RESUMO
A wide range of microbial pathogens can enter the gastrointestinal tract, causing mucosal inflammation and infectious colitis and accounting for most cases of acute diarrhea. Severe cases of infectious colitis can persist for weeks, and if untreated, may lead to major complications and death. While the molecular pathogenesis of microbial infections is often well-characterized, host-associated epithelial factors that affect risk and severity of infectious colitis are less well-understood. The current study characterized functions of the zinc (Zn) transporter ZnT2 (SLC30A2) in cultured HT29 colonocytes and determined consequences of ZnT2 deletion in mice on the colonic response to enteric infection with Citrobacter rodentium. ZnT2 in colonocytes transported Zn into vesicles buffering cytoplasmic Zn pools, which was important for Toll-like receptor 4 (TLR4) expression, activation of pathogen-stimulated NF-κß translocation and cytokine expression. Additionally, ZnT2 was critical for lysosome biogenesis and bacterial-induced autophagy, both promoting robust host defense and resolution mechanisms in response to enteric pathogens. These findings reveal that ZnT2 is a novel regulator of mucosal inflammation in colonocytes and is critical to the response to infectious colitis, suggesting that manipulating the function of ZnT2 may offer new therapeutic strategies to treat specific intestinal infections.
Assuntos
Proteínas de Transporte de Cátions , Colite , Inflamação , Mucosa Intestinal , Receptor 4 Toll-Like , Animais , Proteínas de Transporte de Cátions/genética , Proteínas de Transporte de Cátions/metabolismo , Colite/etiologia , Colite/genética , Colite/metabolismo , Citocinas/genética , Citocinas/metabolismo , Células HT29 , Humanos , Inflamação/genética , Inflamação/metabolismo , Mucosa Intestinal/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo , Zinco/metabolismoRESUMO
Bradynema listronoti is an insect-parasitic nematode known to infect the carrot weevil, Listronotus oregonensis. We present the first sequence for this species and for any Allantonematidae, produced with a combination of short and long reads. The draft genome of B. listronoti is 80.6 Mb in size, assembled in 152 scaffolds.
RESUMO
The opportunistic pathogen Pseudomonas aeruginosa (P. aeruginosa) is associated gastrointestinal (GI) inflammation and illness; however, factors motivating commensal-to-pathogen transition are unclear. Excessive zinc intake from supplements is common in humans. Due to the fact that zinc exposure enhances P. aeruginosa colonization in vitro, we hypothesized zinc exposure broadly activates virulence mechanisms, leading to inflammation and illness. P. aeruginosa was treated with excess zinc and growth, expression and secretion of key virulence factors, and biofilm production were determined. Effects on invasion, barrier function, and cytotoxicity were evaluated in Caco-2 cells co-cultured with P. aeruginosa pre-treated with zinc. Effects on colonization, mucosal pathology, inflammation, and illness were evaluated in mice infected with P. aeruginosa pre-treated with zinc. We found the expression and secretion of key virulence factors involved in quorum sensing (QS), motility (type IV pili, flagella), biosurfactants (rhamnolipids), toxins (exotoxin A), zinc homeostasis (CzcR), and biofilm production, were all significantly increased. Zinc exposure significantly increased P. aeruginosa invasion, permeability and cytotoxicity in Caco-2 cells, and enhanced colonization, inflammation, mucosal damage, and illness in mice. Excess zinc exposure has broad effects on key virulence mechanisms promoting commensal-to-pathogen transition of P. aeruginosa and illness in mice, suggesting excess zinc intake may have adverse effects on GI health in humans.
Assuntos
Translocação Bacteriana/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Mucosa Intestinal/microbiologia , Infecções por Pseudomonas , Pseudomonas aeruginosa , Fatores de Virulência/biossíntese , Zinco/efeitos adversos , Animais , Células CACO-2 , Humanos , Masculino , Camundongos , Infecções por Pseudomonas/induzido quimicamente , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/patogenicidade , Pseudomonas aeruginosa/fisiologia , Zinco/farmacologiaRESUMO
Suboptimal lactation is a common, yet underappreciated cause for early cessation of breastfeeding. Molecular regulation of mammary gland function is critical to the process lactation; however, physiological factors underlying insufficient milk production are poorly understood. The zinc (Zn) transporter ZnT2 is critical for regulation of mammary gland development and maturation during puberty, lactation, and postlactation gland remodeling. Numerous genetic variants in the gene encoding ZnT2 (SLC30A2) are associated with low milk Zn concentration and result in severe Zn deficiency in exclusively breastfed infants. However, the functional impacts of genetic variation in ZnT2 on key mammary epithelial cell functions have not yet been systematically explored at the cellular level. Here we determined a common mutation in SLC30A2/ZnT2 substituting serine for threonine at amino acid 288 (Thr288Ser) was found in 20% of women producing low milk volume (n = 2/10) but was not identified in women producing normal volume. Exploration of cellular consequences in vitro using phosphomimetics showed the serine substitution promoted preferential phosphorylation of ZnT2, driving localization to the lysosome and increasing lysosome biogenesis and acidification. While the substitution did not initiate lysosome-mediated cell death, cellular ATP levels were significantly reduced. Our findings demonstrate the Thr288Ser mutation in SLC30A2/ZnT2 impairs critical functions of mammary epithelial cells and suggest a role for genetic variation in the regulation of milk production and lactation performance.
Assuntos
Proteínas de Transporte de Cátions/metabolismo , Metabolismo Energético , Células Epiteliais/metabolismo , Lactação/metabolismo , Lisossomos/metabolismo , Glândulas Mamárias Humanas/metabolismo , Leite Humano/metabolismo , Mutação , Trifosfato de Adenosina/metabolismo , Adulto , Estudos de Casos e Controles , Proteínas de Transporte de Cátions/genética , Linhagem Celular , Metabolismo Energético/genética , Feminino , Humanos , Concentração de Íons de Hidrogênio , Lactação/genética , Lisossomos/genética , Biogênese de Organelas , Fosforilação , Adulto JovemRESUMO
Bradynema listronoti is a parasitic nematode described from infected specimens of the carrot weevil Listronotus oregonensis. Prevalence of infection by B. listronoti under field conditions was followed over a period of 16 years in an untreated carrot field. Susceptibility of different carrot weevil life stages was evaluated as well as the impact of infection on fecundity and mortality. Gene expression in infected and uninfected carrot weevils was also compared to evaluate the impact of the parasite on the host transcriptome. Prevalence of B. listronoti in carrot weevil populations was sustained over the years ranging from 20 to 63%. All the weevil stages exposed to B. listronoti inoculum were susceptible to infection, larvae being more vulnerable (59 ± 8% infected) compared with pupae (4 ± 3% infected) and adults (7 ± 3% infected). The fecundity of infected female weevils was greatly reduced (60-fold) due to an inhibition of the maturation of the reproductive system. Transcriptomic analyses revealed that this parasitic castration may have been triggered by the inhibition of reproductive hormone production. The B. listronoti-L. oregonensis interaction represents a case of parasitic castration with a unique potential for biological control of an important pest of carrots.
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Root lesion nematodes are very common plant-parasitic nematodes that affect a wide range of plants. More than one species can be found simultaneously in a field, and each has a different impact on crop yield. Unfortunately, identifying them using classical morphometric criteria is very difficult and time consuming. The species Pratylenchus alleni was recently observed for the first time in Canada, associated with severe damage in a soybean field in the province of Quebec. The major species, P. penetrans, is also known to be endemic in Quebec but no data exist on its distribution in field crops. This prompted the development of a multiplex quantitative polymerase chain reaction (PCR) assay for the simultaneous detection and quantification of P. alleni and P. penetrans. The method was found to be specific and sensitive, systematically detecting a single larva in a 100-cm3 soil sample with no cross-amplification with other species, even when they outnumbered the target species. An exogenous internal positive control was included in the test to avoid false negatives due to the presence of PCR inhibitors. This assay was used to study the distribution of P. alleni and P. penetrans in 185 soybean fields in the major soybean-producing areas of Quebec during a 3-year survey. Overall, P. penetrans was found in 42% of the fields, P. alleni in 8%, and both species in 4%. The population density of P. alleni in positive fields was still very low, with only a few larvae detected. However, densities of P. penetrans were much higher: the provincial mean was 51.7 nematodes per 100 cm3 of soil (in positive samples), and 8% of the fields (15 of 185) exceeded the theoretical economic threshold. The presence of P. penetrans was also strongly correlated with soil texture, with lighter soil being the most favorable.
Assuntos
Glycine max/parasitologia , Nematoides/genética , Nematoides/isolamento & purificação , Doenças das Plantas/parasitologia , Reação em Cadeia da Polimerase/métodos , Animais , Raízes de Plantas/parasitologia , QuebequeRESUMO
The impacts of climate change on the development of insects are of great concern due to potential alterations in population dynamics and pest pressure. The carrot weevil, Listronotus oregonensis, is a major agricultural pest, and its development is influenced by temperature and photoperiod. In this study, our aim was to investigate the impact of temperature increases on the voltinism and reproductive diapause of the carrot weevil under field conditions and bioclimatic models. Field observations were conducted over two growing seasons using structures that allowed for temperature increases. The developmental stages of the carrot weevil, including female reproductive status, oviposition and larval stage, were monitored weekly to measure the proportion of individuals undergoing an additional generation. Concurrently, bioclimatic models were used to simulate the probability of a second generation under current (1981-2010) and future (2041-2070) climates, considering a lower and a higher change in emission scenarios. Results showed that rising temperatures led to an increase in the proportion of carrot weevils undergoing inhibition of the reproductive diapause and a higher number of eggs laid in the field. The models indicated a substantial rise in the probability of a second generation developing, from 24% to 37% to 62%-99% under current and future climates, respectively. These findings demonstrate the potential for significant alterations in carrot weevil population dynamics, resulting in increased pest pressure on crops. Further research is needed to fully understand the implications of these findings and to develop effective adaptation measures to mitigate the negative impacts of global warming on insect populations and agriculture.
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Mudança Climática , Diapausa de Inseto , Gorgulhos , Animais , Gorgulhos/fisiologia , Gorgulhos/crescimento & desenvolvimento , Diapausa de Inseto/fisiologia , Feminino , Reprodução , Oviposição , Temperatura , Larva/crescimento & desenvolvimento , Larva/fisiologiaRESUMO
Onion thrips (Thrips tabaci) can pose a significant threat to onion crops, causing leaf damage, reduced bulb size and quality, and yield loss during severe infestations. Conventional insecticide use has been the primary method for managing this pest species, but the efficacy of this approach is inconsistent. Furthermore, emerging pest resistance is a growing concern in some regions. This two-year field study aimed to assess the effectiveness of several pest management strategies in controlling onion thrips populations and limiting their impact on onion yields. The strategies tested consisted of habitat manipulations (including flower strips and straw mulch), biological control agents (Stratiolaelaps scimitus, Neoseiulus cucumeris, Amblyseius swirskii, and Beauveria bassiana), as well as physical barrier control methods (exclusion nets, kaolin, and mineral oil). Habitat manipulation techniques, particularly the use of flower strips, reduced thrips populations by up to 50% and increased onion yields by 25%. In contrast, exclusion nets had a detrimental effect on onion yields, and the other alternative control methods produced results comparable to those obtained for untreated controls. When used alone, biological control agents were not effective at maintaining thrips populations below economically damaging levels. This study offers valuable insights into effective and sustainable pest management practices for the onion industry.
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Population genetic studies of insect pests enhance our ability to anticipate problems in agroecosystems, such as pest outbreaks, insecticide resistance, or expansions of the host range. This study focuses on geographic distance and host plant selection as potential determinants of genetic differentiation of the carrot weevil Listronotus oregonensis, a major pest of several apiaceous crops in North America. To undertake genetic studies on this species, we assembled the first complete genome sequence for L. oregonensis. Then, we used both haplotype discrimination with mitochondrial DNA (mtDNA) and a genotyping-by-sequencing (GBS) approach to characterize the genetic population structure. A total of 220 individuals were sampled from 17 localities in the provinces of Québec, Ontario, Nova Scotia (Canada), and the state of Ohio (USA). Our results showed significant genetic differences between distant populations across North America, indicating that geographic distance represents an important factor of differentiation for the carrot weevil. Furthermore, the GBS analysis revealed more different clusters than COI analysis between Québec and Nova Scotia populations, suggesting a recent differentiation in the latter province. In contrast, we found no clear evidence of population structure associated with the four cultivated apiaceous plants tested (carrot, parsley, celery, and celeriac) using populations from Québec. This first characterization of the genetic structure of the carrot weevil contributes to a better understanding of the gene flow of the species and helps to adapt local pest management measures to better control this agricultural pest.
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Agricultural soil pests, including wireworms (Coleoptera: Elateridae), are managed primarily with pesticides applied directly to seeds before sowing. Seeds coated with neonicotinoids have been used widely in Quebec (Canada) for several years. To assess the agronomic and economic value of neonicotinoid seed treatments in soybeans and corn in Quebec, trials were conducted from 2012 to 2016 in 84 fields across seven regions in Quebec. We evaluated the effect of neonicotinoid seed treatments on soil pest densities, crop damage and yield. The results showed that 92.6% of corn fields and 69.0% of soybean fields had less than 1 wireworm per bait trap. However, no significant differences in plant stand or yield were observed between treated and untreated corn or soybeans during the study. This study shows that neonicotinoid seed treatments in field crops in Quebec are useful in less than 5% of cases, given the very low level of pest-associated pressure and damage, and that they should not be used prophylactically. Integrated pest management (IPM) strategies need to be developed for soil insect pests to offer effective alternative solutions to producers.
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Glycine max/efeitos dos fármacos , Inseticidas/efeitos adversos , Neonicotinoides/efeitos adversos , Sementes/efeitos dos fármacos , Solo , Zea mays/efeitos dos fármacos , Animais , Quebeque , Plântula/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Glycine max/crescimento & desenvolvimento , Zea mays/crescimento & desenvolvimentoRESUMO
Studies in humans and pre-clinical animal models show milk-derived miRNAs reflect mammary gland function during lactation. The zinc transporter SLC30A2/ZnT2 plays a critical role in mammary gland function; ZnT2-null mice have profound defects in mammary epithelial cell (MEC) polarity and secretion, resulting in sub-optimal lactation. Non-synonymous genetic variation in SLC30A2 is common in humans, and several common ZnT2 variants are associated with changes in milk components that suggest breast dysfunction in women. To identify novel mechanisms through which dysfunction might occur, milk-derived miRNA profiles were characterized in women harboring three common genetic variants in SLC30A2 (D103E, T288S, and Exon 7). Expression of ten miRNAs differed between genotypes, and contributed to distinct spatial separation. Studies in breast milk and cultured MECs confirmed expression of ZnT2 variants alters abundance of protein levels of several predicted mRNA targets critical for breast function (PRLR, VAMP7, and SOX4). Moreover, bioinformatic analysis identified two novel gene networks that may underlie normal MEC function. Thus, we propose that genetic variation in genes critical for normal breast function such as SLC30A2 has important implications for lactation performance in women, and that milk-derived miRNAs can be used to identify novel mechanisms and for diagnostic potential.
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Proteínas de Transporte de Cátions/genética , MicroRNAs/metabolismo , Leite Humano/metabolismo , Adolescente , Adulto , Animais , Proteínas de Transporte de Cátions/deficiência , Proteínas de Transporte de Cátions/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Feminino , Genótipo , Humanos , Glândulas Mamárias Animais/citologia , Glândulas Mamárias Animais/metabolismo , Camundongos , Camundongos Knockout , Polimorfismo Genético , Mapas de Interação de Proteínas/genética , Proteínas R-SNARE/genética , Proteínas R-SNARE/metabolismo , Fatores de Transcrição SOXC/genética , Fatores de Transcrição SOXC/metabolismo , Adulto JovemRESUMO
European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), is the main pest causing damage to sweet corn in North America. Conventional management with multiple use of insecticides is a common practice for processing sweet corn. In Canada, the use of Trichogramma spp. began in the 1990s, but the adoption of this approach for European corn borer management is still limited to the fresh market of sweet corn. Trichogramma ostriniae (Peng & Chen) has great potential as a biological control agent for large areas such as in processing sweet corn. The objective of this study was to evaluate an economically and environmentally sustainable alternative to insecticides for controlling European corn borer populations in processing sweet corn. During the growing season, the mean number of larvae decreased after insecticide (0.07 ± 0.04) and Trichogramma (1.32 ± 0.59) treatments compared with the control (2.42 ± 0.72). At harvest, damages associated with European corn borer were similar after Trichogramma (1.0 ± 0.7%) and insecticide (1.0 ± 0.6%) treatments, but significantly lower than the control (8.7 ± 3.3%). This study showed that the use of T. ostriniae can significantly decrease the pressure exerted by European corn borer and its damage on corn ears. This outcome is particularly interesting considering that it was achieved with lower doses of Trichogramma, a lower number of releases, and on large crop areas, compared with what is actually done to protect fresh market corn from European corn borer. Under these conditions, the use of Trichogramma is an economically and competitive alternative to insecticide applications.
Assuntos
Controle de Insetos/economia , Mariposas/parasitologia , Controle Biológico de Vetores/economia , Vespas/fisiologia , Animais , Inseticidas/economia , Larva/crescimento & desenvolvimento , Larva/parasitologia , Larva/fisiologia , Mariposas/crescimento & desenvolvimento , Quebeque , Zea maysRESUMO
Intraguild predation (IGP) occurs when one predator species attacks another predator species with which it competes for a shared prey species. Despite the apparent omnipresence of intraguild interactions in natural and managed ecosystems, very few studies have quantified rates of IGP in various taxa under field conditions. We used molecular analyses of gut contents to assess the nature and incidence of IGP among four species of coccinellid predators in soybean fields. Over half of the 368 predator individuals collected in soybean contained the DNA of other coccinellid species indicating that IGP was very common at our field site. Furthermore, 13.2% of the sampled individuals contained two and even three other coccinellid species in their gut. The interaction was reciprocal, as each of the four coccinellid species has the capacity to feed on the others. To our knowledge, this study represents the most convincing field evidence of a high prevalence of IGP among predatory arthropods. The finding has important implications for conservation biology and biological control.