Table of Content

20 October 2023, Volume 48 Issue 5

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Genomic􀆼wide SNP Genotyping Methods and the Application of Genomic Selection in Livestock and Poultry

WANG Yiqiu, JIANG Ziqin, WANG Yuzhe, XU Zhenqiang, HU Xiaoxiang,

2023, 48(5):  1-6.  DOI: 10.19978/j.cnki.xmsy.2023.05.01

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Genomic selection is a method that uses genome ??wide markers to predict the breeding value of an individual. Single nucleotide polymorphisms（SNPs）are widely used in the field of genetics. In recent years，with the development of SNP genotyping methods，researchers have been able to obtain a large number of genetic markers at a low cost，thereby greatly advancing the technology of genomic selection. This study reviews three main strategies for genome??wide SNP genotyping，summarizes the application of genomic selection in livestock and poultry breeding，and discusses its future prospects. This review will serve as a reference for selecting an appropriate SNP genotyping strategy in the progress of genomic selection.

Research Progress on Biological Function and Application of Methionine in Ducks

ZHANG Yanan , ZHUANG Zhiwei , JIN Yongyan , CHEN Wei, WANG Shuang, XIA Weiguang, JIN Chenglong, ZHU Xiaoping, ZHENG Chuntian

2023, 48(5):  7-14.  DOI: 10.19978/j.cnki.xmsy.2023.05.02

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Methionine is the first limiting amino acid for poultry when fed with corn?soybean meal diet. It plays an important role in maintaining the normal growth and laying performance of poultry. However，the research progress of methionine is mainly focused on chickens and few studies are on ducks. So far，the DL ? methionine，methionine hydroxyl analogue and L?methionine are commonly used in poultry. But they are absorbed and transferred in different ways to form L?methionine，and the efficiency is different. Methionine and its hydroxy analogues are absorbed through the front end of digestive tract，duodenum or ileum，and L ? methionine is produced by rate ? limiting enzymes andtransaminases. They participate in a variety of metabolic pathways in liver and other tissues，promote protein synthesis，provide methyl donors for methionine cycle，and improve antioxidant properties. Therefore，this article summarized the function of methionine and its application in ducks，including the apparent digestibility of methionine in duck，the optimal requirement and bioefficiency between resources in meat ? and egg ? ducks，in order to provide scientific basis for the application of methionine in duck production.

Present and Prospective View of Intestinal Organoids in Swine Basic Research

XIE Zeen, ZHANG Yongliang

2023, 48(5):  15-22.  DOI: 10.19978/j.cnki.xmsy.2023.05.03

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Intestinal organoids are cell ? based structures that mimic the functions of organs. They are three ? dimensional structures composed of various types of cells derived from intestinal stem cells via laboratory cultivation. In recent years，their applications in medical research，drug testing，and animal sciences have drawn significant attention. Intestinal organoids offer a promising tool for studying the physiology of the gut as they closely resemble the in vivo conditions and as reliable alternatives to traditional cell?based models in vitro research. In swine?based studies， these organoids have been successfully employed to simulate viral infections and investigate intestinal functions， including nutrient absorption and viral diseases.The use of intestinal organoids has also gained attention in the field of veterinary medicine. Their potential and advantages have been increasingly recognized，leading to a rise in related publications. However，there are still unresolved challenges that need to be addressed to further expand their applications in this field.In this review article，we provided an overview of the applications for intestinal organoids in animal sciences，with a specific focus on swine research. Furthermore，we proposed several recommendations for the development and future directions of this technology in the field of livestock and veterinary medicine.

A Review of Research on Zoonotic Porcine Deltacoronavirus: Etiology, Epidemiology, Prevention and Control

GUO Yifan, YAN Quanhui, XING Jiabao, LI Zijie, ZENG Weijun, FAN Shuangqi

2023, 48(5):  23-26.  DOI: 10.19978/j.cnki.xmsy.2023.05.04

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In recent years，zoonotic coronaviruses have been emerging or re ?emerging，posing a great threat to public health and the farming economy. Porcine deltacoronavirus，which was first reported in Hong Kong，is one of them. Its etiological characteristics were first identified in the United States in 2014，followed by extensive reports in various countries in North America and Asia. The virus has shown frequent inter ? lineages and intra ? lineages recombination and high genetic diversity，infecting birds and several mammals，including humans.In addition， different lineages of strains infecting humans also have the characteristics of convergent evolution，which also brings severe challenges to human health. In this paper，we reviewed the research status of zoonotic porcine deltacoronavirus， and analyzed the etiology and epidemiology of porcine deltacoronavirus，so as to further understand the origin and cross?species transmission route of porcine deltacoronavirus，and put forward prevention and control suggestions.

Research Progress of Infectious Bursal Disease Vaccines

ZHANG Xuzhi，SUN Caiyi，XU Shaozhu，ZHANG Guoli，QI Dongmei*

2023, 48(5):  27-32.  DOI: 10.19978/j.cnki.xmsy.2023.05.05

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Infectious Bursal Disease（IBD）is an acute，highly contagious infectious disease caused by infectious bursal disease virus（IBDV）mainly affecting young chickens. It is characterized by severe injury of bursa of Fabricius and immunosuppression. Since 1980s，IBDV has been found in many countries and regions. In addition，IBDV variants with increased virulence，namely variant IBDV（vIBDV）and variant/very virulent IBDV（vvIBDV）had emerged. IBD cause significant economic losses to the poultry industry worldwide. At present，comprehensive prevention and control measures combining vaccination and biosafety are mainly adopted. In this paper，IBD attenuated live vaccines，inactivated vaccines，immune complex vaccines，recombinant vector live vaccines，subunit vaccines and DNA vaccines that have been commercialized or are under development are reviewed，in order to provide theoretical reference for better prevention and control of IBD.

Research Progress of Emulsion Adjuvants

XU Shaozhu，ZHANG Guoli，SHI Daqing，QIU Chengwen，KUANG Zhenjie，QI Dongmei*

2023, 48(5):  33-38.  DOI: 10.19978/j.cnki.xmsy.2023.05.06

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Emulsion immune adjuvant，also known as oil adjuvant，is a kind of immune adjuvant that encapsulates antigens in various oil phases to provide a reservoir for immunogens and enhances the body 's specific immune response to antigens by presenting antigens to immune cells. Since the application of emulsion adjuvants was first reported in 1916，emulsion adjuvants have become an indispensable factor in vaccines research and development for more than 100 years，providing a material guarantee for safer and more efficacy vaccines against various infectious diseases in humans and animals. With the continuous development of modern vaccinology，the research and development of new vaccines have put forward higher requirements for adjuvants. Knowing the composition and understanding the immune mechanism of adjuvants are the premise for the application of adjuvants in vaccine research and development. In order to provide a theoretical reference for emulsion adjuvants in clinical use，in this paper， commonly used and commercialized emulsion adjuvants，such as white oil adjuvants，complete Freund 's adjuvant （CFA），incomplete Freund 's adjuvant（IFA），MF59?，AS adjuvants and Montanides adjuvants were chosen，and their components，immune mechanism and clinical application were reviewed.

Effects of Chinese Herbal Preparations on Growth Performance，Serum Antioxidant Capacity and Liver Health of Largemouth Bass Cultured with Low Fish Meal

HU Junru1 ，CHEN Xiaoying1 ，WU Haomin1 ，DENG Jingming2 ，YUAN Minggui3

2023, 48(5):  39-45.  DOI: 10.19978/j.cnki.xmsy.2023.05.07

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This experiment was conducted to evaluate the effects of Chinese herbal preparations on growthperformance，serum antioxidant capacity and liver health of largemouth bass. A total of 840 juvenile fish with an average initial weight of about 4.40 g were randomly divided into 6 groups with 4 replicates per group and 35 fish per replicate. The basic diet was supplemented with 0，0.05%，0.1%，0.2%，0.4% Chinese herbal extracts and 0.4% fermented Chinese herbal，respectively. Six isonitrogenous and isoenergetic experimental diets were recorded as G0， G0.05，G0.1，G0.2，G0.4 and GF0.4，respectively. The experiment was lasted 51 days. The results showed，there were no significant differences in average weight，weight gain rate，specific growth rate，survival rate and feed intake of largemouth bass among all groups（P>0.05）. Compared with G0 group，the VSI and HSI in G0.05 group and HSI in GF0.4 group were the lowest，but the differences were not significant（P>0.05）. The activities of AST and ALT in G0.1 group，the activities of GPX，SOD and CAT in G0.05 group were significantly increased（P<0.05），while the MDA content in G0.1 and GF0.4 was significantly decreased（P<0.05）. The liver tissue sections showed that the cytoplasm of G0 group was loose and lightly stained，and a few lymphocytes were focal infiltration，while G0.05，G0.1 and G0.2 groups only occasionally showed small focal lymphocytes infiltration. G0.4 group showed more watery degeneration of hepatocytes，loose cytoplasm and light staining，and a small amount of lymphocyte infiltration. No obvious abnormality was observed in GF0.4 group. In conclusion，supplementation of Chinese herbal extract and fermented Chinese herbal in low fish meal diet has no significant effect on the growth performance of largemouth bass， but supplementation of 0.05% Chinese herbal extract and 0.4% fermented Chinese herbal can improve the antioxidant capacity and liver tissue health of largemouth bass which fed low fish meal diet.

The Effect of Compound Qinggan Decoction on Acute Liver Injury in Canines Based on Cell Pyroptosis

XIE Zimao1 ，SU Yiman2 ，QIU Wenyue2 ，HUANG Jianjia2 ，BAI Yuman2 ，XIE Wenting2 ， ZHOU Shuilian2 ，WANG Rongmei3*

2023, 48(5):  46-52.  DOI: 10.19978/j.cnki.xmsy.2023.05.08

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（Objective） The purpose of this study was to explore the therapeutic effect and mechanism ofnamely the control group，the model group and the Chinese medicine treatment group. Acute liver injury model was established by intraperitoneal injection of 1 mg/kg of carbon tetrachloride，and the treatment group was given 1 ml/kg of Compound Qinggan Decoction every day for 14 days. At the end of the experiment，blood samples were collected for liver function test and liver tissue samples were taken out. H&E staining was used to observe the pathological changes of liver tissue，qRT ? PCR was used to detect the mRNA expression levels of hepatocyte pyroptosis ? related genes NLRP3，ASC，GSDMD，Caspase ? 1，IL ? 1β and IL ? 18，Western Blot was used to detect the expression levels of hepatocyte pyroptosis ? related proteins NLRP3，GSDMD，Caspase ?1，IL ?1β and IL ?18，and immunohistochemical method was used to detect the localization and expression of pyroptosis executive protein GSDMD.（Results）The experimental results showed that the activities of AST，ALT and γ?GT in the model group increased significantly（P< 0.05）；the arrangement of hepatocytes is disordered，and hepatocyte necrosis and inflammatory cell infiltration occur； the expression of pyroptosis?related genes NLRP3，ASC，IL?1β and IL?18 increased significantly（P<0.05），and the expression of pyroptosis?related proteins NLRP3，GSDMD，Caspase?1，IL?1β and IL?18 increased. After treatment with Compound Qinggan Decoction，the activities of AST，ALT and γ ? GT decreased clearly（P<0.05），and the pathological damage of liver tissue improved obviously. The mRNA expression levels of pyroptosis ? related genes NLRP3，ASC and IL ? 18 decreased evidently（P<0.05），and the expression levels of pyroptosis ? related proteins NLRP3，GSDMD，Caspase?1，IL?1β and IL?18 decreased.（Conclusion）Compound Qinggan Decoction can alleviate acute liver injury induced by carbon tetrachloride in canines by inhibiting the pyroptosis of hepatocytes. Compound Qinggan Decoction on acute liver injury induced by carbon tetrachloride in canines.（Methods）Eighteen mongrel canines aged from 12 to 15 months were randomly divided into three groups，with 6 canines in each group，

Effects of E627K Mutation in the PB2 Protein of H9N2 Avian Influenza Virus on the Expression and Distribution of RIP3, Slit2 and Robo4 in the Lungs of Mice

XU Dan，DENG Zhile，LIU Ruidong，HU Qiming，NING Zhangyong，FAN Xiaolong*

2023, 48(5):  53-58.  DOI: 10.19978/j.cnki.xmsy.2023.05.09

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The site 627 of PB2 protein has been confirmed to be a key factor in determining the host range andC mice，in this study，BALB/C mice were infected with H9N2 AIV（VK627）and its amino acid substitution in PB2 residue 627 strain（rVK627E）. At 1，3，5，and 6 days post ?inoculation，the lungs of three euthanized mice in each group were collected，respectively. The specific expression and histological distribution of RIP3，Slit2 and Robo4 in lung tissue were detected by qRT?PCR and immunohistochemistry. The results showed that the expression of RIP3 in the lungs significantly increased in challenge group compared with the control group，meanwhile，the expression level of RIP3 in VK627 group was significantly higher than that in rVK627E group. These two H9N2 AIV strains inhibited the expression of Slit2 and Robo4 in lung at 1 ? day post infection. After that，the expression level of Slit2 and Robo4 increased dramatically in rVK627E group than VK627 infected group and the control. Immunohistochemical detection showed that the epithelial cells of alveolar wall and pulmonary arterial vascular endothelial cells exhibited mild positive staining for RIP3，and it was also moderately to strongly expressed in the submucosa cells of trachea of the challenge group. The alveolar epithelial cells and mucosal epithelial cells of trachea were medium to strong positive staining for Slit2 protein expression in the control group and rVK627E group，while there was relatively weak positive staining of the same cells in VK627 groups. For Robo4，the location of positive cells in the lung had consistency to Slit2. In summary，H9N2 AIVs infected mice significantly affected the expression and distribution of RIP3 and Slit2/Robo4， which were closely related to the pathogenicity of H9N2 AIVs in mice. Mutations in PB2 protein locus 627 play an important role in the influence of H9N2 AIVs on RIP3，Slit2 and Robo4 expression and distribution. This study will provide basic data for further exploration of the molecular mechanism of H9N2 AIVs and its point mutant strain ? mediated host pathogenesis and the selection of anti?inflammatory strategies. pathogenicity of influenza A virus（IAV）. To further investigate the effects of E627K mutation in the PB2 protein of H9N2 Avian Influenza Virus（AIV）on the expression and distribution of RIP3，Slit2 and Robo4 in the lungs of BALB/

Preparation, Identification and Preliminary Application of Canine Parvovirus Positive Serum

CHEN Yanfei，CHEN Jian，XUE Qinghong，LI Ling，SUN Miao

2023, 48(5):  59-63.  DOI: 10.19978/j.cnki.xmsy.2023.05.10

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Canine parvovirus（CPV）positive serum is indispensable in the efficacy test，exogenous virus testThe results showed that the neutralizing antibody titer of CPV positive serum was as high as 1∶32768，the neutralizing index was higher than 106.2 ，the neutralizing antibody titers of CPV positive serum against canine distemper virus， canine parainfluenza virus and canine adenovirus were all lower than 1∶4，and the neutralizing antibody titer of rabies virus was lower than 0.03 IU/mL. The CPV positive serum can completely neutralize 10 doses of vaccine virus in exogenous virus test for three CPV related live vaccines. The CPV positive serum was used for identification test of two CPV related live vaccines，and the results met the quality standards. IFA for CPV was used to determine the viral titer of three CPV vaccine strains，and the results showed that the IFA method established had cross immune response to the three vaccine strains，which had little difference with the sensitivity of commercial CPV fluorescent monoclonal antibody. All above show that CPV positive serum has strong neutralizing ability and good specificity，and can be used for the efficacy test，exogenous virus test and identification test of canine live vaccines. and identification test of CPV ? related vaccines，and is crucial for the quality control of CPV ? related biological products. In order to prepare CPV positive serum with high neutralizing antibody titer and good specificity，two healthy and susceptible beagles were immunized with CPV DD strain for the test. After four times of immunization，we collected blood and separated serum. The serum was identified by virus neutralization test，immunofluorescence assay，and sterility test，mycoplasma test，and exogenous virus test in the third part of the Chinese Veterinary Pharmacopoeia 2020. And then the serum was used in the exogenous virus test and identification test in canine vaccines. An indirect immunofluorescence assay（IFA）for CPV was established and applied to potency test of vaccine.

A Case Report of An Alpaca Infected with Mycoplasma

LAN Jianyuan1# ，LI Chi2# ，BA Juan1 ，ZHANG Xiandong1 ，LUO Lixia2 ，LIU Jinfang2 ，HOU Yuee1*

2023, 48(5):  64-68.  DOI: 10.19978/j.cnki.xmsy.2023.05.11

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In order to find out the causes of the death of the alpaca in Zhuhai Taichuangyuan alpaca breeding base and accumulate the experience of epidemic prevention and control，the dead alpaca was examined by pathological examination，histopathological examination，hematological examination，molecular biology examination. The results of the complete blood count and blood biochemistry indicated inflammation. Post ? mortem examination and histopathology revealed multiple organ congestion and bleeding. Candidatus Mycoplasma ravipulmonis was confirmed by PCR sequencing.

Recent Advances in Etiological Analysis and Diagnosis of Young Pigeon Disease Syndrome

LIU Mengfan1 ，KE Junhong2 ，JIANG Hanyu1 ，YANG Huihu1 ，XIE Zimin1 ，LUO Rui1 ， HUANG Shujian1，2* ，MEI Kun1*

2023, 48(5):  69-74.  DOI: 10.19978/j.cnki.xmsy.2023.05.12

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In recent years，young pigeon disease syndrome（YPDS）has widely spread in the world，causing huge economic losses to the pigeon breeding industry. At present，there are few studies on YPDS，diagnostic criteria， and specific drugs and vaccines，which brings great challenges to the prevention and control of YPDS. In order to better understand the syndrome，this article systematically reviews the etiology，prevalence，diagnosis and prevention and control measures of YPDS. Referring to the relevant literature at home and abroad in the past 30 years，the pathogens that may cause YPDS were systematically described，and each diagnosis method and control measures were summarized. YPDS is a multifactorial disease mainly caused by Pigeon circovirus （PiCV），Pigeon herpesvirus （PiHV）and Pigeon adenovirus（PiADV）. It can cause the death of young pigeons aged 7 to 15 weeks. The main clinical symptoms included depression，disordered feathers，anorexia，vomiting and diarrhea. At present，the mainprevention and control measures of this syndrome are to strengthen feeding management. This paper summarized the etiology，diagnosis and prevention and control measures of YPDS，which provides reference for production practice.

Comparison the Efficacy of Inactivated Porcine Reproductive and Respiratory Syndrome Virus（PRRSV）Vaccines with that of Commercial Vaccines Against Highly Pathogenic PRRSV Challenges

WANG Pengjiang，CAO Jian，ZHANG Wei，HOU Feng LI Junhui，LIU Qiangde，YUAN Ke， SUN Yuanjun，GAO Yangyi，ZHANG Pan，ZHOU Tao，HE Sun*

2023, 48(5):  75-79.  DOI: 10.19978/j.cnki.xmsy.2023.05.13

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To comparison the protective efficacy of inactivated porcine reproductive and respiratory syndrome virus（PRRSV）vaccines and commercial vaccines against highly pathogenic PRRSV（HP?PRRSV）challenge，twenty four ?weeks ? old piglets were selected from a PRRSV ?free pig farm. Piglets negative for all examined pathogens and antibodies against PRRSV and ASFV were selected for this study. Piglets were randomly divided into four groups（n= 5）and housed in separate rooms. Three groups of piglets were intramuscularly with（A，B and C），2 mL/pig. The piglets in the control group were inoculated with saline，2 mL/pig. All piglets were inoculated again by equal mannerand dose after three weeks. At 10 weeks of age，all pigs were challenged intranasally and intramuscularly with PRRSV XJL03（3×104.5 TCID50）in saline（1.5 ml，respectively）. Blood samples and nasal swab were collected at indicated times and rectal temperatures and deaths were recorded. All surviving piglets were necropsied at 21 days post ? challenge （dpc） for pathological examination. The result showed that four of five pigs of the control group （unvaccinated）were caused seven pathological lesions by HP?PRRSV challenge，three inactivated vaccines could not significantly relieved viremia and clinical symptom，vaccine A，vaccine B and vaccine C were provided only with 20% ，20% and 60% protection against piglets，respectively. In conclusion，our data suggest that the vaccine C vaccination protected 60% of piglets from challenging with HP?PRRSV strain and also provided comparable efficacy to that of the commercially licensed HP?PRRSV vaccine and Classical PRRSV vaccine.

Comparing Detection Effects of Several Domestic Avian Leukosis Antigen Test Kits

LIU Yang1 ，CHEN Shunyan2 ，WANG Zhanxin1 ，LIAO Qiusheng2 ，WU Zhiqiang1*

2023, 48(5):  80-84.  DOI: 10.19978/j.cnki.xmsy.2023.05.14

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At present，there is no effective drug and vaccine to control the avian leukosis，so it is very important to detect and eliminate the positive individuals to purify the breeding flocks in a short time. Therefore，it is significant to select antigen detection kits with strong specificity，high sensitivity and good stability. Due to the impact of the epidemic novel coronavirus，some foreign antigen test kits can not be imported into China，it is necessary to find domestically?made kits with great sensitivity and stability. Therefore，in order to guarantee the continuous purification work，it is necessary to look for the domestic kits with great sensitivity and stability that is closest to the imported kit. The results showed that the sensitivity and stability of the different kits for the detection of the cell supernatant were different，the positive detection rate of the four kits was B>D>C=A. Among the four Avian Leukosis antigen test kits， the kit B had the highest detection sensitivity，the kit C had the best stability and closest to the imported kits. This study provides reference and data support for the kit selection in the process of Avian Leukosis purification.