Fowl adenovirus serotype 11 (FAdV-11) is an emerging pathogen associated with inclusion body hepatitis (IBH) and high mortality in commercial broilers. In this study, we evaluated the protective efficacy of maternal antibody transfer against FAdV-11 in neonatal chicks. A hypervirulent FAdV-11 strain (FAdV-11/HB2022/1), isolated from an IBH outbreak in Hubei Province, China, caused 90% mortality in 1-day-old specific-pathogen-free (SPF) chickens; 7-day-old chicks exhibited no mortality but developed mild hepatic lesions and sustained viral replication. A soluble, trimeric Fiber subunit vaccine (Fiber282–569) was developed through rational truncation of the transmembrane domain, retaining both the shaft and head domains, with its homotrimeric conformation confirmed by size-exclusion chromatography coupled with multi-angle light scattering (SEC-MALS). Immunization of breeder hens with Fiber282–569 elicited high titers of FAdV-11-specific IgY and neutralizing antibodies that were efficiently transferred to progeny via the egg yolk. Upon lethal challenge with FAdV-11/HB2022/1, 1-day-old progeny chicks from vaccinated breeders exhibited 100% survival with no clinical signs, no gross pathological lesions, and undetectable viral loads in liver tissues, demonstrating complete protection. In contrast, all chicks from unvaccinated breeders succumbed within 5 days post-infection. These results demonstrate that maternal antibody transfer of the fiber-based subunit vaccine confers complete protection against lethal FAdV-11 infection in neonatal chicks, providing a safe and effective candidate strategy for controlling FAdV-11 in the poultry industry.
This study investigated the impact of altitude on gut microbiota composition and diversity in Eurasian tree sparrows (Passer montanus), comparing 39 sparrows from a high-altitude site (3400 m) and 43 from a low-altitude site (30 m). Adult male sparrows of similar body mass were selected, and high-throughput sequencing of the 16S rRNA gene was used to analyse gut microbiota from 20 faecal samples. The results showed that high-altitude sparrows had larger body mass and wing length. The gut microbiota diversity of Eurasian tree sparrows at low altitude is significantly higher than that of high-altitude populations. Low-altitude sparrows were dominated by Firmicutes (46.88 ± 21.85%), while high-altitude sparrows had a predominance of Proteobacteria (83.11 ± 16.37%). Significant differences in microbial community structure were confirmed by both alpha and beta diversity analyses. Functional predictions suggested enhanced nutrient synthesis capabilities in high-altitude sparrows, with a higher relative abundance of biosynthesis-related pathways. Additionally, low-altitude sparrows had a higher prevalence of potentially pathogenic bacteria such as Staphylococcus. Linear discriminant analysis effect size analysis identified significant microbial differences across multiple taxonomic levels, with three genera (Rhodococcus, Turicibacter, and Diaphorobacter) demonstrating significantly elevated relative abundance in the high-altitude group. These findings indicate that altitude significantly shapes the gut microbiota and functional potential in tree sparrows, influencing their ecological and physiological adaptations. This understanding aids in avian ecology and conservation strategies across varied altitudinal ranges.
The chestnut-winged cuckoo (Clamator coromandus) is a bird species known for its brood parasitism, laying eggs in the nests of other bird species. However, there is a paucity of genetic information available for this species and their genus Clamator. In this study, we present the first complete mitochondrial genome sequence of C. coromandus and compare it with other species within the Cuculidae family. The mitogenome is a closed circular molecule consisting of 17,082 bp with an organization typical of the mitochondrial genomes of Cuculidae. Alignment of the control regions across Cuculidae species revealed substantial genetic variation and a significant abundance of AT content. A significant difference was detected in AT-skews between brood-parasitic and parental-care species. A distinctive long poly-C sequence was located at the 5′ end of domain I. Phylogenetically, C. coromandus is more closely related to Piaya cayana than Ceuthmochares aereus. The phylogenetic analysis indicated a general divergence between species with brood parasitism and those with parental care, with transitions between these behaviors within brood parasitism branches, suggesting multiple evolutionary occurrences of these traits. The complete mitogenome of C. coromandus serves as a valuable resource for further investigation into the taxonomic status and phylogenetic history of Clamator species.
MicroRNAs are short, noncoding RNA molecules that exert pivotal roles in cancer development and progression by modulating various target genes. There is growing evidence that miR-138-5p is significantly involved in cervical cancer (CC). However, its precise molecular mechanism has yet to be fully understood. In the current investigation, a quantitative proteomics approach was utilized to detect possible miR-138-5p targets in HeLa cells systematically. In total, 364 proteins were downregulated, and 150 were upregulated after miR-138-5p overexpression. Bioinformatic analysis of these differentially expressed proteins (DEPs) revealed significant enrichment in several cancer-related pathways. Zinc finger protein 385A (ZNF385A) was determined as a novel direct target of miR-138-5p and discovered to facilitate the proliferation, migration, and cell cycle progression of HeLa cells. SFN and Fas cell surface death receptor(FAS) were then identified as functional downstream effectors of ZNF385A and miR-138-5p. Moreover, a tumor xenograft experiment was conducted to validate the association of miR-138-5p-ZNF385A-SFN/FAS axis with the development of CC in vivo. Our findings have collectively established a catalog of proteins mediated by miR-138-5p and have provided an in-depth comprehension of the molecular mechanisms responsible for the inhibitory effect of miR-138-5p on CC. The miR-138-5p-ZNF385A-SFN/FAS axis could also be beneficial to the identification of new therapeutic targets.
Recombinant human acidic fibroblast growth factor (rh-aFGF) is a widely used biological product, but it is unstable and its biological activity is easy to decrease. In order to maintain the long-term stability and biological activity of rh-aFGF, based on the response surface method, the freeze-drying characterization and cell proliferation rate of rh-aFGF freeze-dried powder were evaluated by scoring and Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay in this study. The optimal concentrations of trehalose, glycine and BSA were optimized, and the optimal formulation was verified by regression experiment. The results showed that trehalose, glycine and BSA had significant effects on the characterization of lyophilized rh-aFGF and cell proliferation. The optimal formulation of 5.7% trehalose, 2.04% glycine and 1.98%BSA combined with rh-aFGF could achieve the optimal freeze-dried characterization and biological activity. Using the best formulation to verify, the freeze-dried formability index of the freeze-dried powder was 23.35, and the rate of cell proliferation was 43.59%, which was close to the expected 23 and 41.69%. This study determined a freeze-dried formulation of rh-aFGF that meets the requirements of freeze-dried formalization integrity and maintains biological activity, providing reliable support for the subsequent development of related drugs.
MicroRNAs (miRNAs) represent a class of short, non-coding RNAs that are widely acknowledged as crucial participants in virus-host interactions. MiR-184, a highly conserved and abundant miRNA in insects, has yet to be extensively studied for its involvement in baculovirus infection. In this study, we investigated how miR-184 affects the infection and replication of Autographa californica multiple nucleopolyhedrovirus (AcMNPV). The results indicated that after AcMNPV infection, there was an initial increase in the expression of miR-184 within 24 h, followed by a subsequent decrease. MiR-184 can inhibit AcMNPV's DNA replication and budded virus production by directly targeting four viral genes, namely ie1, ac66, p49, and lef9. Moreover, suppressing miR-184 expression enhanced the insecticidal efficacy of AcMNPV against Spodoptera exigua larvae and markedly elevated the host ATPase gene expressions. These findings showed that miR-184 had a substantial impact on the interactions between baculoviruses and insects, presenting a prospective candidate for developing highly effective miRNA-based biopesticides.
禽腺病毒是全球家禽常见的传染病病原之一,目前对禽腺病毒进行流行病学调查,已给全球家禽业造成了巨大的经济损失.为了调查禽腺病毒在湖北地区的流行情况,在2021~2022年从湖北市场收集了224份鸡肝样本,为FAdV疫苗研制提供参考.基因测序结果显示:其中14株携带FAdV,Hexon L1环基因序列系统发育,12株为C型,2株为D型.在鸡肝癌细胞株(LMH)中培养并分离得到FAdV-11 HB200株,对分离物的Hexon基因进行了扩增和测序.遗传进化分析表明:FAdV-11 HB200与多个FAdV-11毒株处于同一分支.FAdV-C的流行率仍然占主导地位,相关部门应采取预防FAdV感染的措施.
Superoxide dismutase (SOD) is an antioxidant enzyme with multiple metal cofactors that can specifically clear reactive oxygen species (ROS), which plays an important role in a variety of ultraviolet-induced lesions. Therefore, SOD has the anti-ultraviolet radiation effect. The objective of this study was to compare the differences in the anti-ultraviolet radiation effect of SOD with distinct metal cofactors: Cu/Zn-SOD and Mn-SOD. SOD was first purified using hydrophobic interaction chromatography and ion-exchange chromatography. Second, the Methylthiazolyldiphenyl-tetrazolium bromide method and cell senescence kits were used to study the protective effect of SOD on ultraviolet-induced cell damage. Finally, the protective effect of SOD on ultraviolet -induced skin damage was histopathologically evaluated, and the expression levels of malondialdehyde (MDA) and matrix metalloproteinases (MMPs) in tissues were detected. The results showed that Cu/Zn-SOD was superior to Mn-SOD in promoting cell proliferation, alleviating cell damage, protecting skin structure, and regulating the expression levels of MDA and MMPs, and it has no side effects. In conclusion, Cu/Zn-SOD had a better anti-ultraviolet radiation effect than Mn-SOD, and it can be used in anti-aging and anti-ultraviolet skin-care products.
Aldehyde reductase (ALR) plays key roles in the detoxification of toxic aldehyde. In this study, the authors cloned the swamp eel ALR gene using rapid amplification of cDNA ends-PCR (RACE-PCR). The recombinant protein (rALR) was expressed in Escherichia coli and purified using a Ni2+-NTA chelating column. The rALR protein exhibited efficient reductive activity towards several aldehydes, ketones and S-nitrosoglutathione (GSNO). A spot assay suggested that the recombinant E. coli strain expressing rALR showed better resistance to formaldehyde, sodium nitrite and GSNO stress, suggesting that swamp eel ALR is crucial for redox homeostasis in vivo. Consequently, the authors investigated the effect of rALR on the oxidative parameters of the liver in swamp eels challenged with Aeromonas hydrophila. The hepatic glutathione (GSH) content significantly increased, and the hepatic NO content and levels of reactive oxygen species and reactive nitrogen species significantly decreased when rALR was administered. In addition, the mRNA expression of hepatic Alr, HO1 and Nrf2 was significantly upregulated, whereas the expression levels of NF-kappa B, IL-1 beta and NOS1 were significantly downregulated in the rALR-administered group. Collectively, these results suggest that ALR is involved in the response to nitrosative stress by regulating GSH/NO levels in the swamp eel.
Amniotes differ substantially in absolute and relative brain size after controlling for allometry, and numerous hypotheses have been proposed to explain brain size evolution. Brain size is thought to correlate with processing capacity and the brain's ability to support complex manipulation such as nest-building skills. The increased complexity of nest structure is supposed to be a measure of an ability to manipulate nesting material into the required shape. The degree of nest-structure complexity is also supposed to be associated with body mass, partly because small species lose heat faster and delicate and insulated nests are more crucial for temperature control of eggs during incubation by small birds. Here, we conducted comparative analyses to test these hypotheses by investigating whether the complexity of species-typical nest structure can be explained by brain size and body mass (a covariate also to control for allometric effects on brain size) across 1353 bird species from 147 families. Consistent with these hypotheses, our results revealed that avian brain size increases as the complexity of the nest structure increases after controlling for a significant effect of body size, and also that a negative relationship exists between nest complexity and body mass.
为了调查鸡传染性贫血病病毒(CAV)在肉鸡群中的流行情况及CAV毒株的致病性,于2021年在湖北荆州地区收集鸡肝样品224份,采用PCR方法进行检测,并对CAV的全基因扩增片段克隆测序,用生物信息学软件对测序结果进行遗传进化及分子特征分析;通过动物回归试验探究分离毒株JZ2118的致病性.结果显示,18份样品为CAV阳性,CAV阳性率为8.04%.CAV全基因序列与CAV VP1氨基酸序列的遗传进化树分组基本相同,说明CAV的遗传进化主要与VP1蛋白氨基酸的突变有关;1日龄SPF鸡感染JZ2118毒株14 d后致死率高达82.6%,在体重、病毒载量、血常规检测与病理组织切片等方面都具有显著性变化,表明JZ2118分离株是具有较强致病性的强毒株.
The variation in egg and clutch mass in sympatric species at high altitudes is poorly understood, and the potential causes of variation are rarely investigated. This study aimed to describe the interspecific variation in avian egg and clutch mass among 22 sympatric bird species at an altitude of 3430 m. Our objective was to reduce potential confounding effects of biotic/abiotic factors and investigated hypotheses concerning allometry, clutch size, parental care, nest predation, and lifespan as possible correlates and explanations for the observed variation. Our findings indicated that both egg and clutch mass evolve with body mass across species. We found that egg mass variation was not explained by clutch size when controlling for allometric effects, which contrasts the "egg mass vs. clutch size trade-off" hypothesis. Additionally, we found that clutch mass was positively associated with parental care (reflected by development period) but negatively associated with predation rate. By substituting egg mass and clutch size into the models, we found that clutch size was significantly correlated with parental care, predation rate, and lifespan, while egg mass was only significantly associated with development period. Overall, these findings support life-history theories suggesting that reduced clutch size or mass is associated with a higher risk of predation, reduced parental care, but longer adult lifespan. Interestingly, our results indicate that clutch size has a greater influence on these factors compared to egg mass. This could be attributed to the fact that smaller clutch sizes result in a more notable decrease in energetic allocation, as they require a reduced effort in terms of offspring production, incubation, and feeding, as opposed to solely reducing egg size. These findings contribute to the growing evidence that life-history and ecological traits correlate with egg and clutch mass variation in sympatric species. However, further research is needed to explore the potential evolutionary causes underlying these patterns.
The brain is among the most energetically costly organs in vertebrates, and thus trade-offs have been hypothesized to exert constraints on brain size evolution. The energy trade-off hypothesis (ETH) predicts that reducing the energy consumption of reproduction or other costly tissues should compensate for the cost of a large brain. Egg production in birds requires a large proportion of the total energy budget, and a clutch mass in some bird species can outweigh the body mass of the female. To date, this hypothesis has mainly been tested in mammals and ectothermic animals such as anurans and fishes. We collated data on adult brain size, body mass and egg-production traits such as clutch size, egg mass and annual broods from published studies, and conducted a phylogenetic comparative test of the interplay between egg-production investment and brain size evolution across bird species. After controlling for phylogenetic relationships and body size, we find a negative correlation between brain size and clutch size across 1395 species, which favored ETH. However, when egg mass was integrated in models, positive associations were detected between brain size and mass of eggs (via egg mass, clutch mass and annual total egg mass). Our results suggest that brain size trades off against egg-production only via certain aspects (e.g., clutch size). By contrast, a positive relationship between brain size and total egg reproduction (e.g., clutch mass and annual total egg mass) implied increased total energy budget outweighing energy allocation across bird species. Our study shows that there is no general energy trade-off between brain size and egg-reproduction investment, and suggests that brain size evolution follows mixed strategies across bird species.
制备禽腺病毒4型(FAdV-4)Fiber2蛋白的特异性单克隆抗体,将重组蛋白FAdV-4 Fiber2纯化后作为免疫原免疫BALB/c小鼠,分离脾脏中的淋巴细胞,与小鼠骨髓瘤细胞SP2/0进行融合,筛选得到10株能稳定分泌抗FAdV-4 Fiber2蛋白单克隆抗体的阳性杂交瘤细胞株1B5F1、1B5G11、1B5C1、1B5D2、1B5E3、1B5F3、1B5E4、1B5G4、1B5A8、1B5G8,取分泌抗体水平最高的1B5F1细胞株制备腹水,利用斑点杂交(Dot blot)和间接免疫荧光试验(IFA)进行特异性检测.结果表明,成功制备10株杂交瘤细胞株;制备的腹水能与FAdV-4 Fiber2蛋白及FAdV-4攻毒后的鸡肝组织产生特异性反应,证明成功制备了FAdV-4 Fiber2蛋白特异性单克隆抗体.
为了 分析猪圆环病毒2b 型病毒样颗粒(virus-like particles of Porcine circovirus type 2b,PCV-2b VLPs)表面具有外源肽展示潜力的位点,试验通过PyMol软件分析了 PCV-2b VLPs表面的位点分布,并以高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)Gp5的中和表位B(Epitope B,EpB)为外源肽,通过重叠PCR方法将表面位点替换为EpB,构建EpB嵌合的PCV-2b衣壳蛋白(Cap),利用原核表达系统诱导表达后进行凝胶层析纯化和Western-blot检测,并利用透射电子显微镜分析VLPs的组装状态.结果表明:在PCV-2b VLPs表面共筛选到8个可用于外源肽展示的位点,分别为S1(Y50TIKRTTVKTP60)、S2(L75PPGGGSN82)、S3(F124VTKATAL131)、S4(V161LDSTIDY168)、S5(Q183TAGNVD189)、S6(1201 YDQE205)、S7(K222DPPL226)、S8(S104PITQGDRGVGS115)位点;将EpB 替换展示到S1~S8位点后,构建的EpB嵌合的PCV-2b Cap能够在原核表达系统中获得不同程度的可溶性表达,其中S1~S7位点被EpB替换后能够实现较高水平的可溶性表达;8个目的蛋白均能被抗EpB抗体特异性识别,也均能够自组装形成VLPs,但粒径大小和形态与PCV-2b VLPs有不同程度的差异.说明试验筛选出的8个位点均有一定的外源肽展示能力.
Altitudinal gradients create environmental variation that can strongly affect avian life history strategies. To fully understand this issue, a comparison of the demography of populations of the same species over large altitudinal differences is required. Here, we investigated the breeding ecology of two populations of Eurasian tree sparrow ( Passer montanus ), one in the temperate zone of central China at an altitude of 30 m and another in the alpine zone of the northeastern Tibetan plateau at an altitude of 3430 m. The breeding ecology of this species was rarely studied at such a high altitude before. Our result revealed that high-altitude tree sparrows started breeding later, had a shorter breeding period (80 versus 140 days), produced smaller broods (1.2 versus 2.5 chicks) and had significantly smaller clutches (4.0 versus 4.9 eggs) but larger eggs (4272 versus 3443 mm 3 ) when compared with their low-altitude counterparts. Besides, tree sparrows at our high-altitude site had longer nestling periods (14.5 versus 13.4 days) than those at the low-altitude site. High-altitude tree sparrows tend to have reduced fecundity but allocate more energy into each offspring to confront the stressful conditions at high altitudes. The observed patterns are consistent with adaptive life history strategies.
Abstract Nest‐site characteristics are thought to play an important role in reproductive performance in birds (e.g., influencing reproductive success and predation risk). Nest‐site characteristics such as concealment may be particularly critical at high elevation where nests are exposed to challenging environmental conditions. In this study, we conducted both conventional and phylogenetically controlled analyses to investigate whether nest concealment affected several reproductive traits across 21 sympatric bird species living on Tibet Plateau (3,400 m altitude). Qualitatively equivalent results were reached in analyses, regardless of phylogenetic controls. We found that clutch size, incubation period, nestling period, and nest success were strongly and positively associated with nest concealment across species. Our study addressed such a high‐elevation bird community that is lacking in the previous studies. This study adds to theory that while there are a few exceptions, overall evidence supports a positive effect of nest concealment on reproductive performance across coexisting alpine species.
Hepatitis-hydropericardium syndrome (HHS) is widespread in China and causes high chicken mortality that results in great economic losses. A safe and effective vaccine is needed, and a subunit vaccine has potential for development. In this study, a truncated region of the FAdV-4 fiber 2 fused with coding sequence of one epitope of hexon was expressed in a prokaryotic expression system, and the immune protective effects of different doses of recombinant fiber 2 subunit vaccine on SPF chickens were compared. The recombinant fiber2 (Gly275- Pro479 aa)-hexon (Met21-Val51 aa) protein (rFH) obtained in Escherichia coli showed good solubility. The chicken survival rate at the lowest dose (2.5 mu g/bird) was 75% (6/8), and at higher doses (>5 mu g/bird) was 100% (8/8) in challenge experiment. Two chickens in the 2.5 mu g/bird treatment showed severe lesions, while birds in the higher dose treatments showed no obvious tissue damage as determined by histopathologic analysis of liver and spleen. Absolute quantitative real-time PCR showed no viral load in the >5 mu g/bird treatments, but two chickens in the 2.5 mu g/bird treatment had high viral loads. The challenge experience demonstrated that the rFH vaccine provided 100% protection at >5 mu g/bird. These results suggested that rFH protein as an effective vaccine to protect against FAdV-4 and provided a new idea for the development of vaccine against HHS.
AbstractBackgroundGaining extrapair copulations (EPCs) is a complicated behavior process. The interaction between males and females to procure EPCs may be involved in brain function evolution and lead to a larger brain. Thus, we hypothesized that extrapair paternity (EPP) rate can be predicted by relative brain size in birds. Past work has implied that the EPP rate is associated with brain size, but empirical evidence is rare.MethodsWe collated data from published references on EPP levels and brain size of 215 bird species to examine whether the evolution of EPP rate can be predicted by brain size using phylogenetically generalized least square (PGLS) models and phylogenetic path analyses.ResultsWe found that EPP rates (both the percentage EP offspring and percentage of broods with EP offspring) are negatively associated with relative brain size. We applied phylogenetic path analysis to test the causal relationship between relative brain size and EPP rate. Best‐supported models (ΔCICc < 2) suggested that large brain lead to reduced EPP rate, which failed to support the hypothesis that high rates of EPP cause the evolution of larger brains.ConclusionThis study indicates that pursuing EPCs may be a natural instinct in birds and the interaction between males and females for EPCs may lead to large brains, which in turn may restrict their EPC level for both sexes across bird species.
目的 对重组猪尿酸氧化酶(recombinant porcine urate oxidase,rPUOX)进行突变改造,以增加rPUOX上可供聚乙二醇(polyethylene glycol,PEG)耦合的赖氨酸(Lys)残基数,探究PEG修饰对改造前后的酶液在SPF鸡体内的药效和抗原屏蔽作用的影响.方法 对纯化的基因改造前后的rPUOX进行PEG化修饰,修饰蛋白静脉注射SPF鸡,共4次,每次间隔7d.实验分为20×5K PEG-rPUOX-215组、20×5K PEG-rPUOX组和空白对照组(0.9%生理盐水).于鸡翼根静脉处采血,用血糖尿酸测试仪及尿酸测试条测定尿酸浓度,酶反应-紫外分光光度法检测尿酸酶活性,间接ELISA法检测免疫原性.结果 20×5K PEG-rPUOX-215和20×5K PEG-rPUOX均获得充分修饰,且前者修饰效果更好.第1、2次注射,降低鸡血尿酸幅度20×5K PEG-rPUOX-215组高于20×5K PEG-rPUOX组约11%和6%.3次注射药效维持时间20×5K PEG-rPUOX组相较于20×5K PEG-rPUOX-215组逐次降低0、24和48 h.20×5K PEG-rPUOX-215组抗体产生速率相对较慢,且14 d抗体阳性率低于20×5K PEG-rPUOX组约14.3%.结论 20×5K PEG-rPUOX-215修饰后药效学作用更显著,药物半衰期延长,且免疫原性相对较低,为研制低免疫原性的rPUOX及实现临床上多次给药提供了新的方法和思路.