Cell entry of bovine respiratory syncytial virus through clathrin-mediated endocytosis is regulated by PI3K-Akt and Src-JNK pathways.

Bovine respiratory syncytial virus (BRSV) is an RNA virus with envelope that causes acute, febrile, and highly infectious respiratory diseases in cattle. However, the manner and mechanism of BRSV entry into cells remain unclear. In this study, we aimed to explore the entry manner of BRSV into MDBK cells and its regulatory mechanism. Our findings, based on virus titer, virus copies, western blot and IFA analysis, indicate that BRSV enters MDBK cells through endocytosis, relying on dynamin, specifically via clathrin-mediated endocytosis rather than caveolin-mediated endocytosis and micropinocytosis. We observed that the entered BRSV initially localizes in early endosomes and subsequently localizes in late endosomes. Additionally, our results of western blot, virus titer and virus copies demonstrate that BRSV entry through clathrin-mediated endocytosis is regulated by PI3K-Akt and Src-JNK signaling pathways. Overall, our study suggests that BRSV enters MDBK cells through clathrin-mediated endocytosis, entered BRSV is trafficked to late endosome via early endosome, BRSV entry through clathrin-mediated endocytosis is regulated by PI3K-Akt and Src-JNK signaling pathways.

Bovine herpesvirus-1 gE protein inhibits IFN-ß production to enhance replication by promoting MAVS ubiquitination and interfering with the interaction between IRF3 and CBP/p300.

Bovine herpesvirus-1 (BoHV-1) can infect all breeds of cattle and cause respiratory and genital tract diseases. In the process of viral infection, viruses can use their own proteins to suppress the innate immunity of the host and promote its replication; however, the mechanism by which BoHV-1 evades the innate immune response is not fully understood. In this study, we found that rabbits inoculated with the live gene deletion vaccine BoHV-1-△gI/gE/TK generated higher interferon-ß (IFN-ß) production in the serum, liver, lung and kidney than rabbits inoculated with wt BoHV-1, which led to milder lesions in the lung and kidney. We performed gene deletion and ectopic expression experiments on viral proteins and found that gE was the major protein that inhibited IFN-ß expression. Further studies showed that MAVS and IRF3 were the targets of gE, and the specific mechanism was that gE inhibited IFN-ß production by promoting MAVS ubiquitination and interfering with the interaction between IRF3 and CBP/p300. These results suggest a new way of BoHV-1 inhibition of IFN-ß production to evade the host innate immunity.

Benefit-cost analysis of a H7N9 vaccination program in poultry in Guangxi, China.

China launched a H7N9 vaccination program in poultry, starting from the Guangxi and Guangdong provinces in July 2017, followed by other provinces in September 2017, as a response to a steep increase of H7N9 influenza human infections from September 2016. Since then, H5-H7 bivalent vaccine has been used in the nationwide avian influenza compulsory vaccination program to replace the existing H5N1 vaccine. However, the economic returns of the H7N9 vaccination program in China have never been adequately assessed. This study was designed to evaluate the economic value of the H7N9 vaccination program in Guangxi by assessing the benefits and costs of the program compared to not vaccinating against H7N9. A benefit-cost analysis (BCA) was undertaken to evaluate the adoption of a vaccination program against H7N9 in each of three consecutive years from July 2017 to June 2020 with the baseline scenario (the absence of H7N9 vaccination in the 12-month period July 2016 to June 2017). Both animal and public health perspectives were included in the BCA framework and took account of both the private and public sectors. Benefit-Cost Ratio (BCR) of the three-year H7N9 vaccination program was 18.6 (90 %PI: 15.4; 21.8), and total Net Present Values reached to CNY 1.63 billion (90 %PI: 1.37 billion; 1.89 billion). The extra revenue generated by the yellow broiler industry comprised 93.8 % of the total benefits after adoption of H7N9 vaccination program in Guangxi. While cost-savings in public health and animal health expenditure avoided were 3.6 % and 2.6 %, respectively. Total costs arising from adoption of the revised vaccination program over the three years were CNY 12.46 million (90 %PI: 11.49 million; 14.14 million), CNY 34.87 million (90 %PI: 31.88 million; 40.06 million), and CNY 44.28 million (90 %PI: 39.66 million; 52.27 million), respectively. Sensitivity analysis found the yellow broiler wholesale prices contributed 97.7 % of the variance of the total NPV of three vaccination years. The study results demonstrate the significant economic advantage of implementing a vaccination program against H7N9 in Guangxi. It also offers a new set of evidence to China's H7N9 vaccination policy and debates around economic values of conducting routine avian influenza vaccination.

Data-driven dynamical modelling of the transmission of African swine fever in a few places in China.

Since the outbreak of African swine fever (ASF) in Shengyang, it has continued spreading in China. In the early stage of the epidemic, multi-point and concentrated outbreaks were mainly in the swill feeding areas. In this paper, we developed compartmental models to investigate the transmission of ASF in several raising units including Guquan, Jinba and Liancheng. Using the data collected from these three infected premises, we calibrated the models to estimate that the average incubation period was between 8 and 11 days, the onset period was about 2-3 days and the basic reproductive number was about 4.83-11.90. We also estimated the infection on the day before culling to be 45.24% (Guquan), 89.20% (Jinba) and 16.35% (Liancheng), respectively. The infection rate of Guquan could reach about 74.8% if culling were postponed by 2 days. We found that the infection was significantly higher than the morbidities (22.11% (Guquan), 49.35% (Jinba) and 12.94% (Liancheng)) calculated by actual statistical data. Besides, we simulated and compared the control effect of stopping transport, disinfecting, stopping swill and culling. Our findings suggest that any single measure was not enough to prevent the spread of ASF on a regional level but the combined measures is the key. Under the current situation, fully culling was recognized as most effective in controlling the epidemic, despite the culling of uninfected pigs.

Epidemic situation and control measures of African Swine Fever Outbreaks in China 2018-2020.

As of 21 April 2020, 176 ASF outbreaks have occurred in China. For each outbreak, an investigation was conducted, including historical data retrieval and traceability of potential contacts. The purpose of this study is to conduct a preliminary analysis of the data obtained from the outbreak investigations, including an investigation of the possible contributing factors of the spread of ASF in China. Based on the epidemic situation and the policies issued, the entire epidemic can be divided into three phases. 71 outbreaks were reported between 3 August 2018 and 17 November 2018; 44 outbreaks between 19 November 2018 and 30 March 2019; and 61 outbreaks between 4 April 2019 and 12 April 2020. Based on the reported outbreaks, the proportional rate of outbreaks in small farms (livestock ≤ 500, 127/168) is significantly higher than that of medium (501 ≤ livestock < 2,000, 14/168; 2001 ≤ livestock ≤ 5,000, 9/168) and large farms (livestock ≥ 5,001, 18/168). The odds of infection related to swill feeding (OR = 2.5, 95% CI, 1.5-4.3) and the mechanical dissemination of vehicles and personnel (OR = 2.7, 95% CI, 1.6-4.5) are significantly higher than those of pigs and pig production transportation. Swill feeding is the major contributing factor for small farms while mechanical dissemination of vehicles and personnel is the major contributing factor for large farms. The average duration from the beginning of the infection to the official outbreak report is gradually decreasing, which means that response speed of industry entities and the animal husbandry and veterinary departments from the beginning of the infection to the outbreak report is gradually increasing. Based on the analysis for ASF outbreaks, some policies and suggestions were put forward, such as improving the biosecurity level of the farms, as well as strengthening the supervision of breeding, transportation and slaughter.

Closure of live bird markets leads to the spread of H7N9 influenza in China.

Following the emergence of H7N9 influenza in March 2013, local animal and public health authorities in China have been closing live bird markets as a measure to try to control the H7N9 influenza epidemic. The role of live bird market (LBM) closure on the spread of N7N9 influenza following the closure of LBMs during March to May 2013 (the first wave) and October 2013 to March 2014 (the second wave) is described in this paper. Different provinces implemented closure actions at different times, and intensive media reports on H7N9 in different provinces started at different times. Local broiler prices dropped dramatically in places with outbreaks and more live chickens were transported to other LBMs in neighboring areas without human cases from infected areas when live bird markets were being closed. There were six clusters of human infection from March to May 2013 and October 2013 to March 2014 and there may have been intensive poultry transportation among cluster areas. These findings provide evidence that the closure of LBMs in early waves of H7N9 influenza had resulted in expansion of H7N9 infection to uninfected areas. This suggests that provincial authorities in inland provinces should be alert to the risks of sudden changes in movement patterns for live birds after LBM closure or increased publicity about LBM closure.

Community-based interventions to enhance knowledge, protective attitudes and behaviors towards canine rabies: results from a health communication intervention study in Guangxi, China.

BACKGROUND: In China canine rabies poses a serious public health problem in that human mortality ranks the second highest globally. While rabies health education interventions are advocated by WHO to be critical components of modern rabies control and prevention programs, available studies have not adequately investigated the relative efficacy of their implementation in at-risk populations. This study aims to measure and compare the effect on knowledge and protective behavior towards rabies of health education interventions that include a novel Short Messaging Service via cell phone (SMS) and rabies health information sessions (IS). METHODS: The study used a between-subject design involving repeated measures of rabies-related KAP (knowledge, attitude and practice). A total of 350 randomly selected villagers were randomly allocated into three intervention (SMS, IS and SMS + IS) and one control group. The content of SMS and IS covered topics about rabies prevention and route of transmission. The SMS intervention consisted of ten separate messages delivered three times two weeks after the pretest; the IS intervention was conducted once immediately after the pretest. A validated questionnaire was used to capture demographic information and KAP information. Ordinary Least Squares regression was used to contrast the effects of interventions. RESULTS: Our results indicate that overall SMS outperforms IS at improving knowledge and protective behavior against rabies. Our results suggest that a combined intervention of SMS and IS can result in higher scores than any of the two in isolation. The impact of SMS, IS and SMS + IS is greatest on knowledge, followed by attitude and practice scores. CONCLUSION: This study demonstrated that health communication modes based on SMS, IS and a combination of the two are all effective to improve rabies-related KAP in the short term. These findings highlight the potential usefulness of SMS as an additional tool for public health communication and promotion; further studies are needed to investigate the long term benefits of these interventions on the reduction of dog bites and resulting human rabies incidence.

Comparative genomic analysis between newly sequenced Brucella suis Vaccine Strain S2 and the Virulent Brucella suis Strain 1330.

BACKGROUND: Brucellosis is a bacterial disease caused by Brucella infection. In the late fifties, Brucella suis vaccine strain S2 with reduced virulence was obtained by serial transfer of a virulent B. suis biovar 1 strain in China. It has been widely used for vaccination in China since 1971. Until now, the mechanisms underlie virulence attenuation of S2 are still unknown. RESULTS: In this paper, the whole genome sequencing of S2 was carried out by Illumina Hiseq2000 sequencing method. We further performed the comparative genomic analysis to find out the differences between S2 and the virulent Brucella suis strain 1330. We found premature stops in outer membrane autotransporter omaA and eryD genes. Single mutations were found in phosphatidylcholine synthase, phosphorglucosamine mutase, pyruvate kinase and FliF, which have been reported to be related to the virulence of Brucella or other bacteria. Of the other different proteins between S2 and 1330, such as Omp2b, periplasmic sugar-binding protein, and oligopeptide ABC transporter, no definitive implications related to bacterial virulence were found, which await further investigation. CONCLUSIONS: The data presented here provided the rational basis for designing Brucella vaccines that could be used in other strains.

Genetic polymorphism characteristics of Brucella canis isolated in China.

In China, brucellosis is an endemic disease typically caused by Brucella melitensis infection (biovars 1 and 3). Brucella canis infection in dogs has not traditionally recognized as a major problem. In recent years however, brucellosis resulting from Brucella canis infection has also been reported, suggesting that infections from this species may be increasing. Data concerning the epidemiology of brucellosis resulting from Brucella canis infection is limited. Therefore, the purpose of this study was to assess the diversity among Chinese Brucella canis strains for epidemiological purposes. First, we employed a 16-marker VNTR assay (Brucella MLVA-16) to assess the diversity and epidemiological relationship of 29 Brucella canis isolates from diverse locations throughout China with 38 isolates from other countries. MLVA-16 analysis separated the 67 Brucella canis isolates into 57 genotypes that grouped into five clusters with genetic similarity coefficients ranging from 67.73 to 100%. Moreover, this analysis revealed a new genotype (2-3-9-11-3-1-5-1:118), which was present in two isolates recovered from Guangxi in 1986 and 1987. Second, multiplex PCR and sequencing analysis were used to determine whether the 29 Chinese Brucella canis isolates had the characteristic BMEI1435 gene deletion. Only two isolates had this deletion. Third, amplification of the omp25 gene revealed that 26 isolates from China had a T545C mutation. Collectively, this study reveals that considerable diversity exists among Brucella canis isolates in China and provides resources for studying the genetic variation and microevolution of Brucella.

Clinical studies on treatment of chronic prostatitis with acupuncture and mild moxibustion.

To observe the therapeutic effect of acupuncture and mild moxibustion on chronic prostatitis and to probe into the mechanism of the therapy. Two hundred patients with chronic prostatitis were randomly divided into two groups so as to observe respectively the changes in clinical symptoms, count of WBC and lecithin corpuscles in succus prostaticus, prostatic capcules and internal echo, tenderness and elasticity of prostate by palpation before and after treatment. After treatment, a remarkable improvement was found in clinical symptoms, succus prostaticus test and ultrasonic examination in the treatment group with a statistically significant difference (P < 0.05) as compared to the control group. The treatment of chronic prostatitis with acupuncture and mild moxibustion can remove the stagnation of succus prostaticus, improve the blood circulation in prostate, inhibit or kill the pathogenic micro-organisms, strengthen or regulate the immune function of the patients, improve local blood circulation, eliminate the accumulation of secretion and relieve the obstruction of the prostatic ducts.