Spatio-temporal analysis and risk modeling of foot-and-mouth disease outbreaks in China.

FMD is an acute contagious disease that poses a significant threat to the health and safety of cloven-hoofed animals in Asia, Europe, and Africa. The impact of FMD exhibits geographical disparities within different regions of China. The present investigation undertook an exhaustive analysis of documented occurrences of bovine FMD in China, spanning the temporal range from 2011 to 2020. The overarching objective was to elucidate the temporal and spatial dynamics underpinning these outbreaks. Acknowledging the pivotal role of global factors in FMD outbreaks, advanced machine learning techniques were harnessed to formulate an optimal prediction model by integrating comprehensive meteorological data pertinent to global FMD. Random Forest algorithm was employed with top three contributing factors including Isothermality(bio3), Annual average temperature(bio1) and Minimum temperature in the coldest month(bio6), all relevant to temperature. By encompassing both local and global factors, our study provides a comprehensive framework for understanding and predicting FMD outbreaks. Furthermore, we conducted a phylogenetic analysis to trace the origin of Foot-and-mouth disease virus (FMDV), pinpointing India as the country posing the greatest potential hazard by leveraging the spatio-temporal attributes of the collected data. Based on this finding, a quantitative risk model was developed for the legal importation of live cattle from India to China. The model estimated an average probability of 0.002254% for FMDV-infected cattle imported from India to China. TA sensitivity analysis identified two critical nodes within the model: he possibility of false negative clinical examination in infected cattle at destination (P5) and he possibility of false negative clinical examination in infected cattle at source(P3). This comprehensive approach offers a thorough evaluation of FMD landscape within China, considering both domestic and global perspectives, thereby augmenting the efficacy of early warning mechanisms.

Significant association of mutations close to LCORL gene with growth performance in Zhedong white geese (Anser cygnoides).

This study aimed to investigate the role of the LCORL gene in regulating the growth performance of Zhedong white (ZDW) geese, belonging to the swan geese (Anser cygnoides), and identify possible selective signatures in diverse goose breeds. Single nucleotide polymorphisms around LCORL were genotyped, and their associations with body-size-related (BSR) traits were estimated. The results showed that the genotyped loci upstream of LCORL were significantly related to the body weight and breast width of ZDW geese aged 10 weeks (p < 0.05). A genome scan comparing expected heterozygosity among different breeds identified a ~150 kb long genomic region with extremely low heterozygosity downstream of LCORL among swan geese. Further, significant associations of variants within the low heterozygosity region among ZDW geese with BSR traits, including body weight, body length and breast width (p < 0.05) were also detected. Overall, mutations adjacent to LCORL were related to the growth performance of swan geese, and the significant effects of variants in a low-heterozygosity region on BSR traits provided valuable insights into the molecular mechanism of artificial selection reshaping body stature in swan geese.

Spatio-temporal and trade export risk analysis of bluetongue disease in France: A case study of China.

Bluetongue disease (BT) is a viral disease that can be introduced through imported animals and animal products, affecting local animal husbandry. In this study, the spatial and temporal patterns of BT outbreaks (outbreak: a BT infection in cattle, sheep, or goats on a farm, involving at least one infected animal) in France were analyzed and the risk of introducing bluetongue virus (BTV) into countries through trade was assessed. A spatiotemporal analysis of BT reported during the study period (2015-2018) showed that there were clustered outbreaks of BT in France in 2016 and 2017, with outbreaks concentrated from August to December. The outbreak moved eastward from the center of mainland France to surrounding countries. A semi-quantitative risk analysis framework was established by combining the likelihood assessment and consequence analysis of introducing BTV into trading countries through trade. Exemplified by China, the research showed that in the analysis of the likelihood of BTV from France being introduced into trading countries through live cattle trade, China imports a large number of live cattle, bringing high risks. The likelihood of introducing bovine semen into trading countries was similar to that of live cattle, but the harm caused by the trade in live cattle was higher than that caused by the trade in bovine semen. This risk analysis framework can provide a reference for other countries to quickly assess the risk of bluetongue transmission in import and export trade.

Epidemiologic and import risk analysis of Peste des petits ruminants between 2010 and 2018 in India.

BACKGROUND: Peste des petits ruminants (PPR) is a serious disease that affects goats, sheep and other small ruminants. As one of the earliest and most serious countries, PPR has seriously threatened India's animal husbandry economy. RESULTS: In this study, the spatiotemporal characteristics of the PPR in India outbreaks were analyzed. Between 2010 and 2018, the epidemic in India broke out all over the country in a cluster distribution. Epidemic clusters in northern and southern India are at higher risk, and the outbreak time of PPR has significant seasonality. The results of the analysis of the development and transmission of PPR under the natural infection conditions showed that the PPR outbreak in India reached a peak within 15 days. Finally, the quantitative risk analysis results based on scenario tree show showed that the average probability of infecting PPRV in live sheep exported from India was 1.45 × 10-4. CONCLUSIONS: This study analyzed the prevalence of PPR in India. The analysis of transmission dynamics on the development of the epidemic provides a reference for the prevention and control of the epidemic. At the same time, it provides risk analysis and suggestions on trade measures for the trading countries of India.

Risk assessment and early warning of the presence of heavy metal pollution in strawberries.

Heavy metal pollution is a major threat to agricultural produce and it can pose potential ecological risks which subsequently impacts on human health. Strawberries are an economically important produce of China. The intrinsic link of heavy metal pollution risk in the soil-strawberry ecosystem is of concern. In this study, the pollution index of heavy metal pollutants in farmlands of different provinces were evaluated, and the results showed significantly high levels of cadmium. In addition, Nemerow integrated pollution index analysis showed that low-pollution farmlands only accounted for 14.07% of the total arable land area. Then, the transfer factors were used to calculate the migration of heavy metals from the soil into strawberries. The results showed that cadmium and nickel were relatively high in strawberries from the Guangxi province. Similar results were found for mercury in Jiangxi Province. The pollution index of single food pollution also showed that mercury in strawberries from Jiangxi Province was at a moderate pollution level. The comprehensive pollution index indicated that heavy metal pollution in strawberries in Central China may be severe. In addition, spatial clustering analysis showed that cadmium, chromium, lead, arsenic and zinc in strawberries had significant hotspot clustering in central, south and southwest China. Finally, our studies also suggested that the risk of carcinogenic and non-carcinogenic diseases was higher in the (2, 4] years age group than in other age groups. People in Yunnan Province were also found to have a higher non-carcinogenic risk than those in other provinces and cities in China. This study provides a comprehensive view of the potential risks of heavy metal contamination in strawberries, which could provide assistance in the design of regulatory and risk management programs for chemical pollutants in strawberries, thus ensuring the safety of consumption of these edible fruits.

Using comparative genomics to detect mutations regulating plumage variations in graylag (A. anser) and swan geese (A. cygnoides).

Although graylag geese (A. anser) showed similar plumages of white, grey, and white with grey patches compared to those in swan geese (A. cygnoides), it was believed the substantial molecular mechanism for plumage variations were different. To date, studies on genes responsible for diverse plumages among graylag geese were limited and causal mutations remain unknown. In this study, genomes from 57 individuals belonging to six breeds showing different plumages were sequenced at ∼10X depth. Firstly, the allele frequency differences (AFD) of variants on the scaffold394 (NW_013185915.1) between grey and white goose breeds (A. anser) was calculated and a genomic region between 768,290-779,889 bp was detected to carry candidate variants associated with plumages, including one SNP (g. 775,151G > T, ∼18.6 kb upstream of EDNRB2) found to be fixed in white geese. This region was overlapped with the one detected by the haplotype-based sweep analysis, in which significant signals defined a candidate region of 736,610-820,622 bp on the same scaffold. Results from the transcriptomic data showed that expression levels of EDNRB2 and many other melanogenesis-related genes were significantly decreased among white geese compared to that in grey geese, especially at late embryonic stages (>E15). Modifications at transcriptional levels might result in abnormal melanocyte developments and thus the white plumages when they grow up. In addition, a frameshift mutation (C > -) in exon4 of MLANA gene on scaffold176 (NW_013185876.1) was suggested as the causal mutation for sex-linked dilution phenotype in graylag geese although this requires more demonstration experiments. Together with observed white plumages caused by EDNRB2 mutations in coding regions among swan geese and chicken, our study provided new examples to study the parallel evolution.

Spatiotemporal characteristics analysis and potential distribution prediction of peste des petits ruminants (PPR) in China from 2007-2018.

Peste des petits ruminants (PPR) is a highly infectious disease that mainly infects small ruminants. To date, PPR has been confirmed in more than 70 countries. In China, PPR has occurred in more than 20 provinces and cities. In this study, based on geographic information system (GIS), spatial analysis was used to examine the occurrence of PPR in China from 2007 to 2018. The results showed that PPR first occurred in Tibet and gradually spread to other provinces. The outbreaks of PPR were concentrated in 2014, 2015 and 2018. Combining climate factors with the maximum entropy (MaxEnt), the results also suggested that the potential risk areas of PPR outbreaks in China were mainly Jiangsu, Yunnan and Anhui in Southeast China. Finally, a phylogenetic tree was used to analyse the evolutionary relationship between the peste des petits ruminants virus (PPRV) in China and the global ones, and it was found that the one in China had a close genetic relationship with the one in Mongolia, India and Bangladesh. Understanding and forecasting the distribution of PPR in China will help policymakers develop targeted monitoring plans. Likewise, analysing the global PPRV epidemic trends will play an important role in the elimination and prevention of PPR.

A High-Resolution Melting Analysis with an Unlabeled Probe for CRISPR/Cas9-Induced ZBED6 Knockout Pigs Detection.

BACKGROUND: The zinc finger BED-type containing six knockout (ZBED6-KO) pigs were created to improve economic traits by increasing the expression of insulin-like growth factor 2. They were generated by CRISPR/CRISPR-associated protein 9 (Cas9) technology and a single-base deletion of ZBED6 was found. An efficient and rapid method was needed to detect this type of pig. OBJECTIVE: This study aimed to develop a high-resolution melting (HRM) method to detect ZBED6-KO pigs. METHODS: An unlabeled probe and two primers were designed to develop the HRM method. The limit of detection, specificity, and accuracy of the established method were tested by the constructed plasmid and DNA extracts of tissue specimens. RESULTS: The limit of detection by the established method was 102 copies/µL. The HRM method with an unlabeled probe showed good specificity and high accuracy. CONCLUSIONS: The established HRM analysis with an unlabeled probe showed it to be a highly effective, rapid, and reliable method to distinguish ZBED6-KO pigs from wild-type pigs. HIGHLIGHTS: It is the first time that HRM analysis with an unlabeled probe has been used in the detection of genome editing pigs by the CRISPR/Cas9 technology.

Comparative evaluation of two commercial ELISA kits for detection of antibodies against Akabane virus in cattle serum.

BACKGROUND: Akabane disease (AD), a barrier to international trade for endemic areas with far economic impact on the countries, is caused by Akabane virus (AKAV). Commercial enzyme-linked immunosorbent assay (ELISA) is a commonly used diagnostic technique for AKAV infection, including the IDEXX and IDVET ELISA kits. However, the comparative evaluation of the IDEXX and IDVET ELISA kits has not been published. The object of this study was to evaluate the test performance of the two commercial ELISA kits in detecting serum anti-AKAV antibodies in cattle. RESULTS: With virus neutralization test (VNT) as the "relative gold standard", the diagnostic sensitivity (DSe) was 80.39% (123/153) and 93.46% (143/153) for the IDEXX and IDVET ELISA kit, when suspect samples were included. The diagnostic specificity (DSp) for the IDEXX and IDVET ELISA kit was 93.48% (502/537) and 82.31% (442/537), respectively. CONCLUSION: Both of the tested ELISA kits could be applied to detect antibodies against AKAV in cattle serum. The IDVET ELISA kit had a higher DSe. The IDEXX ELISA kit possessed the higher DSp. These results have important implications if the kits are used to screen herds or individual cattle in surveillance programs, or at border crossings for import-export inspection and quarantine.

Establishment of a Decaplex PCR-Capillary Gel Electrophoresis Method for the Simultaneous Detection of Six Kinds of Genetically Modified Animals.

This study aimed to establish an event-specific multiplex PCR system using microsatellite markers and fluorescently labeled primers to detect six different genetically modified (GM) animal lines, including human lactoferrin GM cattle, human lysozyme GM cattle, human α-lactalbumin GM cattle, myostatin knockout pigs, phytase GM pigs, and ω-3 fatty acid desaturase gene GM pigs. Four different microsatellite loci for species identification, along with six GM animal-specific fragments, were selected as targets for primer design. The capillary gel electrophoresis results of multiplex PCR showed that the target fragments were amplified successfully. This high-throughput multiplex PCR detection system can be applied for the inspection and quarantine of GM animals.

[Preparation and identification of monoclonal antibodies against African horse sickness virus VP7].

Objective To prepare the monoclonal antibody (mAb) against the African horsefever virus (AHSV) VP7 protein and to identify it. Methods mAbs were prepared by using baculovirus expressed VP7 protein in BALB/c mice, and the effect of mAb was detected by ELISA, indirect immunofluorescence assay (IFA), and AHSV positive serum blockade. Results Four mAb strains were selected, including 20A8, 28B3, 30G8 and 47E6, among which 47E6 had the best blocking effect. Conclusion mAbs were successfully prepared against VP7 protein.

Serum metabolomic profile in genetically modified cows carrying human α­lactalbumin gene.

The present study aimed to investigate the serum metabolomic profiles in genetically modified cows carrying and expressing human lactalbumin α (LALBA) and non­LALBA cows, and identify altered metabolic characteristics following the genetic modification. Serum biochemistry indexes were measured according to protocols recommended by International Federation of Clinical Chemistry. The metabolomic profiles were determined using the serum samples collected from LALBA (n=6) and non­LALBA cows (n=6). Welch's two­sample t­test was used to identify the metabolites that significantly differed between the LALBA and non­LALBA groups (fold­change ≠ 1 and P<0.05), followed by random forest and pathway analysis. The serum biochemistry indexes of LALBA and non­LALBA cows were within the normal ranges of healthy cows. A total of 273 metabolites were detected, among which 79 metabolites, including 46 increased and 33 decreased metabolites, differed significantly between the LALBA and non­LALBA groups. Random forest analysis identified 30 potential key metabolites, including 14 elevated and 16 reduced metabolites. These metabolites were primarily involved in pathways concerning the metabolism of leucine, isoleucine, valine, tryptophan and lipids, such as myristate and eicosapentaenoate. However, the serum in LALBA cow had unique metabolomic signature compared with non­LALBA cows. The accumulation of polyunsaturated fatty acids and amino acids, and the reduced levels of long chain saturated fatty acids in serum may benefit LALBA cows. However, further investigations are required to validate these benefits and the corresponding mechanisms.

Infection prevalence and phylogenetic analysis of Perkinsus olseni in Ruditapes philippinarum from East China.

The prevalence of Perkinsus sp. infection in Manila clam Ruditapes philippinarum was investigated in the coastal areas of east China. Thirteen groups of clams were collected from 5 sites: Dandong and Qingdao Bays (Yellow Sea), Weifang Bay (Bohai Sea), and Ningbo and Fuzhou Bays (East China Sea). The clams were tested for perkinsosis infection using Ray's fluid thioglycollate medium culture assay. Perkinsus sp. was found in samples from all 5 sites from May 2008 to May 2009. Infection prevalence ranged from 43.75 to 95.83%, and was significantly higher in October than in May. The only 3 uninfected groups of clams were collected from Weifang Bay, the site farthest from the ocean. There was no difference in the prevalence of infection among the remaining 4 sites. The conserved internal transcribed spacer regions of the ribosomal RNA gene complex in each of the Perkinsus sp. isolates were amplified by PCR. The resulting amplicons were sequenced and phylogenetically analyzed. All the Perkinsus isolates were identified as Perkinsus olseni.