Anaplasma phagocytophilum is an obligate intracellular, tick-borne bacterial pathogen capable of causing disease and even mortality in various mammals, including humans. Non-coding RNAs play important regulatory roles in multicellular organisms, including innate and adaptive immune pathways, which control bacterial, parasitic, and viral infections. However, the global transcriptomic landscape encompassing both ncRNAs and mRNAs in HL-60 cells invaded by A. phagocytophilum remains unexplored. Cell apoptosis was evaluated by flow cytometry at multiple time points after HL-60 cell infection with A. phagocytophilum. Total RNA was extracted and analyzed by RNA sequencing (RNA-seq) to delineate expression alterations of long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and messenger RNAs (mRNAs) at 24 h post-infection (hpi). Bioinformatics methods were employed for gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses to elucidate the potential functions of these differentially expressed genes. Furthermore, an integrated bioinformatics approach was applied to systematically construct a competing endogenous RNA (ceRNA) network involving lncRNAs, miRNAs, and mRNAs. A. phagocytophilum infection accelerated HL-60 cell apoptosis at multiple time points, with the most significant effect observed at 24 hpi. Transcriptome profiling at 24 hpi identified substantial differential expression, including 487 lncRNAs, 550 mRNAs, and 22 miRNAs with statistically significant changes in expression. Then, expression patterns of eight lncRNAs, eight mRNAs, and seven miRNAs were experimentally validated through reverse transcription quantitative polymerase chain reaction (RT-qPCR), demonstrating strong correlation with RNA-seq results. Bioinformatics analyses revealed significant enrichment of differentially expressed mRNAs in three key pathways: the PI3K/Akt signaling pathway, the actin cytoskeleton regulation pathway and the p53 signaling pathway. Differentially expressed lncRNAs were largely related to the phospholipase D signaling pathway and pathways related to cortisol and aldosterone synthesis/secretion. The altered miRNAs showed predominant enrichment in Rap1 and NF-κB signaling pathways. Notably, computational reconstruction of the lncRNA–miRNA–mRNA ceRNA network identified hsa–miR–4518 and hsa–miR–3609 as central regulatory nodes. This comprehensive transcriptome study elucidates complex gene regulatory networks activated in HL-60 cells after A. phagocytophilum invasion, with particular emphasis on pathogen-modulated miRNA signatures that coordinate critical pathways governing host immune responses and microbial survival strategies. These findings elucidate previously uncharacterized molecular mechanisms underlying A. phagocytophilum pathogenesis and may provide actionable targets for novel therapeutics.

Anaplasma phagocytophilum is a zoonotic pathogen transmitted by arthropod vectors. The pathogen infects various vertebrate hosts, causing mild to severe illness. Molecular studies have demonstrated that A. phagocytophilum exhibits a high level of genetic diversity, with two A. phagocytophilum-related variants identified in several countries. This study represents the first application of PCR amplification and restriction fragment length polymorphism (PCR-RFLP) in conjunction with DNA sequencing to investigate the frequency and phylogenetic relationships of A. phagocytophilum and its related strains in cattle from China. A total of 662 bovine blood samples were collected from diverse regions within Henan Province, China, and pathogen DNA was detected in 75 samples, comprising 11.33% of the total. PCR-RFLP analysis identified three strains with frequency rates of 2.87% (19/662) for A. phagocytophilum, 11.33% (75/662) for A. phagocytophilum-like 1, and 3.22% (22/662) for A. phagocytophilum-like 2. Additionally, co-infections involving A. phagocytophilum and A. phagocytophilum-like 1 were observed as well as between A. phagocytophilum-like 1 and A. phagocytophilum-like 2. Anaplasma phagocytophilum-like strains 1 and 2 should be considered when diagnosing bovine anaplasmosis. Despite recent molecular studies of A. phagocytophilum-related strains, there remains a shortage of data concerning vector capability, the epidemiology of the disease, clinical signs, and genetic diversity of the pathogens. Thus, large-scale investigations involving animals and tick vectors are necessary to obtain more detailed information concerning the etiology of anaplasmosis.

Fermented traditional Chinese medicines (TCMs) have been identified as a low-cost and promising feed additive to to alleviate weaning stress in young livestock and poultry effectively. This study investigated the impact of probiotic fermentation on the metabolite content of BanQi (Radix Isatidis and Astragalus membranaceus) extract while also examined the effects of both fermented-BanQi (FBQ) and unfermented-BanQi (UBQ) on growth performance, serum biochemistry, intestinal villi, and gut microbiota in weaned lambs. This study demonstrated that compared with UBQ, FBQ contained significantly higher levels of free amino acids (e.g., phenylalanine and isoleucine), short peptides (e.g., Val-Leu-Pro-Val-Pro-Gln and Gly-Leu), and the active ingredients (e.g., vindesine and reserpine) (P < 0.05). The addition of FBQ to the diet significantly increased the final body weight and average daily gain of weaned lambs (P < 0.05). In addition, FBQ significantly increased the total protein level in the serum and the villus length of the jejunum and ileum in lambs, while significantly reduced the levels of aspartate aminotransferase (AST) and urea (P < 0.05). Sequencing of the intestinal flora showed that FBQ improved the diversity of intestinal flora and promoted the enrichment of beneficial bacteria in the lamb intestine, such as Mogibacterium and Butyrivibrio, compared to NC or UBQ groups (P < 0.05). Fermentation with Bacillus subtilis can enhance the content of free amino acids, peptides, and active ingredients in BanQi extract, making it an effective method to improve the efficacy of traditional Chinese medicine. Adding FBQ to the diet can improve the growth performance of weaned lambs, and its mechanism may be related to increasing the height of intestinal villi and increasing the diversity of intestinal flora.

Cryptosporidium spp. are significant zoonotic intestinal parasites that induce diarrhea and even death across most vertebrates, including humans. Previous studies showed that sheep are important hosts for Cryptosporidium and that its distribution in sheep is influenced by geography, feeding patterns, age, and season. Environmental factors also influence the transmission of Cryptosporidium. Molecular studies of Cryptosporidium in sheep have been conducted in only a few regions of China, and studies into the effect of sheep-housing environments on Cryptosporidium transmission are even rarer. To detect the prevalence of Cryptosporidium in large-scale sheep-housing farms, a total of 1241 fecal samples were collected from sheep, 727 environmental samples were taken from sheep housing, and 30 water samples were collected in six regions of China. To ascertain the existence of the parasite and identify the species of Cryptosporidium spp., we conducted nested PCR amplification of DNA extracted from all samples using the small-subunit (SSU) rRNA gene as a target. For a more in-depth analysis of Cryptosporidium spp. subtypes, C. xiaoi-and C. ubiquitum-positive samples underwent separate nested PCR amplification targeting the 60 kDa glycoprotein (gp60) gene. The amplification of the Cryptosporidium spp. SSU rRNA gene locus from the whole genomic DNA of all samples yielded a positive rate of 1.2% (20/1241) in fecal samples, 0.1% (1/727) in environmental samples, and no positive samples were found in water samples. The prevalence of Cryptosporidium spp. infection in large-scale housed sheep was 1.7%, which was higher than that in free-ranging sheep (0.0%). The highest prevalence of infection was found in weaning lambs (6.8%). Among the different seasons, the peaks were found in the fall and winter. The most prevalent species were C. xiaoi and C. ubiquitum, with the former accounting for the majority of infections. The distribution of C. xiaoi subtypes was diverse, with XXIIIc (n = 1), XXIIId (n = 2), XXIIIe (n = 2), and XXIIIl (n = 4) identified. In contrast, only one subtype, XIIa (n = 9), was found in C. ubiquitum. In this study, C. xiaoi and C. ubiquitum were found to be the predominant species, and Cryptosporidium was found to be present in the environment. These findings provide an important foundation for the comprehensive prevention and management of Cryptosporidium in intensively reared sheep. Furthermore, by elucidating the prevalence of Cryptosporidium in sheep and its potential role in environmental transmission, this study deepens our understanding of the intricate interactions between animal health, environmental contamination, and public health dynamics.

Abstract Background The parasitic protozoan Giardia duodenalis is an important cause of diarrheal disease in humans and animals that can be spread by fecal–oral transmission through water and the environment, posing a challenge to public health and animal husbandry. Little is known about its impact on large-scale sheep farms in China. In this study we investigated G. duodenalis infection of sheep and contamination of the environment in large-scale sheep farms in two regions of China, Henan and Ningxia. Methods A total of 528 fecal samples, 402 environmental samples and 30 water samples were collected from seven large-scale sheep farms, and 88 fecal samples and 13 environmental samples were collected from 12 backyard farms. The presence of G. duodenalis was detected by targeting the β-giardin (bg) gene, and the assemblage and multilocus genotype of G. duodenalis were investigated by analyzing three genes: bg, glutamate dehydrogenase (gdh) and triphosphate isomerase (tpi). Results The overall G. duodenalis detection rate was 7.8%, 1.4% and 23.3% in fecal, environmental and water samples, respectively. On the large-scale sheep farms tested, the infection rate of sheep in Henan (13.8%) was found to be significantly higher than that of sheep in Ningxia (4.2%) (P < 0.05). However, the difference between the rates of environmental pollution in Henan (1.9%) and Ningxia (1.0%) was not significant (P > 0.05). Investigations of sheep at different physiological stages revealed that late pregnancy ewes showed the lowest infection rate (1.7%) and that young lambs exhibited the highest (18.8%). Genetic analysis identified G. duodenalis belonging to two assemblages, A and E, with assemblage E being dominant. A total of 27 multilocus genotypes were identified for members of assemblage E. Conclusions The results suggest that G. duodenalis is prevalent on large-scale sheep farms in Henan and Ningxia, China, and that there is a risk of environmental contamination. This study is the first comprehensive examination of the presence of G. duodenalis on large-scale sheep farms in China. Challenges posed by G. duodenalis to sheep farms need to be addressed proactively to ensure public health safety. Graphical Abstract

Giardia duodenalis is a common zoonotic intestinal parasitic protozoan and sheep are among its hosts; however, limited information is available on sheep kept in large-scale housing. The Hu sheep is a first-class protected local livestock breed in China. In this study, we investigated the seasonal dynamics of G. duodenalis infection in Hu sheep and the environmental contamination of large-scale sheep farms. We collected 474 fecal samples and 312 environmental samples from Hu sheep on a large-scale sheep farm in Henan, China. The prevalence of G. duodenalis was determined by nested PCR targeting the β‑giardin (bg) gene. The assemblages and multilocus genotypes (MLGs) were investigated based on analyses of three genetic loci, i.e. bg, glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi). To detect mixed infections of different assemblages, assemblage A/E-specific PCRs were performed to amplify the tpi gene. The prevalence of G. duodenalis infection in sheep was 17.9% (81/474) and the positivity rate in environmental samples was 0.96% (3/312). Genetic analysis revealed the presence of two assemblages (assemblages A and E), with assemblage E being detected in both fecal and environmental samples, and assemblage A detected only in fecal samples. A total of 23 MLGs were obtained in fecal and environmental samples, all of which belonged to assemblage E. These results indicate the seasonal dynamics of G. duodenalis infection in sheep and environmental contamination on large-scale housing sheep farms and provide an important reference for the prevention and control of G. duodenalis on large-scale housing sheep farms.

[Objective] The purpose of this study was to investigate the digestive tract infection and environmental pollution parasites of sheep in large-scale sheep farms and free-range sheep farms,and provide reference for the comprehensive prevention and control of digestive tract parasites of sheep in large-scale sheep farms in China.[Method] 1 025 sheep feces samples and712 environmental samples collected from 4 scale sheep farms and 12 free-range households in Henan and Ningxia were examined for parasite infection by centrifugation,Lugol’s iodine staining and saturated sucrose solution floatation methods,and the number of Coccidia oocysts per gram of feces（OPG） in coccidia-positive samples was counted by McMaster’s method.The seasonal dynamic survey was conducted in large-scale sheep farms of Ruzhou to understand the digestive tract infection and environmental pollution parasites of sheep farm in different seasons.[Result]The total infection rate of digestive tract parasites in sheep was 82.83 %（849/1 025）,and 8 species（classes） of parasites were detected,including Coccidia,Amoeba,Giardia,Cryptosporidium,Moniezia,Whipworm,Nematodirus and Other Nematodes.The dominant worm species was Coccidia（71.90%）,followed by Amoeba（49.07%）,and Coccidia and Amoeba were more likely to have mixed infections.The total parasite positivity rate of environmental samples was 38.48%（274/712）,and 6 species（classes） of parasites were detected,including Coccidia,Amoeba,Giardia,Whipworm,Nematodirus and other Nematodes,with the dominant species being Nematodes（22.75%）.Infection rates of digestive tract parasites and parasite detection rates of environmental samples in Henan scale sheep farms were significantly lower than Ningxia scale sheep farms（P＜0.05）.Compared with free-ranging sheep,sheep were infected with fewer worm species and lower infection rate.The infection rate of digestive tract parasites in lactating lambs was significantly lower than that of other physiological stages（P＜0.05）.In Ruzhou large-scale sheep farms,the infection rate of digestive tract parasites of sheep in winter was significantly lower than that in other seasons（P＜0.05）,The infection rate of Coccidia in suckling lambs was the lowest（14.95%）,but the infection intensity was the highest（the average OPG was 105 203）,the infection rate of fattening lambs was the highest（88.99%）.[Conclusion]The infection rate of digestive tract parasitic in sheep on large-scale sheep farms was higher,the mixed infections of Coccidia and Amoeba were common,and environmental parasitic contamination was serious.Hygiene measures should be strengthened to prevent environmental parasitic contamination and pathogen dispersal,and planned deworming should be done.

吉尔吉斯斯坦位于中亚东北部,天山山脉西麓,属于内陆国家.在地理位置上北边与哈萨克斯坦相接,西边为乌兹别克斯坦,西南为塔吉克斯坦,东边紧邻中国.吉尔吉斯斯坦划分为7州(省)2市:楚河州、塔拉斯州、奥什州、贾拉拉巴德州、纳伦州、伊塞克湖州、巴特肯州和比什凯克市、奥什市.吉尔吉斯斯坦是一个以农业和畜牧业为主的国家,拥有九万多平方公里的牧场和天然割草场,全年光照充足,气候适宜,这使吉尔吉斯斯坦的畜牧业拥有得天独厚的资源优势.吉尔吉斯斯坦主要以养殖牛、羊、马为主,活动物出口、牛羊肉、皮、奶等具有极强的出口优势,近几年,每年出口量都有小幅上涨.吉尔吉斯斯坦的农产品加工业是经济重点领域之一,每年的农产品及其加工品的贸易额都超过10亿美元.本文通过对吉尔吉斯斯坦养殖业和疫病防控科技现状进行分析,为中、吉两国农业合作提供理论依据.

无浆体是一种重要的蜱媒病原体,随着国内外学者对其关注度的提高,其对动物和人类健康的危害性也逐渐凸显,高效、准确的诊断技术显得尤其重要.实际工作中,多种分子生物学及血清学方法被用于无浆体的检测,检出率和准确性显著提高.论文综述了近5年无浆体的检测技术,对无浆体的感染诊断提供参考.

为了解我国部分地区近10年羊肠道寄生虫感染情况,探讨羊肠道寄生虫流行特征,采用显微镜检查法对2012年1月至2021年12月来自全国17个省份46个地区的6 745份羊粪便样品进行肠道寄生虫感染情况调查.结果显示,在这些样品中共检测出原虫(艾美耳球虫、隐孢子虫、贾第虫及阿米巴等)、线虫(毛尾线虫也称为鞭虫、毛细线虫、羊仰口线虫、细颈线虫和毛圆线虫等)、绦虫、吸虫(前后盘吸虫、歧腔吸虫及绵羊斯克里亚宾吸虫等)共4纲寄生虫.肠道寄生虫总感染率为82.15％,其中球虫感染率最高(73.73％),其次是阿米巴(37.49％);2012年感染率最高(97.58％),2018年最低(50.77％);放牧羊感染率(87.85％)高于舍饲羊(80.59％).说明我国羊肠道寄生虫感染情况普遍存在,10年来寄生虫感染率无显著下降,应加强肉羊寄生虫病的防控.

The genus Anaplasma comprises eight bacterial species that are obligate intracellular pathogens that affect human and animal health. The zoonotic species A. phagocytophilum is the causative agent of tick-borne fever in ruminants, and of granulocytic anaplasmosis in horses, dogs, and humans. Recently, novel strains related to A. phagocytophilum (A. phagocytophilum-like 1/Japanese variant and A. phagocytophilum-like 2/Chinese variant) have been identified. The aim of this study was to reveal the prevalence and phylogeny of A. phagocytophilum and related stains in small ruminants and ticks in China based on sequences of the 16S rRNA combined restriction fragment length polymorphism (RFLP) and groEL genes. PCR-RFLP and phylogenetic analyses based on the 16S rRNA gene showed the presence of A. phagocytophilum-like 1 and 2 variants in sampled animals from China, with prevalence rates of 22.6% (303/1338) and 0.7% (10/1338), respectively. Only A. phagocytophilum-like 1 DNA was found in Haemaphysalis longicornis. The phylogeny based on the groEL gene showed inclusion of A. phagocytophilum-like 1 and some A. phagocytophilum-like 2 strains in two unique clades distinct from, but related to, Japanese and Chinese strains of related A. phagocytophilum, respectively. One noteworthy result was that the SSAP2f/SSAP2r primers detected Ehrlichia spp. strains. Moreover, the A. phagocytophilum-like 1 and 2 strains should be considered in the differential diagnosis of caprine and ovine anaplasmosis. Further investigations should be conducted to provide additional epidemiological information about A. phagocytophilum and A. phagocytophilum-like variants in animals and ticks.

目的 研究大蒜素对长角血蜱幼蜱的驱避效果及其长效驱避剂的作用时间.方法 以长角血蜱幼蜱为测试对象,利用滤纸片体外驱避装置比较研究0.5、1.0、1.5、2.0、3.0、4.0 mg/ml 6种浓度大蒜素溶液对长角血蜱幼蜱的驱避效果,并计算半数驱避浓度(RC50)、全数驱避浓度(RC100);同时使用Kruskal-Wallis H检验研究不同浓度不同剂型大蒜素与3种市售驱避剂(1％驱蚊酯长效驱避剂、OFF驱蚊液、六神驱蚊花露水)每48 h的驱避效果,确定其作用时间.结果 大蒜素浓度在0.5～4.0mg/ml内,随着浓度的增加,其对长角血蜱幼蜱的驱避率也逐渐上升,在4.0mg/ml时达到最大,为100％,RC50及RC100分别为1.79和3.52 mg/ml;0.5％、1.0％、3.0％大蒜素长效驱避剂的驱避活性在10 d内显著大于各自浓度水乳液处理组(均P＜0.05),与市售驱避剂相比,1.0％、3.0％大蒜素长效驱避剂在12 d内对长角血蜱幼蜱的驱避活性更好.结论 0.5％、1.0％、3.0％大蒜素长效驱避剂的最长作用时间分别为12、14和16d,有效保护时间分别为2、4和6 d,驱避效果优于市售驱避剂.

Ornithonyssus sylviarum (Acari: Macronyssidae) is a common ectoparasite that feeds on the blood of poultry. Following infestation, this mite will cause symptoms such as weight loss, anemia, and decreased egg production. To explore green and safe drugs for the prevention and treatment of O. sylviarum, this study evaluated the effects of ethanol extracts of seven Chinese medicinal herbs-Leonurus artemisia (motherwort), Illicium verum (star anise), Cinnamomum cassia (cinnamon), Hibiscus syriacus, Artemisia argyi (Chinese mugwort), Taraxacum sp. (dandelion), and Syzygium aromaticum (clove)-on O. sylviarum at different life stages. The results showed that different methods of administration affected the acaricidal efficacy of these plant extracts on O. sylviarum. After 6 h of administration with the fumigation method, the acaricidal efficacy of S. aromaticum on adults, nymphs and larvae of O. sylviarum reached 100%. 30 min after administration with the infiltration method, S. aromaticum, H. syriacus and L. artemisia showed acaricidal effects on adults and nymphs of O. sylviarum reaching 100%. In another experiment evaluating the inhibition of egg hatching of O. sylviarum with alcohol extracts of these seven herbs, at 48 h after treatment, A. argyi and C. cassia showed inhibition rates of 19.4%. The results of this study indicate that S. aromaticum induced mortality at all stages of O. sylviarum, whereas A. argyi was found to be the most effective at inhibiting the mite's egg hatching among the seven herbs. These herbs can therefore be used as potential substitutes for chemical pesticides to prevent and control O. sylviarum. These results provide practical knowledge for the control of O. sylviarum.

目的 研制一种节肢动物体外驱避试验装置,用于硬蜱驱避剂的研究.方法 分别使用木条法及滤纸片法制备成蜱及幼蜱的体外驱避试验装置,并与文献报道的硬蜱驱蜱装置进行驱避效果比较,使用SPSS 18.0软件Kruskal-Wallis H检验将不同驱避装置对硬蜱的驱避率及无效区硬蜱占比进行差异性分析.结果 在木条攀爬法驱避装置试验中,自制驱避试验装置(自制装置)(试验组)驱避率为(97.44±2.56)％,文献中装置驱避率为(87.67±6.23)％,二者之间差异无统计学意义(F=2.105,P=0.220);自制装置处于无效区硬蜱占比试验组为(4.45±2.22)％,对照组为(2.22±2.22)％,小于文献中装置[试验组(53.33±3.85)％,对照组(60.00±3.85)％],且差异有统计学意义(试验组F=121.004,P＜0.001;对照组F=168.825,P=0.001);在滤纸片法驱避装置试验中,自制装置(试验组)驱避率为(100.00±0.00)％,文献中装置为(95.83±4.17)％,二者之间差异无统计学意义(F=1.000,P=0.374);自制装置处于无效区的硬蜱试验组和对照组占比均为(0.00±0.00)％,小于文献中装置[试验组(41.67±7.26)％,对照组(35.00±5.77)％)],且差异有统计学意义(试验组F=32.895,P=0.005;对照组F=36.750,P=0.004).结论 研制的硬蜱驱避试验装置结构简单、使用方便、实验结果可信度高,为节肢动物驱避剂的研发和效果测定、药物筛选提供了一种技术手段.

Cryptosporidium spp. can cause diarrhea and even death in humans and animals. Host microRNAs (miRNAs) play an important role in the post-transcriptional regulation of the innate immune response to Cryptosporidium infection. To study host miRNA activity in the innate immune response to C. parvum infection, we examined the expression of miR-181d in HCT-8 cells infected with C. parvum and found that it was significantly downregulated, while TLR2, TLR4, NF-κB, and myD88 involved in the TLR/NF-κB signaling pathway were significantly upregulated at the early stages of C. parvum infection. We transfected cells with short-interfering RNAs (siRNA) as TLR2, TLR4, and NF-κB inhibitors. Analysis by quantitative real-time polymerase chain reaction (qPCR) and western blot confirmed that C. parvum downregulates miR-181d expression via the p50 subunit-dependent TLR2/TLR4-NF-κB signaling pathway in HCT-8 cells. This study provides a new theoretical foundation to elucidate the regulatory mechanism of host miRNAs against Cryptosporidium infection.

为了研究不同配方的驱蚊酯长效硬蜱驱避剂的缓释性能及其与普通驱蚊酯水溶液的差异,对影响羊毛中驱蚊酯检出的萃取温度、萃取时间、萃取液体积以及羊毛在萃取液中的浸泡时间通过单因素试验进行了筛选,建立了羊毛中驱蚊酯含量的气相色谱检测方法;并对浸泡过相应驱避剂的羊毛分别于0、1、2、3、5、8、12、18 d检测其中的驱蚊酯载药量,得到了不同配方的驱蚊酯在羊毛中的释放曲线,进而对4种驱蚊酯制剂的缓释性能进行了评价.结果表明,浸泡过驱蚊酯的羊毛在3 mL无水乙醇中浸泡15 min,并在35℃条件下萃取20 min后,所测得的萃取液中驱蚊酯峰面积最大,利于其定量检测;在配方缓释性能试验中,A、C、D组驱避剂在1～8 d内羊毛中驱蚊酯载药量均大于同时期的对照组,存在显著差异(P<0.05),且D组在1～8 d内每克羊毛中驱蚊酯载药量显著大于其他处理组,其相对释放率分别为35.5％、52.0％、64.3％、73.3％、81.1％;B组则在整个试验过程中每克羊毛中驱蚊酯载药量与对照组均没有显著差异,各时间点相对释放率也较为接近;另外,不同处理组驱蚊酯相对释放曲线中显示一级方程拟合度最好,D组在0～18 d内的积分面积为1349.35 mAU,小于其他处理组.综上,在4组驱蚊酯制剂中,A、C、D组均可有效延缓驱蚊酯的释放,其中D组制剂的缓释效果最好,而B组的缓释效果不明显.

Additional file 7: Table S7. The GO and KEGG enrichment analysis of sponging miRNA targets for the dif-circRNA at 12 hpi.

本试验旨在研究菌酶复合制剂对育肥湖羊生长性能、瘤胃发酵和细菌菌群的影响.试验选取156只60日龄体重相近、健康状况良好的雄性育肥湖羊,随机分为4个组,每组3个重复,每个重复13只羊.对照组(A组)饲喂基础饲粮,试验组饲喂在基础饲粮的基础上分别添加0.1％(B组)、0.2％(C组)和0.3％(D组)菌酶复合制剂的饲粮.试验预试期7 d,正试期60 do结果表明:1)与A组相比,D组第31～60天及第1～60天湖羊平均日增重(ADG)显著提高(P＜0.05),料重比(F/G)显著降低(P＜0.05).2)与A组相比,B组、C组和D组酸性洗涤纤维(ADF)表观消化率显著提高(P＜0.05),B组和C组中性洗涤纤维(NDF)表观消化率显著提高(P＜0.05).3)D组瘤胃pH显著低于A组、B组和C组(P＜0.05);与A组相比,B组、C组和D组瘤胃氨态氮(NH3-N)含量显著提高(P＜0.05),瘤胃总挥发性脂肪酸、丙酸和异丁酸含量显著降低(P＜0.05);B组和D组瘤胃丁酸含量显著降低(P＜0.05).4)16S rRNA测序结果表明,与A组相比,在门水平上,D组拟杆菌门(Bacteroidetes)和TM7相对丰度显著降低(P＜0.05),放线菌门(Actinobacteria)和蓝藻菌门(Cyanobacteria)相对丰度显著提高(P＜0.05);在属水平上,D组欧陆森氏菌属(Olsenella)相对丰度显著提高(P＜0.05),YRC22相对丰度显著降低(P＜0.05)o 5)密螺旋体属(Treponema)相对丰度与NDF表观消化率和瘤胃NH3-N含量呈极显著正相关(P＜0.01);欧陆森氏菌属相对丰度与ADF表观消化率呈显著正相关(P＜0.05),与瘤胃pH和丙酸含量呈显著负相关(P＜0.05).综上所述,在饲粮中添加0.3％的菌酶复合制剂可以提高育肥湖羊ADG,降低F/G,提高饲粮ADF表观消化率,降低瘤胃拟杆菌门相对丰度,同时提高瘤胃欧陆森氏菌属相对丰度,有助于提高对饲粮中纤维的消化率.

Additional file 3: Table S3. The cis- and trans- target genes predicted for dif-lncRNAs at 3 hpi and 12 hpi.

Cryptosporidium species are zoonotic protozoans that are important causes of diarrhoeal disease in both humans and animals. Non-coding RNAs (ncRNAs) play an important role in the innate immune defense against Cryptosporidium infection, but the underlying molecular mechanisms in the interaction between human ileocecal adenocarcinoma (HCT-8) cells and Cryptosporidium species have not been entirely revealed. The expression profiles of messenger RNAs (mRNAs), long non-coding RNAs (lncRNAs), microRNAs (miRNAs) and circular RNAs (circRNAs) in the early phase of infection of HCT-8 cells with Cryptosporidium parvum and at 3 and 12 h post infection were analyzed using the RNA-sequencing technique. The biological functions of differentially expressed RNAs (dif-RNAs) were discovered through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The targeting relationships between three ncRNAs and mRNAs were analyzed using bioinformatics methods, followed by building a competing endogenous RNA (ceRNA) regulatory network centered on miRNAs. After strictly filtering the raw data, our analysis revealed 393 dif-lncRNAs, 69 dif-miRNAs and 115 dif-mRNAs at 3 hpi, and 450 dif-lncRNAs, 129 dif-miRNAs, 117 dif-mRNAs and one dif-circRNA at 12 hpi. Of these, 94 dif-lncRNAs, 24 dif-miRNAs and 22 dif-mRNAs were detected at both post-infection time points. Eleven dif-lncRNAs, seven dif-miRNAs, eight dif-mRNAs and one circRNA were randomly selected and confirmed using the quantitative real-time PCR. Bioinformatics analyses showed that the dif-mRNAs were significantly enriched in nutritional absorption, metabolic processes and metabolism-related pathways, while the dif-lncRNAs were mainly involved in the pathways related to the infection and pathogenicity of C. parvum (e.g. tight junction protein) and immune-related pathways (e.g. cell adhesion molecules). In contrast, dif-miRNAs and dif-circRNA were significantly enriched in apoptosis and apoptosis-related pathways. Among the downregulated RNAs, the miRNAs has-miR-324-3p and hsa-miR-3127-5p appear to be crucial miRNAs which could negatively regulate circRNA, lncRNA and mRNA. The whole transcriptome profiles of HCT-8 cells infected with C. parvum were obtained in this study. The results of the GO and KEGG pathway analyses suggest significant roles for these dif-RNAs during the course of C. parvum infection. A ceRNA regulation network containing miRNA at its center was constructed for the first time, with hsa-miR-324-3p and hsa-miR-3127-5p being the crucial miRNAs. These findings provide novel insights into the responses of human intestinal epithelial cells to C. parvum infection.