk urea nitrogen (p < 0.05), with a tendency for higher total solids (p = 0.08) in primiparous buffaloes' colostrum compared with multiparous. No parity effect was observed in colostrum immunoglobulin G, fat, lactose, and yields of colostrum and composition (p > 0.05). There was no difference in mature milk composition and yield by parity affected (p > 0.05). Compared with mature milk composition, colostrum had a higher content protein, total solids, solid not fat, and milk urea nitrogen (p < 0.05); however, fat and lactose were lower than that of mature milk (p < 0.05). For minerals, multiparous buffaloes' colostrum had a higher concentration of Fe (p = 0.05), while the mature milk had higher concentrations of K and P compared with primiparous. Buffalo colostrum had higher concentrations of Na, Mg, Co, Fe, and K with a lower concentration of Ca relative to mature milk (p < 0.05). It was observed that parity affected colostrum characteristics rather than mature milk and caused subtle variations in minerals in colostrum and mature milk of buffaloes. As lactation proceeded, both milk composition and minerals in the milk changed drastically.

Effects of Different Bedding Materials on Production Performance, Lying Behavior and Welfare of Dairy Buffaloes.

Different bedding materials have important effects on the behavioristics, production performance and welfare of buffalo. This study aimed to compare the effects of two bedding materials on lying behavior, production performance and animal welfare of dairy buffaloes. More than 40 multiparous lactating buffaloes were randomly divided into two groups, which were raised on fermented manure bedding (FMB) and chaff bedding (CB). The results showed that the application of FMB improved the lying behavior of buffaloes, the average daily lying time (ADLT) of buffaloes in FMB increased by 58 min compared to those in CB, with a significant difference (p < 0.05); the average daily standing time (ADST) decreased by 30 min, with a significant difference (p < 0.05); and the buffalo comfort index (BCI) increased, but the difference was not significant (p > 0.05). The average daily milk yield of buffaloes in FMB increased by 5.78% compared to buffaloes in CB. The application of FMB improved the hygiene of buffaloes. The locomotion score and hock lesion score were not significantly different between the two groups and all buffaloes did not show moderate and severe lameness. The price of FMB was calculated to be 46% of CB, which greatly reduced the cost of bedding material. In summary, FMB has significantly improved the lying behavior, production performance and welfare of buffaloes and significantly reduce the cost of bedding material.

Summer Buffalo Milk Produced in China: A Desirable Diet Enriched in Polyunsaturated Fatty Acids and Amino Acids.

The objective of the study was to compare and reveal differences in basic chemical parameters, fatty acids, amino acids, and lipid quality indices of crossbred buffalo (swamp x river type) milk produced in summer and winter. The buffalo milk samples were collected in summer (Jul-Aug) and winter (Dec-Jan) from Hubei province, China. The samples were detected by using CombiFoss apparatus, gas chromatography, and an automated specialized amino acid analyzer. The results showed that the basic chemical parameters, fatty acid profiles, lipid quality indices, and amino acid profiles of crossbred buffalo milk differed between summer and winter. Specifically, summer buffalo milk exhibited a higher content of MUFA (monounsaturated fatty acids) and PUFA (polyunsaturated fatty acids) than winter buffalo milk. Summer buffalo milk had a lower content of major SFA (saturated fatty acids), a higher content of ω-3 and DFA (hypocholesterolemic fatty acids), a lower ω-6/ω-3 ratio, a higher value of 3 unsaturated fatty acid indices (C14, C16, C18), and a lower value of IA (index of atherogenicity) and IT (index of thrombogenicity) than winter buffalo milk. Additionally, 17 amino acids, including 8 EAA (essential amino acids) and 9 NEAA (non-essential amino acids) were higher in summer buffalo milk. These results indicated that summer buffalo milk was more health-beneficial than winter buffalo milk. Therefore, summer buffalo milk might be a desirable diet option for human nutrition and health. Our findings provide valuable information for the research and development of buffalo dairy products in China or other Asian countries.

Effects of capsicum oleoresin supplementation on rumen fermentation and microbial abundance under different temperature and dietary conditions in vitro.

This study aimed to determine the effect of capsicum oleoresin (CAP) on rumen fermentation and microbial abundance under different temperature and dietary conditions in vitro. The experimental design was arranged in a 2 × 2 × 3 factorial format together with two temperatures (normal: 39°C; hyperthermal: 42°C), two forage/concentrate ratios (30:70; 70:30), and two CAP concentrations in the incubation fluid at 20 and 200 mg/L with a control group. Regarding the fermentation characteristics, high temperature reduced short-chain fatty acids (SCFA) production except for molar percentages of butyrate while increasing acetate-to-propionate ratio and ammonia concentration. The diets increased total SCFA, propionate, and ammonia concentrations while decreasing acetate percentage and acetate-to-propionate ratio. CAP reduced acetate percentage and acetate-to-propionate ratio. Under hyperthermal condition, CAP could reduce acetate percentage and increase acetate-to-propionate ratio, lessening the negative effect of high heat on SCFA. Hyperthermal condition and diet altered the relative abundance of microbial abundance in cellulose-degrading bacteria. CAP showed little effect on the microbial abundance which only increased Butyrivibrio fibrisolvens. Thus, CAP could improve rumen fermentation under different conditions, with plasticity in response to the ramp of different temperature and dietary conditions, although hardly affecting rumen microbial abundance.

Icariin Alleviates Escherichia coli Lipopolysaccharide-Mediated Endometritis in Mice by Inhibiting Inflammation and Oxidative Stress.

Icariin (ICA) is a naturally occurring phytochemical agent primarily extracted from Epimedium Brevicornum Maxim (Family Berberidaceae) with a broad spectrum of bioactivities. Endometritis is a uterine disease that causes enormous losses in the dairy industry worldwide. In this study, anti-inflammatory and anti-oxidant properties of ICA were investigated against lipopolysaccharide (LPS)-induced endometritis in mice to investigate possible underlying molecular mechanisms. Sixty heathy female Kunming mice were randomly assigned to four groups (n = 15), namely control, LPS, LPS + ICA, and ICA groups. The endometritis was induced by intrauterine infusion of 50 µL of LPS (1 mg/mL). After 24 h of onset of LPS-induced endometritis, ICA groups were injected thrice by ICA intraperitoneally six hours apart. Histopathological examination, enzyme linked immunosorbent assay (ELISA), real time quantitative polymerase chain reaction (RT-qPCR), western blotting, and immunohistochemistry were used in this study. Histological alterations revealed that ICA markedly mitigated uterine tissue injury caused by LPS. The results showed that the ICA inhibited the production of pro-inflammatory cytokines (IL-1ß, IL-6, and TNF-α) and boosted the production of anti-inflammatory cytokines (IL-10). Additionally, ICA modulated the expression of malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase 1 (SOD1), catalase (CAT), and glutathione peroxidase 1 (Gpx1) induced by LPS. The administration of ICA significantly (p < 0.05) improved the mRNA and protein expression of Toll-like receptor (TLR) 4. The western blotting and ELISA finding revealed that the ICA repressed LPS-triggered NF-κB pathway activation. Moreover, ICA improved the antioxidant defense system via activation of the Nrf2 pathway. The results revealed that ICA up-regulated the mRNA and protein expression of Nuclear erythroid-2-related factor (Nrf2), NAD(P)H: quinone oxidoreductase 1 (NQO1), heme oxygenase-1 (HO-1), and glutamate-cysteine ligase catalytic subunit (GCLC) under LPS exposure. Conclusively, our findings strongly suggested that ICA protects endometritis caused by LPS by suppressing TLR4-associated NF-κB and Nrf2 pathways. Altogether, these innovative findings may pave the way for future studies into the therapeutic application of ICA to protect humans and animals against endometritis.

The expression of IGFBP-5 in the reproductive axis and effect on the onset of puberty in female rats.

Insulin-like growth factor-binding protein-5 (IGFBP-5) has recently been shown to alter the reproductive capacity by regulating insulin-like growth factor (IGF) bioavailability or IGF-independent effects. The present study aimed to investigate the effect and mechanism of IGFBP-5 on the onset of puberty in female rats. Immunofluorescence and real-time quantitative PCR were used to determine the expression and location of IGFBP-5 mRNA and protein distribution in the infant's hypothalamus-pituitary-ovary (HPO) axis prepuberty, peripuberty, puberty and adult female rats. Prepubertal rats with IGFBP-5 intracerebroventricular (ICV) were injected to determine the puberty-related genes expression and the concentrations of reproductive hormones. Primary hypothalamic cells were treated with IGFBP-5 to determine the expression of puberty-related genes and the Akt and mTOR proteins. Results showed that Igfbp-5 mRNA and protein were present on the HPO axis. The addition of IGFBP-5 to primary hypothalamic cells inhibited the expression of Gnrh and Igf-1 mRNAs (P < 0.05) and increased the expression of AKT and mTOR protein (P < 0.01). IGFBP-5 ICV-injection delayed the onset of puberty, reduced Gnrh, Igf-1, and Fshß mRNAs, and decreased the concentrations of E2, P4, FSH,serum LH levels and the ovaries weight (P < 0.05). More corpus luteum and fewer primary follicles were found after IGFBP-5 injection (P < 0.05).

Nutritional Modulation, Gut, and Omics Crosstalk in Ruminants.

Ruminant nutrition has significantly revolutionized a new and prodigious molecular approach in livestock sciences over the last decade. Wide-spectrum advances in DNA and RNA technologies and analysis have produced a wealth of data that have shifted the research threshold scheme to a more affluent level. Recently, the published literature has pointed out the nutrient roles in different cellular genomic alterations among different ruminant species, besides the interactions with other factors, such as age, type, and breed. Additionally, it has addressed rumen microbes within the gut health and productivity context, which has made interpreting homogenous evidence more complicated. As a more systematic approach, nutrigenomics can identify how genomics interacts with nutrition and other variables linked to animal performance. Such findings should contribute to crystallizing powerful interpretations correlating feeding management with ruminant production and health through genomics. This review will present a road-mapping discussion of promising trends in ruminant nutrigenomics as a reference for phenotype expression through multi-level omics changes.

Ginsenoside Rb1 Mitigates Escherichia coli Lipopolysaccharide-Induced Endometritis through TLR4-Mediated NF-κB Pathway.

Endometritis is the inflammatory response of the endometrial lining of the uterus and is associated with low conception rates, early embryonic mortality, and prolonged inter-calving intervals, and thus poses huge economic losses to the dairy industry worldwide. Ginsenoside Rb1 (GnRb1) is a natural compound obtained from the roots of Panax ginseng, having several pharmacological and biological properties. However, the anti-inflammatory properties of GnRb1 in lipopolysaccharide (LPS)-challenged endometritis through the TLR4-mediated NF-κB signaling pathway has not yet been researched. This study was planned to evaluate the mechanisms of how GnRb1 rescues LPS-induced endometritis. In the present research, histopathological findings revealed that GnRb1 ameliorated LPS-triggered uterine injury. The ELISA and RT-qPCR assay findings indicated that GnRb1 suppressed the expression level of pro-inflammatory molecules (TNF-α, IL-1ß and IL-6) and boosted the level of anti-inflammatory (IL-10) cytokine. Furthermore, the molecular study suggested that GnRb1 attenuated TLR4-mediated NF-κB signaling. The results demonstrated the therapeutic efficacy of GnRb1 in the mouse model of LPS-triggered endometritis via the inhibition of the TLR4-associated NF-κB pathway. Taken together, this study provides a baseline for the protective effect of GnRb1 to treat endometritis in both humans and animals.

Knockdown of Ptprn-2 delays the onset of puberty in female rats.

In the present study, we evaluated how Ptprn-2 (encoding tyrosine phosphatase, receptor type, N2 polypeptide protein) affects the onset of puberty in female rats. We evaluated the expression of Ptprn-2 mRNA and protein in the hypothalamus-pituitary-ovary axis of female rats using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunofluorescence at infancy, prepuberty, puberty, peripuberty, and adulthood. We evaluated the effects of Ptprn-2 gene knockdown on different aspects of reproduction-related biology in female rats, including the expression levels of puberty-related genes in vivo and in vitro, the time to onset of puberty, the concentration of serum reproductive hormones, the morphology of ovaries, and the ultrastructure of pituitary gonadotropin cells. Our results demonstrated that PTPRN-2 was primarily distributed in the arcuate nucleus (ARC), periventricular nucleus (PeN), adenohypophysis, and the ovarian follicular theca, stroma, and granulosa cells of female rats at various stages. Ptprn-2 mRNA levels significantly varied between peripuberty and puberty (P < 0.05) in the hypothalamus and pituitary gland. In hypothalamic cells, Ptprn-2 knockdown decreased the expression of Ptprn-2 and Rfrp-3 mRNA (P < 0.05) and increased the levels of Gnrh and Kiss-1 mRNA (P < 0.05). Ptprn-2 knockdown in the hypothalamus resulted in delayed vaginal opening compared to the control group (n = 12, P < 0.01), and Ptprn-2, Gnrh, and Kiss-1 mRNA levels (P < 0.05) all decreased, while the expression of Igf-1 (P < 0.05) and Rfrp-3 mRNA (P < 0.01) increased. The concentrations of FSH and P4 in the serum of Ptprn-2 knockdown rats were lower than in control animals (P < 0.05). Large transverse perimeters and longitudinal perimeters (P < 0.05) were found in the ovaries of Ptprn-2 knockdown rats. There were fewer large secretory particles from gonadotropin cells in adenohypophysis tissue of the Ptprn-2 knockdown group compared to the control group. This indicates that Ptprn-2 knockdown can regulate levels of Gnrh, Kiss-1, and Rfrp-3 mRNA in the hypothalamus, regulate the concentration of serum FSH and P4, and alter the morphology of ovarian and gonadotropin cells, delaying the onset of puberty in female rats.

Effect of GABA-T on Reproductive Function in Female Rats.

This study explored the role of γ-aminobutyric acid transaminase (GABA-T) in the puberty and reproductive performance of female rats. Immunofluorescence technique, quantitative real-time PCR (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect the distribution of GABA-T and the expression of genes and hormones in female rats, respectively. The results showed that GABA-T was mainly distributed in the arcuate nucleus (ARC), paraventricular nucleus (PVN) and periventricular nucleus (PeN) of the hypothalamus, and in the adenohypophysis, ovarian granulosa cells and oocytes. Abat mRNA level at 28 d was lowest in the hypothalamus and the pituitary; at puberty, it was lowest in the ovary. Abat mRNA level was highest in adults in the hypothalamus; at infancy and puberty, it was highest in the pituitary; and at 21 d it was highest in the ovary. After vigabatrin (GABA-T irreversible inhibitor) was added to hypothalamus cells, the levels of Abat mRNA and Rfrp-3 mRNA were significantly reduced, but Gnrh mRNA increased at the dose of 25 and 50 µg/mL; Kiss1 mRNA was significantly increased but Gabbr1 mRNA was reduced at the 50 µg/mL dose. In prepubertal rats injected with vigabatrin, puberty onset was delayed. Abat mRNA, Kiss1 mRNA and Gnrh mRNA levels were significantly reduced, but Rfrp-3 mRNA level increased in the hypothalamus. Vigabatrin reduced the concentrations of GABA-T, luteinizing hormone (LH) and progesterone (P4), and the ovarian index. Lactation performance was reduced in adult rats with vigabatrin treatment. Four hours after vigabatrin injection, the concentrations of GABA-T and LH were significantly reduced in adult and 25 d rats, but follicle-stimulating hormone (FSH) increased in 25 d rats. In conclusion, GABA-T affects the reproductive function of female rats by regulating the levels of Gnrh, Kiss1 and Rfrp-3 in the hypothalamus as well as the concentrations of LH and P4.

Identification and characterization of microRNAs in the pituitary of pubescent goats.

BACKGROUND: Puberty is the period during a female mammal's life when it enters estrus and ovulates for the first time; this indicates that a mammal is capable of reproduction. The onset of puberty is a complex and tightly coordinated biological event; it has been reported that microRNAs (miRNAs) are involved in regulating the initiation of puberty. METHODS: We performed miRNA sequencing on pituitary tissue from prepubescent and pubescent goats to investigate differences in miRNA expression during the onset of puberty in female goats. The target genes of these miRNAs were evaluated by GO enrichment and KEGG pathway analysis to identify critical pathways regulated by these miRNAs during puberty in goats. Finally, we selected four known miRNA and one novel miRNAs to evaluate expression patterns in two samples via qRT-PCR to validate the RNA-seq data. RESULTS: In this study, 476 miRNAs were detected in goat pituitary tissue; 13 of these were specifically expressed in the pituitary of prepubescent goats, and 17 were unique to the pituitary of pubescent goats. Additionally, 73 novel miRNAs were predicted in these two libraries. 20 differentially expressed miRNAs were identified in this study. KEGG pathway enrichment analysis revealed that the differentially expressed miRNA target genes were enriched in pathways related to ovary development during puberty, including the GABAergic synapse, oxytocin signaling pathway, the cAMP signaling pathway, progesterone-mediated oocyte maturation. In this study, differential miRNA expression in the pituitary tissue of prepubescent and pubescent goats were identified and characterized. CONCLUSION: These results provide important information regarding the potential regulation of the onset of goat puberty by miRNAs, and contribute to the elucidation of miRNA regulated processes during maturation and reproduction.

Effect of active immunization using a novel GnRH vaccine on reproductive function in rats.

To investigate the effect of gonadotropin-releasing hormone 2-multiple antigen peptide (GnRH2-MAP) on reproductive function. In our study, 20-day-old male rats (n = 90) were randomly allocated to one of three treatment groups: GnRH2-MAP immunization, GnRH2 immunization, and non-immunized control groups. The immunized animals were administered three doses of GnRH2-MAP or GnRH2 vaccines from 0 to 6 weeks at 2-week intervals. The control group only received oil adjuvant. Blood and right testis samples were collected, and the left testis was weighed and its volume was measured at 0, 2, 4, 6, 8, 10 and 12 weeks after the first immunization. The serum antibody titer and testosterone concentration were determined by ELISA, and the right testis samples were collected for histological analysis. The results revealed that the serum of vaccinated rats elicited a significantly higher antibody titer and a lower T concentration compared with the control group two weeks after the first immunization (P < 0.05), but the highest antibody titer and lowest T concentration were found in animals treated with GnRH2-MAP (P < 0.05). The second immunization resulted in a significant decrease in testicular weight and volume (P < 0.05) in both immunized groups compared to the control, but these values were significantly lower in the GnRH2-MAP group than in the GnRH2 group. Furthermore, seminiferous tubules revealed more significant atrophy in the GnRH2-MAP group than in the GnRH2 group, and no sperm were observed in rats of the GnRH2-MAP group. Thus, GnRH2-MAP may be an effective antigen and a potential immunocastration vaccine with higher effectiveness.

Identification of differential genomic DNA Methylation in the hypothalamus of pubertal rat using reduced representation Bisulfite sequencing.

BACKGROUND: There are many variables affecting the onset of puberty in animals, including genetic, nutritional, and environmental factors. Recent studies suggest that epigenetic regulation, especially DNA methylation, plays a majorrole in the regulation of puberty. However, there have been no reports on DNA methylation of the pubertal genome. METHODS: We investigated DNA methylation in the female rat hypothalamus at prepuberty and puberty using reduced representation bisulfite sequencing technology. The identified genes and signaling pathways exhibiting changes to DNA methylation in pubertal rats were determined by Gene Ontogeny and Kyoto Encyclopedia of Genes and Genomes analysis. RESULTS: The distribution of the three types of methylated C bases in promoter and CpG island (CGI) regions in the hypothalamus was as follows: 87.79% CG, 3.05% CHG, 9.16% CHH for promoters, and 88.35% CG, 3.21% CHG, 88.35% CHH for CGI in prepubertal rats; and 90.78% CG, 2.13% CHG, 7.09% CHH for promoters, and 88.59% CG, 88.59% CHG, 8.35% CHH for CGI in pubertal animals. CG showed the highest percentage of methylation, and was the highest methylation state in CGI. Compared to prepubertal hyoyhalamus samples, we identified ten genes with altered methylation in promoter regions in the pubertal hypothalamus samples, and 43 genes with altered methylation in the CGI. Changes in DNA methylation were found in gonadotropin-releasing hormone signaling pathways, and the oocyte meiosis pathway. CONCLUSION: Our results demonstrate changes in DNA methylation occur in female rats from prepuberty to puberty suggestng DNA methylation may play a crucial role in the regulation of puberty onset. This study provides essential information for future studies on the role of epigenetics in the regulation of puberty.