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1.
Dis Aquat Organ ; 144: 107-115, 2021 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-33884959

RESUMO

Scuticociliatosis, caused by ciliated protozoa of the subclass Scuticociliatia, has been associated with high mortalities in marine fish. Environmental factors such as an increase in water temperature can enhance this disease. The aim of the present report is to describe the occurrence of a cluster of cases of scuticociliatosis in a multispecies marine cold-water system in a public aquarium. Philasterides dicentrarchi was identified by PCR in formalin-fixed tissues of some of the fish showing meningitis or meningoencephalitis, dermatitis and myositis with intralesional protozoa. An increase in water temperature of approximately 2°C was identified as a potential contributing factor for this cluster of infections. Higher temperature may have enhanced the propagation or pathogenicity of scuticociliates or increased host susceptibility of some species of fish, especially wolf-eel Anarrhichthys ocellatus and spotted ratfish Hydrolagus colliei. This report also highlights the complexity of dealing with mixed species systems housing fish from different natural ecozones.


Assuntos
Infecções por Cilióforos , Cilióforos , Doenças dos Peixes , Linguados , Oligoimenóforos , Animais , Infecções por Cilióforos/veterinária , Doenças dos Peixes/epidemiologia , Oligoimenóforos/genética , Reação em Cadeia da Polimerase/veterinária
2.
Exp Parasitol ; 223: 108081, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33549536

RESUMO

Artificial breeding of small yellow croaker (Larimichthys polyactis) was recently achieved, providing a bright future for its commercial farming. In May 2019, a disease outbreak occurred among small yellow croakers in an aquaculture farm near Xiangshan Bay, charactering by white spots spotted on the surface of fish skin, gills and fins. The parasite was preliminarily identified as Cryptocaryon irritans based on morphological feature of the parasite and the symptoms on fish. However, the previously published specific primer pairs failed to confirm the existence of C. iriitans. Six nucleotides mismatches were discovered after mapping specific forward primer back to targeted gene. Therefore, an updated PCR specific primer was developed within the 9th highly variable region of 18S rRNA gene and conserved in all C. irritans sequences available in GenBank database. The specificity was verified in silico by Primer-BLAST against GenBank nucleotide. Laboratory cultured ciliates (Mesanophrys, Pseudokeronopsis and Uronema) as well as natural microbial community samples collected from sea water and river water was used as negative control to verify the specificity of the primer in situ. Besides, tank transfer method was used to evaluate the treatment of the parasite infection. By tank transfer method, 2.00 ± 0.61 out of 10 fish that already sever infected were successfully survived after 8 days treatment, meanwhile the control group died out at d 6. More loss to the treatment group during first five days was observed and may attribute to the combined effect from infection and stress the recent domesticated fish suffered during rotation. Therefore, tank transfer method was also effective to prevent small yellow croaker from further infection, however the loss of the small yellow croaker suffered from stress during rotation also needs to be carefully concerned. In conclusion, this study reported the first diagnose of C. irritans infection on small yellow croaker, provided updated specific primer to detect C. irritans infection on fish body and reported the effect of tank transfer on small yellow croaker treatment.


Assuntos
Infecções por Cilióforos/veterinária , Cilióforos/isolamento & purificação , Doenças dos Peixes/parasitologia , Perciformes/parasitologia , Nadadeiras de Animais/parasitologia , Nadadeiras de Animais/patologia , Animais , China/epidemiologia , Cilióforos/classificação , Cilióforos/genética , Infecções por Cilióforos/diagnóstico , Infecções por Cilióforos/epidemiologia , Infecções por Cilióforos/parasitologia , Surtos de Doenças/veterinária , Doenças dos Peixes/diagnóstico , Doenças dos Peixes/epidemiologia , Doenças dos Peixes/terapia , Pesqueiros , Brânquias/parasitologia , Brânquias/patologia , Músculo Esquelético/parasitologia , Músculo Esquelético/patologia , Filogenia , RNA Ribossômico 18S/genética , Pele/parasitologia , Pele/patologia , Especificidade da Espécie
3.
J Fish Dis ; 44(3): 249-262, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33314157

RESUMO

Takifugu rubripes and Dicentrarchus labrax are important commercial fish in China that are under serious threat from Cryptocaryon irritans. C. irritans is a ciliated obligate parasite that causes marine white spot disease and leads to heavy economic losses. We analysed the transcriptome in the gills of T. rubripes and D. labrax to compare differentially expressed genes (DEGs) and pathways during infection with C. irritans. In total, we identified 6,901 and 35,736 DEGs from T. rubripes and D. labrax, respectively. All DEGs were annotated into GO terms; 6,901 DEGs from T. rubripes were assigned into 991 sub-categories, and 35,736 DEGs from D. labrax were assigned into 8,517 sub-categories. We mapped DEGs to the KEGG database and obtained 153 and 350 KEGG signalling pathways from T. rubripes and D. labrax, respectively. Immune-related categories included Toll-like receptors, MAPK, lysosome, C-type lectin receptor and NOD-like receptor signalling pathways were significantly enriched pathways. In immune-related signalling pathways, we found that AP-1, P38, IL-1ß, HSP90 and PLA were significantly up-regulated DEGs in T. rubripes, but P38 and PLA were significantly down-regulated in D. labrax. In this study, transcriptome was used to analyse the difference between scaly and non-scaly fish infection by C. irritans, which not only provided a theoretical basis for the infection mechanism of C. irritans, but also laid a foundation for effectively inhibiting the occurrence of this disease. Our work provides further insight into the immune response of host resistance to C. irritans.


Assuntos
Infecções por Cilióforos/veterinária , Doenças dos Peixes/parasitologia , Perfilação da Expressão Gênica , Animais , Bass , Infecções por Cilióforos/genética , Infecções por Cilióforos/imunologia , Doenças dos Peixes/genética , Doenças dos Peixes/imunologia , Brânquias/imunologia , Brânquias/parasitologia , Hymenostomatida/fisiologia , Transdução de Sinais , Takifugu
4.
PLoS One ; 15(8): e0238167, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32857792

RESUMO

The ciliated protozoan Cryptocaryon irritans infects a wide range of marine fish and causes the highly lethal white spot disease. This parasite possesses three morphologically and physiologically distinct life stages: an infectious theront, a parasitic trophont, and an asexually reproductive tomont. In the past few years, several attempts have been made to help elucidate how C. irritans transforms from one stage to another using transcriptomic or proteomic approaches. However, there has been no research studying changes in transcription profiles between different time points of a single C. irritans life stage-the development of this parasite. Here we use RNA-seq and compare gene expression profiles of theront cells collected by 1 and 10 hrs after they emerged from tomonts. It has been shown that infectivity of theront cells declines 6-8 hours post-emergence, and we used this characteristic as a physiological marker to confirm the aging of theront cells. We identified a total of 41 upregulated and 90 downregulated genes that were differentially expressed between young and aging theront cells. Using Blast2Go to further analyze functions of these genes, we show that genes related to energy production are downregulated, but quite surprisingly many genes involved in transcription/translation processes are upregulated. We also show that expression of all nine detectable agglutination/immobilization antigen genes, with great sequence divergence, is invariably downregulated. Functions of other differentially expressed genes and indications are also discussed in our study.


Assuntos
Envelhecimento/metabolismo , Cilióforos/metabolismo , Cilióforos/patogenicidade , Animais , Cilióforos/genética , Cilióforos/crescimento & desenvolvimento , Infecções por Cilióforos/veterinária , Doenças dos Peixes/parasitologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Perciformes , Análise de Sequência de RNA , Transcriptoma
5.
Parasitol Res ; 119(9): 3075-3081, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32656656

RESUMO

Balantioides coli (syn. Balantidium coli) is an important zoonotic but usually neglected protozoa infecting human and a great number of animals, and the pig was considered to be the most important natural host and reservoir. However, no information about the infection of B. coli in pigs in northwestern China was available. In the present study, the prevalence and genetic diversity of B. coli in pigs in Shaanxi province were investigated. A total of 560 fecal samples were collected from pigs of four age groups in five different geographical regions and analyzed by using PCR targeting the ITS1-5.8S rRNA-ITS2 gene fragment. The infection of B. coli was detected in all age groups and regions, with the total prevalence of 16.8% (94/560). Significant differences (P < 0.01) in prevalence were found among four investigated age groups, with the highest in fatteners (38.8%) and the lowest in adults (5.7%). The prevalence was also significantly (P < 0.01) different among pigs from five sampling regions. Sequence analysis revealed two genetic variants, namely, A and B, in these investigated pigs, and both of them were detected in all age groups and regions, with the latter as the predominant one. Further, sixty-eight different haplotypes were found, with 19 and 49 belonged to genetic variants A and B, respectively. The findings in the present study indicated wide distribution and high diversity of B. coli in pigs in Shaanxi province and provided fundamental data for implementing control strategies on B. coli infection in pigs as well as other hosts in this province.


Assuntos
Infecções por Cilióforos/veterinária , Doenças dos Suínos/parasitologia , Trichostomatida/genética , Animais , China/epidemiologia , Infecções por Cilióforos/epidemiologia , Infecções por Cilióforos/parasitologia , Fezes/parasitologia , Prevalência , Suínos , Doenças dos Suínos/epidemiologia , Trichostomatida/classificação , Trichostomatida/isolamento & purificação
6.
Ann Parasitol ; 66(2): 227-230, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32592544

RESUMO

A total of 578 specimens of common carp (Cyprinus carpio) from eight the most significant and larger cyprinid aquaculture facilities in Macedonia (fish farms and cage culture systems) were examined for parasitological investigation. Protozoa Apiosoma piscicola was found in cage culture system on Globochica reservoir. In this fish farm, a total of 127 fish samples were examined for parasitological investigation, in which parasite infestation with A. piscicola was found on fins and gills in 79 specimens of common carp, in winter season. The prevalence of A. piscicola in common carp was 62.20%, while the mean intensity was 17.58. Our findings of A. piscicola in common carp (C. carpio) are first recorded in Macedonia. At the same time, common carp represent new host for A. piscicola in Macedonian waters.


Assuntos
Carpas , Infecções por Cilióforos/veterinária , Cilióforos , Doenças dos Peixes , Animais , Aquicultura , Carpas/parasitologia , Cilióforos/classificação , Cilióforos/fisiologia , Infecções por Cilióforos/epidemiologia , Doenças dos Peixes/epidemiologia , Doenças dos Peixes/parasitologia , República da Macedônia do Norte/epidemiologia
7.
Parasitol Int ; 76: 102093, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32120050

RESUMO

Miamiensis avidus causes scuticociliatosis in cultured olive flounders (Paralichthys olivaceus), leading to economic losses in aquaculture in Korea. Quantitative evaluation of the viability of M. avidus is important to develop an effective vaccine or chemotherapeutic agent against it. We used a colorimetric assay based on the reduction of 2-(4-Iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium (WST-1) to quantify the viability of M. avidus. Using this method, we investigated the effect of protease inhibitors on the viability of M. avidus. The assay showed a clear difference in the optical density (OD) of over 104 ciliates, and the metalloprotease inhibitors 1, 10-phenanthroline and ethylenediaminetetraacetic acid (EDTA) reduced the viability of M. avidus by more than 90% when used at concentration of 5 mM and 100 µM, respectively. However, different morphological changes in the parasite were observed when exposed to these two inhibitors. These results indicate that the WST-1 assay is a simple and reliable method to quantify the viability of M. avidus, and metalloproteases are excellent targets for the development of agents and vaccines to control M. avidus infection.


Assuntos
Antiparasitários/farmacologia , Infecções por Cilióforos/veterinária , Colorimetria/veterinária , Doenças dos Peixes/parasitologia , Oligoimenóforos/efeitos dos fármacos , Inibidores de Proteases/farmacologia , Sais de Tetrazólio/química , Animais , Infecções por Cilióforos/parasitologia , Colorimetria/instrumentação , Viabilidade Microbiana/efeitos dos fármacos , Oligoimenóforos/fisiologia
8.
Fish Shellfish Immunol ; 99: 291-300, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32058095

RESUMO

Phagocytic cells are activated to produce a large amount of reactive oxygen species (ROS) that kill pathogens quickly and efficiently through oxidation. NADPH oxidase is the main source of intracellular ROS. In the present study, five subunits of the phagocytic NADPH oxidase complex were identified in orange-spotted grouper (Epinephelus coioides). The open reading frame of grouper gp91phox, p22phox, p67phox, p47phox, and p40phox were 1,698 bp, 564 bp, 1,497 bp, 1,290 bp, and 1,050 bp, respectively, and encoded 565, 187, 498, 429, and 349 amino acids. Evolutionary analysis indicated that these proteins are evolutionarily homologous to the corresponding proteins of other fish and mammals, and contain conserved functional domains and sites that are important in mammals. In addition, real-time polymerase chain reaction analysis showed that the expression of these five genes was higher in immune-related tissues in normal grouper, and that these genes were up-regulated in gill and spleen after C. irritans infection, which suggests that these genes may be involved in the defense against C. irritans infection.


Assuntos
Infecções por Cilióforos/veterinária , Doenças dos Peixes/parasitologia , NADPH Oxidases/metabolismo , Perciformes/metabolismo , Sequência de Aminoácidos , Animais , Cilióforos , Infecções por Cilióforos/imunologia , Infecções por Cilióforos/metabolismo , Clonagem Molecular , Biologia Computacional , Doenças dos Peixes/imunologia , Doenças dos Peixes/metabolismo , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Regulação Enzimológica da Expressão Gênica , NADPH Oxidases/genética , Filogenia
9.
Rev Bras Parasitol Vet ; 29(1): e018319, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32049149

RESUMO

Scuticociliatosis, caused by an opportunistic ciliate protozoan, is responsible for significant economic losses in marine ornamental fish. This study reports the occurrence of Uronema spp., parasitizing ten species of marine reef fish at an ornamental fish wholesaler: Blue green damselfish (Chromis viridis), Vanderbilt's Chromis (Chromis vanderbilti), Pennant coralfish (Heniochus acuminatus), Threespot angelfish (Apolemichthys trimaculatus), Goldspotted angelfish (Apolemichthys xanthopunctatus), Sea goldie (Pseudanthias squamipinnis), Orchid dottyback (Pseudochromis fridmani), Threadfin butterflyfish (Chaetodon auriga), Vagabond butterflyfish (Chaetodon vagabundus), and Bluecheek butterflyfish (Chaetodon semilarvatus). Diseased fish showed disorders such as hemorrhages and ulcerative lesions on the body surface. Histopathological analysis of the muscle, liver, gut, kidney, spleen, gills, and stomach revealed hemorrhages and degeneration of muscle fiber, vacuolar degeneration of hepatocyte, inflammatory process and granuloma in the liver, atrophy of intestinal villi, inflammatory process and granuloma in the kidney, melanomacrophage centers, as well as inflammatory process in the spleen, epithelial cells hyperplasia and granuloma formation in the gills, and vacuolar degeneration and eosinophils in the stomach. Due to the severity of the disease, it is necessary to implement biosecurity measures with rapid and accurate diagnosis to minimize the risk of economic losses caused by Uronema spp.


Assuntos
Infecções por Cilióforos/veterinária , Cilióforos/classificação , Doenças dos Peixes/parasitologia , Perciformes/parasitologia , Animais , Brasil , Infecções por Cilióforos/parasitologia , Infecções por Cilióforos/patologia , Doenças dos Peixes/patologia
10.
Fish Shellfish Immunol ; 99: 462-472, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32070786

RESUMO

Large yellow croaker (Larimichthys crocea) is one of the most important mariculture fish in China. In the past decades, cryptocaryonosis caused by Cryptocryon irritans has led to huge economic losses, posing great threat to the healthy and sustainable development of L. crocea mariculture industry. As the largest immunologically active mucosal organ in fish, skin provides the first defense line against external pathogens. To better understand the gene expression dynamics, the large yellow croakers were artificially infected with C. irritans and their skin tissues were collected at 0 h, 24 h, 48 h, 72 h and 96 h post infection. The total RNA in the skin tissues were extracted and the transcriptome were sequenced. After sequencing, a total of 1,131, 311, 140 million high quality RNA-seq reads were collected. A set of 215, 473, 968, 1055 differentially expressed genes were identified at 24 h, 48 h, 72 h and 96 h post infection respectively. Further analysis clustered these DEGs into six profiles and 75 hub genes for six profiles were identified. Among these hub genes, 18 immune related genes including TLR5, TOPK, NFKBIZ, MAPK14A were identified post C. irritans infection. Cytokine-cytokine receptor interaction was the only pathway that significantly enriched at four timepoints post infection. This study provides an in-depth understanding of skin transcriptome variance of large yellow croaker after C. irritans infection, which would be helpful for further understanding of the molecular mechanism of L. crocea in response to C. irritans infection.


Assuntos
Infecções por Cilióforos/veterinária , Proteínas de Peixes/genética , Hymenostomatida/imunologia , Perciformes/parasitologia , Pele/parasitologia , Transcriptoma , Animais , Infecções por Cilióforos/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/parasitologia , Proteínas de Peixes/imunologia , Expressão Gênica , Perfilação da Expressão Gênica , Doenças Parasitárias em Animais/imunologia , Perciformes/imunologia , Análise de Sequência de DNA , Pele/imunologia
11.
Fish Shellfish Immunol ; 99: 52-58, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31935553

RESUMO

Cryptocaryon irritans infection could cause huge economic losses to the marine fish industry. Larimichthys crocea, a special economic species in China, suffered from the threat of serious infection, and L. crocea could enhance the level of piscidin 5-like to defense against the infection. This study set out to observe the main histopathological changes of some key tissues caused by infection, and determineed how an ectoparasite affected the expression of piscidin-5 like in its hosts. Pathological changes and immune response were assessed using histological and in situ hybridization (ISH) technologies. The infection induced inflammation occurring, especially in the gill where epithelium cells swell, hyperplasia, necrosis shedding adjacent to the parasites attachment sites. Infected hepatic cells grown big vacuoles in the cytoplasm. The boundary between red pulp and white pulp turned indistinct, splenic corpuscle lost the normal structure, the number and size of melano-macrophage centers increased apparently in the infected spleen. The whole structure of head kidney became loose. Immunostaining with RNA probes against piscidin 5-like showed subpopulations of mast cells (MCs) were positive. Piscidin 5-like-positive MCs existed mainly in the head kidney where they distributed around melano-macrophage center, followed in the gill located at different positions they also distributed in the margin of spleen, and randomly and sparsely existed in the liver. After being infected by C. irritans, the gill arch arose positive MCs groups, and they also migrated to spleen, while the positive staining deepen in other detected tissues. Therefore, organism enhanced the expression level through improving expression ability of positive MCs, or increasing the number of positive MCs.


Assuntos
Peptídeos Catiônicos Antimicrobianos/genética , Infecções por Cilióforos/veterinária , Ectoparasitoses/veterinária , Doenças dos Peixes/parasitologia , Proteínas de Peixes/genética , Perciformes/parasitologia , Animais , China , Infecções por Cilióforos/patologia , Ectoparasitoses/patologia , Doenças dos Peixes/patologia , Hymenostomatida/patogenicidade , Mastócitos/imunologia
12.
Fish Shellfish Immunol ; 96: 107-113, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31805410

RESUMO

In fish, interferon (IFN) regulatory factor 2 (IRF2) is a regulator of the type I IFN-dependent immune response, thereby playing a crucial role in innate immunity. However, the specific mechanism by which IRF2 regulates type II IFN in fish remains unclear. In the present study, first, to analyse the potential role of golden pompano (Trachinotus ovatus) IRF2 (ToIRF2) in the immune response, the mRNA level of ToIRF2 was detected by quantitative real-time polymerase chain reaction (qRT-PCR) after parasite infection. ToIRF2 was upregulated at early time points in both local infection sites (skin and gill) and system immune tissues (liver, spleen, and head-kidney) after stimulation with Cryptocaryon irritans. Second, to investigate the modulation effect of ToIRF2 on type II IFN (interferon gamma, IFNγ) expression, a promoter analysis was performed using progressive deletion mutations of ToIFNγ. The expression level of IFNγ-5 was highest among the five truncated mutants in response to ToIRF2, indicating that the core promoter region was located from -189 bp to +120 bp, which included the IRF2 binding sites. Mutation analyses showed that the activity of the ToIFNγ promoter dramatically decreased after the targeted mutation of the M1, M2 or M3 binding sites. Additionally, electrophoretic mobile shift assay (EMSA) confirmed that IRF2 interacted with the M1 binding site in the ToIFNγ promoter region to dominate ToIFNγ expression. Finally, overexpressing ToIRF2 in vitro notably increased ToIFNγ and the transcription of several type II IFN/IRF-based signalling pathway genes. These results suggested that ToIRF2 might be involved in the host defence against C. irritans infection and contribute to a better understanding of the transcriptional mechanisms by which ToIRF2 regulates type II IFN in fish.


Assuntos
Doenças dos Peixes/imunologia , Peixes/genética , Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Fator Regulador 2 de Interferon/genética , Fator Regulador 2 de Interferon/imunologia , Animais , Sequência de Bases , Cilióforos/fisiologia , Infecções por Cilióforos/imunologia , Infecções por Cilióforos/parasitologia , Infecções por Cilióforos/veterinária , Doenças dos Peixes/parasitologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Interferon gama/genética , Interferon gama/metabolismo , Alinhamento de Sequência/veterinária
13.
Fish Shellfish Immunol ; 98: 819-831, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31751659

RESUMO

Massive infection caused by Cryptocaryon irritans is detrimental to the development of marine aquaculture. Recently, our lab found that Nibea albiflora has low sensitivity and low mortality to C. irritans infection. The present study was designed to investigate the mechanisms of the N. albiflora response to C. irritans infection by analyzing transcriptome changes in the skin. Skin samples of control and experimental groups with C. irritans infection were collected at 24 and 72 h (24 h control, 24 h post-infection, 72 h control, and 72 h post-infection). Three parallels were set for each group and sample time, and a total of 12 skin samples were collected for sequencing. Overall, 297,489,843 valid paired-end reads and 48,817 unigenes were obtained with an overall length of 59,010,494 nt. In pairwise comparisons, changes in expression occurred in 1621 (764 upregulated and 857 downregulated), 285 (180 upregulated and 105 downregulated), 993 (489 upregulated and 504 downregulated), and 37 (8 upregulated and 29 downregulated) genes at 24 h control vs 24 h post-infection, 72 h control vs 72 h post-infection, 24 h post-infection vs 72 h post-infection, and 24 h control vs 72 h control, respectively. Gene Ontology (GO) analysis of differentially expressed genes (DEGs) indicated that the number of genes enriched in GO sub-categories were ordered 24 h control vs 24 h post-infection > 24 h post-infection vs 72 h post-infection >72 h control vs 72 h post-infection > 24 h control vs 72 h control. Further analysis showed that immune-related GO terms (including immune system process, complement activation, and humoral immunity) were significantly enriched at both 72 h control vs 72 h post-infection and 24 h post-infection vs 72 h post-infection, but no immune-related GO terms were significantly enriched in the 24 h control vs 72 h control and at 24 h control vs 24 h post-infection, indicating that C. irritans infection mainly affected the physiological metabolism of N. albiflora at an early stage (24 h), and immune-related genes play an important role at a later stage (72 h) of infection. In KEGG pathway analysis, the complement and coagulation cascade pathway are involved in early infection. Hematopoietic cell lineage, natural killer (NK) cell-mediated cytotoxicity, and the intestinal immune network for IgA production are involved in later infection. Further analysis showed that the alternative pathway of complement and coagulation cascades plays an important role in the resistance of N. albiflora to early C. irritans infection. During late infection, CD34, IgM, and IgD were significantly upregulated in the hematopoietic cell lineage pathway. CCR9 was significantly downregulated, and IGH and PIGR were significantly upregulated in the intestinal immune network for IgA production. GZMB and IGH were significantly downregulated in NK cell-mediated cytotoxicity. These findings indicate that acquired immunity at the mRNA level was initiated during later infection. In addition, the IL-17 signaling pathway was enriched by downregulated DEGs at 24 h post-infection vs 72 h post-infection, suggesting the inflammatory response at 24 h was stronger than at 72 h and the invasion of the parasite has a greater impact on the host.


Assuntos
Infecções por Cilióforos/veterinária , Cilióforos/fisiologia , Doenças dos Peixes/imunologia , Perciformes , Dermatopatias/veterinária , Transcriptoma , Animais , Infecções por Cilióforos/imunologia , Infecções por Cilióforos/parasitologia , Doenças dos Peixes/parasitologia , Perfilação da Expressão Gênica/veterinária , Pele/metabolismo , Dermatopatias/imunologia , Dermatopatias/parasitologia
14.
Parasite Immunol ; 42(8): e12675, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-31587318

RESUMO

The parasitic ciliate Ichthyophthirius multifiliis causes white spot disease in freshwater fish worldwide. The theront penetrates external surfaces of the naïve fish where it develops into the feeding trophont stage and elicits a protective immune response both at the affected site as well as at the systemic level. The present work compiles data and presents an overall model of the protective reactions induced. A wide spectrum of inflammatory reactions are established upon invasion but the specific protection is provided by adaptive factors. Immunoglobulin IgT is involved in protection of surfaces in several fish species and is thereby one of the first adaptive immune molecules reacting with the penetrating theront. IgT producing lymphocytes occur in epithelia, dispersed or associated with lymphoid cell aggregations (skin epidermis, fins, gills, nostrils and buccal cavities) but they are also present in central immune organs such as the head kidney, spleen and liver. When theronts invade immunized fish skin, they are encountered by host factors which opsonize the parasite and may result in complement activation, phagocytosis or cell-mediated killing. However, antibody (IgT, IgM and IgD) binding to parasite cilia has been suggested to alter parasite behaviour and induce an escape reaction, whereby specific IgT (or other classes of immunoglobulin in fish surfaces) takes a central role in protection against the parasite.


Assuntos
Anticorpos Antiprotozoários/imunologia , Infecções por Cilióforos/veterinária , Hymenostomatida/imunologia , Imunoglobulinas/imunologia , Animais , Infecções por Cilióforos/imunologia , Infecções por Cilióforos/parasitologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/parasitologia , Peixes , Brânquias/imunologia , Imunização , Baço/imunologia
15.
Parasitol Res ; 119(2): 755-758, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31811426

RESUMO

The ciliate species Balantioides coli can be cross-transmitted between humans and several animal species. Usually harmless, sometimes it can be pathogenic and cause the death of the host. In birds, B. coli has been confirmed in ostriches by genetic analysis, but the identification from South American greater rheas (Rhea americana) and lesser rheas (Rhea pennata pennata) is tentative. Since these species are reared for commercial purposes and for reintroduction into the wild, it is necessary to elucidate whether the ciliate from rheas is B. coli to minimize health risks for humans and for other domestic and wild animals. Individual parasite cells are collected from Argentinean isolates of reared greater rheas and of wild and reared lesser rheas, and their ITS region was PCR amplified; the cloning products were sequenced and compared with sequences available in public databases. The results have shown that several sequence types are expressed at the same time in the parasite cells, and all correspond to B. coli, confirming the possibility of cross-transmission of the parasite between wild and reared South American rheas and several mammal species and humans.


Assuntos
Infecções por Cilióforos/veterinária , Cilióforos/genética , Cilióforos/isolamento & purificação , Reiformes/parasitologia , Animais , Animais Selvagens , Aves , Infecções por Cilióforos/epidemiologia , Infecções por Cilióforos/parasitologia , Humanos , América do Sul/epidemiologia
16.
Rev. bras. parasitol. vet ; 29(1): e018319, 2020. graf
Artigo em Inglês | LILACS | ID: biblio-1058008

RESUMO

Abstract Scuticociliatosis, caused by an opportunistic ciliate protozoan, is responsible for significant economic losses in marine ornamental fish. This study reports the occurrence of Uronema spp., parasitizing ten species of marine reef fish at an ornamental fish wholesaler: Blue green damselfish (Chromis viridis), Vanderbilt's Chromis (Chromis vanderbilti), Pennant coralfish (Heniochus acuminatus), Threespot angelfish (Apolemichthys trimaculatus), Goldspotted angelfish (Apolemichthys xanthopunctatus), Sea goldie (Pseudanthias squamipinnis), Orchid dottyback (Pseudochromis fridmani), Threadfin butterflyfish (Chaetodon auriga), Vagabond butterflyfish (Chaetodon vagabundus), and Bluecheek butterflyfish (Chaetodon semilarvatus). Diseased fish showed disorders such as hemorrhages and ulcerative lesions on the body surface. Histopathological analysis of the muscle, liver, gut, kidney, spleen, gills, and stomach revealed hemorrhages and degeneration of muscle fiber, vacuolar degeneration of hepatocyte, inflammatory process and granuloma in the liver, atrophy of intestinal villi, inflammatory process and granuloma in the kidney, melanomacrophage centers, as well as inflammatory process in the spleen, epithelial cells hyperplasia and granuloma formation in the gills, and vacuolar degeneration and eosinophils in the stomach. Due to the severity of the disease, it is necessary to implement biosecurity measures with rapid and accurate diagnosis to minimize the risk of economic losses caused by Uronema spp.


Resumo Scuticociliatose, causada pelo protozoário ciliado oportunista, é responsável por significativas perdas econômicas em peixes marinhos ornamentais. O estudo relata a ocorrência de Uronema spp., parasitando dez espécies de peixes de recife em um distribuidor de peixes ornamentais: "Blue green damselfish" (Chromis viridis), "Vanderbilt's Chromis" (Chromis vanderbilti), "Pennant coralfish" (Heniochus acuminatus), "Threespot angelfish" (Apolemichthys trimaculatus), "Goldspotted angelfish" (Apolemichthys xanthopunctatus), "Sea goldie" (Pseudanthias squamipinnis), "Orchid dottyback" (Pseudochromis fridmani), "Threadfin butterflyfish" (Chaetodon auriga), "Vagabond butterflyfish" (Chaetodon vagabundus), e "Bluecheek butterflyfish" (Chaetodon semilarvatus). Peixes doentes apresentaram distúrbios como hemorragias e lesões ulcerativas na superfície do corpo. A análise histopatológica do músculo, fígado, intestino, rim, baço, brânquias e estômago revelou hemorragias e degeneração das fibras musculares, degeneração vacuolar de hepatócitos, processo inflamatório e granuloma no fígado, atrofia das vilosidades intestinais, processo inflamatório e granuloma no rim, centros de melanomacrófagos e processo inflamatório no baço, hiperplasia das células epiteliais, bem como formação de granuloma nas brânquias e degeneração vacuolar e eosinófilos no estômago. Devido à gravidade da doença, é necessário implementar medidas de biossegurança com diagnóstico rápido e preciso para minimizar o risco de perdas econômicas causadas por Uronema spp.


Assuntos
Animais , Perciformes/parasitologia , Infecções por Cilióforos/veterinária , Cilióforos/classificação , Doenças dos Peixes/parasitologia , Brasil , Infecções por Cilióforos/parasitologia , Infecções por Cilióforos/patologia , Doenças dos Peixes/patologia
17.
Rev. bras. parasitol. vet ; 28(4): 750-753, Oct.-Dec. 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1057970

RESUMO

Abstract Cryptocaryon irritans is an obligate parasitic ciliate protozoan of wild and cultured marine fish. It causes white spot disease, and infections with this pathogen can cause significant losses for aquarists and commercial marine cultures worldwide. This study reports the occurrence of C. irritans parasitizing the ornamental reef fish, yellowtail tang, Zebrasoma xanthurum. Six days after being introduced to a new environment, 11 yellowtail tangs had white spots scattered across their bodies and fins. Suspicion of infection with C. irritans was evaluated by scraping the skin to confirm clinical diagnosis. After confirmation, the yellowtail tangs were transferred to a hospital aquarium and treated with the therapeutic agent Seachem Cupramine® for 15 days. During the treatment period, the copper concentration was monitored daily. At the end of the treatment, none of the yellowtail tangs showed clinical signs of white spots on their bodies, and skin scraping confirmed the yellowtail tangs were no longer infected. Subsequently, the yellowtail tangs were released for sale.


Resumo Cryptocaryon irritans é um protozoário ciliado parasita obrigatório de peixes marinhos selvagens e cultivados. Causa a doença dos "pontos brancos". Infecções por esse patógeno pode causar perdas significativas para aquaristas e para marinocultura comercial em todo o mundo. Este estudo relata a ocorrência de C. irritans parasitando a espécie de peixe ornamental de recife Zebrasoma xanthurum. Seis dias após serem introduzidos no novo ambiente, onze peixes apresentaram pontos brancos espalhados pelo corpo e nadadeiras. Suspeitando de infecção por C. irritans foi realizado raspado de pele para confirmar o diagnóstico clínico. Após a confirmação, os peixes foram transferidos para um aquário hospital e tratados com o agente terapêutico Seachem Cupramine® por 15 dias. Durante o período de tratamento o monitoramento da concentração de cobre foi diário através da utilização de um fotômetro. Após o final do tratamento, todos os peixes já não apresentavam mais sinais clínicos de pontos brancos pelo corpo, e foram negativos no raspado de pele, sendo foram encaminhados para comercialização.


Assuntos
Animais , Perciformes/parasitologia , Infecções por Cilióforos/veterinária , Cilióforos/isolamento & purificação , Doenças dos Peixes/parasitologia , Perciformes/classificação , Cilióforos/classificação , Aquicultura
18.
Fish Shellfish Immunol ; 94: 661-674, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31521785

RESUMO

The yellow drum Nibea albiflora is less susceptible to Cryptocaryon irritans infection than is the case with other marine fishes such as Larimichthys crocea, Lateolabrax japonicus, and Pagrus major. To investigate further their resistance mechanism, we infected the N. albiflora with the C. irritans at a median lethal concentration of 2050 theronts/g fish. The skins of the infected and the uninfected fishes were sampled at 24 h and 72 h followed by an extensive analysis of metabolism. The study results revealed that there were 2694 potential metabolites. At 24 h post-infection, 12 metabolites were up-regulated and 17 were down-regulated whereas at 72 h post-infection, 22 metabolites were up-regulated and 26 were down-regulated. Pathway enrichment analysis shows that the differential enriched pathways were higher at 24 h with 22 categories and 58 subcategories (49 up, 9 down) than at 72 h whereby the differential enriched pathways were 6 categories and 8 subcategories (4 up, 4 down). In addition, the principal component analysis (PCA) plot shows that at 24 h the metabolites composition of infected group were separately clustered to uninfected group while at 72 h the metabolites composition in infected group were much closer to uninfected group. This indicated that C. irritans caused strong metabolic stress on the N. albiflora at 24 h and restoration of the dysregulated metabolic state took place at 72 h of infection. Also, at 72 h post infection a total of 17 compounds were identified as potential biomarkers. Furthermore, out of 2694 primary metabolites detected, 23 metabolites could be clearly identified and semi quantified with a known identification number and assigned into 66 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Most of the enriched KEGG pathways were mainly from metabolic pathway classes, including the metabolic pathway, biosynthesis of secondary metabolites, taurine and hypotaurine metabolism, purine metabolism, linoleic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis. Others were glyoxylate and dicarboxylate metabolism, glutathione metabolism, and alanine, aspartate, and glutamate metabolism. Moreover, out of the identified metabolites, only 6 metabolites were statistically differentially expressed, namely, L -glutamate (up-regulated) at 24 h was important for energy and precursor for other glutathiones and instruments of preventing oxidative injury; 15-hydroxy- eicosatetraenoic acid (15-HETE), (S)-(-)-2-Hydroxyisocaproic acid, and adenine (up-regulated) at 72 h were important for anti-inflammatory and immune responses during infection; others were delta-valerolactam and betaine which were down-regulated compared to uninfected group at 72 h, might be related to immure responses including stimulation of immune system such as production of antibodies. Our results therefore further advance our understanding on the immunological regulation of N. albiflora during immune response against infections as they indicated a strong relationship between skin metabolome and C. irritans infection.


Assuntos
Infecções por Cilióforos/veterinária , Doenças dos Peixes/imunologia , Imunidade Inata/imunologia , Metaboloma/imunologia , Perciformes , Pele/imunologia , Animais , Cilióforos/fisiologia , Infecções por Cilióforos/imunologia , Infecções por Cilióforos/parasitologia , Doenças dos Peixes/parasitologia
19.
Fish Shellfish Immunol ; 93: 863-870, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31422178

RESUMO

Interleukins (ILs) are a subgroup of cytokines, which are molecules involved in the intercellular regulation of the immune system. These cytokines have been extensively studied in mammalian models, but systematic analyses of fish are limited. In the current study, 3 IL genes from golden pompano (Trachinotus ovatus) were characterized. The IL-1ß protein contains IL-1 family signature motif, and four long helices (αA - αD) in IL-11 and IL-34, which were well conserved. All 3 ILs clustered phylogenetically with their respective IL relatives in mammalian and other teleost species. Under normal physiological conditions, the expression of IL-1ß, IL-11, and IL-34 were detected at varied levels in the 11 tissues examined. Most of the 3 ILs examined were highly expressed in liver, spleen, kidney, gill, or skin. Following pathogenic bacterial, viral, or parasitic challenge, IL-1ß, IL-11, and IL-34 exhibited distinctly different expression profiles in a time-, tissue-, and pathogen-dependent manner. In general, IL-1ß was expressed at higher levels following challenge with all pathogens examined than was observed for IL-11 and IL-34. Furthermore, Streptococcus agalactiae and Cryptocaryon irritans caused higher levels of IL-1ß and IL-11 expression than Vibrio harveyi and viral nervous necrosis virus (VNNV). The increased expression of IL-34 caused by VNNV and C. irritans were higher than that caused by V. harveyi and S. agalactiae. These results suggest that these 3 ILs in T. ovatus may play different effect pathogen type specific responses.


Assuntos
Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Animais , Cilióforos/fisiologia , Infecções por Cilióforos/imunologia , Infecções por Cilióforos/veterinária , Perfilação da Expressão Gênica/veterinária , Interleucina-11/genética , Interleucina-11/imunologia , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Interleucinas/genética , Interleucinas/imunologia , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/veterinária , Streptococcus agalactiae/fisiologia , Vibrio/fisiologia , Vibrioses/imunologia , Vibrioses/veterinária
20.
Fish Shellfish Immunol ; 94: 308-317, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31470140

RESUMO

The channel catfish (Ictalurus punctatus) immune response against Ichthyophthirius multifiliis (Ich) after vaccination using plasmid DNA vaccines pcDNA3.1-IAg52a and pcDNA3.1-IAg52b, encoding Ich immobilization antigen genes was studied. Parasite infection level, serum anti-Ich antibodies level, fish mortality after theront challenge, and immune-related gene expression were measured. After in vitro transfection of walking catfish gill cells (G1b) with both pcDNA3.1-IAg52a and pcDNA3.1-IAg52b, antigens IAG52A and IAG52B were detected. During the vaccination trial, 76-fold increase in the Iag52b gene expression was observed in the vaccinated fish group h4 post vaccination. Administration of DNA vaccines by IM injection induced significant gene up-regulation in the head kidney, including immunoglobulin M (IgM), cluster of differentiation 4 (CD4), major histocompatibility I (MHC I), and T cell receptor α (TcR-α) from h4 to d5 post immunization. Fish vaccinated with DNA vaccines or theronts showed increased gene expression of the cytokine interferon (IFN-γ), complement component 3 (C3), and toll-like receptor-1 (TLR-1). Anti-Ich antibodies were detected in fish received pcDNA3.1-IAg52a, pcDNA3.1-IAg52b and the combination of both vaccines d10 post vaccination. Fish vaccinated with pcDNA3.1-IAg52b showed mild parasite infection level, partial survival (20%) and longer mean day to death (MDD) after theront challenge. By contrast, a heavy parasite load, 0% survival and short MDD were observed in the sham vaccinated control fish that received pcDNA3.1 (plasmid without genes encoding Ich immobilization antigen). Further research is needed to improve DNA vaccines for Ich that can induce strong protective immunity in fish. Suggested studies include improved transfection efficiency, use of appropriate adjuvants and including additional parasite antigen genes in the plasmid.


Assuntos
Infecções por Cilióforos/veterinária , Doenças dos Peixes/prevenção & controle , Hymenostomatida/imunologia , Ictaluridae , Imunidade Inata , Vacinas Protozoárias/farmacologia , Vacinação/veterinária , Imunidade Adaptativa , Animais , Antígenos de Protozoários/farmacologia , Infecções por Cilióforos/imunologia , Infecções por Cilióforos/prevenção & controle , Doenças dos Peixes/imunologia , Proteínas de Protozoários/farmacologia , Vacinas de DNA/farmacologia
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