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1.
Toxicon ; 184: 167-174, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32565098

RESUMO

Only a few work have been done for peptides from non-venom gland tissues of venomous animals. Here, with the help of the whole body transcriptomic and the hemolymph proteomic data of the Chinese scorpion Buthus martensii Karsch, we identified the first Ascaris-type peptide BmHDP from scorpion hemolymph. The precursor of BmHDP has 80 residues, including a 16 residue signal peptide and a 64 residue mature peptide. The mature peptide has 10 conserved cysteines and adopts a conserved Ascaris-type fold. Using combined inclusion body refolding and biochemical identification strategies, recombinant BmHDP was obtained successfully. Protease inhibitory assays showed that BmHDP inhibited chymotrypsin apparently at a concentration of 8 nM. Patch-clamp experiments showed that BmHDP inhibited the Kv1.3 potassium channel apparently at a concentration of 1000 nM. Coagulation experiment assays showed that BmHDP inhibited intrinsic coagulation pathway apparently at a concentration of 500 nM. To the best of our knowledge, BmHDP is the first Ascaris-type peptide from scorpion hemolymph. Our work highlighted a functional link between scorpion non-venom gland peptides and venom gland toxin peptides, and suggested that scorpion hemolymph might be a new source of bioactive peptides.


Assuntos
Ascaris , Hemolinfa/química , Venenos de Escorpião/química , Escorpiões , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA Complementar , Biblioteca Gênica , Peptídeos , Proteômica
2.
J Appl Microbiol ; 129(1): 3-16, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32395854

RESUMO

AIMS: Brown ring disease (BRD) is an infection of the Manila clam Ruditapes philippinarum due to the pathogen Vibrio tapetis. During BRD, clams are facing immunodepression and shell biomineralization alteration. In this paper, we studied the role of pH on the growth of the pathogen and formulated hypothesis on the establishment of BRD by V. tapetis. METHODS AND RESULTS: In this study, we monitored the evolution of pH during the growth of V. tapetis in a range of pH and temperatures. We also measured the pH of Manila clam haemolymph and extrapallial fluids (EPFs) during infection by V. tapetis. We highlighted that V. tapetis modulates the external pH during its growth, to a value of 7·70. During the development of BRD, V. tapetis also influences EPFs and haemolymph pH in vitro in the first hours of exposure and in vivo after 3 days of infection. CONCLUSIONS: Our experiments have shown a close interaction between V. tapetis CECT4600, a pathogen of Manila clam that induces BRD, and the pH of different compartments of the animals during infection. These results indicate that the bacterium, through a direct mechanism or as a consequence of physiological changes encountered in the animal during infection, is able to interfere with the pH of Manila clam fluids. This pH modification might promote the infection process or at least create an imbalance within the animal that would favour its persistence. This last hypothesis should be tested in future experiment. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first observation of pH modifications in the context of BRD and might orient future research on the fine mechanisms of pH modulation associated with BRD.


Assuntos
Bivalves/microbiologia , Vibrio/fisiologia , Vibrio/patogenicidade , Animais , Hemolinfa/química , Interações Hospedeiro-Patógeno , Concentração de Íons de Hidrogênio , Alimentos Marinhos/microbiologia , Temperatura , Vibrio/crescimento & desenvolvimento
3.
Subcell Biochem ; 94: 35-62, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32189295

RESUMO

Lipoproteins mediate the transport of apolar lipids in the hydrophilic environment of physiological fluids such as the vertebrate blood and the arthropod hemolymph. In this overview, we will focus on the hemolymph lipoproteins in Crustacea that have received most attention during the last years: the high density lipoprotein/ß-glucan binding proteins (HDL-BGBPs), the vitellogenins (VGs), the clotting proteins (CPs) and the more recently discovered large discoidal lipoproteins (dLPs). VGs are female specific lipoproteins which supply both proteins and lipids as storage material for the oocyte for later use by the developing embryo. Unusual within the invertebrates, the crustacean yolk proteins-formerly designated VGs-are more related to the ApoB type lipoproteins of vertebrates and are now termed apolipocrustaceins. The CPs on the other hand, which are present in both sexes, are related to the (sex specific) VGs of insects and vertebrates. CPs serve in hemostasis and wound closure but also as storage proteins in the oocyte. The HDL-BGBPs are the main lipid transporters, but are also involved in immune defense. Most crustacean lipoproteins belong to the family of the large lipid transfer proteins (LLTPs) such as the intracellular microsomal triglyceride transfer protein, the VGs, CPs and the dLPs. In contrast, the HDL-BGBPs do not belong to the LLTPs and their relationship with other lipoproteins is unknown. However, they originate from a common precursor with the dLPs, whose functions are as yet unknown. The majority of lipoprotein studies have focused on decapod crustaceans, especially shrimps, due to their economic importance. However, we will present evidence that the HDL-BGBPs are restricted to the decapod crustaceans which raises the question as to the main lipid transporting proteins of the other crustacean groups. The diversity of crustaceans lipoproteins thus appears to be more complex than reflected by the present state of knowledge.


Assuntos
Crustáceos , Hemolinfa , Lipoproteínas , Animais , Crustáceos/química , Crustáceos/imunologia , Crustáceos/metabolismo , Hemolinfa/química , Hemolinfa/imunologia , Hemolinfa/metabolismo , Lipoproteínas/biossíntese , Lipoproteínas/química , Lipoproteínas/imunologia , Lipoproteínas/metabolismo
4.
Subcell Biochem ; 94: 195-218, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32189300

RESUMO

Instead of the red blood of vertebrates, most molluscs have blue hemolymph containing hemocyanin, a type-3 copper-containing protein. The hemoglobin of vertebrate blood is replaced in most molluscs with hemocyanin, which plays the role of an  oxygen transporter. Oxygen-binding in hemocyanin changes its hue from colorless deoxygenated hemocyanin into blue oxygenated hemocyanin. Molecules of molluscan hemocyanin are huge, cylindrical multimeric proteins-one of the largest protein molecules in the natural world. Their huge molecular weight (from 3.3 MDa to more than 10 MDa) are the defining characteristic of molluscan hemocyanin, a property that has complicated structural analysis of the molecules for a long time. Recently, the structural analysis of a cephalopod (squid) hemocyanin has succeeded using a hybrid method employing both X-ray crystallography and cryo-EM. In a biochemical breakthrough for molluscan hemocyanin, the first quaternary structure with atomic resolution is on the verge of solving the mystery of molluscan hemocyanin. Here we describe the latest information about the molecular structure, classification and evolution of the molecule, and the physiology of molluscan hemocyanin.


Assuntos
Hemocianinas/química , Hemocianinas/metabolismo , Animais , Cristalografia por Raios X , Hemolinfa/química , Modelos Moleculares , Estrutura Molecular , Moluscos/química
5.
PLoS One ; 15(2): e0228556, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32027696

RESUMO

Invertebrates are becoming more popular models for research on the immune system. The innate immunity possessed by insects shows both structural and functional similarity to the resistance displayed by mammals, and many processes occurring in insect hemocytes are similar to those that occur in mammals. The humoral immune response in insects acts by melanization, clotting and the production of reactive oxygen species and antimicrobial peptides, while the cellular immunity system is based on nodulation, encapsulation and phagocytosis. An increasingly popular insect model in biological research is Galleria mellonella, whose larvae are sensitive to infection by the entomopathogenic fungus Conidiobolus coronatus, which can also be dangerous to humans. One group of factors that modulate the response of the immune system during infection in mammals are heat shock proteins (HSPs). The aim of this study was to investigate whether infection by C. coronatus in G. mellonella hemolymph is accompanied by an increase of HSP90, HSP70, HSP60 and HSP27. Larvae (five-day-old last instar) were exposed for 24 hours to fully-grown and sporulating fungus. Hemolymph was collected either immediately after termination of exposure (F24) or 24 hours later (F48). The concentration of the HSPs in hemolymph was determined using ELISA. Immunolocalization in hemocytes was performed using fluorescence microscopy and flow cytometry. HSP90, HSP70, HSP60 and HSP27 were found to be present in the G. mellonella hemocytes. HSP60 and HSP90 predominated in healthy insects, with HSP70 and HSP27 being found in trace amounts; HSP60 and HSP27 were elevated in F24 and F48, and HSP90 was elevated in F48. The fungal infection had no effect on HSP70 levels. These findings shed light on the mechanisms underlying the interaction between the innate insect immune response and entomopathogen infection. The results of this innovative study may have a considerable impact on research concerning innate immunology and insect physiology.


Assuntos
Conidiobolus/patogenicidade , Proteínas de Choque Térmico/metabolismo , Hemolinfa/química , Lepidópteros/química , Zigomicose/imunologia , Animais , Proteínas de Choque Térmico/análise , Proteínas de Choque Térmico/imunologia , Hemócitos/metabolismo , Hemolinfa/microbiologia , Humanos , Imunidade Inata , Larva/microbiologia , Lepidópteros/imunologia , Lepidópteros/microbiologia , Fatores de Tempo , Regulação para Cima
6.
J Chem Ecol ; 46(2): 186-197, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31953704

RESUMO

The horseradish flea beetle Phyllotreta armoraciae exclusively feeds on Brassicaceae, which contain glucosinolates as characteristic defense compounds. Although glucosinolates are usually degraded by plant enzymes (myrosinases) to toxic isothiocyanates after ingestion, P. armoraciae beetles sequester glucosinolates. Between and within brassicaceous plants, the glucosinolate content and composition can differ drastically. But how do these factors influence sequestration in P. armoraciae? To address this question, we performed a five-day feeding experiment with three Arabidopsis thaliana lines that differ four-fold in glucosinolate content and the composition of aliphatic and indolic glucosinolates. We quantified the amounts of ingested, sequestered, and excreted glucosinolates, and analyzed the changes in glucosinolate levels and composition in beetles before and after feeding on Arabidopsis. P. armoraciae accumulated almost all ingested glucosinolate types. However, some glucosinolates were accumulated more efficiently than others, and selected glucosinolates were modified by the beetles. The uptake of new glucosinolates correlated with a decrease in the level of stored glucosinolates so that the total glucosinolate content remained stable at around 35 nmol/mg beetle fresh weight. Beetles excreted previously stored as well as ingested glucosinolates from Arabidopsis, which suggests that P. armoraciae regulate their endogenous glucosinolate level by excretion. The metabolic fate of ingested glucosinolates, i.e. the proportions of sequestered and excreted glucosinolates, depended on glucosinolate type, content, and composition in the food plant. Overall, P. armoraciae sequestered and excreted up to 41% and 31% of the total ingested aliphatic and indolic glucosinolates from Arabidopsis, respectively. In summary, we show that glucosinolate variability in Brassicaceae influences the composition but not the level of sequestered glucosinolates in P. armoraciae beetles.


Assuntos
Brassicaceae/química , Besouros/metabolismo , Glucosinolatos/metabolismo , Animais , Arabidopsis/química , Arabidopsis/metabolismo , Brassicaceae/metabolismo , Fezes/química , Glucosinolatos/química , Hemolinfa/química , Hemolinfa/metabolismo , Interações Hospedeiro-Parasita
7.
Artigo em Inglês | MEDLINE | ID: mdl-31931329

RESUMO

The silkworm, Bombyx mori, is a promising expression system for the production of recombinant proteins, but the purification of these proteins is not easy because of the large amount of host proteins present. To investigate purity, recovery and scale-up ability of the purification of recombinant proteins expressed in silkworm larval hemolymph without any affinity tags, we used mCherry, a red fluorescence protein, as a model. The host cell proteins could be greatly reduced using a three-step chromatography protocol consisting of hydrophobic interaction chromatography (HIC), size exclusion chromatography (SEC) and heparin chromatography after heat pretreatment. The thermal treatment had the greatest impact on the removal of host cell extracellular proteins and increasing purity. There were still some minor traces of host cell proteins in the purified sample, which showed that the purification of recombinant proteins from the silkworm hemolymph was still challenging. The proposed protocol and affinity tag purification reduced the overall protein content by 99.84% and 99.95%, respectively, while the amount of DNA was reduced by 98.41% and 99.53%, respectively. Purities of our proposed protocol based on SDS-PAGE and capillary electrophoresis (CE) analyses were 85.45% and 43.60%, respectively, while those of Strep-tag affinity purification were 100% or 63.69%, respectively. Using densitometry, the overall recovery was calculated was 5.78%, which was higher than 4.09% using Strep-tag affinity purification. This proposed protocol, mainly based on thermal treatment, HIC, SEC and HiTrap Heparin HP column chromatography, is applicable to an upscalable purification for the silkworm expression system without employing affinity tag chromatography process.


Assuntos
Bombyx/química , Cromatografia de Afinidade/métodos , Hemolinfa/química , Larva/química , Proteínas Recombinantes de Fusão/isolamento & purificação , Animais , Bombyx/metabolismo , Eletroforese em Gel de Poliacrilamida , Larva/metabolismo , Oligopeptídeos/genética , Oligopeptídeos/metabolismo , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética
8.
Dev Comp Immunol ; 102: 103470, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31430486

RESUMO

Special innate immune mechanisms against pathogens are developed in marine invertebrates such as mud crab, which is also an economically important aquaculture species in many coastal countries. Hemolymph is a critical site in host immune response, but its source of microorganisms is less known. In this study, we provided a detailed investigation of the microorganisms inhabiting various body sites of healthy mud crabs, including hemolymph, midgut, gill, subcuticular epidermis and hepatopancreas. By using fluorescence microscopy and high-throughput sequencing of the bacterial 16S rRNA genes, various abundances and kinds of microorganisms were observed in the healthy mud crabs, of which some are potential pathogens to mud crab and human. The SourceTracker analysis and oral injection experiment confirm the hypothesis that hemolymph microorganisms are derived from the digestive systems of invertebrates with open circulatory systems, indicating that these microorganisms play vital roles in crab immune response. Moreover, physiological differences (gut length), behavioral characteristics (foraging behavior), diet preferences (herbivory), and/or sex hormones (testosterone) possibly determine the unique features of the crab-associated microbiota for both sexes. These findings also contribute to the development of appropriate microbial immunoenhancers, which has potential applications for improving quality and yield during crab aquaculture.


Assuntos
Braquiúros/imunologia , Braquiúros/microbiologia , Hemolinfa/microbiologia , Microbiota , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/patogenicidade , Feminino , Hemolinfa/química , Masculino , Especificidade de Órgãos , Filogenia , RNA Ribossômico 16S/genética , Alimentos Marinhos/microbiologia , Água do Mar/microbiologia
9.
Naturwissenschaften ; 107(1): 1, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31797054

RESUMO

Larvae of most Pergidae and Argidae (Symphyta: Tenthredinoidea) species contain toxic peptides such as pergidin and lophyrotomin. Here, larval hemolymph and organs of the pergid Lophyrotoma zonalis and the argid Arge pagana were analysed by liquid chromatography-tandem mass spectrometry. The major identified peptides were pergidin and 4-valinepergidin in L. zonalis, whereas pergidin and lophyrotomin in A. pagana. The storage period prior to chemical analysis was longer for the samples of the pergid than the argid species, which influenced peptide concentrations. In both species, however, the peptides occurred in decreasing order of concentration, first in the hemolymph, then in the integument, while minor amounts of the peptides were detected in other organs such as gut and fat body. By separating the cuticle of the pergid from the remaining integument, the peptides were found in equivalent amounts in each of these two body structures. The results suggest that the peptides play an important role in the defence of these sawfly larvae against predators.


Assuntos
Himenópteros/metabolismo , Peptídeos/metabolismo , Toxinas Biológicas/metabolismo , Animais , Cromatografia Líquida , Corpo Adiposo/química , Trato Gastrointestinal/química , Hemolinfa/química , Himenópteros/química , Larva/metabolismo , Espectrometria de Massas , Distribuição Tecidual , Toxinas Biológicas/química
10.
Parasit Vectors ; 12(1): 485, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31619284

RESUMO

BACKGROUND: Parasite traits associated with transmission success, such as the number of infective stages released from the host, are expected to be optimized by natural selection. However, in the trematode parasite Schistosoma mansoni, a key transmission trait, i.e. the number of cercariae larvae shed from infected Biomphalaria spp. snails, varies significantly within and between different parasite populations and selection experiments demonstrate that this variation has a strong genetic basis. In this study, we compared the transmission strategies of two laboratory schistosome population and their consequences for their snail host. METHODS: We infected inbred Biomphalaria glabrata snails using two S. mansoni parasite populations (SmBRE and SmLE), both isolated from Brazil and maintained in the laboratory for decades. We compared life history traits of these two parasite populations by quantifying sporocyst growth within infected snails (assayed using qPCR), output of cercaria larvae and impact on snail host physiological response (i.e. hemoglobin rate, laccase-like activity) and survival. RESULTS: We identified striking differences in virulence and transmission between the two studied parasite populations. SmBRE (low shedder (LS) parasite population) sheds very low numbers of cercariae and causes minimal impact on the snail physiological response (i.e. laccase-like activity, hemoglobin rate and snail survival). In contrast, SmLE (high shedder (HS) parasite population) sheds 8-fold more cercariae (mean ± SE cercariae per shedding: 284 ± 19 vs 2352 ± 113), causes high snail mortality and has strong impact on snail physiology. We found that HS sporocysts grow more rapidly inside the snail host, comprising up to 60% of cells within infected snails, compared to LS sporocysts, which comprised up to 31%. Cercarial production is strongly correlated to the number of S. mansoni sporocyst cells present within the snail host tissue, although the proportion of sporocyst cells alone does not explain the low cercarial shedding of SmBRE. CONCLUSIONS: We demonstrated the existence of alternative transmission strategies in the S. mansoni parasite consistent with trade-offs between parasite transmission and host survival: a "boom-bust" strategy characterized by high virulence, high transmission and short duration infections and a "slow and steady" strategy with low virulence, low transmission but long duration of snail host infections.


Assuntos
Biomphalaria/parasitologia , Schistosoma mansoni/fisiologia , Schistosoma mansoni/patogenicidade , Esquistossomose mansoni/parasitologia , Esquistossomose mansoni/transmissão , Animais , Biomphalaria/fisiologia , Brasil , Cercárias , Estudos de Coortes , Cricetinae , DNA de Helmintos/química , DNA de Helmintos/isolamento & purificação , Vetores de Doenças , Feminino , Hemoglobinas/análise , Hemolinfa/química , Hemolinfa/enzimologia , Humanos , Lacase/análise , Masculino , Mesocricetus , Reação em Cadeia da Polimerase Multiplex , Reação em Cadeia da Polimerase em Tempo Real , Schistosoma mansoni/crescimento & desenvolvimento , Razão de Masculinidade , Virulência
11.
Parasit Vectors ; 12(1): 472, 2019 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-31604479

RESUMO

BACKGROUND: The parasitic dinoflagellates of the genus Hematodinium represent the causative agent of so-called bitter or pink crab disease in a broad range of shellfish taxa. Outbreaks of Hematodinium-associated disease can devastate local fishing and aquaculture efforts. The goal of our study was to examine the potential role of the common shore (green) crab Carcinus maenas as a reservoir for Hematodinium. Carcinus maenas is native to all shores of the UK and Ireland and the North East Atlantic but has been introduced to, and subsequently invaded waters of, the USA, South Africa and Australia. This species is notable for its capacity to harbour a range of micro- and macro-parasites, and therefore may act as a vector for disease transfer. METHODS: Over a 12-month period, we interrogated 1191 crabs across two distinct locations (intertidal pier, semi-closed dock) in Swansea Bay (Wales, UK) for the presence and severity of Hematodinium in the haemolymph, gills, hepatopancreas and surrounding waters (eDNA) using PCR-based methods, haemolymph preparations and histopathology. RESULTS: Overall, 13.6% were Hematodinium-positive via PCR and confirmed via tissue examination. Only a small difference was observed between locations with 14.4% and 12.8% infected crabs in the Dock and Pier, respectively. Binomial logistic regression models revealed seasonality (P < 0.002) and sex (P < 0.001) to be significant factors in Hematodinium detection with peak infection recorded in spring (March to May). Male crabs overall were more likely to be infected. Phylogenetic analyses of the partial ITS and 18S rRNA gene regions of Hematodinium amplified from crabs determined the causative agent to be the host generalist Hematodinium sp., which blights several valuable crustaceans in the UK alone, including edible crabs (Cancer pagurus) and langoustines (Nephrops norvegicus). CONCLUSIONS: Shore crabs were infected with the host generalist parasite Hematodinium sp. in each location tested, thereby enabling the parasite to persist in an environment shared with commercially important shellfish.


Assuntos
Alveolados/patogenicidade , Braquiúros/parasitologia , Reservatórios de Doenças/parasitologia , Alveolados/classificação , Alveolados/genética , Alveolados/fisiologia , Animais , Distribuição Binomial , DNA Espaçador Ribossômico/genética , Dinoflagelados/classificação , Dinoflagelados/genética , Dinoflagelados/patogenicidade , Dinoflagelados/fisiologia , Feminino , Brânquias/parasitologia , Hemolinfa/química , Hemolinfa/parasitologia , Modelos Logísticos , Masculino , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/análise , Estações do Ano , Água do Mar/parasitologia , País de Gales
12.
Fish Shellfish Immunol ; 94: 896-906, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31533083

RESUMO

The study is carried out to understand the antimicrobial and immunological response of a potential immune molecule lectin, MmLec isolated from haemolymph of Speckled shrimp, Metapenaeus monoceros. MmLec was purified using mannose coupled Sepharose CL-4B affinity chromatography, which was further subjected on SDS-PAGE to ascertain the distribution of their molecular weight. Sugar binding specificity assay was conducted at various pH and temperatures to investigate the binding affinity of MmLec towards the specific carbohydrate molecule. Functional analysis of immune molecule MmLec included haemagglutination assays performed using human erythrocytes and yeast agglutination activity against Saccharomyces cerevisiae which, were analyzed using light microscopy. In order to study the antimicrobial activity, two Gram-negative (Vibrio parahaemolyticus and Aeromonas hydrophila) and two Gram-positive (Staphylococcus aureus and Enterococcus faecalis) bacteria were treated with purified MmLec. Moreover, these bacterial species were also treated at different concentration of the MmLec to speculate the antibiofilm properties of MmLec which was analyzed under Light Microscopy and Confocal Laser Scanning Microscopy. In addition, other functional characterization of MmLec showed the uniqueness of MmLec in agglutination of human erythrocyte as well as the cells of yeast Saccharomyces cerevisiae. Also, the phenoloxidase activity and encapsulation assay was evaluated. MTT assay displayed that MmLec are potent in anticancer activity. The study will help to understand the immunological interference and antimicrobial nature of MmLec which would be supportive in establishing a potential therapeutic tool and to develop better and novel disease control strategies in shrimp and farmed aquaculture industries as well as in health management.


Assuntos
Biofilmes/efeitos dos fármacos , Lectinas/imunologia , Penaeidae/imunologia , Aeromonas hydrophila/fisiologia , Animais , Enterococcus faecalis/fisiologia , Hemolinfa/química , Lectinas/farmacologia , Penaeidae/fisiologia , Staphylococcus aureus/fisiologia , Vibrio parahaemolyticus/fisiologia
13.
Mar Pollut Bull ; 149: 110537, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31466014

RESUMO

Plastic waste is ubiquitous in marine environments. Despite the sheer volume of plastic waste, it remains relatively unknown how marine invertebrates will interact with microplastics (plastic <1 mm). Microplastics (<2 µm) were ingested by the economically and ecologically significant Sydney rock oyster Saccostrea glomerata and translocated to the haemolymph, perhaps via phagocytosis. The presence of microplastics in the haemolymph indicates that filter feeding S. glomerata can ingest and accumulate microplastics which are prevalent in the environment. This research shows microplastics can enter marine molluscs and highlights the need to monitor microplastics in the marine environment and aquaculture to safeguard the seafood industry.


Assuntos
Hemolinfa/química , Microplásticos/farmacocinética , Ostreidae/química , Poluentes Químicos da Água/farmacocinética , Animais , Exposição Dietética , Ecotoxicologia , Microplásticos/análise , Frutos do Mar , Poluentes Químicos da Água/análise
14.
Comp Immunol Microbiol Infect Dis ; 66: 101343, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31446196

RESUMO

Invertebrates have been instrumental in understanding the mechanisms involved in infectious diseases, considering the idea to replace, reduce and refine the use of mammals as well as to understand the basic principles of immune response in insect. We evaluated the consequences of Staphylococcus aureus-induced sepsis in the last instar nymphs of Nauphoeta cinerea injected with different concentrations of bacteria preserved in two culture media. Infected groups had a decrease in hemolymph metabolites (glucose, amino acids, total proteins, and cholesterol), in contrast to the proteins in the fat body. Higher concentrations of S. aureus caused permanent morphological alterations in adults, decrease in food consumption, increase in isolation, and increase in CFU until death of the cockroaches. Survival and protection of nymphs against a repeated and stronger challenge with the same bacteria varied according to the medium they were conserved. N. cinerea proves to be a suitable and promising model for studies related to bacterial infections.


Assuntos
Baratas/microbiologia , Hemolinfa/química , Ninfa/microbiologia , Sepse/microbiologia , Infecções Estafilocócicas/sangue , Staphylococcus aureus/crescimento & desenvolvimento , Animais , Contagem de Colônia Microbiana , Comportamento Alimentar , Sepse/imunologia , Infecções Estafilocócicas/imunologia , Staphylococcus aureus/patogenicidade
15.
J Insect Physiol ; 118: 103913, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31302015

RESUMO

Silkworms are economically important insects because of the value of their silk. After finishing silk spinning, silkworms begin another important physiological process, vitellogenesis. In this study, we explored the relationship between silk spinning and vitellogenin (BmVg) expression in silkworms. In silkworms with the silk fibroin heavy chain gene knocked-out, the concentration of amino acids in the hemolymph was found to be significantly higher than that in the wild type, and the expression of BmVg was advanced at day 7 of the fifth instar stage and 0 h after spinning. Furthermore, through culturing fat body in vitro with different substances treatment including glucose, trehalose, amino acids, 20-hydroxyecdysone, and insulin, we found that only amino acids could induce BmVg expression. RNA interference of BmTOR1 in female silkworms could down-regulate BmVg transcription, resulting in shortened egg ducts and smaller eggs relative to the control. Therefore, these results showed that amino acids could induce BmVg expression through the TOR signaling pathway. Fat body cultured with amino acids in vitro and experiments involving amino acids injected into the silkworm showed that the majority of main amino acids of silk protein could induce BmVg expression. These results suggested that BmVg expression is related to silk spinning and this study would lay a foundation for elucidating the stage specificity expression of BmVg.


Assuntos
Aminoácidos/metabolismo , Bombyx/metabolismo , Vitelogeninas/metabolismo , Aminoácidos/farmacologia , Animais , Bombyx/genética , Bombyx/crescimento & desenvolvimento , Corpo Adiposo/efeitos dos fármacos , Corpo Adiposo/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Genitália Feminina/metabolismo , Hemolinfa/química , Proteínas de Insetos/metabolismo , Estágios do Ciclo de Vida , Interferência de RNA , Seda/química , Técnicas de Cultura de Tecidos , Vitelogeninas/genética
16.
Pol J Vet Sci ; 22(2): 61-65, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31269337

RESUMO

Edible snails are kept in farms in many countries worldwide. As farm animals, they are an object of interest of veterinary studies and applied biology. There is a large demand for tests which would help identify their health and well-being. The objective of this study was to assess the usefulness of determining the concentration of urea in hemolymph as a marker of health of the Lissachatina fulica and Cornu aspersum edible snails. The observation covered snails from four farms marked from A to D, in which numerous deaths (farm A) and decreased body weight gain (farms B and C) were observed. In experimental farm D we observed a group of snails subjected to stress and a control group maintained in correct conditions. High concentrations of urea were found in the hemolymph of all farm animals from farms A, B and C, as well as in those subjected to food deprivation in farm D (on average from 96 mg/dl in farm D to 320 mg/dl in farm A). On the other hand, in controls from group D, the concentration of the parameter in question was much lower (⟨ 2.0 mg/dl). The results obtained indicate that the urea concentration is a non-specific marker of pathological conditions in snails, and that the continuous monitoring of this parameter makes it possible to demonstrate irregularities in farming and introduce appropriate and early measures to eliminate such disturbances.


Assuntos
Hemolinfa/química , Caramujos/fisiologia , Ureia/química , Criação de Animais Domésticos , Animais
17.
J Environ Sci Health B ; 54(8): 676-680, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31230517

RESUMO

Members of TRP receptor family are involved in response to acidification. Here, we determined the effect of capsaicin, one of the TRP receptor activators, on hemolymph acid-base status in the American cockroach. Periplaneta americana adult individuals were injected with lactic acid (5% or 10%) and exposed to 100 µM capsaicin solution. Hemolymph pH was measured 15 min, 1, 4, 8 and 24 h after lactic acid and capsaicin application with a glass microelectrode. The results demonstrated that cockroaches recover from acidosis within 4 h from acid injection. Capsaicin impaired the buffering capacity of insects' hemolymph, resulting in significant drop of hemolymph pH observed even 24 h after application. Joint action of capsaicin and acidosis reveals new insight into possible mechanism of capsaicin action on TRP receptors in insects.


Assuntos
Capsaicina/farmacologia , Hemolinfa/efeitos dos fármacos , Periplaneta/efeitos dos fármacos , Equilíbrio Ácido-Base/efeitos dos fármacos , Animais , Feminino , Hemolinfa/química , Hemolinfa/metabolismo , Concentração de Íons de Hidrogênio , Inseticidas/farmacologia , Ácido Láctico/farmacologia , Masculino , Mortalidade , Periplaneta/metabolismo , Canais de Receptores Transientes de Potencial/metabolismo
18.
Fish Shellfish Immunol ; 93: 752-762, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31163297

RESUMO

The West Coast rock lobster (WCRL), Jasus lalandii, inhabits highly variable environments frequented by upwelling events, episodes of hypercapnia and large temperature variations. Coupled with the predicted threat of ocean acidification and temperature change for the coming centuries, the immune response in this crustacean will most likely be affected. We therefore tested the hypothesis that chronic exposure to hypercapnia and elevated seawater temperature will alter immune function of the WCRL. The chronic effects of four combinations of two stressors (seawater pCO2 and temperature) on the total number of circulating haemocytes (THC) as well as on the lobsters' ability to clear (inactivate) an injected dose of Vibrio anguillarum from haemolymph circulation were assessed. Juvenile lobsters were held in normocapnic (pH 8.01) or hypercapnic (pH 7.34) conditions at two temperatures (15.6 and 18.9 °C) for 48 weeks (n = 30 lobster per treatment), after which a subsample of lobsters (n = 8/treatment), all at a similar moult stage, were selected from each treatment for the immune challenge. Baseline levels of haemocytes (THC ml-1) and bacteria (CFU ml-1) in their haemolymph were quantified 24 h prior to bacterial challenge. Lobsters were then challenged by injecting 4 × 104V. anguillarum per g body weight directly into the cardiac region of each lobster and circulating haemocyte and culturable bacteria were measured at 20 min post challenge. No significant differences in THC ml-1 (p < 0.05) were observed between any of the treatment groups prior to the bacterial challenge. However lobsters chronically exposed to a combination of hypercapnia and low temperature had significantly higher (p < 0.05) THCs post-challenge in comparison with lobsters chronically exposed to hypercapnia and high temperature. A significant interactive effect was recorded between temperature and pH for the post-challenge THC data (two-way ANOVA, p = 0.0025). Lobster were very efficient at rendering an injected dose of bacteria non-culturable, with more than 83% of the theoretical challenge dose (∼1.7 × 105Vibrio ml-1 haemolymph) inactivated within the first 10 min following injection. Although differences in the inactivation of V. anguillarum were observed between treatment groups, none of these differences were significant. Clearance efficiency was in the following order: Hypercapnia/low temperature > normocapnia/high temperature > normocapnia/low temperature > hypercapnia/high temperature. This study demonstrated that despite chronic exposure to combinations of reduced seawater pH and high temperature, the WCRL was still capable of rapidly rendering an injected dose of bacteria non-culturable.


Assuntos
Dióxido de Carbono/sangue , Hemolinfa/química , Imunidade Inata/fisiologia , Palinuridae/imunologia , Animais , Temperatura Alta/efeitos adversos , Concentração de Íons de Hidrogênio , Palinuridae/química , Palinuridae/fisiologia , Água do Mar/química
19.
J Zoo Wildl Med ; 50(1): 111-122, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-31120669

RESUMO

Large arachnids are commonly managed under professional care, and anesthesia is occasionally required for physical examination and diagnostic and therapeutic procedures. Anesthetic responses and hemolymph gas analysis have been studied previously in spiders, but scorpions have yet to be investigated. This study measured hemolymph gas values with an i-STAT point of care blood gas analyzer in healthy adult Asian forest scorpions (Heterometrus longimanus = HL, n = 8) and dictator scorpions (Pandinus dictator = PD, n = 12) breathing: 1) room air (RA), 2) 100% oxygen for 10 min in a chamber (OX), and 3) 5% isoflurane and oxygen (ISO) in a chamber until induction or loss of righting reflex. All scorpions recovered without complications, and there were no cartridge failures. Analysis of hemolymph gas values revealed that pH was lower in OX compared with RA and ISO and was lower in PD compared with HL scorpions. The partial pressure of carbon dioxide did not differ between inhaled gases but was higher in PD compared with HL. The partial pressure of oxygen (pO2) was higher in ISO compared with OX, and both were higher than when breathing RA. Despite a lack of species difference in pO2, PD had a more dramatic increase in pO2 in ISO compared with HL (significant species and inhalant interaction). PD had a significantly shorter induction time than HL, but recovery time (return of righting reflex) did not differ between species. Subjectively, HL exhibited rough inductions compared with PD, characterized by violent whole-body and tail movements. The unexpected increase in pO2 in ISO compared with OX, along with the species-specific differences and anesthetic effects, emphasizes the unique respiratory physiology of scorpions and demonstrates that further species-specific studies of anesthetics are warranted.


Assuntos
Anestésicos Inalatórios/efeitos adversos , Isoflurano/efeitos adversos , Oxigênio/efeitos adversos , Escorpiões/efeitos dos fármacos , Aranhas/efeitos dos fármacos , Anestésicos Inalatórios/administração & dosagem , Animais , Gasometria/veterinária , Hemolinfa/química , Isoflurano/administração & dosagem , Oxigênio/administração & dosagem , Reflexo de Endireitamento/efeitos dos fármacos , Escorpiões/fisiologia , Aranhas/fisiologia
20.
J Zoo Wildl Med ; 50(1): 127-136, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-31120671

RESUMO

Tarantulas are a commonly kept species that are occasionally presented to veterinarians in exotic practice. A recent study on Grammastola rosea hemolymph biochemistry has been performed with nonanesthetized adult theraphosids. The objective of this study was to produce reference intervals for biochemistry biomarkers in hemolymph of chemically restrained G. rosea for use diagnostically by exotic veterinarians. Cardiac hemolymph collection was performed on 20 subadult tarantulas under general anesthesia with isoflurane. Samples were processed by a commercial laboratory. Statistics performed on the data include outlier exclusion, descriptive statistics, normality tests, and Pearson correlations. Reference intervals were made for total protein, creatine kinase (CK), aspartate aminotransferase (AST), glucose, uric acid (UA), calcium, and phosphorus. No cortisol was detected. The majority of the intervals produced were normally distributed with the exceptions of UA, phosphorus, and CK. Pearson correlation tests found several significant (P = <0.05) correlations between variables. The majority of the data displayed a normal distribution, unlike the previous study, with a greater number of replicates. The total protein, glucose, UA, calcium, and AST values generated were similar to those reported in the previous study. Conversely several variables such as phosphorus, CK, and albumin were not consistent with those previously reported. Evidence is presented for a lack of albumin, CK, and AST in Arachnida and thus previous data for these proteins is likely to be artifactual.


Assuntos
Anestésicos Gerais/administração & dosagem , Hemolinfa/química , Isoflurano/administração & dosagem , Aranhas/química , Anestesia Geral/veterinária , Animais , Análise Química do Sangue/veterinária , Valores de Referência , Aranhas/efeitos dos fármacos
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