Integrated Analysis of Transcriptome and Metabolome Profiles in the Longissimus Dorsi Muscle of Buffalo and Cattle.

Buffalo meat is gaining popularity for its nutritional properties, such as its low fat and cholesterol content. However, it is often unsatisfactory to consumers due to its dark color and low tenderness. There is currently limited research on the regulatory mechanisms of buffalo meat quality. Xinglong buffalo are raised in the tropical Hainan region and are undergoing genetic improvement from draught to meat production. For the first time, we evaluated the meat quality traits of Xinglong buffalo using the longissimus dorsi muscle and compared them to Hainan cattle. Furthermore, we utilized a multi-omics approach combining transcriptomics and metabolomics to explore the underlying molecular mechanism regulating meat quality traits. We found that the Xinglong buffalo had significantly higher meat color redness but lower amino acid content and higher shear force compared to Hainan cattle. Differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) were identified, with them being significantly enriched in nicotinic acid and nicotinamide metabolic and glycine, serine, and threonine metabolic pathways. The correlation analysis revealed that those genes and metabolites (such as: GAMT, GCSH, PNP, L-aspartic acid, NADP+, and glutathione) are significantly associated with meat color, tenderness, and amino acid content, indicating their potential as candidate genes and biological indicators associated with meat quality. This study contributes to the breed genetic improvement and enhancement of buffalo meat quality.

Immune Cells in the Spleen of Mice Mediate the Inflammatory Response Induced by Mannheimia haemolytica A2 Serotype.

(1) Background: Mannheimia haemolytica (M. haemolytica) is an opportunistic pathogen and is mainly associated with respiratory diseases in cattle, sheep, and goats. (2) Methods: In this study, a mouse infection model was established using a M. haemolytica strain isolated from goats. Histopathological observations were conducted on various organs of the mice, and bacterial load determination and RNA-seq analysis were specifically performed on the spleens of the mice. (3) Results: The findings of this study suggest that chemokines, potentially present in the spleen of mice following a M. haemolytica challenge, may induce the migration of leukocytes to the spleen and suppress the release of pro-inflammatory factors through a negative feedback regulation mechanism. Additionally, an interesting observation was made regarding the potential of hematopoietic stem/progenitor cells congregating in the spleen to differentiate into immune cells, which could potentially collaborate with leukocytes in their efforts to counteract M. haemolytica invasion. (4) Conclusions: This study revealed the immune regulation mechanism induced by M. haemolytica in the mouse spleen, providing valuable insights into host-pathogen interactions and offering a theoretical basis for the prevention, control, and treatment of mannheimiosis.

Activation of MyD88-Dependent TLR Signaling Modulates Immune Response of the Mouse Heart during Pasteurella multocida Infection.

Pasteurella multocida (P. multocida) is an important zoonotic pathogen. In addition to lung lesions, necropsies have revealed macroscopic lesions in the heart in clinical cases. However, most previous studies focused on lung lesions while ignoring heart lesions. Therefore, to investigate the immune response of the P. multocida-infected heart, two murine infection models were established by using P. multocida serotype A (Pm HN02) and D (Pm HN01) strains. Histopathological examination revealed heterogeneous inflammatory responses, including immune cell infiltration in the epicardial and myocardial areas of the heart. Transcriptome sequencing was performed on infected cardiac tissues. To explore the traits of immune responses, we performed the functional enrichment analysis of differentially expressed genes, gene set enrichment analysis and gene set variation analysis. The results showed that the innate immune pathways were significantly regulated in both groups, including the NOD-like receptor signaling pathway, the complement and coagulation cascade and cytokine-cytokine receptor interaction. The Toll-like receptor signaling pathway was only significantly activated in the Pm HN02 group. For the Pm HN02 group, immunohistochemistry analysis further verified the significant upregulation of the hub component MyD88 at the protein level. In conclusion, this study reveals critical pathways for host heart recognition and defense against P. multocida serotypes A and D. Moreover, MyD88 was upregulated by P. multocida serotype A in the heart, providing a theoretical basis for future prevention, diagnosis and treatment research.

First Report and Comparative Genomic Analysis of Mycoplasma capricolum subsp. capricolum HN-B in Hainan Island, China.

Mycoplasma capricolum subsp. Capricolum (Mcc) is an important member of the Mycoplasma mycoides cluster (Mm cluster) and causes caprine contagious agalactia. Mcc can infect goats of all age groups, especially pregnant ewes and kids. It can cause the abortion in pregnant ewes and the death of goat kids, leading to enormous losses in the goat breeding industry. To date, the prevalence of epidemic Mcc strains on Hainan Island, China, remains unclear. This study aimed to isolate and identify Mcc strains endemic to Hainan Island, China. Genome sequencing and comparative genomic analysis were performed to reveal the molecular characteristics and evolutionary relationships of the isolated strain. Mcc HN-B was isolated and identified in Hainan Island, China. The Mcc HN-B genome consists of a 1,117,925 bp circular chromosome with a 23.79% G + C content. It contains 912 encoding genes, 3 gene islands, and 14 potential virulence genes. The core genome with the features of the Mm cluster and the specific genes of Mcc HN-B were identified by comparative genomic analysis. These results revealed the evolutionary relationship between Mcc HN-B and other members of the Mm cluster. Our findings provide a reference for further studies on the pathogenic mechanism and local vaccine development of Mcc.

First Report and Comparative Genomic Analysis of a Mycoplasma mycoides Subspecies capri HN-A in Hainan Island.

Mycoplasma mycoides subspecies capri (Mmc) is one of the six Mycoplasma mycoides cluster (Mm cluster) members, which can cause "MAKePS" (Mastitis, Arthritis, Keratoconjunctivitis, Pneumonia, Septicemia) syndrome in ruminants. These symptoms can occur alone or together in individuals or flocks of goats. However, little is known about the epidemic Mmc strains in Hainan Island, China. We aimed to isolate the endemic Mmc strains in Hainan Island and reveal their molecular characteristics by genomic sequencing and comparative genomics to mitigate the impact of Mmc on local ruminant farming. Here, the Mmc HN-A strain was isolated and identified for the first time in Hainan Island, China. The genome of Mmc HN-A was sequenced. It contains a 1,084,691 bp-long circular chromosome and 848 coding genes. The genomic analysis of Mmc HN-A revealed 16 virulence factors, 2 gene islands, and a bacterial type IV secretion system protein VirD4. Comparative genomics showed that the core genome of the five Mycoplasma mycoides contained 611 genes that could be exploited to develop drugs and endemic vaccines. Additionally, 36 specific genes were included in the Mmc HN-A genome, which could provide the possibility for the further control and prevention of the Mmc effects on local ruminants and enrich the information on Mmc strains.

Up-Regulation of Interleukin-10 in Splenic Immune Response Induced by Serotype A Pasteurellamultocida.

Pasteurella multocida (P. multocida) is an opportunistic pathogen that is common in livestock and poultry and leads to massive economic losses in the animal husbandry sector. In this study, we challenged mice with P. multocida strain HN02 by intraperitoneal injection and collected spleens to measure bacterial loads. We also performed histopathological analysis by hematoxylin and eosin (H&E) staining. Then we used RNA-sequencing (RNA-seq) to detect the mRNA expression levels in the mouse spleen and quantitative real-time PCR (qRT-PCR) to verify the sequencing data. Finally, we examined the effect of HN02 on anti-inflammatory cytokine interleukin-10 (IL-10) protein expression in the spleen through immunohistochemical analysis. The results showed that compared to those in the control group, the mouse spleens in the challenge group had lesions, and the average bacteria loads was (3.07 ± 1.09) × 106 CFU (colony-forming unit)/g. The RNA-seq results determined 3653 differentially expressed genes (DEGs), and the qRT-PCR analysis revealed immune-related genes consistent with the expression trend in the sequencing data. The number and area of IL-10 positive cells substantially increased to resist inflammation in the challenge group. In conclusion, we analyzed the spleens of mice infected with P. multocida from multiple perspectives, and our findings lay a foundation for subsequent studies on the mechanism of pathogen-host interactions.

Microbiological Characteristics and Pathogenesis of Klebsiella pneumoniae Isolated from Hainan Black Goat.

K. pneumoniae is an opportunistic pathogen that leads to widespread infection in humans and animals, seriously threatening human health and animal husbandry development. In our research, we investigated the biological characteristics of the isolate by using a 16S rRNA gene sequencing, biochemical assay, and drug sensitivity test. Moreover, the pathogenicity study, including the bacteria load determination, the histopathology examination, and the RNA sequencing was carried out to explore whether the isolate could cause lung injury in mice through intraperitoneal injection. The results indicated that the isolate was identified as K. pneumoniae and named as KPHN001. The drug susceptibility test showed that KPHN001 was only sensitive to polymyxin B and furazolidone, and was resistant to other 28 antibiotics. In the bacteria load determination, the highest bacterial load of the organs was found in the spleen, and abundant bacterial colonization was also found in the lung. The histopathology showed the mainly acute inflammations in the lung were due to congestion, edema, and exudation. RNA-seq analysis revealed that the differentially expressed genes (DEGs) of inflammatory cytokines and chemokines were expressed massively in mice. In the present research, the biological characteristics and pathogenesis of clinically isolated K. pneumoniae were systematically studied, revealing the pathogenic mechanism of K. pneumoniae to animals, and providing a theoretical basis for the following prevention, control, and diagnosis research.

[Clinical study of Yihuo Qingxia method in treating hyperlipoidemia-related severe acute pancreatitis in early stage].

OBJECTIVE: To investigate the therapeutic effect of Yihuo Qingxia method, a traditional Chinese medicine therapeutic method for replenishing qi to activate blood, clearing away heat and dredging intestines, in treating hyperlipoidemia-related severe acute pancreatitis (SAP) in early stage. METHODS: One hundred and four patients with hyperlipoidemia-related SAP were divided into two groups: early group (admitted to hospital within 3 days after onset) and late group (admitted to hospital from 3 days to 7 days after onset). There were 52 cases in each group. All the patients were treated by Yihuo Qingxia method. RESULTS: There were no statistical differences in 48-hour Ranson scores, CT scores, 24-hour acute physiology and chronic heath evaluation II scores (APACHE II scores), and the levels of 24-hour serum triglyceride (TG) and serum glucose in the two groups (P>0.05). At the 10th day after onset, the serum TG level in early group was lower than that in late group (P<0.01). The incidences of acute respiratory distress syndrome, acute renal failure, hepatic inadequacy, congestive heart failure, shock, encephalopathy, infection and alimentary tract hemorrhage in early group were higher than those in late group (P<0.05). The mortality in early group was lower than that in the late group (P<0.05). The length of hospital stay in early group was shorter than that in late group (P<0.05). CONCLUSION: Yihuo Qingxia method has a good efficacy in treating hyperlipoidemia-related SAP in early stage.

Comparison of integrated Chinese and Western medicine with and without somatostatin supplement in the treatment of severe acute pancreatitis.

AIM: To evaluate the therapeutic effect of the combined use of early short-term somatostatin and conventional integrated Chinese and Western medicine in treating severe acute pancreatitis. METHODS: Sixty patients with severe acute pancreatitis were divided at random into a somatostatin group and a basic treatment group. Both groups received integrated traditional Chinese and Western medicine without surgery. For patients in the somatostatin group, somatostatin was infused intravenously 250 microg/h for 72 h; other medications were the same as in the basic treatment group. In both groups, comparisons of therapeutic effectiveness were made in terms of morbidity of organic dysfunction and mortality rate, and severity of the disease according to serum levels of C-reaction protein, scores of acute physiology and chronic health evaluation (APACHE II), and scores of Balthazar-CT. RESULTS: The indexes for C-reaction protein levels on the fourth and seventh days, and APACHE II scores on the seventh day after treatment, were significantly improved in the somatostatin group than in the basic treatment group. The morbidity of organic dysfunction was lower in the somatostatin group than in the basic treatment group, although the difference was not statistically significant. There was no significant difference in mortality between the two groups. CONCLUSION: We conclude that combined traditional Chinese and Western medicines with an early short-term use of somatostatin can improve the condition of patients with severe acute pancreatitis.

[The effects of traditional Chinese medicine on distribution of F-actin of leukocytes in acute edematous pancreatitis].

OBJECTIVE: To explore the distribution of F-actin of leukocytes in local circulation of pancreas and system circulation and assess the effects of traditional Chinese medicine WPY complex there on in the case of acute edematous pancreatitis(AEP). METHODS: AEP model of Wistar rats was induced with subcutaneous injection of cerulein. In the treatment group, rats were given 50% WPY (1.5 ml every time) at 0, 2, 4, 6 and 8 hours during the experiment. The leukocytes of blood from the splenic vein and inferior vena cava were incubated with TRITC-Phalloidin for F-actin, and the images were taken with confocal laser scanning microscope. The The distribution of F-actin at the border or central part of leukocyte was analyzed by using Mias system. Results ratio of F-actin border/center gray value of leukocytes increased at the 2nd, 4th and 6th hour, then declined at the 8th hour in AEP rats, but the ratio of F-actin border/center gray value of leukocytes in the blood from the inferior vena cava declined significantly at the 6th hour in the WPY-treated AEP rats(P < 0.01). At 6 hours following the onset of experimental acute edematous pancreatitis, F-actin polymerization and redistribution in the border region of leukocytes were prominent. CONCLUSION: These evidences indicate that leukocytes play an important role in the development of acute pancreatitis. Traditional Chinese medicine WPY can regulate actin redistribution of leukocytes; moreover, WPY may attenuate the stiffness of leukocytes and render them more flexible and transformable in microcirculation.

[Impact of traditional Chinese medicine WPY on ICAM-1 expression and MPO activity in pancreas and lungs of rats with acute necrotizing pancreatitis].

OBJECTIVE: To explore the character of the ICAM-1 induced change of polymorphonuclear neutrophil (PMN) in the pancreas and lung of rats with acute necrotizing pancreatitis (ANP) and to evaluate the effects of Chinese medicine WPY on ANP. METHODS: ANP model was induced by retrograde injection of 3.5% sodium taurocholic acid into the biliopancreatic duct of Wistar rats. Expression of ICAM-1 at different time points in pancreas and lung was assessed by immunohistochemistry. The quantity of polymorphonuclear neutrophil (PMN) in tissues was measured by the level of myeloperoxidase activity (MPO). In addition, the indices above were also observed after the administration of WPY. RESULTS: In the ANP group, ICAM-1 expression and MPO activity were evidently up-regulated at 3 h in pancreas and at 6 h in lung, and both reached their highest level at 12 h. In the WPY treated group, ICAM-1 expression and MPO activity in pancreas and lung were significantly decreased at 12 h. CONCLUSION: The numerous PMNs infiltration mediated by ICAM-1 plays an important role in the damage to pancreas and lung caused by ANP. The traditional Chinese medicine WPY can attenuate ICAM-1 expression and MPO activity in pancreas and lung during ANP.

[The change of cryptdin-4 mRNA expression in the intestine of mouse with experimental pancreatitis].

OBJECTIVE: experimental pancreatitis and explore the mechanism of bacterial translocation at the molecular level in mice with severe acute pancreatitis. METHODS: Thirty-six ICR mice were randomly divided into 6 groups with 6 mice in each group. The animals in the experimental groups (from A to E) received seven intraperitoneal injections of caerulein (50 microg/kg body weight) at hourly intervals over 6 hours, and were killed at 9, 18, 24, 48 and 72 hours after the first injection. The control group (F) received intraperitoneal injection of the same volume of saline, and the animals were killed at the 18th hour after the first injection. Blood and pancreatic tissue samples were obtained after the animals were killed. Amylase and pancreatic pathological alterations were observed. The ileum sequential segments were removed from each mouse. The amplification products of RT-PCR were electrophoresed and the images were analyzed by UVI software. RESULTS: Acute necrotizing pancreatitis in ICR mice were induced by the intraperitoneal injection of caerulein at large doses. Markedly pathological lesions were observed at the 18th hour after the first injection of caerulein. And, intestinal cryptdin-4 mRNA expression was down regulated slightly at the 9th hour and most markedly at the 18th hour (P < 0.05). After 24 hours, the cryptdin-4 mRNA expression recovered to the normal level gradually. CONCLUSION: Acquired cryptdin-4 deficiency may play an important role in the pathogenesis of bacterial tanslocation in acute necrotizing pancreatits.

[Improvement of the model of pancreas-duodenal transplantation in swines].

OBJECTIVE: To improve the procedures of pancreas-duodenal transplantation in swines in order to establish a technically stable model that accords with the physiological situation. METHODS: The whole pancreatoduodenal graft was harvested with in situ flush using 4 degrees C preservation solution. Type 1 diabetes was induced surgically by total pancreatectomy. By end-to-side fashion, the segment of abdominal aorta containing the celiac artery and the anterior mesenteric artery (equal to superior mesenteric artery in human) was anastomosed to the abdominal aorta of recipient which is behind the renal artery, the portal vein was anastomosed to the anterior mesenteric vein of the recipient, the duodenum was anastomosed to the jejunum of the recipient. The graft was placed in the anterior right peritoneal cavity. RESULTS: A total of 28 swines were used as recipients. Of them, two died of anesthetic accident, another two died of hyperacute rejection 2 hours, 19 hours after operation respectively; the others underwent the operation uneventfully. Their survival time was 5-21 days, blood sugar was (11.33+/-1.66) mmol/L on the first day after operation, and (5.65+/-0.89) mmol/L on the third day. CONCLUSION: This kind of model is stable, and the operative method and procedures are relatively simple and practicable.

[Clinical study on the treatment of severe acute pancreatitis by integrated traditional Chinese medicine and Western medicine].

OBJECTIVE: To sum up clinical experiences and formulate a new treatment program for the effective managment of severe acute pancreatitis(SAP) by integrated TCM and western medicine. METHODS: The therapeutic regimens and end results of 1376 cases of SAP in three periods were reviewed and analyzed. Period I (1980-1990) was characterized by early surgical intervention supplemented with integrated traditional Chinese and western medical therapy, period II (1991-1993) by integrated traditional Chinese and western medical therapy supplemented with surgery for early complications, and period III (1994-2003) by integrated traditional Chinese and western medical therapy supplemented with surgery for local infectious complications at a late stage in the course of disease. RESULTS: With use of integrated traditional Chinese and western medical therapy in the treatment of SAP, the proportion of surgical operation and the case-fatality rate kept declining in the three consecutive periods, the proportions of operation being 77.59%, 54.54% and 19.38%, and the case-fatality rates being 40.52%, 17.17% and 10.77%, respectively. CONCLUSION: Integrated TCM and western medicine is effective for most SAP cases, and surgery is indicated mainly for late complications such as necrotizing infection. Early surgery should be handled with more circumspection and strictness in most of the patients with SAP.

[Mollusicidal effects of some species of plants in hilly and mountainous areas].

OBJECTIVE: To valuate the mollusicidal effects of some plants cultivated in the hilly and mountainous areas on Oncomelania hupensis and screen the plants against O. hupensis snails. METHODS: The mollusicidal effects of water extracts of plant materials were tested in laboratory. RESULT: The LC50 of all the species in 24 h, 48 h, and 120 h reached the middle noxious level against O. hupensis snails. According to the taxis of LC50(s) in 120 h, Eucalyptus camaldulensis was best with a LC50 of 0.013%, and Ailanthus altissima, Zanthoxylum bungeanum, Juglans regia, Melia azedarach, Nerium indicum, Pterocarva stenobtera, Cinnamomum camphora, Vetiveria zizanioides, and Citrus reticulata were the second with LC50 (s) between 0.020% and 0.370%, and Zingiber officinale had a lower effect on snail control with a LC50 of above 0.500%. There was a correlation between the death rate of snails and soak time. CONCLUSION: The resources of snail-restrained plants are very abundant, and four plants, such as Eucalyptus camaldulensis, Ailanthus altissima, Cinnamomum camphora, and Vetiveria zizanioides have some mollusicidal effect.

Experimental study of Tong Xia purgative method in ameliorating lung injury in acute necrotizing pancreatitis.

AIM:To investigate the role of tumor necrosis factor (TNF) in lung injury during acute necrotizing pancreatitis (ANP), and the therapeutic effect of Tong Xia purgative method in minimizing the severity of lung injury.METHODS:Fourteen canines were randomly divided into 3 groups:the Tong Xia treatment group (n = 5) using Dachengqitang; saline control group (n = 5), and the sham operation group (n =4). TNF activity in serum and in bronchoalveolar lavage fluid (BALF), the serum endotoxin levels were measured,and the severity of lung injury evaluated.RESULTS:Elevation of TNF activity was more prominent in BALF than in serum. TNF activity in serum at 6 and 12 hours and in BALF was significantly decreased in the Tong Xia treatment group than in the saline control one (q = 21.11, q = 12.07, q = 9.03, respectively, P <0.01) and the lung injury was significantly alleviated at 12 hours as compared with that in the saline group, manifested as amelioration of the lung wet/dry weight ratio, decrease in protein concentration and neutrophils count in BALF, and improvement of pulmonary inflammatory changes. A positive correlation was demonstrated between serum TNF activity and endotoxin level.CONCLUSION:Hypersecretion of TNF is shown to be one of the major causes of lung injury during ANP; Tong Xia purgative method could alleviate the degree of lung injury mediated by TNF.