RESUMO
Extrahepatic injury, particularly neurologic dysfunctions such as Guillain-Barré syndrome, neurologic amyotrophy, and encephalitis/meningoencephalitis/myositis were associated with HEV infection, which was supported by both clinical and laboratory studies. Thus, it is crucial to figure out how the virus invades into the central nervous system (CNS). In this study, CNS lesions were determined in rabbits and Mongolian gerbils inoculated with genotype 4 HEV. Junctional proteins were detected in HEV infected primary human brain microvascular cells (HBMVCs). Viral encephalitis associated perivascular cuffs of lymphocytes and microglial nodules were observed in HEV infected rabbits. Both positive- and negative-strand of HEV RNA was detected in brain and spinal cord in rabbits intraperitoneally infected with HEV at 28 dpi (days postinoculation), but not in rabbits gavaged with HEV. HEV ORF2 protein was further examined in both brain and spinal cord sections of infected rabbits, with positive signals located mainly in neural cells and perivascular areas. Ultrastructural study showed thickened and reduplicated basement membranes of capillary endothelium in HEV RNA positive brain tissues. In vitro study showed loss of tight junction proteins including Claudin5, Occludin, and ZO-1 (zonula occludens-1) in HBMVCs inoculated with HEV for 48 h. These findings indicated that disruption of the blood-brain barrier (BBB) might be potential mechanisms of HEV invasion into the CNS. It provides new insights to further study HEV associated neurologic disorders and will be helpful for seeking potential therapeutics for HEV infection in the future.
Assuntos
Barreira Hematoencefálica/patologia , Encefalite Viral/patologia , Encefalite Viral/virologia , Vírus da Hepatite E/patogenicidade , Hepatite E/patologia , Hepatite E/virologia , Proteínas de Junções Íntimas/análise , Animais , Encéfalo/virologia , Células Cultivadas , Modelos Animais de Doenças , Células Endoteliais/virologia , Gerbillinae , Vírus da Hepatite E/isolamento & purificação , Humanos , Coelhos , Medula Espinal/virologiaRESUMO
The aim of this study was to investigate the occurrence of hepatitis E virus (HEV) in sewage samples in Shen Zhen, China. Sewage samples were collected from 152 sewage plants including livestock sewage, domestic sewage and treated sewage from May to July of 2015. Two of 152 samples were HEV positive (1.32%) from the livestock sewage plants. Partial ORF2 fragments of HEV were sequenced and a phylogenetic tree was constructed using MEGA5.1. Blast and phylogenetic analyses showed that both of these two sequences belonged to HEV Genotype 4. To the best of our knowledge, this is the first study on the molecular characterization of HEV in wastewater in China and the first time to detect Genotype 4 in the sewage. Results from this study indicate that the possibilities of sporadic infections of HEV should be emphasized because virus still has the possibility to be circulating in the sewage in China.
Assuntos
Vírus da Hepatite E/isolamento & purificação , Esgotos/virologia , Animais , China , Genótipo , Vírus da Hepatite E/classificação , Vírus da Hepatite E/genética , Gado , Filogenia , Águas Residuárias/virologiaRESUMO
BACKGROUND: Hemoglobin is a rich source of biological peptides. As a byproduct and even wastewater of poultry-slaughtering facilities, chicken blood is one of the most abundant source of hemoglobin. RESULTS: In this study, the chicken hemoglobin antimicrobial peptides (CHAP) were isolated and the antimicrobial and bactericidal activities were tested by the agarose diffusion assay, minimum inhibitory concentration (MIC) analysis, minimal bactericidal concentration (MBC) analysis, and time-dependent inhibitory and bactericidal assays. The results demonstrated that CHAP had potent and rapid antimicrobial activity against 19 bacterial strains, including 9 multidrug-resistant bacterial strains. Bacterial biofilm and NaCl permeability assays, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were further performed to detect the mechanism of its antimicrobial effect. Additionally, CHAP showed low hemolytic activity, embryo toxicity, and high stability in different temperatures and animal plasma. CONCLUSION: CHAP may have great potential for expanding production and development value in animal medication, the breeding industry and environment protection.
Assuntos
Anti-Infecciosos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Hemoglobinas/farmacologia , Fragmentos de Peptídeos/farmacologia , Animais , Antibacterianos/efeitos adversos , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Anti-Infecciosos/isolamento & purificação , Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Bacteriólise/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Atividade Bactericida do Sangue , Permeabilidade da Membrana Celular/efeitos dos fármacos , Galinhas , Farmacorresistência Bacteriana Múltipla , Hemoglobinas/química , Hemolíticos/isolamento & purificação , Hemolíticos/farmacologia , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Fragmentos de Peptídeos/química , Estabilidade Proteica , Cloreto de Sódio/metabolismoRESUMO
We examined the effect of hepatitis E virus (HEV) on the renal tissue pathogenesis, morphological damages and related molecular mechanisms following swine HEV suspension intraperitoneally inoculation in Mongolian gerbils. The microscopic and ultramicroscopic analyses of kidney tissue structure were carried out at different points after inoculation of HEV. The immunohistochemistry, real-time PCR and Western blot were performed to explore the molecular mechanisms associated with HEV presence in the renal tissues. Real-time PCR revealed that the copies of HEV RNA in the kidney were detected at 7 dpi, and peaked at 14 dpi at a concentration was 7.18 logs g(-1), with detection of HEV ORF2 antigen by immunohistochemistry. Hematoxylin and eosin (HE) staining showed pathological lesions including glomerular atrophy, degeneration, edema and necrosis of renal tubular epithelial cells and Mallory and Sirius red staining indicated the presence of collagen fibers and fibrosis in kidney tissues of inoculated gerbils. Ultrastructural studies of basal membrane of renal tubules demonstrated the rough and uneven with mitochondria swelling and vacuolation in the tissues of HEV inoculated animals. Similarly, significantly higher number of (TUNEL)-positive cells were seen in renal tubule tissues compared to control group. Moreover, immuno histochemical results indicated that significant increase expression of the B-cell lymphoma 2 (Bcl-2), Bcl-2 associated X protein (Bax), FAS and Caspase-3 in HEV inoculated Mongolian gerbils at each time points. Relative mRNA expression by real-time PCR revealed a significantly higher (P<0.05) mRNA level of BAX, Bcl-2 and caspase-3 transcription in HEV inoculated Mongolian gerbils. Our results demonstrates that activation of mitochondria and Caspase-3 protease might be induced the apoptosis which subsequently cause the necrosis and cell death of renal epithelial cells during acute phase of HEV infection in HEV inoculated Mongolian gerbils.
Assuntos
Antígenos Virais/análise , Apoptose , Gerbillinae/virologia , Vírus da Hepatite E/isolamento & purificação , Hepatite E/patologia , Rim/patologia , Rim/virologia , Animais , Western Blotting , Modelos Animais de Doenças , Hepatite E/virologia , Histocitoquímica , Imuno-Histoquímica , Injeções Intraperitoneais , Microscopia , Reação em Cadeia da Polimerase em Tempo Real , Suínos , Fatores de TempoRESUMO
This study involved a death which occurred in four Himalayan griffons housed in Beijing zoo, China. Based on pathogen identification and the pathological changes observed, we did characterize the fungi and Hepatitis E virus (HEV) in four dead Himalayan griffons. Pathological changes were severe. Membranous-like material was observed on the surface of the internal organs. Spleen was necrotic. Focal lymphocyte infiltration in the liver and many sunflower-like fungi nodules were evident in the tissues, especially in the kidney. PCR was used to identify the pathogen. Based on the 18SrRNA genomic sequence of known fungi, the results confirmed that all four dead Himalayan griffons were infected with Aspergillus. At the same time the detection of HEV also showed positive results. To the best of our knowledge, this work appears to be the first report of concurrent presence of Aspergillosis and Hepatitis E virus in rare avian species.
Assuntos
Aspergilose , Coinfecção , Falconiformes , Hepatite E , Hepevirus , Animais , Aspergilose/veterinária , Aspergilose/virologia , Doenças das Aves/microbiologia , Doenças das Aves/virologia , China , Hepatite E/microbiologia , Hepatite E/veterináriaRESUMO
Previous studies have shown that hepatitis E virus (HEV) can be transmitted between rats, pigs, cattle, rabbits, chicken, cats, and deer. Because wild and domestic rodents have anti-HEV antibodies, they are considered potential reservoirs of HEV. In the current study, Mongolian gerbils were experimentally infected with swine hepatitis E virus and the effects of this infection were investigated. After inoculation with HEV, the liver-to-body weight ratio increased at 7 dpi. Mongolian gerbils demonstrated significant increase (p<0.05) in Aspartate Transaminase (AST), alanine transaminase (ALT) and total bilirubin (T-BIL) concentrations in the sera, and HEV IgG was detected at 21 days post-inoculation (dpi). Real-time PCR revealed that the copies of HEV RNA in the liver were detected at 7 dpi, and peaked at 28 dpi at a concentration of 7.73 logs g(-1). Using both light and electron microscopy, hepatic lesions were observed in the HEV inoculated animals. In the experimental group, characteristic viral hepatitis lesions were prominent in the liver. HEV antigen was detected in the liver by immunohistochemistry, and HEV ORF3 antigen was detectable in liver by Western blot. These results clearly demonstrate that viral load of HEV in livers was dynamic, and ultrastructural hepatic injury in HEV infected Mongolian gerbils and anti-HEV IgG positive seroconversion were observed during infection.
Assuntos
Modelos Animais de Doenças , Gerbillinae , Vírus da Hepatite E/fisiologia , Hepatite E/veterinária , Fígado/virologia , Doenças dos Suínos/virologia , Animais , Anticorpos Anti-Hepatite , Hepatite E/patologia , Hepatite E/virologia , Vírus da Hepatite E/genética , Fígado/patologia , Masculino , Suínos , Doenças dos Suínos/patologiaRESUMO
BACKGROUND: In recent decades, Porcine circovirus 2 (PCV2) infection has been recognized as the causative agent of postweaning multisystemic wasting syndrome, and has become a threat to the swine industry. Hepatitis E virus (HEV) is another high prevalent pathogen in swine in many regions of the world. PCV2 and HEV are both highly prevalent in pig farms in China. CASE PRESENTATION: In this study, we characterized the HEV and PCV2 co-infection in 2-3 month-old piglets, based on pathogen identification and the pathological changes observed, in Hebei Province, China. The pathological changes were severe, and general hyperemia, hemorrhage, inflammatory cell infiltration, and necrosis were evident in the tissues of dead swine. PCR was used to identify the pathogen and we tested for eight viruses (HEV, Porcine reproductive and respiratory syndrome virus, PCV2, Classical swine fever virus, Porcine epidemic diarrhea virus, Transmissible gastroenteritis coronavirus, Porcine parvovirus and Pseudorabies virus) that are prevalent in Chinese pig farms. The livers, kidneys, spleens, and other organs of the necropsied swine were positive for HEV and/or PCV2. Immunohistochemical staining showed HEV- and PCV2-antigen-positive signals in the livers, kidneys, lungs, lymph nodes, and intestine. CONCLUSION: HEV and PCV2 co-infection in piglets was detected in four out of seven dead pigs from two pig farms in Hebei, China, producing severe pathological changes. The natural co-infection of HEV and PCV2 in pigs in China has rarely been reported. We speculate that co-infection with PCV2 and HEV may bring some negative effect on pig production and recommend that more attention should be paid to this phenomenon.
Assuntos
Infecções por Circoviridae/veterinária , Circovirus , Vírus da Hepatite E , Hepatite E/veterinária , Doenças dos Suínos/virologia , Animais , China/epidemiologia , Infecções por Circoviridae/complicações , Infecções por Circoviridae/epidemiologia , Infecções por Circoviridae/patologia , Circovirus/genética , Coinfecção/patologia , Coinfecção/veterinária , Coinfecção/virologia , Hepatite E/complicações , Hepatite E/epidemiologia , Hepatite E/patologia , Vírus da Hepatite E/genética , Filogenia , Reação em Cadeia da Polimerase/veterinária , Suínos/virologia , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/patologiaRESUMO
Rabbit hepatitis E virus (HEV) is a novel genotype of HEV, and is considered to pose a risk of zoonotic transmission. Research into the systemic distribution of rabbit HEV in rabbits during different periods of infection has rarely been reported. To better understand this virus, we infected rabbits with second-passage rabbit HEV via an intraperitoneal route. After inoculation, the infection showed two types, temporary and constant infection. The detection of HEV RNA in the feces varied with time, and serum antigen correlated with fecal HEV RNA. Viremia only appeared 72 days after inoculation. The rabbits remained antibody negative throughout the experimental period. When HEV was localized, several organs besides the liver were HEV RNA positive. Tissue antigen was observed immunohistochemically in the different cells of various organs, especially in parts of the small intestine and the characteristic rabbit gut-associated lymphoid tissue. These data provide valuable information for future research into the pathogenesis of HEV.
Assuntos
Antígenos de Hepatite/imunologia , Vírus da Hepatite E/imunologia , Hepatite E/imunologia , Hepatite E/virologia , Especificidade de Órgãos/imunologia , Animais , Ensaio de Imunoadsorção Enzimática , Fezes/virologia , Hepatite E/sangue , Imuno-Histoquímica , Injeções Intraperitoneais , RNA Viral/sangue , CoelhosRESUMO
Eight-week-old male Kunming mice were administered either melamine (MA, 30, 140, or 700 mg/kg/day), a melamine and cyanuric acid mixture (MC, each at 15, 70, or 350 mg/kg/day), or vehicle (control) for 3 consecutive days. Testicular toxicity was evaluated on days 1 and 5 after the final exposure. The testicular and epididymal weights and serum testosterone level were significantly decreased in the highest MC group (350 mg/kg/day). Histopathologically, both MA and MC caused obvious lesions in the testis and epididymis, with significant increases in sperm abnormalities. By TEM, the blood-testis barrier was damaged dose dependently. TUNEL staining showed that both MA and MC induced increases in germ cell apoptosis. The Sertoli cell vimentin was collapsed in the treated animals as detected by immunohistochemical staining and Western blotting. This study demonstrated that both MA and MC treatments could disrupt the blood-testis barrier and cause a clear testicular toxicity.