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1.
Fish Shellfish Immunol ; 138: 108835, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37236552

RESUMO

Atlantic salmon (Salmo salar) are highly susceptible to infestations with the ectoparasite Lepeophtheirus salmonis, the salmon louse. Infestations elicit an immune response in the fish, but the response does not lead to parasite clearance, nor does it protect against subsequent infestations. It is, however, not known why the immune response is not adequate, possibly because the local response directly underneath the louse has been poorly evaluated. The present study describes the transcriptomic response by RNA sequencing of skin at the site of copepodid attachment. Analysing differentially expressed genes, 2864 were higher and 1357 were lower expressed at the louse attachment site compared to uninfested sites in the louse infested fish, while gene expression at uninfested sites were similar to uninfested control fish. The transcriptional patterns of selected immune genes were further detailed in three skin compartments/types: Whole skin, scales only and fin tissue. The elevation of pro-inflammatory cytokines and immune cell marker transcripts observed in whole skin and scale samples were not induced in fin, and a higher cytokine transcript level in scale samples suggest it can be used as a nonlethal sampling method to enhance selective breeding trials. Furthermore, the immune response was followed in both skin and anterior kidney as the infestation developed. Here, newly moulted preadult 1 stage lice induced a higher immune response than chalimi and adult lice. Overall, infestation with salmon louse induce a modest but early immune response with an elevation of mainly innate immune transcripts, with the response primarily localized to the site of attachment.


Assuntos
Copépodes , Doenças dos Peixes , Salmo salar , Animais , Transcriptoma , Salmo salar/genética , Salmo salar/metabolismo , Pele , Imunidade/genética , Citocinas/genética
2.
Parasitology ; 150(11): 990-1005, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37705306

RESUMO

Atlantic salmon (Salmo salar) is repeatedly exposed to and infected with ectoparasitic salmon lice (Lepeophtheirus salmonis) both in farms and in nature. However, this is not reflected in laboratory experiments where fish typically are infected only once. To investigate if a previous lice infection affects host response to subsequent infections, fish received 4 different experimental treatments; including 2 groups of fish that had previously been infected either with adult or infective salmon lice larvae (copepodids). Thereafter, fish in all treatment groups were infected with either a double or a single dose of copepodids originating from the same cohort. Fish were sampled when lice had developed into the chalimus, the pre-adult and the adult stage, respectively. Both the specific growth rate and cortisol levels (i.e. a proxy for stress) of the fish differed between treatments. Lice success (i.e. ability to infect and survive on the host) was higher in naïve than in previously infected fish (pre-adult stage). The expression of immune and wound healing transcripts in the skin also differed between treatments, and most noticeable was a higher upregulation early in the infection in the group previously infected with copepodids. However, later in the infection, the least upregulation was observed in this group, suggesting that previous exposure to salmon lice affects the response of Atlantic salmon towards subsequent lice infections.


Assuntos
Copépodes , Ectoparasitoses , Doenças dos Peixes , Salmo salar , Humanos , Animais , Copépodes/fisiologia , Ectoparasitoses/veterinária , Ectoparasitoses/parasitologia , Pele/parasitologia , Doenças dos Peixes/parasitologia
3.
J Fish Dis ; 45(8): 1133-1147, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35612902

RESUMO

The effect of different intensities of the ectoparasitic salmon lice (Lepeophtheirus salmonis) on stress, growth and the expression of immune and wound healing transcripts in the skin of Atlantic salmon (Salmo salar) was investigated. Lice infection success and survival were similar at the chalimus and preadult stage in the low and high dose group, but infection success and survival were significantly lower in the high than in the low dose group at the adult stage. The expression of investigated transcripts was not correlated to lice intensities, but several of them were significantly differently expressed locally in the skin at the site of lice attachment in infected fish compared to controls. This included an up-regulation of pro-inflammatory markers at the site of lice attachment (e.g., interleukin 1-beta, interleukin 8 and the acute phase protein serum amyloid A), a reduction of markers of adaptive immunity (cluster of differentiation 8-alpha and immunoglobulin M) and decreased expression of the anti-inflammatory cytokine interleukin 10.


Assuntos
Copépodes , Ectoparasitoses , Doenças dos Peixes , Salmo salar , Animais , Copépodes/fisiologia , Ectoparasitoses/metabolismo , Ectoparasitoses/veterinária , Doenças dos Peixes/parasitologia , Pele
4.
J Fish Dis ; 45(7): 975-990, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35397139

RESUMO

In this study, the effect of temperature on Atlantic salmon (Salmo salar) stress and immune response to the ectoparasitic salmon lice (Lepeophtheirus salmonis) was investigated. We found that infestation affected the expression of several immune and wound healing transcripts in the skin especially at the site of lice attachment compared to un-infested control fish. Moreover, expression patterns in the skin of infested fish suggest that host immune responses towards salmon lice are impaired at low temperatures. However, reduced lice infestation success and survival at the lowest investigated temperatures suggest that cold water temperatures are more detrimental to the lice than their fish hosts. Finally, temperature affected the stress response of the fish and infected fish had a higher increase in cortisol levels in response to handling (a stressor) than un-infested controls.


Assuntos
Copépodes , Doenças dos Peixes , Salmo salar , Animais , Copépodes/fisiologia , Doenças dos Peixes/parasitologia , Pele , Temperatura
5.
Genomics ; 113(6): 3666-3680, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34403763

RESUMO

Copepods encompass numerous ecological roles including parasites, detrivores and phytoplankton grazers. Nonetheless, copepod genome assemblies remain scarce. Lepeophtheirus salmonis is an economically and ecologically important ectoparasitic copepod found on salmonid fish. We present the 695.4 Mbp L. salmonis genome assembly containing ≈60% repetitive regions and 13,081 annotated protein-coding genes. The genome comprises 14 autosomes and a ZZ-ZW sex chromosome system. Assembly assessment identified 92.4% of the expected arthropod genes. Transcriptomics supported annotation and indicated a marked shift in gene expression after host attachment, including apparent downregulation of genes related to circadian rhythm coinciding with abandoning diurnal migration. The genome shows evolutionary signatures including loss of genes needed for peroxisome biogenesis, presence of numerous FNII domains, and an incomplete heme homeostasis pathway suggesting heme proteins to be obtained from the host. Despite repeated development of resistance against chemical treatments L. salmonis exhibits low numbers of many genes involved in detoxification.


Assuntos
Copépodes , Doenças dos Peixes , Parasitos , Aclimatação , Animais , Copépodes/genética , Copépodes/parasitologia , Doenças dos Peixes/genética , Parasitos/genética , Transcriptoma
6.
J Fish Dis ; 44(7): 863-879, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33586246

RESUMO

Monitoring of planktonic salmon louse (Lepeophtheirus salmonis salmonis) abundance and parameterization of key life-history traits has been hindered by labour-intensive and error-prone quantification using traditional light microscopy. Fluorescence illumination has been proposed as a means of improving visualization, but prior to this study adequate investigation of the relevant fluorescence profiles and measurement conditions has not been undertaken. We investigated the fluorescence profiles of L. salmonis and non-target copepod spp. with excitation and emission matrices (200-600 nm) and identified unique fluorescence signals. Fluorescence microscopy using excitation wavelengths of 470 ± 40 nm, and emission wavelengths of 525 ± 50 nm, showed that after 90 days of formalin storage salmon lice have a mean fluorescence intensity that is 2.4 times greater than non-target copepods (copepodid and adult stages). A 7-day heat treatment of 42°C in formalin increased the difference between salmon louse copepodids and non-target copepods to a factor of 3.6, eliminating the need for prolonged storage. Differences in the fluorescence signal and endogenous fluorophores were investigated with respect to variation in sea lice species, age, stage and host fish origin. Under the conditions outlined in this paper, the fluorescence signal was found to be a reliable means of visualizing and differentiating salmon lice from non-target zooplankters. Adaptation of the fluorescence signal would greatly expedite traditional methods of enumerating salmon louse larvae in plankton samples and could provide a means of automated detection.


Assuntos
Copépodes/fisiologia , Ectoparasitoses/veterinária , Doenças dos Peixes/parasitologia , Estágios do Ciclo de Vida/fisiologia , Imagem Óptica/métodos , Zooplâncton , Animais , Ectoparasitoses/parasitologia , Salmão/parasitologia
7.
Fish Shellfish Immunol ; 103: 200-210, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32422189

RESUMO

The marine crustacean Lepeophtheirus salmonis (salmon louse) is a common ectoparasite of wild and farmed salmonids. The parasite has a complex ontogeny comprising eight instars. The planktonic copepodid stage settles on host skin and pass through five instars to reach the adult stage. The present study comprises an experimental infestation of Oncorhynchus mykiss (rainbow trout) with salmon lice and describes histopathology and host immune responses in skin beneath the louse at multiple time points encompassing all louse developmental stages. Each fish was exposed to 80 infective copepodids, a mean no. of 32 parasites reached the preadult I stage whereas a mean no. of 11 parasites reached the adult stage. A progression in the severity of cutaneous lesions was observed, and levels of immune gene transcripts at the attachment site revealed a dynamic response, initially related to innate immunity. Later, immune cells accumulated in the dermis concomitant with a moderate decrease in levels of transcripts characteristic of both innate and adaptive immune responses. The present study also demonstrates that the cutaneous immune response was mainly induced at lice affected sites, while non-affected skin resembled the skin of untreated control. This indicates that the skin cannot be regarded as a uniform organ and requires careful sampling at all salmon louse stages.


Assuntos
Copépodes/fisiologia , Ectoparasitoses/veterinária , Doenças dos Peixes/imunologia , Interações Hospedeiro-Parasita , Oncorhynchus mykiss , Pele/imunologia , Imunidade Adaptativa , Animais , Ectoparasitoses/imunologia , Ectoparasitoses/parasitologia , Doenças dos Peixes/parasitologia , Imunidade Inata , Pele/parasitologia
8.
J Fish Dis ; 43(12): 1519-1529, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32882750

RESUMO

The salmon louse (Lepeophtheirus salmonis) is an ecologically and economically important parasite of salmonid fish. Temperature is a strong influencer of biological processes in salmon lice, with development rate increased at higher temperatures. The successful attachment of lice onto a host is also predicted to be influenced by temperature; however, the correlation of temperature with parasite survival is unknown. This study describes the effects of temperature on infection success, and survival on the host during development to the adult stage. To accurately describe infection dynamics with varying temperatures, infection success was recorded on Atlantic salmon (Salmo salar) between 2 and 10°C. Infection success ranged from 20% to 50% and was strongly correlated with temperature, with the highest success at 10°C. Parasite loss was monitored during development at eight temperatures with high loss of lice at 3 and 24°C, whilst no loss was recorded in the temperature range from 6 to 21°C. Sea temperatures thus have large effects on the outcome of salmon louse infections and should be taken into account in the management and risk assessment of this parasite. Improving understanding of the infection dynamics of salmon lice will facilitate epidemiological modelling efforts and efficiency of pest management strategies.


Assuntos
Copépodes/fisiologia , Doenças dos Peixes/parasitologia , Salmo salar/parasitologia , Temperatura , Animais , Ectoparasitoses/parasitologia
9.
Artigo em Inglês | MEDLINE | ID: mdl-30326269

RESUMO

Animals with exoskeleton need to molt to grow and develop. Molting is well described in some arthropods especially insects. Chitin is a polymer of N-acetylglucosamine, and one of the major components of the exoskeleton of arthropods. Chitin is synthesized and degraded by a series of enzymes during the molting cycle. However, the presence and function of these enzymes are largely unknown in copepods such as the ectoparasite salmon louse (Lepeophtheirus salmonis) a major pest in salmonid aquaculture. Here we describe six genes found in the L. salmonis genome (LsCHS1, LsCHS2, LsGFAT, LsGNA1, LsAGM, and LsUAP) with high homology to enzymes in the chitin synthesis pathway. The transcription profiles of these enzymes together with three chitinases enzymes (LsChi1, LsChi2, and LsChi4), which have been characterized before, were examined during the synthesis of a new exoskeleton and revealed a dynamical expression concurrent with the morphological changes during the molt cycle. Further understanding of chitin metabolism and its regulation may prove useful tool to develop new pesticides.


Assuntos
Quitina/biossíntese , Copépodes/metabolismo , Perfilação da Expressão Gênica , Animais , Quitina/metabolismo , Copépodes/genética , Hidrólise , Filogenia , RNA Mensageiro/genética
10.
Exp Parasitol ; 188: 83-92, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29625096

RESUMO

The salmon louse Lepeophtheirus salmonis (Copepods, Caligida) is a marine ectoparasite infecting salmonid fishes in the northern hemisphere. At present, salmon lice infections are the most severe disease problem in the salmon farming industry causing significant economic losses. Due to development of resistance towards available chemotherapeutants, it is clear that new chemotherapeutants or non-chemical control methods are essential to manage the parasite in the future. The TOR signaling pathway is present in all metazoans and is a major regulator of cellular activity according to nutrient availability. In this study, we identified the TOR pathway genes in salmon louse; LsTSC1, LsTSC2, LsRheb, LsTOR, LsRaptor and LsRictor. RNA interference mediated gene silencing was performed to elucidate the functional role of each member of the pathway. Our results show that interference of the TOR signaling pathway either directly or indirectly inhibits many biological processes including egg maturation. In addition, the effect of gene knock-down results in more comprehensive physiological defects when targeting TORC1 and the upstream regulator Rheb. This is the first report on the TOR pathway in the salmon louse and that our research contributes to the basic knowledge of the parasite that could lead to development of novel treatment methods.


Assuntos
Copépodes/fisiologia , Ectoparasitoses/veterinária , Doenças dos Peixes/parasitologia , Salmo salar/parasitologia , Serina-Treonina Quinases TOR/metabolismo , Animais , Copépodes/anatomia & histologia , Copépodes/enzimologia , Copépodes/genética , Ectoparasitoses/parasitologia , Feminino , Pesqueiros , Técnicas de Silenciamento de Genes , Inativação Gênica , Interferência de RNA , RNA de Cadeia Dupla/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Reprodução/genética , Água do Mar , Análise de Sequência , Transdução de Sinais , Serina-Treonina Quinases TOR/genética , Vitelogênese/genética
11.
J Lipid Res ; 58(8): 1613-1623, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28601811

RESUMO

The salmon louse, Lepeophtheirus salmonis, is an endemic ectoparasite on salmonid fish that is challenging for the salmon farming industry and wild fish. Salmon lice produce high numbers of offspring, necessitating sequestration of large amounts of lipids into growing oocytes as a major energy source for larvae, most probably mediated by lipoproteins. The microsomal triglyceride transfer protein (MTP) is essential for the assembly of lipoproteins. Salmon lice have three L. salmonis MTP (LsMTP) transcript variants encoding two different protein isoforms, which are predicted to contain three ß-sheets (N, C, and A) and a central helical domain, similar to MTPs from other species. In adult females, the LsMTPs are differently transcribed in the sub-cuticular tissues, the intestine, the ovary, and in the mature eggs. RNA interference-mediated knockdown of LsMTP in mature females gave offspring with significantly fewer neutral lipids in their yolk and only 10-30% survival. The present study suggests the importance of LsMTP in reproduction and lipid metabolism in adult female L. salmonis, a possible metabolic bottleneck that could be exploited for the development of new anti-parasitic treatment methods.


Assuntos
Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Copépodes/genética , Copépodes/metabolismo , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/deficiência , Sequência de Bases , Proteínas de Transporte/química , Copépodes/fisiologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Larva/fisiologia , Lipoproteínas/metabolismo , Modelos Moleculares , Estrutura Secundária de Proteína , Interferência de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Salmão/parasitologia , Análise de Sequência
12.
Artigo em Inglês | MEDLINE | ID: mdl-28087330

RESUMO

Heme peroxidases are the most abundant type of peroxidase catalyzing a H2O2-dependent oxidation of a wide variety of substrates. They are involved in numerous processes like the innate immune response, hormone and prostaglandin synthesis and crosslinking of proteins within extracellular matrixes (ECM) as well as molecules within the cuticle and chorion of arthropods and nematodes. In the present study, a Lepeophtheirus salmonis heme peroxidase (LsHPX) 1 was characterized. Amino acids in the active site of heme peroxidases were conserved, and the predicted protein sequence showed the highest similarity to genes annotated as chorion peroxidases and genes suggested to be involved in cuticle hardening or adhesion. LsHPX1 exhibited a dynamic expression during ontogenesis and during the nauplius molting cycle. Transcripts were localized to muscle cells near the muscle-tendon junction, in nerve tissue especially at neuromuscular junctions, subcuticular epithelium, subepithelial cells facing the hemolymph, exocrine glands within the subepithelial tissue and in isolated cells within the testis. Knock-down of LsHPX1 in nauplius larvae decreased the swimming activity of emerging copepodids. Histological analysis of knock-down animals revealed increased spacing between myofibers and changes in subepithelial and exocrine gland tissue. Considering these results, the potential role of LsHPX1 in crosslinking molecules of salmon louse ECMs is discussed.


Assuntos
Proteínas de Artrópodes/metabolismo , Copépodes/enzimologia , Matriz Extracelular/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Heme/metabolismo , Junção Neuromuscular/metabolismo , Peroxidases/metabolismo , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/antagonistas & inibidores , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Domínio Catalítico , Sequência Conservada , Copépodes/citologia , Copépodes/crescimento & desenvolvimento , Copépodes/fisiologia , Matriz Extracelular/enzimologia , Feminino , Técnicas de Silenciamento de Genes , Estágios do Ciclo de Vida , Masculino , Muda , Junção Neuromuscular/citologia , Junção Neuromuscular/enzimologia , Especificidade de Órgãos , Oxirredução , Estresse Oxidativo , Peroxidases/antagonistas & inibidores , Peroxidases/química , Peroxidases/genética , Interferência de RNA , Especificidade por Substrato , Natação
13.
BMC Genomics ; 16: 81, 2015 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-25765704

RESUMO

BACKGROUND: Nuclear receptors have crucial roles in all metazoan animals as regulators of gene transcription. A wide range of studies have elucidated molecular and biological significance of nuclear receptors but there are still a large number of animals where the knowledge is very limited. In the present study we have identified an RXR type of nuclear receptor in the salmon louse (Lepeophtheirus salmonis) (i.e. LsRXR). RXR is one of the two partners of the Ecdysteroid receptor in arthropods, the receptor for the main molting hormone 20-hydroxyecdysone (E20) with a wide array of effects in arthropods. RESULTS: Five different LsRXR transcripts were identified by RACE showing large differences in domain structure. The largest isoforms contained complete DNA binding domain (DBD) and ligand binding domain (LBD), whereas some variants had incomplete or no DBD. LsRXR is transcribed in several tissues in the salmon louse including ovary, subcuticular tissue, intestine and glands. By using Q-PCR it is evident that the LsRXR mRNA levels vary throughout the L. salmonis life cycle. We also show that the truncated LsRXR transcript comprise about 50% in all examined samples. We used RNAi to knock-down the transcription in adult reproducing female lice. This resulted in close to zero viable offspring. We also assessed the LsRXR RNAi effects using a L. salmonis microarray and saw significant effects on transcription in the female lice. Transcription of the major yolk proteins was strongly reduced by knock-down of LsRXR. Genes involved in lipid metabolism and transport were also down regulated. Furthermore, different types of growth processes were up regulated and many cuticle proteins were present in this group. CONCLUSIONS: The present study demonstrates the significance of LsRXR in adult female L. salmonis and discusses the functional aspects in relation to other arthropods. LsRXR has a unique structure that should be elucidated in the future.


Assuntos
Copépodes/genética , Interações Hospedeiro-Parasita/genética , Receptores do Ácido Retinoico/genética , Animais , Copépodes/patogenicidade , Proteínas de Ligação a DNA/genética , Ecdisterona/genética , Ecdisterona/metabolismo , Feminino , Estágios do Ciclo de Vida , Metabolismo dos Lipídeos/genética , Dados de Sequência Molecular , Ovário/crescimento & desenvolvimento , Ovário/parasitologia , Estrutura Terciária de Proteína , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Receptores do Ácido Retinoico/metabolismo , Reprodução/genética , Salmão/parasitologia
14.
Exp Parasitol ; 159: 79-93, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26348267

RESUMO

The salmon louse (Lepeophtheirus salmonis) is a major parasite of salmonid fish in the marine environment. The interaction between the parasite and the host upon infection is not completely understood. However, it is clear that the parasite influences the host and its immune system. Prostaglandins produced by parasites such as flatworms, roundworms and ticks are documented or assumed to play a role in immunomodulation of the host. In the salmon louse, the effect of prostaglandins on the host is assumed, but remains to be documented. In this study, a salmon louse prostaglandin E2 synthase (LsPGES2) is characterized. Ontogenetic analysis showed that LsPGES2 is relatively stable expressed during development. The highest level of expression was seen in the free living stages, although elevated levels of LsPGES2 were also found in adult females. In copepodids, LsPGES2 is found around muscle cells, while it is observed in the reproductive organs of adult female lice. LsPGES2 expression was knocked-down by RNA interference in nauplii, but emerging copepodids did not display any changes in morphology nor ability to infect and develop to adult stages on fish. Additional knock-down of LsPGES2 in adult female lice did not produce any characteristic changes in phenotype nor reproductive output. It is concluded that under these experimental conditions, knock-down of LsPGES2 did not affect any essential functions of the salmon louse, neither in the free-living nor the parasitic stages.


Assuntos
Copépodes/enzimologia , Ectoparasitoses/veterinária , Doenças dos Peixes/parasitologia , Oxirredutases Intramoleculares/genética , Salmo salar/parasitologia , Sequência de Aminoácidos , Animais , Copépodes/classificação , Copépodes/genética , Ectoparasitoses/parasitologia , Etiquetas de Sequências Expressas/química , Feminino , Técnicas de Silenciamento de Genes , Hibridização In Situ , Oxirredutases Intramoleculares/metabolismo , Masculino , Dados de Sequência Molecular , Filogenia , Prostaglandina-E Sintases , Reação em Cadeia da Polimerase em Tempo Real , Alinhamento de Sequência
15.
Exp Parasitol ; 151-152: 39-48, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25643862

RESUMO

The salmon louse (Lepeophtheirus salmonis spp.) is an economically important parasite on Atlantic salmon reared in aquaculture globally. Production and degradation of chitin, a major component of the exoskeleton, is the target of some pesticides (Di/Teflubenzuron) used in management of lice on farmed fish. These chemicals inhibit molting of the salmon louse leading to the death of the parasite. We found three chitinases (LsChi1, LsChi2 and LsChi4) in the salmon louse genome. Sequence analysis and phylogeny showed that they belong to the GH18 type of chitinase group and show high sequence similarity to chitinases found in other crustaceans and in insects. Expression patterns were different for all three chitinases suggesting different functions during louse development. Furthermore, the function of LsChi2 was further explored through the use of RNA interference and infection trials. Copepodids with knock down of LsChi2 transcripts were deformed and showed a highly reduced infection success.


Assuntos
Quitinases/genética , Copépodes/enzimologia , Ectoparasitoses/veterinária , Doenças dos Peixes/parasitologia , Salmo salar/parasitologia , Sequência de Aminoácidos , Animais , Quitinases/química , Quitinases/classificação , Quitinases/metabolismo , Copépodes/anatomia & histologia , Copépodes/genética , DNA Complementar/biossíntese , Ectoparasitoses/parasitologia , Feminino , Masculino , Filogenia , RNA/genética , RNA/isolamento & purificação , Interferência de RNA , RNA de Cadeia Dupla/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Alinhamento de Sequência
16.
Exp Parasitol ; 140: 44-51, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24632188

RESUMO

The salmon louse (Lepeophtheirus salmonis), an ectoparasitic copepod of salmonid fish, is a major threat to aquaculture in Norway, Ireland, Scotland and Canada. Due to rise in resistance against existing pesticides, development of novel drugs or vaccines is necessary. Posttranscriptional gene silencing by RNA interference (RNAi), when established in a high throughput system is a potential method for evaluation of molecular targets for new medical compounds or vaccine antigens. Successful use of RNAi has been reported in several stages of salmon lice. However, when we employed a previously described protocol for planktonic stages, no reproducible down-regulation of target genes was gained. In the present study, we describe a robust method for RNAi, where nauplius larvae are soaked in seawater added double stranded RNA (dsRNA). In order to test for when dsRNA may be introduced, and for the efficacy and duration of RNAi, we performed a series of experiments on accurately age determined larvae, ranging from the hatching egg to the copepodid with a salmon louse coatomer and a putative prostaglandin E synthase gene. Presumptive knock-down was monitored by real time PCR. Significant gene silencing was obtained only when nauplius I larvae were exposed to dsRNA during the period in which they molted to nauplius II. A knock down effect could be detected 2days after soaking, and it remained stable until the last measurement, on day 12. Soaking nauplius I larvae, knock-down was verified for six additional genes with a putative role in molting. For one chitinase, a loss-of-function phenotype with abnormal swimming was obtained. Hence, RNAi, induced in the nauplius, may facilitate studies of the molecular biology of the louse, such as the function of specific genes in developmental processes and physiology, host recognition, host-parasite interaction, and, in extension, the engineering of novel medicines.


Assuntos
Copépodes/genética , Técnicas de Silenciamento de Genes/métodos , Interferência de RNA , Animais , Copépodes/crescimento & desenvolvimento , Copépodes/fisiologia , DNA Complementar/biossíntese , Regulação para Baixo , Ectoparasitoses/parasitologia , Ectoparasitoses/veterinária , Doenças dos Peixes/parasitologia , Expressão Gênica/genética , Larva , Locomoção , Muda , RNA/genética , RNA/isolamento & purificação , RNA de Cadeia Dupla/administração & dosagem , RNA de Cadeia Dupla/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Salmo salar/parasitologia
17.
Conserv Physiol ; 11(1): coac078, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36655169

RESUMO

The aims of this study were to compare male and female sea trout (Salmo trutta) with respect to their hypo-osmoregulatory ability over a simulated migration to seawater and their clinical and physiological response to salmon louse (Lepeophtheirus salmonis) infection in seawater and over a simulated pre-mature return to freshwater. For this purpose, 2-year-old hatchery-reared male and female brown trout (F1 offspring of wild caught anadromous fish) were infected with salmon lice and measured for changes in plasma ions, glucose, lactate and osmolality and relative heart, liver and gonad sizes during a simulated seawater migration and thereafter a premature return to freshwater after 4 weeks in seawater (pre-adult louse). Un-infected trout served as control. Male trout used longer time to develop full hypo-osmoregulatory ability in seawater and showed a stronger response in plasma glucose and lactate following simulated premature return to freshwater, compared to female trout. Response to salmon louse was stronger in female trout, shown by stronger osmotic stress by chalimus (plasma Cl-) and pre-adult louse (plasma osmolality) and elevated relative liver size (hepatosomatic index) by pre-adult louse in female compared to male trout. Moreover, high plasma cortisol in infected female and low plasma cortisol in infected male trout produced a significant treatment-sex interaction on plasma cortisol. Lice infection intensity was initially higher in male (0.18 lice g-1) compared to female trout (0.11 lice g-1) at the chalimus stage, but equal between sexes at the pre-adult stage (male 0.15 and female 0.17 lice g-1). This study showed that female trout were better adapted for changes in water salinity, while male trout were more robust against salmon louse infection. These results suggests that the elevated salmon louse infection pressure generated by salmon farming have strong and unexplored negative effects on wild sea trout populations. Further research on this topic is vital for the conservation of wild sea trout populations.

18.
Dis Aquat Organ ; 94(3): 211-24, 2011 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-21790068

RESUMO

The salmon louse Lepeophtheirus salmonis Krøyer affects a variety of wild salmonoid hosts, but is also an important pest in aquaculture, which is a globally important and rapidly growing industry. Salmon lice have large reproductive outputs, and knowledge of reproductive processes may be crucial for the control of this parasite. Here, we report on the characterisation of 2 vitellogenins (LsVit1 and LsVit2), which are the precursors of salmon-louse egg-yolk glycoprotein. The structure of LsVit1 and LsVit2 was examined and compared to that in other oviparous animals. Phylogenetic analysis of LsVit1 and LsVit2 confirmed the view that crustaceans are a polyphyletic group. Transcriptional and translational analysis demonstrated production of LsVit1 and LsVit2 in the subcuticular tissue of the adult female lice. LsVit1 and LsVit2 could also be found in maturing oocytes and developing embryos and early larval stages. LsVit2 was found to be processed into 2 smaller fragments, whereas LsVit1 was found to be full length when deposited into the oocytes. Degradation of LsVit1 and LsVit2 was characterised through embryogenesis and the early non-feeding larval stages. Finally, protein content and the level of free amino acids were analysed in embryos and larval stages and their role in nutrition and osmoregulation discussed. In conclusion, our results confirm the role of vitellogenins in reproduction as providers of embryonic and larval nutrition.


Assuntos
Copépodes/metabolismo , Vitelogeninas/metabolismo , Sequência de Aminoácidos , Animais , Copépodes/embriologia , Ectoparasitoses/veterinária , Proteínas do Ovo/metabolismo , Desenvolvimento Embrionário , Feminino , Doenças dos Peixes/parasitologia , Masculino , Óvulo , Filogenia , Salmo salar , Fatores de Tempo , Vitelogeninas/química , Vitelogeninas/genética
19.
Parasit Vectors ; 14(1): 206, 2021 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-33874988

RESUMO

BACKGROUND: The salmon louse (Lepeophtheirus salmonis) is a parasite of salmonid fish. Atlantic salmon (Salmo salar) exhibit only a limited and ineffective immune response when infested with this parasite. Prostaglandins (PGs) have many biological functions in both invertebrates and vertebrates, one of which is the regulation of immune responses. This has led to the suggestion that prostaglandin E2 (PGE2) is important in the salmon louse host-parasite interaction, although studies of a salmon louse prostaglandin E2 synthase (PGES) 2 gene have not enabled conformation of this hypothesis. The aim of the present study was, therefore, to characterize two additional PGES-like genes. METHODS: Lepeophtheirus salmonis microsomal glutathione S-transferase 1 like (LsMGST1L) and LsPGES3L were investigated by sequencing, phylogenetics, transcript localization and expression studies. Moreover, the function of these putative PGES genes in addition to the previously identified LsPGES2 gene was analyzed in double stranded (ds) RNA-mediated knockdown (KD) salmon louse. RESULTS: Analysis of the three putative LsPGES genes showed a rather constitutive transcript level throughout development from nauplius to the adult stages, and in a range of tissues, with the highest levels in the ovaries or gut. DsRNA-mediated KD of these transcripts did not produce any characteristic changes in phenotype, and KD animals displayed a normal reproductive output. The ability of the parasite to infect or modulate the immune response of the host fish was also not affected by KD. CONCLUSIONS: Salmon louse prostaglandins may play endogenous roles in the management of reproduction and oxidative stress and may be a product of salmon louse blood digestions.


Assuntos
Proteínas de Artrópodes/metabolismo , Copépodes/enzimologia , Doenças dos Peixes/parasitologia , Prostaglandina-E Sintases/metabolismo , Animais , Proteínas de Artrópodes/genética , Copépodes/classificação , Copépodes/genética , Copépodes/crescimento & desenvolvimento , Feminino , Interações Hospedeiro-Parasita , Masculino , Filogenia , Prostaglandina-E Sintases/genética , Prostaglandinas/metabolismo , Salmo salar/parasitologia
20.
Biochem Biophys Rep ; 28: 101156, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34729423

RESUMO

The salmon louse, Lepeophtheirus salmonis is an ectoparasite of salmonid fish in the Northern Hemisphere, causing large economical losses in the aquaculture industry and represent a threat to wild populations of salmonids. Like other oviparous animals, it is likely that female lice use lipoproteins for lipid transport to maturing oocytes and other organs of the body. As an important component of lipoproteins, apolipoproteins play a vital role in the transport of lipids through biosynthesis of lipoproteins. Apolipoproteins have been studied in detail in different organisms, but no studies have been done in salmon lice. Two apolipoprotein encoding genes (LsLp1 and LsLp2) were identified in the salmon lice genome. Transcriptional analysis revealed both genes to be expressed at all stages from larvae to adult with some variation, LsLp1 generally higher than LsLp2 and both at their highest levels in adult stages of the louse. In adult female louse, the LsLp1 and LsLp2 transcripts were found in the sub-epidermal tissue and the intestine. RNA interference-mediated knockdown of LsLp1 and LsLp2 in female lice resulted in reduced expression of both transcripts. LsLp1 knockdown female lice produced significantly less offspring than control lice, while knockdown of LsLp2 in female lice caused no reduction in the number of offspring. These results suggest that LsLp1 has an important role in reproduction in female salmon lice.

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