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1.
Artigo em Inglês | MEDLINE | ID: mdl-32656652

RESUMO

Vibrio alginolyticus is posting an increasing threat to survival of grouper. Classical complement cascade can trigger initiation of immunity, while complement 9 (C9) is a major complement molecule involved in final step of membrane attack complex (MAC) formation. In this study, full-length EcC9 contained an ORF sequence of 1779 bp, encoding a polypeptide of 592 amino acids. A high-level expression of EcC9 mRNA was observed in liver. Following vibrio challenge, increased expression levels of EcC1q, EcBf/C2, EcC4, EcC6, EcC7 and EcC9 mRNA were detected in liver and kidney. These results implied that elevated expression level of classical complement pathway (CCP) and terminal complement components (TCCs) may assess toxicological effect of V. alginolyticus.


Assuntos
Bass/genética , Bass/imunologia , Complemento C9/genética , Complemento C9/imunologia , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Complemento C9/química , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Filogenia , Alinhamento de Sequência/veterinária , Vibrio alginolyticus/fisiologia
2.
J Parasitol ; 106(3): 383-391, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32491171

RESUMO

The long-term fidelity of pinniped hosts to their natal rookery site suggests the genetic architecture of their Uncinaria spp. hookworms should be strongly structured by host breeding biology. However, historical events affecting host populations may also shape parasite genetic structure. Sequences of the mitochondrial cytochrome c oxidase 1 (COI) gene of 86 Uncinaria lucasi individuals were obtained to assess genetic variation and structure of nematodes from 2 host species (68 hookworms from northern fur seals; 18 hookworms from Steller sea lions) and rookeries from 3 widely separated geographic regions: the western Bering Sea and Sea of Okhotsk, eastern Bering Sea, and the eastern Pacific Ocean. High COI haplotype (h = 0.96-0.98) and nucleotide (π = 0.014) diversity was found. The haplotype network showed a star-shaped pattern with a large number of haplotypes separated by few substitutions. The network did not show separation of U. lucasi by geographic region or host species. Fst values between U. lucasi individuals representing geographic regions showed no differentiation, consistent with the absence of genetic structure. At face value, this lack of genetic structure in U. lucasi suggests high gene flow but could also be explained by recent (post-glacial) population expansions of northern fur seals and their hookworms.


Assuntos
Ancylostomatoidea/fisiologia , Caniformia/parasitologia , Infecções por Uncinaria/veterinária , Sequência de Aminoácidos , Ancylostomatoidea/genética , Animais , Sequência de Bases , Complexo IV da Cadeia de Transporte de Elétrons/genética , Feminino , Variação Genética , Haplótipos/genética , Infecções por Uncinaria/parasitologia , Infecções por Uncinaria/transmissão , Masculino , Mitocôndrias/enzimologia , Oceano Pacífico , Alinhamento de Sequência/veterinária
3.
J Parasitol ; 106(3): 350-359, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32227224

RESUMO

Thelohanellus magnacysta n. sp. (Bivalvulida: Myxobolidae) infects the skeletal muscle of blacktail shiner, Cyprinella venusta Girard, 1856 (Cypriniformes: Cyprinidae) in Bull Creek, Chattahoochee River Basin, eastern Georgia. Although numerous members of ThelohanellusKudo, 1933 have overlapping myxospore dimensions with the new species, it differs from all nominal congeners by polar filament coil number and polar capsule width as well as by lacking a mucous envelope, iodinophilic vacuole, and sutural markings. With the use of novel primers for Myxozoa, a phylogenetic analysis of the small subunit ribosomal DNA (SSU rDNA) suggests that the new species shares a recent common ancestor with a clade of cyprinid-infecting species of Myxobolus Bütschli, 1882 (Bivalvulida: Myxobolidae) and Thelohanellus. Consistent with other published research concerning the systematics of Thelohanellus, this result suggested that Thelohanellus and Myxobolus are polyphyletic and need revision. Histological sections of infected blacktail shiners confirmed that myxospores were only found within a plasmodium and only infected skeletal muscle and that plasmodia were encapsulated by a granuloma comprising varying degrees of acute granulomatous inflammation. The new species is the fourth of Thelohanellus reported from North America and the first reported from Cyprinella, as well as the first myxozoan described from the blacktail shiner.


Assuntos
Cyprinidae/parasitologia , Doenças dos Peixes/parasitologia , Músculo Esquelético/parasitologia , Myxozoa/classificação , Doenças Parasitárias em Animais/parasitologia , Animais , Sequência de Bases , Teorema de Bayes , Análise de Fourier , Georgia , Microscopia de Interferência , Myxozoa/genética , Myxozoa/isolamento & purificação , Filogenia , Rios , Alinhamento de Sequência/veterinária , Esporos/isolamento & purificação , Esporos/ultraestrutura
4.
J Parasitol ; 106(2): 221-232, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-32164028

RESUMO

Members of the sucking louse genus Pedicinus are ectoparasites of cercopithecid primates in Africa, Asia, and Gibraltar. Pedicinus gabonensis n. sp. is described on the basis of adult male and female specimens collected from the mandrill (Mandrillus sphinx) in Gabon. The new species is compared morphologically with other members of the genus Pedicinus, and a nuclear elongation factor 1 alpha gene sequence is provided. Host associations and geographical distributions of the 18 previously recognized species of the genus and of P. gabonensis n. sp. are reviewed. Updated identification keys are provided for males and females of all known valid species of Pedicinus.


Assuntos
Anoplura/classificação , Infestações por Piolhos/veterinária , Mandrillus/parasitologia , Doenças dos Macacos/parasitologia , Animais , Anoplura/anatomia & histologia , Anoplura/genética , Anoplura/fisiologia , DNA/química , DNA/isolamento & purificação , Feminino , Gabão/epidemiologia , Infestações por Piolhos/epidemiologia , Infestações por Piolhos/parasitologia , Masculino , Doenças dos Macacos/epidemiologia , Alinhamento de Sequência/veterinária , Análise de Sequência de DNA/veterinária
5.
Artigo em Inglês | MEDLINE | ID: mdl-32162033

RESUMO

Seagrass meadows are among the four most productive marine natural ecosystems in the world. Zostera japonica (Z. japonica) is the most widely distributed species of seagrass in China. Nucleotide exchange factors (NEFs) promote the release of ADP during heat stress, accelerating the rate-limiting step of Heat shock protein 70 (Hsp70). Although NEFs play an important role in abiotic stress tolerance of plants, NEFs in seagrass have not been studied. In this study, we cloned Fes1 from Z. japonica (ZjFes1) by rapid amplification of the cDNA ends using RACE, and full length ZjFes1 was 1171 bp. It contained an 81 bp 5'-terminal untranslated region (UTR), 109 bp 3'-UTR and 981 bp open reading frame (ORF). The ORF (ZjFes1) was predicted to encode a polypeptide of 326 amino acids with theoretical molecular weight (MW) of 36.10 kDa and pI of 5.22. ZjFes1 shared 89% amino acid identity with Fes1 from Zostera marina (Z. marina). The transcriptional levels of ZjFes1 increased significantly 1 h after heat treatment. ZjFes1 was localized to the cytoplasm. Taken together, we found that ZjFes1 was a stress-inducible gene that may be involved in heat stress response. This study lays the foundation for further studies on the role of ZjFes1 in heat resistance.


Assuntos
Proteínas de Transporte/genética , Expressão Gênica/fisiologia , Resposta ao Choque Térmico , Proteínas de Plantas/genética , Zosteraceae/fisiologia , Sequência de Aminoácidos , Sequência de Bases , Proteínas de Transporte/química , Proteínas de Transporte/metabolismo , Regulação da Expressão Gênica de Plantas , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Alinhamento de Sequência/veterinária , Zosteraceae/genética
6.
Transbound Emerg Dis ; 67(4): 1730-1738, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32037673

RESUMO

Porcine respiratory and reproductive syndrome virus (PRRSV) causes an economically important disease affecting commercial pork production worldwide. NADC34-like PRRSV has had a strong impact on the U.S. and Peruvian pig industries in recent years and also emerged in northeastern China in 2017. However, the endemic status of NADC34-like PRRSV in China is unclear. In this study, we examined 650 tissue samples collected from 16 Provinces in China from 2018 to 2019. Six NADC34-like PRRSV strains were detected in samples from three Provinces, and the complete genomes of four of these strains were sequenced. Phylogenetic analysis showed that these novel PRRSV strains belong to sublineage 1.5 (or NADC34-like PRRSV), forming two groups in China. Sequence alignment suggested that these novel strains share the same 100-aa deletion in the Nsp2 protein that was identified in IA/2014/NADC34 isolated from the United States in 2014. Recombination analysis revealed that five of eight complete genome sequences are derived from recombination between IA/2014/NADC34 and ISU30 or NADC30. The number and distribution of NADC34-like PRRSVs is increasing in China. Importantly, compared with the currently endemic strain NADC30-like PRRSV, NADC34-like PRRSV has the potential to be an endemic strain in China. This study will help us understand the epidemic status of NADC34-like PRRSV in China and provide data for further monitoring this type of PRRSV in China.


Assuntos
Doenças Endêmicas/veterinária , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , Doenças dos Suínos/epidemiologia , Animais , Sequência de Bases , China/epidemiologia , Variação Genética , Genoma Viral/genética , Filogenia , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Recombinação Genética , Alinhamento de Sequência/veterinária , Suínos , Doenças dos Suínos/virologia , Sequenciamento Completo do Genoma/veterinária
7.
J Parasitol ; 106(1): 157-166, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32053468

RESUMO

This study provides additional taxonomic features based on scanning electron microscopy (SEM) and molecular data for Paracamallanus cyathopharynx ( Baylis, 1923 ) (Nematoda: Camallanidae). Parasites were collected from the posterior end of the intestine of cultured freshwater Sharptooth catfish Clarias gariepinus (Burchell, 1822) from Kibos fish farm, Kisumu County, Kenya, from December 2017 to March 2018. Additional taxonomic features recorded for P. cyathopharynx include the occurrence of 4-5 equal length digitate processes on the caudal end of the female, 4 processes (2 smaller and 2 larger) on the male caudal end, and a description of the shape of the distal tip of the right spicule. The study provides SEM images of the exposed sclerotized buccal capsule. This gives more information on the tridents, the sclerotized plate that extends laterally from the buccal capsule, and the narrow isthmus separating the anterior buccal capsule from the posterior. The prevalence, intensity, mean intensity, and mean abundance was 52.91%, 2-38, 12.37 and 6.54, respectively. 18S rDNA fragments were amplified, sequenced, and compared to other camallanid taxa, and 18S data confirmed the identity of the newly obtained sequences (MN396556) as that of P. cyathopharynx, being identical to sequence DQ813445 from Tanzania. This represents the first geographical record of P. cyathopharynx in Kenya.


Assuntos
Peixes-Gato/parasitologia , Doenças dos Peixes/parasitologia , Infecções por Spirurida/veterinária , Spirurina/genética , Spirurina/ultraestrutura , Animais , DNA Ribossômico/química , Feminino , Doenças dos Peixes/epidemiologia , Pesqueiros , Água Doce , Intestinos/parasitologia , Quênia/epidemiologia , Masculino , Microscopia Eletrônica de Varredura/veterinária , Prevalência , RNA Ribossômico 18S/genética , Alinhamento de Sequência/veterinária , Análise de Sequência de DNA/veterinária , Infecções por Spirurida/epidemiologia , Infecções por Spirurida/parasitologia
8.
Vet Res ; 51(1): 12, 2020 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-32070432

RESUMO

High-mobility group box 1 protein (HMGB1) shows endogenous damage-associated molecular patterns (DAMPs) and is also an early warning protein that activates the body's innate immune system. Here, the full-length coding sequence of HMGB1 was cloned from the spleen of Cherry Valley duck and analyzed. We find that duck HMGB1(duHMGB1) is mostly located in the nucleus of duck embryo fibroblast (DEF) cells under normal conditions but released into the cytoplasm after lipopolysaccharide (LPS) stimulation. Knocking-down or overexpressing duHMGB1 had no effect on the baseline apoptosis rate of DEF cells. However, overexpression increased weakly apoptosis after LPS activation. In addition, overexpression strongly activated the IFN-I/IRF7 signaling pathway in DEF cells and significantly increased the transcriptional level of numerous pattern recognition receptors (PRRs), pro-inflammatory cytokines (IL-6, TNF-α), IFNs and antiviral molecules (OAS, PKR, Mx) starting from 48 h post-transfection. Overexpression of duHMGB1 strongly impacted duck virus replication, either by inhibiting it from the first stage of infection for novel duck reovirus (NDRV) and at late stage for duck Tembusu virus (DTMUV) or duck plague virus (DPV), or promoting replication at early stage for DTMUV and DPV infection. Importantly, data from duHMGB1 overexpression and knockdown experiments, time-dependent DEF cells transcriptional immune responses suggest that duHMGB1 and RIG-I receptor might cooperate to promote the expression of antiviral proteins after NDRV infection, as a potential mechanism of duHMGB1-mediated antiviral activity.


Assuntos
Proteínas Aviárias/genética , Patos/genética , Infecções por Flavivirus/veterinária , Proteína HMGB1/genética , Infecções por Herpesviridae/veterinária , Imunidade Inata/genética , Doenças das Aves Domésticas/prevenção & controle , Transdução de Sinais/genética , Sequência de Aminoácidos , Animais , Antivirais , Proteínas Aviárias/química , Proteínas Aviárias/metabolismo , Patos/metabolismo , Flavivirus , Infecções por Flavivirus/prevenção & controle , Infecções por Flavivirus/virologia , Perfilação da Expressão Gênica/veterinária , Proteína HMGB1/química , Proteína HMGB1/metabolismo , Infecções por Herpesviridae/prevenção & controle , Infecções por Herpesviridae/virologia , Mardivirus , Filogenia , Doenças das Aves Domésticas/virologia , Alinhamento de Sequência/veterinária
9.
BMC Genomics ; 21(1): 97, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32000661

RESUMO

BACKGROUND: Alternative splicing is an important biological process whose precision must be tightly regulated during growth and development. Although there are species, disease (e.g. cancers), or study specific databases available in many organisms, no database exists in livestock animals specifically tailored for alternative splicing. DESCRIPTION: We present in this study the development and implementation of a database for alternative splicing atlas in livestock animals (ASlive.org). Using publicly available RNASeq data sets across many tissues, cell types, and biological conditions totaling 28.6 T bases, we built a database of alternative splicing events in five major livestock and poultry animal species (cattle, sheep, pigs, horses, and chickens). The database contains many types of information on alternative splicing events, including basic information such as genomic locations, genes, and event types, quantitative measurements of alternative splicing in the form of percent spliced in (PSI), overlap with known DNA variants, as well as orthologous events across different lineage groups. CONCLUSIONS: This database, the first of its kind in livestock animals, will provide a useful exploratory tool to assist functional annotation of animal genomes.


Assuntos
Processamento Alternativo , Bases de Dados Genéticas , Gado/genética , Animais , Bovinos , Cavalos , Alinhamento de Sequência/veterinária , Análise de Sequência de RNA/veterinária , Ovinos , Suínos
10.
Artigo em Inglês | MEDLINE | ID: mdl-32056240

RESUMO

The caspase family proteins are aspartate-specific cysteine proteases that transmit extracellular signals to cells, ultimately cause apoptosis and therefore play a key role in cellular immunity. In this study, we cloned and characterized three caspases from Chinese black sleeper (Bostrychus sinensis), Bscasp-1, Bscasp-8 and Bscasp-9. Real-time PCR analysis showed that Bscasp-1, Bscasp-8 and Bscasp-9 were universally expressed in all tested tissues of B. sinensis. Expression analyses showed that after poly(I:C) stimulation and bacterial (Vibrio parahaemolyticus) infection, the three caspases were significantly upregulated. After poly(I:C) stimulation, the change of Bscasp-1 expression in the head kidney was the most obvious; peak expression was about 80.78-fold more than that of the control. In addition, the expression of Bscasp-8 and Bscasp-9 in the peripheral blood and liver was 167.99- and 17.98-fold higher than that in the control group, respectively. After V. parahaemolyticus infection, the expression peaks of Bscasp-1 and Bscasp-8 in the peripheral blood and spleen were 85.82-fold and 280.83-fold that of the control. However, the expression of Bscasp-9 in the peripheral blood was upregulated only 8.33-fold higher than that in the control group. These results indicate that Bscasp-1, Bscasp-8 and Bscasp-9 are likely involved in response to viral and bacterial infection.


Assuntos
Caspases/genética , Caspases/imunologia , Doenças dos Peixes/imunologia , Peixes/genética , Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Caspases/química , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Filogenia , Poli I-C/farmacologia , Alinhamento de Sequência/veterinária , Vibrioses/imunologia , Vibrioses/veterinária , Vibrio parahaemolyticus/fisiologia
11.
Transbound Emerg Dis ; 67(4): 1428-1432, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31968152

RESUMO

Pseudorabies is a disease that seriously endangers the pig industry in China. Recently, we successfully isolated a pseudorabies virus from the brain tissue of piglets at a farm in Sichuan, China, and named it the FJ62 strain. In order to understand the molecular biological characteristics of the strain, primers were designed for glycoproteins gB, gC, gD and gE, which were amplified by a polymerase chain reaction (PCR) and sequenced. After comparing the sequence with the GenBank 22 pseudorabies virus reference strains and establishing the genetic evolutionary tree, it was found that the gB gene of pseudorabies virus was highly homologous (up to 100%) with the MY-1 strain which is isolated from a wild boar in Japan (AP018925) but that homology with other strains in China was low. The gC gene was in the same branch as most of the representative strains in China, with 99.5% homology. The gD gene is in the same branch as the domestic strain LA in China (KU552118), and the homology was 99.9%. The gE gene was in the same branch as the domestic BJ/YT strain in China (KC981239), with 99.9% homology. The results showed that the FJ62 strain of the pseudorabies virus isolated here may be a variant strain of FJ62 isolated from a domestic pig after natural recombination of pseudorabies virus genotype I from wild boar and genotype II from pigs in China. There have been no similar reports in Sichuan. The discovery of the recombinant virus strain provides a reference basis for the prevention and control of pseudorabies and a design strategy for a vaccine in Sichuan, China, in the future.


Assuntos
Evolução Molecular , Herpesvirus Suídeo 1/genética , Pseudorraiva/virologia , Doenças dos Suínos/virologia , Proteínas do Envelope Viral/genética , Vacinas Virais/genética , Animais , China , Fazendas , Genótipo , Glicoproteínas/genética , Herpesvirus Suídeo 1/imunologia , Herpesvirus Suídeo 1/isolamento & purificação , Filogenia , Pseudorraiva/prevenção & controle , Recombinação Genética , Alinhamento de Sequência/veterinária , Sus scrofa , Suínos , Doenças dos Suínos/prevenção & controle
12.
Fish Shellfish Immunol ; 98: 19-24, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31899359

RESUMO

Calpains (CAPNs) belong to the papain superfamily of cysteine proteases, and they are calcium-dependent cytoplasmic cysteine proteases that regulate a variety of physiological processes. We obtained the sequence of CAPN3 from an NGS-based analysis of Pagrus major (PmCAPN3) and confirmed the conserved molecular biological properties in the predicted amino acid sequence. The amino acid sequence and predicted domains of CAPN3 were found to be highly conserved in all of the examined species, and one catalytic domain and four calcium binding sites were identified. In healthy P. major, the PmCAPN3 mRNA was most abundantly expressed in the muscle and skin, and ubiquitously expressed in the other tissues used in the experiment. After artificial infections with fish pathogens, significant changes in its expression levels were found in immune-related tissues, most of showed upregulation. In particular, the highest level of expression was found in the liver, a tissue associated with protease activity. Taken together, these results suggest a physiological activity for PmCAPN3 in P. major and reveal functional possibilities that have not yet been reported in the immune system.


Assuntos
Calpaína/genética , Calpaína/imunologia , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Dourada/genética , Dourada/imunologia , Imunidade Adaptativa/genética , Sequência de Aminoácidos , Animais , Calpaína/química , DNA Complementar/genética , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Filogenia , RNA Mensageiro/genética , Alinhamento de Sequência/veterinária , Análise de Sequência de DNA/veterinária
13.
Fish Shellfish Immunol ; 98: 112-121, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31904542

RESUMO

Apart from mitigating endoplasmic reticulum (ER) stress, vast studies have demonstrated the crucial role of inositol-requiring transmembrane kinase and endonuclease 1α (IRE1α) - spliced X-box binding protein 1 (XBP1s) signaling pathway in inflammatory response in mammals. In addition, palmitic acid (PA)-induced inflammation has been verified in large yellow croaker (Larimichthys crocea). However, whether the IRE1α-XBP1s signaling pathway is involved in inflammatory response caused by PA remains poorly studied in fish. The present study was aimed at elucidating the role of the IRE1α-XBP1s signaling pathway in inflammatory response induced by PA in primary hepatocytes from large yellow croaker. In the present study, the full-length cDNA of ire1α and xbp1s were cloned and comprised 3793 bp and 1789 bp with an open reading frame of 3279 bp and 1170 bp, encoding 1093 and 390 amino acids, respectively. IRE1α protein possessed a protein kinase and endoribonuclease domain and XBP1s protein possessed a basic-leucine zipper domain. The IRE1α protein and XBP1s protein located to the ER membrane and nucleus respectively. The ire1α and xbp1s were widely transcribed in various tissues with the higher level in intestine, liver, adipose and head kidney. The ER stress-inducing agent tunicamycin (Tm) and PA treatment significantly activated the IRE1α-XBP1s signaling pathway and increased the pro-inflammatory genes expression including tumor necrosis factor α (tnfα), interleukin 6 (il-6) and interleukin 1ß (il-1ß) (P < 0.05). When KIRA6, the IRE1α kinase inhibitor, was used to block the IRE1α-XBP1s signaling pathway, the Tm and PA-induced pro-inflammatory genes expression was significantly suppressed (P < 0.05). These data indicated that the IRE1α-XBP1s signaling pathway was involved in the PA-induced inflammatory response in large yellow croaker.


Assuntos
Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Perciformes/genética , Perciformes/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Proteínas de Peixes/química , Perfilação da Expressão Gênica/veterinária , Filogenia , Proteínas Serina-Treonina Quinases/química , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/imunologia , Alinhamento de Sequência/veterinária
14.
Fish Shellfish Immunol ; 98: 167-175, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31917321

RESUMO

Granulocyte colony-stimulating factor (GCSF) is a growth factor that drives the proliferation and differentiation of granulocytes and monocytes/macrophages. Currently, two copies of GCSF, named GCSFa and GCSFb, have been identified in teleost fish, but data on the functions and signal pathways of these fish GCSFs are still limited. In the present study, a GCSFa homologue (LcGCSFa) was identified from large yellow croaker (Larimichthys crocea). The open reading frame (ORF) of LcGCSFa is 636 bp long and encodes a protein of 211 amino acids (aa), with a 19-aa signal peptide and a typical IL-6 domain, conserved in fish GCSF sequences. The phylogenetic analysis showed that LcGCSFa clustered with other fish GCSFa homologues. LcGCSFa was constitutively expressed in all tissues tested and significantly up-regulated in head kidney and spleen by Vibrio alginolyticus or poly(I:C). LcGCSFa transcripts were also detected in primary head kidney leucocytes (PKL), primary head kidney macrophages (PKM), and primary head kidney granulocytes (PKG), and significantly up-regulated in PKL and PKG by LPS or poly(I:C). These data indicated that LcGCSFa may be involved in the immune responses induced by bacterium and virus. The recombinant LcGCSFa protein (rLcGCSFa) produced in Pichia pastoris promoted the proliferation of PKL both in vivo and in vitro. Furthermore, rLcGCSFa significantly increased both transcription and phosphorylation levels of the signal transducers and activators of transcription (STAT) proteins (LcSTAT3 and LcSTAT5) in PKL, which are required for the GCSF-dependent proliferation. These results showed that LcGCSFa may promote the proliferation of PKL via the activation of LcSTAT3 and LcSTAT5, suggesting a conserved role across vertebrate GCSFs.


Assuntos
Imunidade Adaptativa/genética , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Fator Estimulador de Colônias de Granulócitos/genética , Fator Estimulador de Colônias de Granulócitos/imunologia , Imunidade Inata/genética , Perciformes/genética , Perciformes/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Fator Estimulador de Colônias de Granulócitos/química , Filogenia , Alinhamento de Sequência/veterinária
15.
Can J Vet Res ; 84(1): 18-23, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31949325

RESUMO

Feline infectious peritonitis (FIP) is a fatal disease for which no simple antemortem diagnostic assay is available. A new polymerase chain reaction (PCR) test has recently been developed that targets the spike protein region of the FIP virus (FIPV) and can identify specific mutations (M1030L or S1032A), the presence of which indicates a shift from feline enteric coronavirus (FeCV) to FIPV. This test will only be useful in the geographical region of interest, however, if the FIP viruses contain these mutations. The primary objective of this study was to determine the presence of the M1030L or S1032A mutations in FeCV derived from stool samples from a selected group of healthy cats from households and shelters and determine how many of these cats excrete FeCV. The secondary objective was to evaluate how often these specific FIPV mutations were present in tissue samples derived from cats diagnosed with FIP at postmortem examination. Feline enteric coronavirus (FeCV) was detected in 46% of fecal samples (86/185), all were FeCV type 1, with no difference between household or shelter cats. Only 45% of the FIPV analyzed contained the previously reported M1030L or S1032A mutations. It should be noted that, as the pathological tissue samples were opportunistically obtained and not specifically obtained for PCR testing, caution is warranted in interpreting these data.


Assuntos
Doenças do Gato/virologia , Coronavirus Felino/química , Peritonite Infecciosa Felina/diagnóstico , Glicoproteína da Espícula de Coronavírus/genética , Alberta/epidemiologia , Sequência de Aminoácidos , Animais , Doenças do Gato/diagnóstico , Doenças do Gato/epidemiologia , Gatos , Coronavirus Felino/classificação , Coronavirus Felino/genética , Coronavirus Felino/isolamento & purificação , Análise Mutacional de DNA/veterinária , Fezes/virologia , Peritonite Infecciosa Felina/epidemiologia , Peritonite Infecciosa Felina/virologia , Feminino , Funções Verossimilhança , Masculino , Mutação , Filogenia , Reação em Cadeia da Polimerase/veterinária , Prevalência , RNA Viral/genética , RNA Viral/isolamento & purificação , Saskatchewan/epidemiologia , Alinhamento de Sequência/veterinária , Glicoproteína da Espícula de Coronavírus/química
16.
Vet Microbiol ; 241: 108563, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31928703

RESUMO

Complement component 1, q subcomponent binding protein (C1QBP) is a receptor for the globular heads of C1q and modulates various biological processes including infection, inflammation, autoimmunity, and cancer. In our previous study to identify differentially expressed secretory proteins in Marc-145 cells infected with porcine reproductive and respiratory syndrome virus (PRRSV), mass spectrum data showed that C1QBP was secreted after PRRSV infection. However, the biological significance of secreted C1QBP remains unclear. In this study, we confirmed that PRRSV infection promoted C1QBP secretion in Marc-145 cells and porcine alveolar macrophages (PAMs), the target cells of PRRSV in vivo. Knockdown of endogenous C1QBP decreased PRRSV-induced inflammatory responses. The purified recombinant porcine C1QBP (poC1QBP) had proinflammatory effects. The exogenous addition of poC1QBP significantly enhanced PRRSV-induced inflammatory responses and abolished the inhibitory effects mediated by poC1QBP-knockdown. Taken together, these results demonstrate that PRRSV infection promotes poC1QBP secretion that enhances inflammatory responses.


Assuntos
Complemento C1q/metabolismo , Proteínas Mitocondriais/metabolismo , Síndrome Respiratória e Reprodutiva Suína/metabolismo , Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Sequência de Aminoácidos , Animais , Western Blotting/veterinária , Linhagem Celular , Chlorocebus aethiops , Clonagem Molecular , Complemento C1q/genética , Citocinas/metabolismo , DNA Complementar/genética , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Regulação Viral da Expressão Gênica , Técnicas de Silenciamento de Genes , Inflamação/metabolismo , Rim/citologia , Rim/virologia , Macrófagos Alveolares/virologia , Síndrome Respiratória e Reprodutiva Suína/patologia , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Alinhamento de Sequência/veterinária , Suínos
17.
Anim Biotechnol ; 31(1): 17-24, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30570352

RESUMO

GTP binding protein overexpressed in skeletal muscle (GEM) is an important gene with many functions, such as regulating the rearrangement of cytoskeleton and the activity of voltage-dependent calcium channel, and GEM was regarded as a candidate gene for obesity. However, little investigation has been carried out to explore whether GEM affected the intramuscular fat (IMF) deposition of goat. To explore the role of GEM gene in goat, this gene was cloned and its tissue and temporal expression profile were detected. Effect of GEM on adipogenesis was examined by losing function of GEM in vitro. Thereafter, several lipid metabolism-related genes were examined, including CCAAT/enhancing-binding protein α (C/EBPα), CCAAT/enhancing-binding protein ß (C/EBPß), lipoprotein lipase (LPL), preadipocyte factor 1 (Pref-1), peroxisome proliferator activated receptor γ (PPARγ) and sterol regulatory element binding protein 1 (SREBP1). We found that the goat GEM gene consisted of 936 bp, which encoded a protein of 311 amino acids. The expression of GEM was higher in spleen, lung and large intestine and it appeared sharp in the interim stage of differentiation. Furthermore, GEM knockdown blocked adipogenesis and the expression of C/EBPα, C/EBPß, LPL, PPARγ and SREBP1. These results indicated that GEM might promote lipid accumulation and adipogenesis.


Assuntos
Adipogenia/genética , Proteínas de Ligação ao GTP/genética , Cabras/genética , Metabolismo dos Lipídeos/genética , Adipócitos/fisiologia , Sequência de Aminoácidos , Animais , Diferenciação Celular , Técnicas de Silenciamento de Genes/veterinária , Cabras/fisiologia , Intestino Grosso/fisiologia , Pulmão/fisiologia , Masculino , Músculo Esquelético/fisiologia , Alinhamento de Sequência/veterinária , Baço/fisiologia
18.
Fish Shellfish Immunol ; 96: 279-289, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31783148

RESUMO

The interferon-induced GTP-binding protein Mx is responsible for a specific antiviral state against a broad spectrum of viral infections that are induced by type-I interferons (IFN α/ß) in different vertebrates. In this study, the Mx gene was isolated from the constructed mullet cDNA database. Structural features of mullet Mx (MuMx) were analyzed using different in-silico tools. The pairwise comparison revealed that the MuMx sequence was related to Stegastes partitus Mx with an 83.7% sequence identity, whereas MuMx was clustered into the teleost category in the phylogentic analysis. Sequence alignment showed that the dynamin-type guanine nucleotide-binding domain (G_DYNAMIN_2), central interactive domain (CID), and GTPase effector domain (GED) were conserved among Mx counterparts. The transcriptional expression of MuMx was the highest in blood cells from unchallenged fish. The temporal mRNA profile showed that MuMx expression was significantly elevated in all tissues, including blood, spleen, head kidney, liver, and gills after the injection of polyinosinic-polycytidylic acid (poly I:C) at many time points. Moreover, MuMx expression increased slightly, in the blood, spleen, and head kidney at a few time points after the injection of lipopolysaccharide (LPS) and Lactococcus garvieae (L. garvieae). Results of the subcellular localization analysis confirmed that the MuMx protein was highly expressed in the cytoplasm. The analysis of the gene expression of the viral hemorrhagic septicemia virus (VHSV) under conditions of MuMx overexpression confirmed the significant inhibition of viral transcripts. The cell viability (MTT) assay and VHSV titer quantification with the presence of MuMx indicated a significant reduction in virus replication. Collectively, these findings suggest that Mx is a specific immune-related gene that elicits crucial antiviral functions against viral antigens in the mullet fish.


Assuntos
Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Proteínas de Resistência a Myxovirus/genética , Proteínas de Resistência a Myxovirus/imunologia , Smegmamorpha/genética , Smegmamorpha/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Expressão Gênica , Perfilação da Expressão Gênica/veterinária , Infecções por Bactérias Gram-Positivas/imunologia , Infecções por Bactérias Gram-Positivas/veterinária , Lactococcus/fisiologia , Lipopolissacarídeos/farmacologia , Proteínas de Resistência a Myxovirus/química , Novirhabdovirus/fisiologia , Filogenia , Poli I-C/farmacologia , Infecções por Rhabdoviridae/imunologia , Infecções por Rhabdoviridae/veterinária , Alinhamento de Sequência/veterinária
19.
Fish Shellfish Immunol ; 98: 773-787, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31734286

RESUMO

Interleukin (IL)-4 and IL-13 play a central role in T helper 2 immune response in mammals. The cell signalling is mediated by the type I heterodimeric receptor containing the IL-4Rα and γC chains, and the type II receptors formed by IL-4Rα and IL-13Rα1. In salmonid species, three paralogues of the IL-4 and IL-13 cytokines have been reported, il-4/13a, il-4/13b1 and il-4/13b2. In regard to receptors, two paralogues of each IL-4/13 receptor chains have been identified in rainbow trout while five genes named γc1, il-4rα, il-13rα1a, il-13rα1b, and il-13rα2 have identified in Atlantic salmon. Since Atlantic salmon is an important farmed fish species, the aim of this work was to get new insights into distribution, structure and expression regulation of the IL-4/13 receptors in salmon. By using qRT-PCR, it was shown that all γc1, il-4rα, il-13rα1a, il-13rα1b, and il-13rα2 receptor chains were expressed in lymphoid and non-lymphoid tissues of healthy salmon, nonetheless γC expression was higher in lymphoid than non-lymphoid tissues. The in silico structural analysis and homology modelling of the predicted receptor proteins showed that domains and most motifs present in the superior vertebrate chains are conserved in salmon suggesting a conserved role for these receptor chains. Only IL-13Rα1B is a receptor chain with a unique structure that seem not to be present in higher vertebrates but in fish species. In order to determine the regulatory role of IL-4/13 on the expression of receptor chains, Atlantic salmon il-4/13A gene was synthetized and cloned in pET15b. The recombinant IL-4/13A was produced in E. coli and the activity of the purified cytokine was confirmed in vitro. The regulatory role of IL-4/13A on the expression of their potential receptors was tested in salmon receiving the recombinant cytokine and effects were compared with those of the control group. The results showed that IL-4/13A induced the expression of its own gene and GATA-3, in the head kidney of fish but not in the spleen, while IL-10 increased in both lymphoid organs indicating a regulatory role of this cytokine on the induction of Th2 responses in salmon. IFN-γ and MHC class II were also later induced in head kidney. In regard to the expression of the receptor chains, IL-4/13A upregulated the expression of γC, IL-13Rα1A and IL-13Rα2A in the spleen but not in the head kidney of salmon, indicating a role on the modulation of cell signalling for the Th2 response. Furthermore, Piscirickettsia salmonis infection of Atlantic salmon occurred with an increase of γC and IL-13Rα1A suggesting a potential role of the IL-4/13 system in bacterial immunity or pathogenesis. This study contributes to a better understanding of the IL-4/13A system in salmon, which as a key axis for Th2 response may be involved not only in pathogen elimination but also in adaptive immune repair that seems critical tolerance to infectious diseases.


Assuntos
Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Receptores Tipo II de Interleucina-4/genética , Receptores Tipo II de Interleucina-4/imunologia , Salmo salar/genética , Salmo salar/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Família Multigênica , Filogenia , Receptores Tipo II de Interleucina-4/química , Alinhamento de Sequência/veterinária
20.
Transbound Emerg Dis ; 67(2): 476-480, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31536676

RESUMO

Visceral leishmaniasis is an endemic zoonotic disease identified especially in developing territories. Brazil's northeast, southeast and midwest have been endemic for several years; currently, the infection is spreading to the south. Dogs are the main reservoirs; however, other mammal species have also been infected. Herein, we have identified the infecting Leishmania species in dogs and horses from the south of Brazil, a new outbreak of the infection. Blood samples were collected in the urban area of Uruguaiana city. DNA was extracted from peripheral blood, kinetoplast DNA (kDNA) and ribosomal DNA (rDNA) fragments were obtained by polymerase chain reaction (PCR) and sequenced. Out of 123 samples, 25 of them (14 dogs and 11 horses) were positive for Leishmania spp. Sequence alignment and phylogenetic analysis revealed that the kDNA in positive samples was similar to four species previously reported: L. infantum/L. chagasi, L. donovani, L. major. Despite kDNA minicircles regions are very useful due to high sensitivity to Leishmania spp. DNA detection, the sequence polymorphism among minicircles can be an obstacle to interspecific differentiation. Our results suggest that these strains are circulating in Brazil south region cross-border and indicate the susceptibility of new outbreak for visceral leishmaniasis infection in horses domiciled in endemic region for canine and human visceral leishmaniasis.


Assuntos
Reservatórios de Doenças/parasitologia , Doenças do Cão/epidemiologia , Doenças dos Cavalos/epidemiologia , Leishmania donovani/genética , Leishmaniose Visceral/veterinária , Zoonoses/epidemiologia , Animais , Brasil/epidemiologia , DNA de Cinetoplasto/genética , Doenças do Cão/parasitologia , Cães , Feminino , Doenças dos Cavalos/parasitologia , Cavalos , Humanos , Leishmania donovani/isolamento & purificação , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/parasitologia , Masculino , Mamíferos , Filogenia , Reação em Cadeia da Polimerase/veterinária , Alinhamento de Sequência/veterinária , Zoonoses/parasitologia
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