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1.
Exp Parasitol ; 206: 107755, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31493393

RESUMO

The aim of the present study was to assess the expression of cytokines and FCεR1A receptor stimulated by Haemonchus placei larval excretory and secretory (ES) products associated with the pathogenesis in calves. Bovine peripheral blood mononuclear cells (PBMC) were stimulated in in vitro assays with H. placei L4 ES product at 8, 12, 16 and 24 h. ES products were collected in in vitro assays at 48 h with molecular weight of 72/60 kDa and isoelectric point of 7.2 pI. Specific IgG for infected and control calves, positive and negative, were employed to recognise H. placei larval ES products by indirect ELISA, showing a mean of 1.8, 0.83 and 0.28 OD, respectively, (p ≤ 0.001). The quantification of relative gene expression was performed using a set of cytokines (IL-2, IFNγ, TGFß, IL-4, IL-5, IL-6, IL-8, IL-10 and IL-13), FCεR1A receptor and housekeeping (GAPDH, ß-actin and ß-2-microglobulin) by RT-qPCR. An early increased expression, 2.2- to 3.4-fold change, of IL-2 (p ≤ 0.001), IL-5 and TGFß (p ≥ 0.05) was determined, followed by TGFß (30.7 and 14.14), IL-8 (102.8 and 1504.4) and IL-10 (60.4 and 1.7) (p ≤ 0.05) after 12 and 16 h, respectively, and reducing the expression level at 24 h. In addition, IL-6, IL-13 and FCεR1A receptor also displayed mild expression level, 2.1 - to 7.60-fold change, at 24 h (p ≥ 0.05). We conclude that ES products of 72/60 kDa collected in vitro from H. placei larvae are recognised by infected hosts and have the ability to induce diverse immune factors to modulate the nematode damage.


Assuntos
Citocinas/metabolismo , Haemonchus/metabolismo , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/imunologia , Receptores de IgE/metabolismo , Animais , Bovinos , Proliferação de Células/efeitos dos fármacos , Citocinas/genética , Haemonchus/genética , Haemonchus/imunologia , Imunoglobulina G/metabolismo , Larva/genética , Larva/imunologia , Larva/metabolismo , Leucócitos Mononucleares/metabolismo , Masculino , RNA Mensageiro/metabolismo , Regulação para Cima
2.
J Agric Food Chem ; 67(36): 9979-9988, 2019 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-31411878

RESUMO

A delta class glutathione S-transferase gene (BoGSTd2) is identified from Bradysia odoriphaga for the first time. Developmental expression analysis showed that expression of BoGSTd2 is significantly higher in the fourth instar larval stage and the adult stage. Tissue-specific expression analysis found that BoGSTd2 was expressed predominantly in the midgut and Malpighian tubules in the fourth instar larvae and the abdomen of adults. Expression of BoGSTd2 was significantly upregulated following exposure to chlorpyrifos and clothianidin. In vitro inhibition and metabolic assays indicated that recombinant BoGSTd2 could not directly metabolize chlorpyrifos and clothianidin. Nevertheless, disk diffusion assays indicated that BoGSTd2 plays an important role in protection against oxidative stress. RNAi assays showed that BoGSTd2 participates in the elimination of reactive oxygen species induced by chlorpyrifos and clothianidin. These results strongly suggest that BoGSTd2 plays an important role in chlorpyrifos and clothianidin detoxification in B. odoriphaga by protecting tissues from oxidative stress induced by these insecticides.


Assuntos
Dípteros/enzimologia , Glutationa Transferase/metabolismo , Proteínas de Insetos/metabolismo , Inseticidas/metabolismo , Animais , Clorpirifos/metabolismo , Dípteros/genética , Dípteros/crescimento & desenvolvimento , Dípteros/metabolismo , Glutationa Transferase/genética , Guanidinas/metabolismo , Inativação Metabólica , Proteínas de Insetos/genética , Larva/enzimologia , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Neonicotinoides/metabolismo , Estresse Oxidativo , Tiazóis/metabolismo
3.
BMC Evol Biol ; 19(1): 150, 2019 07 24.
Artigo em Inglês | MEDLINE | ID: mdl-31340758

RESUMO

BACKGROUND: Understanding how variation in gene expression contributes to morphological diversity is a major goal in evolutionary biology. Cichlid fishes from the East African Great lakes exhibit striking diversity in trophic adaptations predicated on the functional modularity of their two sets of jaws (oral and pharyngeal). However, the transcriptional basis of this modularity is not so well understood, as no studies thus far have directly compared the expression of genes in the oral and pharyngeal jaws. Nor is it well understood how gene expression may have contributed to the parallel evolution of trophic morphologies across the replicate cichlid adaptive radiations in Lake Tanganyika, Malawi and Victoria. RESULTS: We set out to investigate the role of gene expression divergence in cichlid fishes from these three lakes adapted to herbivorous and carnivorous trophic niches. We focused on the development stage prior to the onset of exogenous feeding that is critical for understanding patterns of gene expression after oral and pharyngeal jaw skeletogenesis, anticipating environmental cues. This framework permitted us for the first time to test for signatures of gene expression underlying jaw modularity in convergent eco-morphologies across three independent adaptive radiations. We validated a set of reference genes, with stable expression between the two jaw types and across species, which can be important for future studies of gene expression in cichlid jaws. Next we found evidence of modular and non-modular gene expression between the two jaws, across different trophic niches and lakes. For instance, prdm1a, a skeletogenic gene with modular anterior-posterior expression, displayed higher pharyngeal jaw expression and modular expression pattern only in carnivorous species. Furthermore, we found the expression of genes in cichlids jaws from the youngest Lake Victoria to exhibit low modularity compared to the older lakes. CONCLUSION: Overall, our results provide cross-species transcriptional comparisons of modularly-regulated skeletogenic genes in the two jaw types, implicating expression differences which might contribute to the formation of divergent trophic morphologies at the stage of larval independence prior to foraging.


Assuntos
Adaptação Fisiológica/genética , Ciclídeos/genética , Comportamento Alimentar , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Variação Genética , Arcada Osseodentária/anatomia & histologia , Animais , Ecossistema , Redes Reguladoras de Genes , Lagos , Larva/genética , Morfogênese/genética , Faringe/metabolismo , Filogenia , Padrões de Referência , Tanzânia
4.
J Agric Food Chem ; 67(32): 8896-8904, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31339308

RESUMO

The mosquito Aedes aegypti is associated with the spread of many viral diseases in humans, including Dengue virus (DENVs), Yellow fever virus (YFV), Zika virus (ZIKV), and Chikungunya virus (CHIKV). Bacillus thuringiensis (Bt) is widely used as a biopesticide, which produces Cry toxins for mosquito control. The Cry toxins bind mainly to important receptors, including alkaline phosphatase (ALP) and aminopeptidase-N (APN). This work investigated the function of a C-type lectin, CTLGA9, in A. aegypti in response to Cry toxins. Our results showed by far-western blot and ELISA methods that the CTLTGA9 protein interacted with brush border membrane vesicles (BBMVs) of A. aegypti larvae and with ALP1, APN, and Cry11Aa proteins. Furthermore, molecular docking showed overlapping binding sites in ALP1 and APN for binding to Cry11Aa and CTLGA9. The toxicity assays further demonstrated that CTLGA9 inhibited the larvicidal activity of Cry toxins. According to the results of molecular docking, CTLGA9 may compete with Cry11Aa for binding to ALP1 and APN receptors and thus decreases the mosquitocidal toxicity of Cry11Aa. Our results provide further insights into better understanding the mechanism of Cry toxins and help improve the Cry toxicity for mosquito control.


Assuntos
Aedes/efeitos dos fármacos , Aedes/metabolismo , Proteínas de Bactérias/toxicidade , Endotoxinas/toxicidade , Proteínas Hemolisinas/toxicidade , Proteínas de Insetos/metabolismo , Aedes/química , Aedes/genética , Animais , Proteínas de Bactérias/química , Sítios de Ligação , Endotoxinas/química , Proteínas Hemolisinas/química , Proteínas de Insetos/química , Proteínas de Insetos/genética , Larva/efeitos dos fármacos , Larva/genética , Larva/metabolismo , Simulação de Acoplamento Molecular , Ligação Proteica/efeitos dos fármacos
5.
Int J Food Microbiol ; 305: 108256, 2019 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-31299548

RESUMO

Acanthocephalans of the genus Corynosoma are known as intestinal parasites, mainly of pinnipeds. Human corynosomiasis has been reported as an infrequent foodborne disease in Hokkaido, the northernmost island of Japan. Potential sources of the human infection are marine fish, because they are paratenic hosts of these parasites. In this study, the prevalence and intensity of larval Corynosoma in commercial fish from 17 fishing ports of Hokkaido were examined from April 2016 to January 2019. Out of a total of 1217 fish examined, 122 (10.0%) were infected with cystacanth larvae. The infected fish assemblage was composed of 7 families and 13 species from all the coastal seas of Hokkaido (the Pacific Ocean, Okhotsk Sea, and Japan Sea), showing that commercial fish can be source of human infection when eaten raw. Flatfish of the family Pleuronectidae showed the highest intensity of cystacanths, ranging from 1 to 56. A DNA barcoding system was developed in this study, based on the standard mitochondrial cox1 sequences of morphologically identified adults of Corynosoma spp. from pinnipeds in Hokkaido. By using the DNA barcoding, most of the fish-derived cystacanths were identified as either C. strumosum or C. villosum, and furthermore, a clinical isolate from human as C. villosum. Both of the species were commonly detected from various fish of Hokkaido, irrespective of the coastal seas. Flatfish frequently harbored C. villosum. Considering the wide range of commercial fish in Hokkaido and the advanced transportation system of fresh fish, there is a possibility that human corynosomiasis will occur everywhere in Japan.


Assuntos
Acantocéfalos/isolamento & purificação , Peixes/parasitologia , Acantocéfalos/classificação , Acantocéfalos/genética , Acantocéfalos/fisiologia , Animais , Código de Barras de DNA Taxonômico , Peixes/classificação , Helmintíase Animal , Ilhas , Japão , Larva/genética , Larva/fisiologia
6.
Environ Monit Assess ; 191(8): 497, 2019 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-31312907

RESUMO

A few studies had determined the effects of silver nanoparticles on the development of Drosophila melanogaster. However, none had addressed its genotoxic effects on specific larval cells of the fly in details. This study was conducted to determine the effects of silver nanoparticle on the development of D. melanogaster with simultaneous evaluation of its genotoxic potential on specific larval cell types that play important roles in immunological defenses as well as growth and development. Five male and five female flies were maintained in standard Drosophila melanogaster culture medium containing varying concentrations of silver nanoparticles, i.e., 25, 50, 100, 200, and 300 mg/l with control culture medium containing no nanoparticle. Total time needed for stage-specific development, population yield, and genotoxic effects on third instar larval polytene chromosomes, hemocytes, and neuroblasts was determined. Body pigmentation of pupae and young adults was examined visually. In comparison with control, silver nanoparticles dose dependently inhibited the metamororphosis and population yields of pupae and young adults of Drosophila melanogaster. Every concentration of the nanoparticles inhibited pupa to adult conversion, with huge reduction under the influence of nanoparticle concentration of 100 mg/ml and above. Developmental inhibition was accompanied by dose-dependent and significant structural aberrations of larval polytene chromosomes and deformities of hemocytes and neuroblasts. Pupae and young adults also exhibited gradual discoloration of body with the increase in exposure to nanoparticle concentration.


Assuntos
Dano ao DNA , Drosophila melanogaster/efeitos dos fármacos , Larva/efeitos dos fármacos , Nanopartículas Metálicas/toxicidade , Células-Tronco Neurais/efeitos dos fármacos , Prata/toxicidade , Animais , Relação Dose-Resposta a Droga , Drosophila melanogaster/genética , Drosophila melanogaster/crescimento & desenvolvimento , Monitoramento Ambiental , Feminino , Larva/genética , Larva/crescimento & desenvolvimento , Masculino , Pupa/efeitos dos fármacos , Pupa/genética , Pupa/crescimento & desenvolvimento
7.
Aquat Toxicol ; 214: 105257, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31336221

RESUMO

The interactions between nanoparticles (NPs) and metals in aquatic environments may modify the bioavailability and toxicity of metals to organisms. In this study, we investigated the effects of titanium dioxide NPs (n-TiO2) on the bioconcentration, depuration, and neurotoxic effects of lead (Pb) in zebrafish larvae. Transmission electron microscopy coupled with energy-dispersive X-ray spectroscopy showed that Pb2+ was adsorbed by n-TiO2 to form NP-Pb complexes in suspension, and these complexes were observed in larval tissues. The bioconcentration of Pb in larvae along with the depuration rates of Pb were higher in the presence of n-TiO2 compared to when n-TiO2 was absent. Exposure to Pb alone induced the expression of the biomarker metallothionein, downregulated neurodevelopment-related genes, and reduced swimming activity of larvae. However, the addition of n-TiO2 to the exposure solution alleviated these effects. The results suggest that n-TiO2 can act as a carrier of Pb to increase its bioconcentration; however, the formation of NP-Pb complexes likely reduces the amount of free Pb2+, thereby reducing toxicity to larvae.


Assuntos
Chumbo/toxicidade , Nanopartículas/toxicidade , Titânio/toxicidade , Peixe-Zebra/metabolismo , Adsorção , Animais , Comportamento Animal/efeitos dos fármacos , Disponibilidade Biológica , Larva/efeitos dos fármacos , Larva/genética , Larva/ultraestrutura , Locomoção/efeitos dos fármacos , Metalotioneína/metabolismo , Nanopartículas/ultraestrutura , Suspensões , Transcrição Genética/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/genética
8.
Nature ; 571(7765): 349-354, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31292549

RESUMO

Ascidian embryos highlight the importance of cell lineages in animal development. As simple proto-vertebrates, they also provide insights into the evolutionary origins of cell types such as cranial placodes and neural crest cells. Here we have determined single-cell transcriptomes for more than 90,000 cells that span the entirety of development-from the onset of gastrulation to swimming tadpoles-in Ciona intestinalis. Owing to the small numbers of cells in ascidian embryos, this represents an average of over 12-fold coverage for every cell at every stage of development. We used single-cell transcriptome trajectories to construct virtual cell-lineage maps and provisional gene networks for 41 neural subtypes that comprise the larval nervous system. We summarize several applications of these datasets, including annotating the synaptome of swimming tadpoles and tracing the evolutionary origin of cell types such as the vertebrate telencephalon.


Assuntos
Linhagem da Célula/genética , Ciona intestinalis/citologia , Ciona intestinalis/genética , Análise de Célula Única , Transcriptoma , Animais , Sequência de Bases , Evolução Biológica , Ciona intestinalis/classificação , Ciona intestinalis/crescimento & desenvolvimento , Gastrulação , Redes Reguladoras de Genes , Larva/citologia , Larva/genética , Sistema Nervoso/citologia , Sistema Nervoso/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Notocorda/citologia , Notocorda/embriologia , Especificidade de Órgãos , Sinapses/genética , Sinapses/metabolismo
9.
Parasitol Res ; 118(7): 2159-2168, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31165239

RESUMO

Terranova pectinolabiata n. sp. is described from the great hammerhead, Sphyrna mokarran, from Australian waters. This represents the first report of a species of Terranova from the host species. The new species is characterised by the morphology of the caudal plates and labia. ITS sequences were obtained for 20 specimens which were identical, despite morphological variation that has traditionally been indicative of separation of species. Additionally, genetic analyses confirmed the identification of the larval Terranova Type II previously reported in Australian and New Caledonian waters as Terranova pectinolabiata n. sp.


Assuntos
Ascaridoidea/classificação , Ascaridoidea/genética , Tubarões/parasitologia , Animais , Ascaridoidea/isolamento & purificação , Austrália , DNA Intergênico/genética , Larva/genética
10.
Arch Insect Biochem Physiol ; 101(4): e21588, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31180585

RESUMO

Boric acid (BA) is widely used in various industrial process and can be accessed to nontarget organisms. This study aimed to investigate the insecticidal effects of BA and its toxic activities with respect to immunologic and genotoxic effects using Galleria mellonella larvae as a model. BA concentrations (78.125-10,000 ppm) were administrated to the larvae using the feeding method. Concentration-dependent mortality was observed in all larval groups. Probit analysis revealed LC30 , LC50 , and LC70 values to be 112.4, 320.1, and 911.4 ppm, respectively. These concentrations were used in all bioassays. Drastic reductions in total hemocyte counts along with changes in differential hemocyte counts were observed following BA treatment. Cell viability assays showed dose-dependent reductions in viable cells and an increase in the necrotic and apoptotic ratios after BA treatment. However, mitotic indices of larval hemocytes did not change at all BA concentrations. The cytotoxic effect of BA led to a significant reduction in cellular immune responses such as encapsulation, melanization, and nodulation activities of treated larvae. While BA increased micronucleus ratios at the highest concentration, comet parameters indicating DNA damage increased in G. mellonella larval hemocytes at all concentrations. These report that BA suppresses the immune system of G. mellonella and also poses risks of genotoxicity at high concentrations.


Assuntos
Ácidos Bóricos/toxicidade , Mariposas/efeitos dos fármacos , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Ensaio Cometa , Relação Dose-Resposta a Droga , Hemócitos/efeitos dos fármacos , Hemolinfa , Inseticidas/toxicidade , Larva/efeitos dos fármacos , Larva/genética , Larva/imunologia , Testes para Micronúcleos , Mariposas/genética , Mariposas/imunologia
11.
Parasite ; 26: 34, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31166909

RESUMO

BACKGROUND: Gnathostoma spinigerum is a harmful parasitic nematode that causes severe morbidity and mortality in humans and animals. Effective drugs and vaccines and reliable diagnostic methods are needed to prevent and control the associated diseases; however, the lack of genome, transcriptome, and proteome databases remains a major limitation. In this study, transcriptomic and secretomic analyses of advanced third-stage larvae of G. spinigerum (aL3Gs) were performed using next-generation sequencing, bioinformatics, and proteomics. RESULTS: An analysis that incorporated transcriptome and bioinformatics data to predict excretory-secretory proteins (ESPs) classified 171 and 292 proteins into classical and non-classical secretory groups, respectively. Proteins with proteolytic (metalloprotease), cell signaling regulatory (i.e., kinases and phosphatase), and metabolic regulatory function (i.e., glucose and lipid metabolism) were significantly upregulated in the transcriptome and secretome. A two-dimensional (2D) immunomic analysis of aL3Gs-ESPs with G. spinigerum-infected human sera and related helminthiases suggested that the serine protease inhibitor (serpin) was a promising antigenic target for the further development of gnathostomiasis immunodiagnostic methods. CONCLUSIONS: The transcriptome and excretory-secretory proteome of aL3Gs can facilitate an understanding of the basic molecular biology of the parasite and identifying multiple associated factors, possibly promoting the discovery of novel drugs and vaccines. The 2D-immunomic analysis identified serpin, a protein secreted from aL3Gs, as an interesting candidate for immunodiagnosis that warrants immediate evaluation and validation.


Assuntos
Gnathostoma/genética , Proteínas de Helminto/genética , Testes Imunológicos , Larva/genética , Proteoma , Transcriptoma , Animais , Antígenos de Helmintos/genética , Biologia Computacional/métodos , Gnatostomíase/tratamento farmacológico , Proteínas de Helminto/isolamento & purificação , Humanos
12.
Exp Appl Acarol ; 78(3): 361-372, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31254229

RESUMO

In insects, the ecdysteroid 20-hydroxyecdysone coordinates with juvenile hormone (JH) to regulate the process of molting, development and metamorphosis; however, this interaction is still unclear in the mites. In this study, we investigated the gene related to ecdysteroid and JH biosynthesis pathways, including four ecdysteroid and 11 JH biosynthesis genes. We examined their expression patterns during molting of different developmental stages of the two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), an important agricultural pest that feeds on more than 1100 plant species. The expression of ecdysteroid biosynthesis Halloween genes exhibited a positive zigzag-like pattern, with a peak after 8 h of molting and a drop 8 h after entering each quiescent stage. In contrast, JH biosynthesis genes expression displayed a negative zigzag-like pattern, with a peak at 8 h after entering each quiescent stage and a drop after 8 h of each molting. These opposite patterns imply that ecdysteroid and JH expression is coordinated during the developmental transition. Our data provide an initial perspective on the co-expression of ecdysteroid and JH biosynthesis genes to regulate this important developmental process in the two-spotted spider mite.


Assuntos
Proteínas de Artrópodes/genética , Ecdisteroides/biossíntese , Expressão Gênica , Hormônios Juvenis/biossíntese , Muda/genética , Tetranychidae/genética , Animais , Proteínas de Artrópodes/metabolismo , Ecdisteroides/genética , Hormônios Juvenis/genética , Larva/genética , Larva/crescimento & desenvolvimento , Ninfa/genética , Ninfa/crescimento & desenvolvimento , Óvulo/crescimento & desenvolvimento , Tetranychidae/crescimento & desenvolvimento
13.
J Fish Dis ; 42(7): 1013-1021, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31037725

RESUMO

A tropicalization phenomenon of ichthyofauna has been described in the last decades in Galicia (north-eastern Atlantic), with increasing reports of tropical and subtropical fishes appearing northward this distribution range. A search for parasites was carried out in the digestive tract of two specimens first captured in Galician waters: the prickly puffer Ephippion guttifer (Tetraodontidae) and the African stripped grunt Parapristipoma octolineatum (Haemulidae). Examination of E. guttifer showed high intensity of nematodes, from three different genera: Cucullanus (Cucullanidae), Hysterothylacium (Raphidascaridae) and Anisakis (Anisakidae), with demonstrated pathogenicity to humans. Molecular identification allowed the identification of Anisakis pegreffii, already described in the area, and first reports for European waters of Cucullanus dodsworthi, Hysterothylacium reliquens and a new Hysterothylacium sp. P. octolineatum showed a far lower level of parasitization, with two Hysterothylacium larvae, genetically identified as Hysterothylacium deardorffoverstreetorum, also its first report in the eastern Atlantic. Thus, possible ecological impact of the occurrence of two non-native individual fishes in a new area could be remarkably higher if we see this issue through the lens of the parasitological perspective, as far as only two individual fish can harbour more of one hundred nematode parasites belonging to different species, most of them also new species for that area.


Assuntos
Anisakis/isolamento & purificação , Ascaridoidea/isolamento & purificação , Doenças dos Peixes/parasitologia , Peixes/parasitologia , Espécies Introduzidas , Animais , Anisakis/genética , Ascaridoidea/genética , Oceano Atlântico , Mudança Climática , Feminino , Larva/genética , Masculino , Temperatura Ambiente , Clima Tropical
14.
J Fish Dis ; 42(7): 1047-1056, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31094002

RESUMO

There are limited reports of infectious agents affecting Australian cowtail stingrays. In the present study, a new species of ascaridoid nematode belonging to the genus Mawsonascaris is described. The most distinct characteristic features were observed in females (the presence of a polar spine in the eggs and a flap-like projection in the vulval area). An identification key for Mawsonascaris spp. is provided. Additionally, internal transcribed spacers (ITS) sequences were obtained for the new species. Alignment of the ITS sequence of the specimens in the present study with those deposited in GenBank showed that there exists no other highly similar sequence. Phylogenetic analyses resulted in a distinct grouping of our specimens supporting morphological distinction from previously described Mawsonascaris spp. Histology was used to investigate the pathology caused by the infection. Necrosis, inflammation and fibrosis were evident at the border of the nodules formed by parasite. A large number of parasites were present in muscularis mucosae and submucosa but not in the muscularis of the stomach. The parasites were associated with an increased inflammatory response, which was also found in the muscularis mucosae and submucosa. Similar pathology has been described in elasmobranchs infected by cestodes, although with more severe lesions.


Assuntos
Anisaquíase/veterinária , Anisakis/anatomia & histologia , Anisakis/genética , Raias/parasitologia , Animais , Anisakis/isolamento & purificação , Austrália , DNA de Helmintos/genética , DNA Espaçador Ribossômico/genética , Feminino , Doenças dos Peixes/parasitologia , Larva/genética , Masculino , Filogenia , Estômago/parasitologia
15.
Environ Sci Pollut Res Int ; 26(19): 19560-19574, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31079296

RESUMO

The current study checks the effect of various concentrations of dietary graphene oxide (GO) nano-sheets on the development of Drosophila melanogaster. GO was synthesized and characterized by XRD, FTIR, FESEM, and TEM analytical techniques. Various concentrations of GO were mixed with the fly food and flies were transferred to the vial. Various behavioral and morphological as well as genetic defects were checked on the different developmental stages of the offspring. In the larval stage of development, the crawling speed and trailing path change significantly than the control. GO induces the generation of oxygen radicals within the larval hemolymph as evidenced by nitroblue tetrazolium assay. GO induces DNA damage within the gut cell, which was detected by Hoechst staining and within hemolymph by comet assay. Adult flies hatched after GO treatment show defective phototaxis and geotaxis behavior. Besides behavior, phenotypic defects were observed in the wing, eye, thorax bristles, and mouth parts. At 300 mg/L concentration, wing spots were observed. Altogether, the current study finds oral administration of GO which acts as a mutagen and causes various behavioral and developmental defects in the offspring. Here for the first time, we are reporting GO, which acts as a teratogen in Drosophila, besides its extensive medical applications.


Assuntos
Drosophila melanogaster/efeitos dos fármacos , Grafite/toxicidade , Mutagênicos/toxicidade , Nanoestruturas/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Teratogênios/toxicidade , Administração Oral , Animais , Comportamento Animal/efeitos dos fármacos , Dano ao DNA , Drosophila melanogaster/genética , Drosophila melanogaster/crescimento & desenvolvimento , Larva/efeitos dos fármacos , Larva/genética , Larva/crescimento & desenvolvimento
16.
Exp Appl Acarol ; 78(1): 133-147, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31093859

RESUMO

A new tick species of the genus Ornithodoros (Acari: Argasidae) was described from larvae collected on the toad Rhinella arenarum in a locality from Argentina belonging to the Monte Biogeographic Province. Ornithodoros montensis n. sp. was described based on morphological traits and sequences of the mitochondrial 16S rRNA gene. The diagnostic characters for this species are a combination of idiosoma oval, dorsal plate pyriform with posterior margin slightly concave, dorsal surface with 17 pairs of setae (7 anterolateral, 4 to 5 central and 5 to 6 posterolateral), ventral surface with 6 pairs of setae and 1 pair on anal valves, three pairs of sternal setae, postcoxal setae absent, and hypostome pointed apically with dental formula 3/3 in the anterior half and 2/2 posteriorly almost to base. Phylogenetic analysis of 16S rDNA sequences and a principal component analysis based on morphometric characters provided additional support to the description of O. montensis as an independent lineage within the genus Ornithodoros. Larvae of O. montensis are phylogenetically closely related to O. puertoricensis, O. rioplatensis, O. talaje s.s., O. guaporensis, O. hasei and O. atacamensis, all of them belonging to the "O. talaje group".


Assuntos
Bufonidae/parasitologia , Interações Hospedeiro-Parasita , Ornithodoros/classificação , Animais , Argentina , Larva/anatomia & histologia , Larva/classificação , Larva/genética , Ornithodoros/anatomia & histologia , Ornithodoros/genética , Ornithodoros/crescimento & desenvolvimento , RNA Ribossômico 16S/análise
17.
Gene ; 710: 387-398, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31136783

RESUMO

Busseola fusca (Fuller) (Lepidoptera: Noctuidae), a major insect pest of maize in sub-Saharan Africa, has developed high levels of non-recessive resistance to Cry1Ab toxin expressed in genetically modified Bt maize. Multiple resistance mechanisms to various Cry toxins have been identified in Lepidoptera, but no study has yet been done to determine the mechanism of Cry1Ab resistance in B. fusca. Therefore, the larval transcriptome of B. fusca was sequenced, de novo assembled and characterized. Differential expression analysis was performed to compare gene expression profiles of Cry toxin challenged and unchallenged neonate larvae to assess the molecular basis of the defence mechanism employed by this insect. Several genes associated with Cry toxin resistance in other lepidopteran pests were detected in B. fusca. Results suggest that differential expression of metabolic and immune-related genes might explain Cry1Ab toxin defence in this pest (supplemental file). Transcript expression profiles of neonates demonstrated that 33.59% and 60.31% of the 131 differentially expressed genes were upregulated and downregulated in the toxin-challenged neonate larvae, respectively. Transcripts were grouped into two subclusters according to the similarity of their expression patterns. Transcripts in subcluster 1 were moderately upregulated in the toxin-challenged neonate larvae, and, conversely, downregulated in the unchallenged neonate larvae. The solute carrier organic anion transporter, which is involved in insecticide detoxification, was upregulated in the toxin-challenged neonate larvae. Conversely, most of the transcripts in subcluster 2 were moderately downregulated in the toxin-challenged neonate larvae, and upregulated for neonates feeding on non-challenged maize. Four unidentified transcripts were extremely down-regulated in the toxin-challenged neonate larvae, and upregulated in the unchallenged neonate larvae. Further studies are recommended to establish if there is a direct correlation between these differentially expressed genes and the observed resistance. Elucidation of such defence mechanisms is crucial for developing insect resistance management strategies to ensure sustainable use of genetically modified maize in Africa. Nevertheless, this is the first study on gene expression profiles of B. fusca strains challenged with Cry toxin. The transcriptome characterized in this study provides a significant resource base for future studies on B. fusca and contributes to understanding some of the gene regulation and signalling networks involved in the defence of B. fusca against Cry1Ab toxin.


Assuntos
Proteínas de Bactérias/farmacologia , Endotoxinas/farmacologia , Perfilação da Expressão Gênica/métodos , Proteínas Hemolisinas/farmacologia , Proteínas de Insetos/genética , Resistência a Inseticidas , Mariposas/genética , Animais , Bacillus thuringiensis/genética , Bacillus thuringiensis/metabolismo , Proteínas de Bactérias/genética , Endotoxinas/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas Hemolisinas/genética , Larva/efeitos dos fármacos , Larva/genética , Redes e Vias Metabólicas , Mariposas/efeitos dos fármacos , Plantas Geneticamente Modificadas/parasitologia , Análise de Sequência de RNA , Zea mays/genética , Zea mays/metabolismo , Zea mays/parasitologia
18.
Parasit Vectors ; 12(1): 256, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31118082

RESUMO

BACKGROUND: RNA interference (RNAi), which has facilitated functional characterization of mosquito neural development genes such as the axon guidance regulator semaphorin-1a (sema1a), could one day be applied as a new means of vector control. Saccharomyces cerevisiae (baker's yeast) may represent an effective interfering RNA expression system that could be used directly for delivery of RNA pesticides to mosquito larvae. Here we describe characterization of a yeast larvicide developed through bioengineering of S. cerevisiae to express a short hairpin RNA (shRNA) targeting a conserved site in mosquito sema1a genes. RESULTS: Experiments conducted on Aedes aegypti larvae demonstrated that the yeast larvicide effectively silences sema1a expression, generates severe neural defects, and induces high levels of larval mortality in laboratory, simulated-field, and semi-field experiments. The larvicide was also found to induce high levels of Aedes albopictus, Anopheles gambiae and Culex quinquefasciatus mortality. CONCLUSIONS: The results of these studies indicate that use of yeast interfering RNA larvicides targeting mosquito sema1a genes may represent a new biorational tool for mosquito control.


Assuntos
Proteínas de Insetos/genética , Controle de Mosquitos/métodos , Interferência de RNA , Saccharomyces cerevisiae/genética , Semaforinas/genética , Aedes/genética , Animais , Anopheles/genética , Bioengenharia , Culex/genética , Feminino , Larva/genética , RNA Interferente Pequeno
19.
Korean J Parasitol ; 57(2): 207-211, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31104416

RESUMO

Anisakiasis is a zoonotic disease induced by anisakid nematodes, and endoscopic inspection is used for a diagnosis or remedy for it. Anisakis simplex, Anisakis physeteris, and Pseudoterranova decipiens had been reported to be the major species causing human infections, particularly, in Japan. However, in Korea, recent studies strongly suggested that Anisakis pegreffii is the major species of human infections. To support this suggestion, we collected anisakid larvae (n=20) from 20 human patients who were undergone gastrointestinal endoscopy at a health check-up center in Korea, and molecular identification was performed on the larvae using PCR-RFLP analysis and gene sequencing of rDNA ITS regions and mtDNA cox2. In addition, anisakid larvae (n=53) collected from the sea eel (Astroconger myriaster) were also examined for comparison with those extracted from humans. The results showed that all human samples (100%) were identified as A. pegreffii, whereas 90.7% of the samples from the sea eel were A. pegreffii with the remaining 9.3% being Hysterothylacium aduncum. Our study confirmed that A. pegreffii is the predominant species causing human anisakiasis in Korea, and this seems to be due to the predominance of this larval type in the fish (sea eels) popularly consumed by the Korean people. The possibility of human infection with H. aduncum in Korea is also suggested.


Assuntos
Anisaquíase/diagnóstico , Anisakis/classificação , Anisakis/isolamento & purificação , Endoscopia Gastrointestinal/métodos , Técnicas de Diagnóstico Molecular/métodos , Adulto , Idoso , Animais , Anisaquíase/veterinária , Anisakis/genética , Ciclo-Oxigenase 2/genética , DNA Mitocondrial/química , DNA Mitocondrial/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Enguias/parasitologia , Feminino , Doenças dos Peixes/parasitologia , Humanos , Larva/classificação , Larva/genética , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição , República da Coreia , Análise de Sequência de DNA
20.
PLoS Negl Trop Dis ; 13(5): e0007422, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31107878

RESUMO

New mosquito control strategies are vitally needed to address established and emerging arthropod-borne infectious diseases. Here we describe the characterization of a yeast interfering RNA larvicide that was developed through the genetic engineering of Saccharomyces cerevisiae (baker's yeast) to express a short hairpin RNA targeting the Aedes aegypti synaptotagmin (Aae syt) gene. The larvicide effectively silences the Aae syt gene, causes defects at the larval neural synapse, and induces high rates of A. aegypti larval mortality in laboratory, simulated-field, and semi-field trials. Conservation of the interfering RNA target site in multiple mosquito species, but not in humans or other non-target species, suggested that it may function as a broad-range mosquito larvicide. In support of this, consumption of the yeast interfering RNA larvicide was also found to induce high rates of larval mortality in Aedes albopictus, Anopheles gambiae, and Culex quinquefasciatus mosquito larvae. The results of these studies suggest that this biorational yeast interfering RNA larvicide may represent a new intervention that can be used to combat multiple mosquito vectors of human diseases.


Assuntos
Proteínas de Insetos/genética , Controle de Mosquitos/métodos , Mosquitos Vetores/genética , Controle Biológico de Vetores/métodos , Interferência de RNA , Saccharomyces cerevisiae/genética , Sinaptotagminas/genética , Aedes/genética , Aedes/metabolismo , Aedes/microbiologia , Animais , Anopheles/genética , Anopheles/metabolismo , Anopheles/microbiologia , Culex/genética , Culex/metabolismo , Culex/microbiologia , Feminino , Engenharia Genética , Proteínas de Insetos/metabolismo , Larva/genética , Larva/metabolismo , Larva/virologia , Masculino , Mosquitos Vetores/metabolismo , Mosquitos Vetores/virologia , Saccharomyces cerevisiae/metabolismo , Sinaptotagminas/metabolismo
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