Introduction: The differentially expressed proteins between healthy cows and those with footrot were identified to explore changes in protein profiles associated with the disease. Material and Methods: Out of 36 cows selected for the experiment, 18 footrot-affected cows were included in the treatment group (group T) and 18 unaffected cows were included in the control group (group C). Plasma samples from groups T and C were subjected to two-dimensional electrophoresis analysis and differentially expressed proteins were identified by matrix-assisted laser desorption/ionisation tandem time-of-flight mass spectrometry. Bioinformatics, including gene ontology analysis and pathway analysis, was used for analysing all proteins. Results: Out of 63 spots identified by 2DE, 33 were selected for mass spectrum analysis, which identified 11 differentially expressed proteins in 26 spots. Footrot led to changes in profiles in plasma proteins that were classified to the pathway of inflammatory response, complement, and blood coagulation, among others. Conclusion: This study provides evidence of the defence mechanisms of cows with footrot to explore strategies for treatment.
This study aimed to characterise the effects of ketosis on milk yield and composition and digestive capacity in transition dairy cows.
Material and Methods
Seven ketotic and seven healthy cows were housed in individual stalls for six days. Samples of plasma, milk, refused total mixed ration, and faeces were collected, and the blood biochemical parameters, milk yield and composition, dry matter intake, and faecal dry matter (FDM) production were determined.
Compared with healthy cows, the ketotic cows had significantly higher concentrations of milk fat and citrate, but lower levels of milk protein and lactose. The cows exhibited a need for acid detergent fibre in forage and better digestion of neutral detergent fibre, starch, crude protein, and phosphorus than healthy cows, but more fat and gross energy were excreted in their faeces. Ketotic cows had higher energy-corrected milk yields and lower FDM than healthy cows.
Lower feed intake coinciding with the requirement to maintain high milk production is considered to be the cause of ketosis in dairy cows. Ketotic cows exhibited lower dry matter fat digestion.
Differential metabolites (DMs) between cows with inactive ovaries (IO) and oestrous (E) cows were screened and metabolic pathways of DMs associated with IO were determined.
Material and Methods
Cows at 50 to 60 days (d) postpartum from an intensive dairy farm were randomly selected and allocated into an E group (n = 16) or an IO group (n = 16) according to a pedometer and rectal examinations. Their plasma samples were analysed by liquid chromatography–mass spectrometry (LC–MS) to compare plasma metabolic changes between the E and IO groups. Multivariate pattern recognition was used to screen the DMs in the plasma of IO cows.
Compared with normal E cows, there were abnormalities in 20 metabolites in IO cows, including a significantly decreased content (VIP > 1, P < 0.05) of cholic acid, p-chlorophenylalanine, and arachidonic acid, and a significantly increased content (VIP > 1, P < 0.05) of tyramine, betaine, L-phenylalanine, L-glutamate, D-proline, L-alanine, and L-pyrophosphate. Five DMs (cholic acid, D-proline, L-glutamate, L-alanine, and L-pyroglutamic acid) with higher variable importance in projection (VIP) values between groups were validated by ELISA with blind samples of re-selected cows (IO, 50 to 60 d postpartum) and the validated results were consistent with the LC–MS results.
The 20 DMs in IO cows during the peak of lactation indicated that the pathogenesis of IO was involved in complex metabolic networks and signal transduction pathways. This study provides a basis for further exploration of the pathogenesis and prevention of IO in cows in the future.
Introduction: A model of fatty liver in postpartum sheep was established to measure blood paraoxonase 1 (PON1) and other biochemical indicators, which were used to predict fatty liver in sheep.
Material and Methods: Sheep were assigned into two experimental groups: a fatty liver group (T, n = 10) and a healthy control group (C, n = 5). PON1 enzyme activity towards paraoxon as a substrate was quantified spectrophotometrically. The results were analysed by t-test and pearson correlation coefficient. Disease was predicted by binary logistic analysis, and diagnostic thresholds were determined by receiver operatingcharacteristic (ROC) analysis.
Results: The activity of serum PON1 in group T was significantly decreased (P < 0.05) when compared with C group, and liver lipid content and the levels of serum BHBA, NEFA, and TG were significantly increased (P < 0.05). Thresholds were lower than 74.0 U/mL for PON1, higher than 0.97 mmol/L for β-hydroxybutyrate, higher than 1.29 mmol/L for non-esterified fatty acids, higher than 0.24 mmol/L for triglycerides, and lower than 71.35 g/L for total protein.
Conclusion: This study verified that PON1, BHBA, NEFA, TG, and TP could be used to predict the risk of fatty liver in sheep.
Introduction: The predictive value of selected parameters in the risk of ketosis and fatty liver in dairy cows was determined.
Material and Methods: In total, 21 control and 17 ketotic Holstein Friesian cows with a β-hydroxybutyrate (BHBA) concentration of 1.20 mmol/L as a cut-off point were selected. The risk prediction thresholds for ketosis were determined by receiver operating characteristic (ROC) curve analysis.
Results: In the ketosis group, paraoxonase-1 (PON-1) activity and concentration of PON-1 and glucose (GLU) were decreased, and aminotransferase (AST) activity as well as BHBA and non-esterified fatty acid (NEFA) contents were increased. The plasma activity and concentration of PON-1 were significantly positively correlated with the level of plasma GLU. The plasma activity and concentration of PON-1 were significantly negatively correlated with the levels of AST and BHBA. According to ROC curve analysis, warning indexes of ketosis were: plasma PON-1 concentration of 46.79 nmol/L, GLU concentration of 3.04 mmol/L, AST concentration of 100 U/L, and NEFA concentration of 0.82 mmol/L.
Conclusion: This study showed that the levels of PON-1, GLU, AST, and NEFA could be used as indicators to predict the risk of ketosis in dairy cows.
Introduction: To identify novel pathways involved in the pathogenesis of ketosis, an isobaric tag for relative and absolute quantitation/mass spectrometry was used to define differences in protein expression profiles between healthy dairy cows and those with clinical or subclinical ketosis.
Material and Methods: To define the novel pathways of ketosis in cattle, the differences in protein expression were analysed by bioinformatics. Go Ontology and Pathway analysis were carried out for enrich the role and pathway of the different expression proteins between healthy dairy cows and those with clinical or subclinical ketosis.
Results: Differences were identified in 19 proteins, 16 of which were relatively up-regulated while the remaining 3 were relatively down-regulated. Sorbitol dehydrogenase (SORD) and glyceraldehyde-3-phosphate dehydrogenase (G3PD) were up-regulated in cattle with ketosis. SORD and G3PD promoted glycolysis. These mechanisms lead to pyruvic acid production increase and ketone body accumulation.
Conclusion: The novel pathways of glycolysis provided new evidence for the research of ketosis.
Early diagnosis of pregnancy is important in livestock production, but there is no reliable technology used for pregnancy diagnosis within the first three weeks after insemination. During early pregnancy, the expression of interferon-stimulating genes (ISGs) in peripheral blood leukocytes (PBL) is significantly increased. However, due to different strains, detection sample types, detection methods, threshold value, etc. the specific effectiveness of early pregnancy diagnosis using ISGs is worth further study. The purpose of this study was to test interferon-stimulated protein 15 (ISG15), 2'–5'-oligoadenylate synthetase 1 (OAS1) and radical S-adenosyl methionine domain containing 2 (RSAD2) for early pregnancy diagnosis in dairy cows. The expression of ISG15, OAS1, and RSAD2 in PBL of pregnant and non-pregnant heifers on days 0, 14, 18, 21 and 28 after artificial insemination (AI) was detected by fluorescence quantitative polymerase chain reaction (PCR). The sensitivity and specificity of the pregnancy diagnosis was analyzed using expression of these three genes separately or in combination by receiver operating characteristic curve. The combination with the highest accuracy used probe primers and duplex fluorescence quantitative PCR. The single quantitative PCR results showed that expression of ISG15, OAS1 and RSAD2 on day 18 after AI was significantly higher in pregnant than in non-pregnant cows. When these three genes were used separately, or in combination, for early pregnancy diagnosis, the sensitivity for the RSAD2 gene was 100%, and the combination of ISG15 with RSAD2 was 94.7%. The duplex quantitative PCR showed that, although the sensitivity of ISG15 alone was 100%, its specificity was only 88.2% (cut-off value 1.402). The sensitivity of RSAD2 alone was 89.5%, and the specificity was 88.2%; however, when the two genes were used in combination, the sensitivity, specificity and diagnostic cut-off value were consistent with the results of single quantitative PCR. These results indicated that a duplex quantitative PCR assay system for early pregnancy diagnosis in cows using ISG15 and RSAD2 was established. Simultaneous detection of expression of ISG15 and RSAD2 by duplex quantitative PCR can effectively improve the diagnostic accuracy for dairy cows.
The present study was carried out to evaluate the hematology, serum biochemistry, immune responses and oxidative damage of growing beagles fed a diet supplemented with housefly (Musca domestica) maggot meal (MM). Weanling beagles (initial body weight 2.69 ± 0.17 kg) were fed a control diet (0% MM) or experimental diet (5% MM) for 42 days. The results indicated that the diet supplemented with 5% MM had no significant effects on the hematology and serum biochemistry of growing beagles (P>0.05). Meanwhile, neither the serum concentrations of lysozyme and C-reactive protein nor the serum antibody responses to canine distemper virus and canine parvovirus were influenced by dietary MM supplementation (P>0.05). However, dogs in the experimental group had lower serum levels of malondialdehyde and protein carbonyl than those in the control group (P<0.05). These findings demonstrated that MM could be used as an alternative protein source in growing beagles without any adverse effects on hematology, serum biochemistry and immune responses. Furthermore, dietary MM could alleviate oxidative damage in growing beagles.