Avian haemosporidian parasites, especially Plasmodium juxtanucleare, Leucocytozoon caulleryi, and Leucocytozoon sabrazesi, represent major threats to poultry health and production. However, there is limited epidemiological information about these pathogens in domestic chickens in Southern China, which hinders effective disease prevention and control. The objective of this study was to conduct a cross-sectional survey to investigate the epidemiological characteristics of these three parasites in Guangxi and Hunan Provinces between June 2024 and December 2025. A total of 941 blood samples were collected from domestic chickens and analyzed using both nested PCR targeting the cytb gene and species-specific PCR assays targeting the coxI gene. The overall detection rate of haemosporidian infection was 25.40% (239/941). P. juxtanucleare was the most commonly detected species, with a detection rate of 23.59% (222/941), followed by L. caulleryi at 1.81% (17/941), while no L. sabrazesi infections were identified. Analysis of risk factors showed that chickens older than 90 days had significantly higher detection rates for both P. juxtanucleare and L. caulleryi compared to younger birds. Additionally, breed-specific differences were noted, with black-bone and partridge chickens showing higher susceptibility to P. juxtanucleare than three-yellow chickens. Genetic analysis of coxI sequences demonstrated high conservation among P. juxtanucleare isolates (99.7-100% similarity) and complete identity among L. caulleryi strains. Phylogenetic analysis confirmed that all sequences clustered with the corresponding reference strains from GenBank. This study presents an epidemiological evaluation of these three haemosporidian parasites in domestic chickens from Guangxi and Hunan Provinces, identifying P. juxtanucleare as a widespread pathogen and highlighting age and breed as important risk factors. These results emphasize the importance of ongoing monitoring and targeted control measures in the area.

Fasciola gigantica can modulate host immune mechanisms through excretory–secretory products (ESP). As one of the components of ESP, it is unknown whether Abelson tyrosine protein kinase (Abl) is involved in parasite–host immune interaction. To investigate the immunoregulatory function of Abl in Fasciola gigantica, we cloned and expressed the Fasciola gigantica Abl protein and assessed its effect on specific immune functions of buffalo peripheral blood mononuclear cells (PBMCs). Recombinant F. gigantica Abelson tyrosine protein kinase (rFgAbl) was expressed in Escherichia coli. Western blot analysis was performed to assess the reactivity of anti-rFgAbl antibodies with rFgAbl, serum from F. gigantica-infected buffalo, and excretion and secretion products of F. gigantica. Immunohistochemical analysis was conducted to determine the localization of FgAbl in tissues from larval stages and adult worms of F. gigantica. Furthermore, immunofluorescence analysis was utilized to evaluate the binding ability of the rFgAbl protein to buffalo peripheral blood mononuclear cells (PBMCs), as well as to investigate the effects of varying concentrations of rFgAbl protein (5, 10, 20, 40, and 80 μg/mL) on the functional responses of PBMCs. Anti-rFgAbl antibodies specifically recognize rFgAbl, serum from buffalo infected with F. gigantica, and FgESP. rFgAbl is localized in the cecum and capsule of juvenile worms, as well as in the testis and viellaria of adult worms. Additionally, rFgAbl enhances cell proliferation, migration, nitric oxide (NO) production, and phagocytosis, while also increasing the transcription levels of cytokines (IFN-γ, IL-12, TNF-α, IL-4, IL-10, and TGF-β). The results indicate that rFgAbl can influence the immune function of PBMCs. Further investigation into the immunomodulatory properties of the rFgAbl protein will enhance our understanding of the immune interaction mechanisms between trematodes and their hosts.

Tox op lasma gondii is a pathogen that poses a serious threat to human health and causes significant economic losses to the global livestock industry. The prevalence of toxoplasmosis infection has been reported to be high in humans and animals around the world, but the occurrence of the disease has not yet been reported in water buffaloes in Guangxi Zhuang Autonomous Region, southern China. To understand the overall seroprevalence of T . gondii infection in Guangxi, a total of 1041 water buffalo and 114 cat serum samples were examined using an indirect enzyme-linked immunosorbent assay (I-ELISA). Of the 1041 water buffaloes analyzed, an overall seroprevalence of 52.9% (551/1041) was obtained, with year, season, and city location being significant factors affecting the rate of T . gondii infection in water buffaloes ( P < 0.001). The results also revealed a high seroprevalence of 57% (65/114) in cats. Given that buffalo milk and meat products are vital food sources, these findings suggest that toxoplasmosis in water buffaloes may be a public health threat. This study provides the first T . gondii seroprevalence data in Guangxi, which could contribute to the prevention and control of toxoplasmosis in the region.

随着社会的发展和人民生活水平的提高,对肉类的需求逐步由传统的猪肉、鸡肉和鱼肉向牛肉等高档肉类消费转变.近年来,肉牛肉羊养殖产业市场行情稳中向好,大力的政策扶持使得大量的社会资本进入到牛羊养殖领域,各地牛羊草食动物养殖蓬勃发展,产业迅速扩大发展也逐渐暴露了制约产业发展的问题,牛羊养殖企业难赚钱、不赚钱、亏本经营的现象愈发普遍.本文调查和跟踪了桂林市牛羊产业发展状况,阐述了广西牛羊产业发展存在的共性问题,总结了养殖盈利运营的一些好的经验做法,为桂林以及广西牛羊产业高质量发展提供参考.

为探讨大片形吸虫外泌体(FgExo)在宿主免疫调节方面的作用,本研究采用聚合物沉淀法和超速离心法从大片形吸虫分泌排泄产物(FgESP)中分离得到FgExo;采用荧光定量PCR(qPCR)法检测不同浓度的FgESP或FgExo作用于水牛外周血单核巨噬细胞(Mψ)后γ-干扰素(IFN-γ)、白介素-10(IL-10)的转录水平;根据 qPCR 结果选择50μg/mLFgExo(+5 μg/mL脂多糖)、200 μg/mL FgESP(+5μg/mL脂多糖)作为试验组,以PBS为阴性对照组、脂多糖为阳性对照组,分别作用于Mψ后采用qPCR检测Mψ分型相关基因诱导型一氧化氮合酶(iNOS)、CD86、精氨酸酶2(Arg-2)、CD206的转录水平;ELISA检测白介素-1β(IL-1β)、转化生长因子-β1(TGF-p1)分泌水平.结果显示:FgExo和FgESP可显著上调MψIL-10mRNA的转录水平,显著抑制IFN-γ mRNA的转录水平;显著上调M2型Mψ标记物CD206及相关因子Arg-2、TGF-β1的表达,显著抑制Ml型Mψ标记物CD86及相关因子iNOS、IL-1β的表达.本研究结果表明,FgExo和FgESP均能够使水牛外周血单核巨噬细胞向M2型极化,利于大片形吸虫在水牛体内生存.

Background Extracellular vesicles (EVs) released by helminths play an important role in parasite-host communication. However, little is known about the characteristics and contents of the EVs of Fasciola gigantica, a parasitic flatworm that causes tropical fascioliasis. A better understanding of EVs released by F. gigantica will help elucidate the mechanism of F. gigantica-host interaction and facilitate the search for new vaccine candidates for the control and treatment of fascioliasis.Methods Two different populations of EVs (15k EVs and 100k EVs) were purified from adult F. gigantica culture media by ultracentrifugation. The morphology and size of the purified EVs were determined by transmission electron microscopy (TEM) and by the Zetasizer Nano ZSP high performance particle characterization system. With the aim of identifying diagnostic markers or potential vaccine candidates, proteins within the isolated 100k EVs were analyzed using mass spectrometry-based proteomics (LC-MS/MS). Mice were then vaccinated with excretory/secretory products (ESPs; depleted of EVs), 15k EVs, 100k EVs and recombinant F. gigantica heat shock protein 70 (rFg-HSP70) combined with alum adjuvant followed by challenge infection with F. gigantica metacercariae. Fluke recovery and antibody levels were used as measures of vaccine protection.Results TEM analysis and nanoparticle tracking analysis indicated the successful isolation of two subpopulations of EVs (15k EVs and 100k EVs) from adult F. gigantica culture supernatants using differential centrifugation. A total of 755 proteins were identified in the 100k EVs. Exosome biogenesis or vesicle trafficking proteins, ESCRT (endosomal sorting complex required for transport) pathway proteins and exosome markers, heat shock proteins and 14-3-3 proteins were identified in the 100k EVs. These results indicate that the isolated 100k EVs were exosome-like vesicles. The functions of the identified proteins may be associated with immune regulation, immune evasion and virulence. Mice immunized with F. gigantica ESPs, 15k EVs, 100k EVs and rFg-HSP70 exhibited a reduction in fluke burden of 67.90%, 60.38%, 37.73% and 56.6%, respectively, compared with the adjuvant control group. The vaccination of mice with F. gigantica 100k EVs, 15k EVs, ESP and rFg-HSP70 induced significant production of specific immunoglobulins in sera, namely IgG, IgG1 and IgG2a.Conclusion The results of this study suggest that proteins within the exosome-like vesicles of F. gigantica have immunomodulatory, immune evasion and virulence functions. This knowledge may lead to new strategies for immunotherapy, vaccination and the diagnosis of fascioliasis.

本研究采用八角益母草醇提液组方对河南某蛋鸡养殖户中暴发的鸡皮刺螨病进行临床杀灭效果观察,以用药期间螨虫聚集点和捕螨器的螨虫变化为评判指标.结果显示,八角益母草醇提液组方临床用药两周后捕螨器中成虫减少率为99.96％、幼虫为99.96％、虫卵为99.93％.用药前聚集堆49个,用药14天后,无明显聚集堆.综上,本研究初步确定八角益母草醇提液组方对鸡皮刺螨具有良好的杀灭作用,可作为潜在的杀灭鸡皮刺螨的中草复方药物.

[目的]制备抗大片形吸虫组织蛋白酶L1(rFgCat L1)特异性单克隆抗体,并构建其双抗体夹心ELISA检测方法.[方法]用1 mg/mL rFgCat L1蛋白分4次对5只BALB/c小鼠进行免疫,分离小鼠脾细胞,与SP2/0细胞融合构建杂交瘤细胞,筛选强阳性杂交瘤细胞株,每只小鼠腹腔注射1×106个细胞制备单克隆抗体.ELISA法检测抗体效价及抗原表位,Western blotting法鉴定抗体亚型和特异性;结合抗rFgCat L1多克隆抗体构建双抗体夹心ELISA检测方法,并检测其敏感性和特异性;用建立的方法对20份阴性血清及阳性血清对照进行检测,筛选其阴阳临界值,并用47份山羊阳性血清及47份奶牛阳性血清对所构建的双抗体夹心ELISA方法进行验证.[结果]免疫后,4只小鼠血清中抗体效价均＞104;取小鼠脾细胞与SP2/0融合后,共筛选到8株阳性杂交瘤细胞株,其中5D5和7G6为可稳定分泌抗体的强阳性株,抗体效价分别达29和210,腹水效价分别达107和108.经Western blotting鉴定2株抗体均为IgG1型,轻链为Kappa型,均可特异性结合大片形吸虫排泄-分泌产物(excretory-secretory product,ESP).由于2株抗体识别相同抗原位点,对比2株单抗的抗体效价,选择以7G6作为捕获抗体,抗rFgCat L1多克隆抗体作检测抗体构建双抗体夹心ELISA法.双抗体夹心ELISA法条件为:7G6以2 μg/mL浓度包被,抗rFgCat L1多克隆抗体检测浓度为25 μg/mL,酶标二抗稀释度为1∶4000,5％脱脂奶粉封闭,显色时间为25 min.经验证,该方法可识别的最低抗原浓度为0.625 μg/mL,并且可特异性识别大片形吸虫抗原.利用构建的双抗体夹心ELISA方法对阳性的奶牛和山羊血清样本进行抗原检测,抗原检出率分别为72.3％及78.7％.[结论]本研究成功制备抗rFgCat L1单克隆抗体并构建大片形吸虫病双抗体夹心ELISA检测方法,结果可为研发低成本、快速诊断试剂盒提供良好的理论依据及物质基础.

Abstract Background: Extracellular vesicles (EVs) released by helminths are important in parasite-host communication. However, little is known about the character and contents of Fasciola gigantica EVs. A better understanding of EVs released by F. gigantica will help reveal the mechanism of F. gigantica-host interaction and aid in the search for new vaccine candidates for fascioliasis control. Methods: We used ultracentrifugation to purify two different populations of EVs (15K EVs and 100K EVs) from adult F. gigantica culture media. The morphology and size of the purified EVs were determined by transmission electron microscopy (TEM) and ZETASIZER NANO ZSP. With the aim of identifying diagnostic markers or potential vaccine candidates, proteins within the isolated 100K EVs were analyzed using mass spectrometry-based proteomics (LC-MS/MS). Then, mice were vaccinated with ESPs, 15K EVs, 100K EVs, and rFg-HSP70 combined with alum adjuvant followed by challenge infection with F. gigantica metacercariae. Fluke recovery and antibody levels were used to evaluate vaccine protection. Results: TEM analysis and Nanoparticle tracking analysis indicated successful isolation of two subpopulations of EVs (15K EVs and 100K EVs) from adult F. gigantica culture supernatants using differential centrifugation. A total of 755 proteins were identified in the 100K EVs. Exosome biogenesis or vesicle trafficking proteins, ESCRT pathways proteins,as well as exosome markers, heat shock proteins and 14-3-3, were identified in the 100K EVs. These results indicate that the isolated 100K EVs were exosome-like vesicles. The functions of the identified proteins may be associated with immune regulation, immune evasion, and virulence. Mice immunized with F. gigantica ESP, 15K EVs, 100K EVs, and rFg-HSP70 exhibited a fluke burden reduction of 67.90%, 60.38%, 37.73%, and 56.6%, respectively, compared with the adjuvant control group. The vaccination of mice with F. gigantica 100K EVs, 15K EVs, ESP, and rFg-HSP70 induced significant production of specific IgG, IgG1, and IgG2a in sera. Conclusion: This study suggests that proteins within the exosome-like vesicles of F. gigantica have immunomodulatory, immune evasion, and virulence functions. This knowledge may lead to new strategies for immunotherapy, vaccination, and the diagnosis of fascioliasis.

近年来,国家出台多项政策推动草食动物产业发展,广西肉牛肉羊产业在自治区党委和政府的支持下也取得了明显成效.本文基于对广西全区14个市辖区牛羊养猪主要县(区)的调研,系统总结了广西肉牛肉羊产业的基本现状,包括广西牛羊养殖的规模、品种、经营模式、产业发展的区域优势、产业扶贫及方式、产业发展遇到的问题等情况,最后提出加快广西牛羊产业发展的对策,为下一步推动广西全区肉牛肉羊产业高质量发展提供参考.

为了优化胆膏中胆汁酸的提取工艺,采用L9(34)正交试验设计,以总胆汁酸含量为考察指标,筛选酸碱纯化工艺提取胆膏中胆汁酸时的1:1盐酸用量、NaOH用量和纯化时间,确定最佳工艺条件,并与传统的乙醇纯化工艺进行比较.结果表明,胆膏中胆汁酸的最佳提取工艺为1:1盐酸用量4 mL、NaOH用量0.2 g、纯化时间12 h.优选的提取工艺具有生产成本低,纯化时间短,所需场地小,设备要求不高,操作简单,对环境无污染,且不使用易燃易爆有机溶剂,安全性好的特点,更符合工业生产的实际需要.

为了建立一种体外诱导培养水牛外周血来源树突状细胞(dendritic cell,DC)的方法,试验分离了水牛外周血来源的单个核细胞(PBMC),在粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素(IL-4)联合作用下进行诱导分化,并以LPS诱导的未成熟树突状细胞发育为成熟树突状细胞作对照组,通过显微镜(光学显微镜、扫描电子显微镜、透射电子显微镜)和流式细胞术分别对获得的细胞进行形态学观察和表型鉴定.结果 表明:PBMC在50 ng/mL GM-CSF和50 ng/mL IL-4体外持续作用6d后,流式细胞术检测表现出CD11chighMHCⅡmed“CD40medCD86low的表型特征,与未成熟树突状细胞的一般表型特征相符.培养至第4天,圆形的PBMC开始呈现出不规则状态;培养至第6天,可在细胞膜上观察到小的微绒毛、突起和细胞聚集等未成熟树突状细胞的形态特征,与表型鉴定结果一致.在LPS的进一步作用下,PBMC呈现出成熟树突状细胞的典型形态.说明GM-CSF和IL-4联合作用可以获得未成熟树突状细胞,并且该细胞可以在LPS的进一步作用下发育成熟,通过该方法可以获得足量的水牛树突状细胞以满足试验需求.

Background Fasciola gigantica infection threatens the health of both humans and animals in the world. The excretory/secretory products (ESPs) of this fluke has been reported to impair the activation and maturation of immune cells. We have previously shown the influence of F. gigantica ESPs (FgESPs) on the maturation of buffalo dendritic cells (DCs). However, the underlying mechanisms remain unclear. The objective of this study was to investigate the potency of FgESPs in shifting the differentiation and immune functions of buffalo DCs. Methods Buffalo DCs were incubated with FgESPs directly or further co-cultured with lymphocytes in vitro. qRT-PCR was employed to determine the gene expression profile of DCs or the mixed cells, and an ELISA was used to measure cytokine levels in the supernatants. Hoechst and Giemsa staining assays, transmission electron microscopy, caspase-3/7 activity test and histone methylation test were performed to determine DC phenotyping, apoptosis and methylation. To investigate the mechanism involved with DNA methylation, a Co-IP assay and immunofluorescent staining assay were performed to observe if there was any direct interaction between FgESPs and DNMT1/TET1 in buffalo DCs, while RNAi technology was employed to knockdown DNMT1 and TET1 in order to evaluate any different influence of FgESPs on DCs when these genes were absent. Results qRT-PCR and ELISA data together demonstrated the upregulation of DC2 and Th2/Treg markers in DCs alone and DCs with a mixed lymphocyte reaction (MLR), suggesting a bias of DC2 that potentially directed Th2 differentiation in vitro. DC apoptosis was also found and evidenced morphologically and biochemically, which might be a source of tolerogenic DCs that led to Treg differentiation. In addition, FgESPs induced methylation level changes of histones H3K4 and H3K9, which correlate with DNA methylation. Co-IP and immunofluorescent subcellular localization assays showed no direct interaction between the FgESPs and DNMT1/TET1 in buffalo DCs. The productions of IL-6 and IL-12 were found separately altered by the knockdown of DNMT1 and TET1 in DCs after FgESPs treatment. Conclusions FgESPs may induce the DC2 phenotype or the apoptosis of buffalo DCs to induce the downstream Th2/Treg response of T cells, possibly through a DNMT1- or TET1-dependent manner(s).

Background: Helminth-induced Th2 responses are essential to modify the structure and diversity of gut microbes. However, observations have come mainly from studies of helminth-infected humans or rodent models. Very little research has been conducted in veterinary animals. Methods: In this study, we searched for links between microbiota and Th2-biased responses during the time course of Fasciala gigantica infection in buffaloes.16S rRNA gene amplicon and metagenome sequencing were applied to analyze the structure and function of the gut microbiota. Results: Both alpha and beta diversities decreased during infection, and gut microbes differed considerably across different sections of the gut at different stages. Immune responses changed when the microbiota traverses the gut wall into the peritoneal cavity, in line with the changes in Th2 response induced by F. gigantica infection. We found that the order Coriobacteriales was greatly decreased at the early stages in which the Peptostreptococcaceae and Family_XIII families are closely linked to the upregulation of IgG1 and IL4, respectively. The F. gigantica infection significantly reduced short-chain fatty acid (SCFAs)-producing microbes, reduced the concentrations of gut SCFAs and downregulated the SCFAs-producing metabolic pathways. In addition, The microbes associated with TLR2 increased and showed similar trend to the TLR2 and Th2 cytokine production during infection, suggesting that bacteria ligands might recognize TLR2 and subsequently induce a Th2-biased response. Conclusions: Our data show that buffalo gut microbes may affect the host Th2 response during F. gigantica infection via the SCFAs metabolism and TLR signaling pathway. These findings provide new insights into the relationship between F. gigantica–microbiota-host, which may provide new potential therapeutic targets for prevention and control Fasciolosis.

动物球虫病是一种流行广泛、危害严重的原虫病,一旦爆发会引起大批动物发病甚至死亡,是养殖业重点防控的对象.治疗球虫病的化学药物广泛存在耐药性、毒副作用和药物残留等缺点.随着国家限抗令的实施和绿色健康养殖的兴起,中药以其天然安全、易吸收、不易产生抗药性和药物残留的特性而逐渐得到养殖业的青睐.有些中药如白龙散、白头翁苦参散、青蒿常山颗粒等已获认可并被广泛应用.就中药防治球虫病的应用进行综述,以期为养殖场、户畜禽健康养殖中球虫病防治提供参考.

为了制备大片形吸虫诊断抗体,试验采用原核表达制备大片形吸虫组织蛋白酶L1重组蛋白(rFgCatL1),用该蛋白免疫Balb/c小鼠制备多克隆抗体,通过间接酶联免疫吸附法(ELISA)检测抗体效价,Western-blot法检测多克隆抗体的特异性.结果 表明:CatL1基因大小为981 bp,蛋白质分子质量为39.0 ku左右,与预期分子质量大小一致.制备的小鼠多克隆抗体血清效价达1∶128 000,能特异性识别大片形吸虫排泄-分泌物(ESP)抗原.说明制备的rFgCatL1多克隆抗体可作为良好的诊断抗体用于研制快速诊断试剂盒.

Background: Toll-like receptors (TLRs) are innate signaling receptors, which sense invading pathogens and play roles in induction of immune responses to counter infection. In this study, we for the first time, cloned, sequenced and characterized the coding sequences of TLRs 1-10 in the Chinese “swamp-type” Guangxi (GX) buffalo breed. Results: Protein sequence analysis of the 10 TLRs showed that GX buffalo share significant sequence similarity with river buffalo (94.7 to 99.9%) and other ruminants (89.6% to 99%), but similarity with non-ruminants was low (48% to 91%). Prediction of functional domains revealed the presence of Toll/IL-1 receptor domains and variable numbers of leucine-rich repeats (LRRs) in all TLRs of GX buffalo. Transmembrane domain and signal peptide were absent in TLRs 7, 9 and TLRs 1, 3, 9, 10, respectively. When comparing GX buffalo and Nili-Ravi (NR) buffalo, the similarity of TIR domains ranged from 95.9% to 100%, while variations were found in the number of LRR motifs between these two breeds. Phylogenetic analysis based on the coding DNA and protein sequences demonstrated that, GX buffalo and NR buffalo were closely related with respect to TLRs 1, 2, 4, 5, 7, 8 and 10. However, regarding TLRs 3, 6, and 9, GX buffalo clustered with genera Bos and Bison, but not NR buffalo. RT-qPCR analysis provided an overview of TLRs transcription in tissues of GX buffalo. Conclusions: Our study obtained and characterized the full sequences of 10 TLR genes of GX buffaloes, and comparatively quantified the mRNA expression of these genes across various tissues of GX buffaloes. These findings lay the groundwork for discovering breed-specific differences in the occurrence and progression of infections in buffalo.

The infection of ruminants by Fasciola spp. always induces a non-protective Th2-type immune response. However, little is known about changes in the local and systemic immune environment during F. gigantica migration in buffalo. In this study, native swamp buffaloes were each infected with 500 viable F. gigantica metacercariae. Mesenteric lymph node (MLN), hepatic lymph node (HLN), spleen, and serum samples were collected from control and infected buffaloes at 3, 10, 28, 42, 70, and 98 days post-infection (DPI). The mRNA expression levels of the Th1- and Th2-related cytokines IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p40, IFN-γ, TNF-α, and CD4 were measured during different infection stages in the MLNs, spleens, and HLNs using quantitative real-time PCR (qRT-PCR). Levels of the specific anti-ESP isotype antibodies IgG, IgG1, and IgG2 were used to reflect changes in humoral immunity. The results of this study indicated that swamp buffaloes were susceptible to F. gigantica infection, and that susceptibility to this infection was closely related to the cytokine environment associated with the Th2-type immune response. The MLNs showed a mixed Th1- and Th2-type immune response during the acute infection stages, after which the production of these cytokines returned to normal. Cytokine expression in the HLNs also expressed a mixed Th1- and Th2-type immune response during the early infection stages. When the infection became chronic, the typical Th2 immune response was induced in the HLNs. At the acute infection stages, the spleen exhibited a Th2 immune response. Nevertheless, cytokines associated with the Th1 and Th2 immune responses were upregulated at 98 DPI. In addition, the total IgG and IgG1 of the parasite-specific antibodies increased. This suggested that the Th2-related cytokines and IgG1 induced by F. gigantica infection might mediate successful F. gigantica infection in the natural host, swamp buffalo.

Liver flukes of animals are parasitic flatworms of major socioeconomic importance in many countries. Particularly, Fasciola gigantica is a leading cause of production losses to the livestock (mainly sheep and cattle) and meat industries due to clinical disease, reduced weight gain and milk production, and deaths. Immune responses induced by helminth have been extensively studied, but there is limited information on this aspect by F. gigantica, especially on macrophages induced with this parasite. Studies have shown that host immune responses induced by parasitic infection is greatly correlated with the macrophage polarization axis. In the present study, we used the murine model of F. gigantica to explore the interaction of host and F. gigantica. We found F. gigantica NEJs promoted pathology and fibrosis of mice liver, and the enlargement of mice spleen. We also showed that macrophages were recruited to mice peritoneal cavity at 5 days post infection. By evaluating the expression of genetic markers of M2 macrophages such as Arg-1, Ym1 and RELMɑ, and genetic marker of M1 macrophages iNOS, we showed that M2 macrophages were induced by F. gigantica. M2 macrophages are central to the immune response during helminth infection, and our findings in this study provided insight into the immune interaction between F. gigantica and host.

Excretory/secretory products (ESPs) released by parasites influence the development and functions of host dendritic cells (DCs). However, little is known about changes of DNA (hydroxy)methylation on DC development during Fasciola gigantica infection. The present study aimed to investigate whether F. gigantica ESPs (FgESPs) affects the development and functions of buffalo DCs through altering the DNA (hydroxy)methylation of DCs. Buffalo DCs were prepared from peripheral blood mononuclear cells (PBMCs) and characterized using scanning and transmission electron microscopy (SEM/TEM) and quantitative reverse transcriptional PCR (qRT-RCR). DCs were treated with 200 μg/ml of FgESPs in vitro, following DNA extraction. The DNA methylome and hydroxymethylome were profiled based on (hydroxy)methylated DNA immunoprecipitation sequencing [(h)MeDIP-Seq] and bioinformatics analyses. qRT-RCR was also performed to assess the gene transcription levels of interest. FgESPs markedly suppressed DC maturation evidenced by morphological changes and downregulated gene expression of CD1a and MHC II. Totals of 5432 and 360 genes with significant changes in the 5-methylcytosine (5-mC) and the 5-hydroxymethylcytosine (5-hmC) levels, respectively, were identified in buffalo DCs in response to FgESPs challenge. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that these differentially expressed genes were highly enriched in pathways associated with immune response. Some cancer-related pathways were also indicated. There were 111 genes demonstrating changes in both 5-mC and 5-hmC levels, 12 of which were interconnected and enriched in 12 pathways. The transcription of hypermethylated genes TLR2, TLR4 and IL-12B were downregulated or in a decreasing trend, while the mRNA level of high-hydroxymethylated TNF gene was upregulated in buffalo DCs post-exposure to FgESPs in vitro. To our knowledge, the present study provides for the first time a unique genome-wide profile of DNA (hydroxy)methylation for DCs that interact with FgESPs, and suggests a possible mechanism of FgESPs in suppressing DC maturation and functions that are involved in TLR signaling.