International journal for parasitology最新文献

{"title":"Effects of Nosema ceranae and Lotmaria passim infections on honey bee foraging behaviour and physiology.","authors":"Courtney I MacInnis, Lien T Luong, Stephen F Pernal","doi":"10.1016/j.ijpara.2024.12.003","DOIUrl":"10.1016/j.ijpara.2024.12.003","url":null,"abstract":"<p><p>Nosema ceranae and Lotmaria passim are two commonly encountered digestive tract parasites of the adult honey bee (Apis mellifera L.). Although these parasites are associated with colony losses, little is known about how they affect individual bee physiology and behaviour at the colony level. Using locally obtained isolates, we investigated the effects of both single and mixed infections of L. passim and N. ceranae on honey bee vitellogenin (Vg) expression and foraging behaviour. At the first instance of foraging, bees inoculated with either parasite had significantly lower Vg expression than uninoculated bees, with bees from the mixed infection treatment having the lowest Vg expression. Bees from the mixed infection treatment also had significantly higher densities of N. ceranae spores and numerically greater densities of L. passim cells per bee compared with bees inoculated with either parasite alone. In addition, bees from the mixed infection treatment had a significantly younger average foraging age compared with uninoculated bees from the same cohort. Although we did not find any effect of treatment on foraging effort, we discovered that bees inoculated with L. passim alone, or together with N. ceranae, had higher returning rates of foragers than control bees or bees inoculated with N. ceranae alone. Our findings indicate that both parasites can alter individual bee physiology, leading to individual changes in behaviour that could alter colony foraging dynamics. These have the potential to result in smaller, less productive colonies, decreased colony survivorship and reduced income for beekeepers.</p>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142907094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Trichinella spiralis excretory/secretory antigens ameliorate porcine epidemic diarrhea virus-induced mucosal damage in porcine intestinal oganoids by alleviating inflammation and promoting tight junction.","authors":"Yinju Liu, Jinlong Tan, Nianzhang Zhang, Zigang Qu, Wenhui Li, Yaodong Wu, Hong Yin, Guangliang Liu, Baoquan Fu","doi":"10.1016/j.ijpara.2024.12.002","DOIUrl":"10.1016/j.ijpara.2024.12.002","url":null,"abstract":"<p><p>Trichinella spiralis and porcine epidemic diarrhea virus (PEDV) are two infectious swine pathogens. Parasite excretory/secretory antigens play critical roles in various disease processes. To explore the coexistence mechanism of two pathogens infecting the same host, the intestinal organoid was utilized to reproduce these biological processes. In this study, we investigated the effects of T. spiralis excretory/secretory antigens (TsES) on PEDV-induced inflammatory regulation, lesion recovery, and mucosal barrier repair in porcine intestinal organoids. The results showed that PEDV effectively infected the porcine intestinal organoids. Next, TsES inhibited pro-inflammatory cytokines and increased the anti-inflammatory cytokines produced by PEDV-infected porcine intestinal organoids. Further, four-dimensional (4D) label-free quantitative proteomics and western blotting confirmed that TsES regulate the inflammation caused by PEDV infection through the nuclear factor kappa-B (NF-κB) pathway. In addition, TsES promoted cell proliferation, inhibited apoptosis, and reduced PEDV-induced lesions in intestinal organoids. The elevated secretory immunoglobulin A (sIgA) levels caused by PEDV infection were downregulated by TsES treatment in intestinal organoids. TsES treatment reversed the mucosal barrier damage caused by PEDV infection in intestinal organoids. Finally, PEDV replication increased after TsES treatment in organoids. We highlight the potential of TsES to ameliorate PEDV-induced inflammation, mucosal lesions, and barrier damage in porcine intestinal organoids. TsES also contribute to PEDV replication. This study presents a novel research model for research on host-virus-parasite interactions, while also providing a theoretical foundation to consider parasite derivatives as a potential adjunctive therapy for intestinal inflammation.</p>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142894453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Widespread occurrence of benzimidazole resistance single nucleotide polymorphisms in the canine hookworm, Ancylostoma caninum, in Australia.","authors":"Swaid Abdullah, Thomas Stocker, Hyungsuk Kang, Ian Scott, Douglas Hayward, Susan Jaensch, Michael P Ward, Malcolm K Jones, Andrew C Kotze, Jan Šlapeta","doi":"10.1016/j.ijpara.2024.12.001","DOIUrl":"10.1016/j.ijpara.2024.12.001","url":null,"abstract":"<p><p>Canine hookworm (Ancylostoma caninum), a gastrointestinal nematode of domestic dogs, principally infects the small intestine of dogs and has the potential to cause zoonotic disease. In greyhounds and pet dogs in the USA, A. caninum has been shown to be resistant to multiple anthelmintics. We conducted a molecular survey of benzimidazole resistance in A. caninum from dogs at veterinary diagnostic centers in Australia and New Zealand. First, we implemented an internal transcribed spacer (ITS)-2 rDNA deep amplicon metabarcoding sequencing approach to ascertain the species of hookworms infecting dogs in the region. Then, we evaluated the frequency of the canonical F167Y and Q134H isotype-1 β-tubulin mutations, which confer benzimidazole resistance, using the same sequencing approach. The most detected hookworm species in diagnostic samples was A. caninum (90%; 83/92); the related Northern hookworm (Uncinaria stenocephala) was identified in 11% (10/92) of the diagnostic samples. There was a single sample with coinfection by A. caninum and U. stenocephala. Both isotype-1 β-tubulin mutations were present in A. caninum, 49% and 67% for Q134H and F167Y, respectively. Mutation F167Y in the isotype-1 β-tubulin mutation was recorded in U. stenocephala for the first known time. Canonical benzimidazole resistance codons 198 and 200 mutations were absent. Egg hatch assays performed on a subset of the A. caninum samples showed significant correlation between 50% inhibitory concentration (IC₅₀) to thiabendazole and F167Y, with an increased IC₅₀ for samples with > 75% F167Y mutation. We detected 14% of dogs with > 75% F167Y mutation in A. caninum. Given that these samples were collected from dogs across various regions of Australia, the present study suggests that benzimidazole resistance in A. caninum is widespread. Therefore, to mitigate the risk of resistance selection and further spread, adoption of a risk assessment-based approach to limit unnecessary anthelmintic use should be a key consideration for future parasite control.</p>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142881999","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Insights into extracellular vesicle biogenesis and secretion of the tapeworm Hymenolepis diminuta: host interaction and cultivation dynamics.","authors":"Hynek Mazanec, Javier Sotillo, Peter Koník, Nikol Bušková, Jiří Kyslík, Zdenko Gardian, Tomáš Bílý, Kateřina Jirků, Roman Kuchta","doi":"10.1016/j.ijpara.2024.11.005","DOIUrl":"10.1016/j.ijpara.2024.11.005","url":null,"abstract":"<p><p>Extracellular vesicles (EVs) from parasites have been identified as potent modulators of host-parasite interactions. However, their biogenesis and secretory activity are still poorly understood. Here we present a comprehensive examination of the secretory dynamics of two distinct EV fractions isolated from the adult tapeworm Hymenolepis diminuta. Additionally, we perform a detailed analysis of changes in proteomic content and morphology during EV secretion, utilising electron tomography to shed light on a previously described novel mechanism of EV biogenesis via bead-like protrusion. Our findings reveal a significant decrease in EV secretion between 24 and 48 h of in vitro cultivation when external host stimuli are no longer present. Finally, this study addresses, for the first known time, the potential bias in EV analysis resulting from extended in vitro cultivation of model parasites.</p>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142785568","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expanding the known haemosporidian parasite diversity in Eurasian bluethroat (Luscinia svecica) subspecies through amplicon sequencing.","authors":"Dragomir Damnjanović, Masoud Nazarizadeh, Václav Pavel, Bohumír Chutný, Arild Johnsen, Milena Nováková, Jan Štefka","doi":"10.1016/j.ijpara.2024.11.007","DOIUrl":"10.1016/j.ijpara.2024.11.007","url":null,"abstract":"<p><p>Monitoring haemosporidian parasites in birds is essential to comprehend the dynamics of avian malaria, a disease that significantly affects bird populations worldwide. This study concentrated on the prevalence and diversity of haemosporidian parasites in 198 specimens from two subspecies of the Eurasian bluethroat (Luscinia svecica), aiming to explore the genetic diversity and species richness of haemosporidian fauna across the host populations. By utilizing next-generation amplicon high-throughput sequencing (NGS), we observed a marked increase in the detection of haemosporidian diversity, revealing cryptic variants and species previously unidentified by Sanger sequencing. A high prevalence of Plasmodium was seen in all studied sites, accompanied by a less frequent Leucocytozoon infection in the red-spotted subspecies and minimal occurrence of Haemoproteus. Both previously known and new, low prevalence cryptic variants were detected, underscoring the complexity of haemosporidian infections in avian hosts. The use of species delimitation tools provided a detailed understanding of haemosporidian species diversity, their coexistence within hosts, and their phylogenetic relationships. Despite the varying ecological characteristics of the study sites, no significant difference in haemosporidian alpha diversity among populations was found. However, significant differences in beta diversity were identified, suggesting that habitat characteristics and geographic distance influence parasite distribution. These findings highlight the importance of advanced molecular techniques in revealing the hidden diversity of parasites, offering valuable insights into the ecology and evolution of haemosporidian infections. Given the threatened status of one of the host's populations, knowledge on local diversity of haemosporidian parasites also has implications for possible conservation strategies.</p>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142785564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Do specialist and generalist parasites differ in their prevalence and intensity of infection? A test of the niche breadth and trade-off hypotheses.","authors":"Sonia Cebrián-Camisón, Josué Martínez-de la Puente, María José Ruiz-López, Jordi Figuerola","doi":"10.1016/j.ijpara.2024.11.009","DOIUrl":"10.1016/j.ijpara.2024.11.009","url":null,"abstract":"<p><p>Studying host specificity is crucial to understanding the ability of parasites to spread to new hosts and trigger disease emergence events. The relationship between host specificity and parasite prevalence and infection intensity, has typically been studied in the context of two opposing hypotheses. According to the trade-off hypothesis generalist parasites, which can infect a broad range of hosts, will reach a lower prevalence and infection intensity than more specialist parasites due to the higher costs to adapt to multiple host immune systems. In contrast, the niche breadth hypothesis proposes that generalists' ability to infect more host species makes them more efficient in colonising host communities and thus they are found at higher prevalences and infection intensities. This study aims to test these hypotheses using the widespread avian malaria parasites of the genera Plasmodium and the related malaria-like parasite Haemoproteus. Overall, 1188 wild house sparrows from 17 localities in southwestern Spain were screened for parasite presence and intensity of infection. For each lineage found infecting house sparrows, we estimated host specificity as i) the number of different bird taxa infected by that lineage according to the MalAvi database and ii) an index that accounts for the phylogenetic relatedness between the host species. Parasite infections were recorded in 419 house sparrows, and eight Plasmodium and three Haemoproteus lineages were identified. Prevalence was positively associated with the number of host species. Lineages found in more localities showed both higher prevalence and host range. Overall, these results support the niche breadth hypothesis in relation to blood parasites infecting house sparrows.</p>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142785562","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production and characterization of monoclonal antibodies for specific detection of Nosema ceranae and Nosema apis in beehive samples.","authors":"Fernando Izquierdo, Carmen Fernández Vadillo, Soledad Fenoy, Carolina Hurtado-Marcos, Angela Magnet, Mariano Higes, Raquel Martín-Hernández, Carmen Del Aguila","doi":"10.1016/j.ijpara.2024.11.008","DOIUrl":"10.1016/j.ijpara.2024.11.008","url":null,"abstract":"<p><p>Two microsporidian species infect honeybees worldwide, Nosema apis and Nosema ceranae. Two different clinical patterns are considered: nosemosis type A (N. apis) and nosemosis type C (N. ceranae). Nosemosis type A is characterized in acute forms and nosemosis type C shows no clear outward clinical signs. The development of a rapid and simple tool for Nosema detection could allow beekeepers or veterinarians to carry out diagnostic tests in situ. Currently, PCR and microscopy are expensive techniques that require qualified staff and may not be available in every laboratory. The present study describes the production and characterization of four monoclonal antibodies (mAbs) against N. ceranae and N. apis, and the development of an IFAT. An IFAT using the mAbs was compared with microscopy and PCR for 180 beehive samples. The diagnostic test revealed similar sensitivity and specificity percentages to IFAT (97.79% and 93.18%, respectively) and microscopy (97.79% and 95.45%), considering 100% for the PCR as the 'gold standard'. A mAb (7D2) was patented for its high specificity for N. ceranae. The IFAT using the mAbs is a good alternative to microscopy and PCR in laboratories where PCR is not available for the detection and identification of both Nosema spp.</p>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142785571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"How do trematode clones differ by fitness-related traits and interact within a host?","authors":"Ekaterina Mironova, Sergei Spiridonov, Danila Sotnikov, Anastasia Shpagina, Kseniia Savina, Mikhail Gopko","doi":"10.1016/j.ijpara.2024.11.006","DOIUrl":"10.1016/j.ijpara.2024.11.006","url":null,"abstract":"<p><p>Polyclonal infections are widespread and provide evidence of facilitation, competition, and neutral interactions between parasite clones, even within the same host-parasite system. The outcome of coinfections is usually assessed by means of parasite infection intensities, while other important fitness-related traits, e.g., larval growth rates, are often ignored. We experimentally infected fish (Salvelinus malma) with different clones of the common trematode Diplostomum pseudospathaceum and pairs of clones. Clones were identified by microsatellite analysis. Their infectivity and growth rates within the fish were investigated in double-clone infections compared with single-clone ones. In total, 3838 parasite larvae (metacercariae) from 325 fish were measured. The growth rates of the D. pseudospathaceum clones were more variable than their infectivity. Relationships of these parasite traits with host mass were clone-specific. Some clones demonstrated higher infection intensities and growth rates in larger fish. Therefore, specialization toward different size groups of fish hosts may occur in this parasite species. Furthermore, we noticed a positive correlation between population density and parasite growth (Allee effect; rarely reported for parasites) but only in mixed infections. In double-clone infections, evidence of both interclonal facilitation and interclonal competition was found. When clones interacted, they either \"cooperated\" during infection of the host or competed while growing. There were no clone pairs in which interactions changed in type with time or were present constantly during development of the parasite.</p>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":" ","pages":""},"PeriodicalIF":3.7,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142785567","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Parasite β-diversity along a stream: effect of distance and environment.","authors":"Verónica Taglioretti, María Alejandra Rossin, Eugenia Levy, Juan Tomás Timi","doi":"10.1016/j.ijpara.2024.09.002","DOIUrl":"10.1016/j.ijpara.2024.09.002","url":null,"abstract":"<p><p>Parasites can provide suitable models for studying β-diversity due to their strict dependence on both the environment and the biology and distribution of their hosts, aiding in the interpretation of any patterns that hosts can display. With the aim of quantifying the relative importance of host features, environmental factors and spatial distances as drivers of fish parasite β-diversity along a unidirectional gradient, the structure of fish parasite assemblages was analysed using generalised dissimilarity models (GDMs). A total of 150 poeciliid fish were examined for larval trematodes, recording host features and physical parameters of each sampling site along the stream. Differences among digenean communities increased when Strahler order changed along the stream, associated with increasing species richness and abundance downstream. Environmental gradient, spatial distance and host features were identified as significant determining factors of species turnover, with conductivity being the most important, followed by spatial distance. In the present study, environmental variables were spatially structured along the stream, their effects as structurers of parasite β-diversity being higher than the pure environmental or the pure distance effect. Such predominance prevents us from establishing at what point on the continuum from niche to neutrality these communities are located. Results from the present research contribute to improving our knowledge of the factors that shape parasite community changes, and underline the importance of considering the pure and shared effects of spatial, environmental and host feature factors in order to determine the real contribution of each one as a determinant of parasite β- diversity.</p>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":" ","pages":"755-765"},"PeriodicalIF":3.7,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142346129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"What shapes a microbiome? Differences in bacterial communities associated with helminth-amphipod interactions.","authors":"Célia Koellsch, Robert Poulin, Priscila M Salloum","doi":"10.1016/j.ijpara.2024.08.005","DOIUrl":"10.1016/j.ijpara.2024.08.005","url":null,"abstract":"<p><p>The fast technological advances of molecular tools have enabled us to uncover a new dimension hidden within parasites and their hosts: their microbiomes. Increasingly, parasitologists characterise host microbiome changes in the face of parasitic infections, revealing the potential of these microscopic fast-evolving entities to influence host-parasite interactions. However, most of the changes in host microbiomes seem to depend on the host and parasite species in question. Furthermore, we should understand the relative role of parasitic infections as microbiome modulators when compared with other microbiome-impacting factors (e.g., host size, age, sex). Here, we characterised the microbiome of a single intermediate host species infected by two parasites belonging to different phyla: the acanthocephalan Plagiorhynchus allisonae and a dilepidid cestode, both infecting Transorchestia serrulata amphipods collected simultaneously from the same locality. We used the v4 hypervariable region of the 16S rRNA prokaryotic gene to identify the hemolymph bacterial community of uninfected, acanthocephalan-infected, and cestode-infected amphipods, as well as the bacteria in the amphipods' immediate environment and in the parasites infecting them. Our results show that parasitic infections were more strongly associated with differences in host bacterial community richness than amphipod size, presence of amphipod eggs in female amphipods, and even parasite load. Amphipods infected by acanthocephalans had the most divergent bacterial community, with a marked decrease in alpha diversity compared with cestode-infected and uninfected hosts. In accordance with the species-specific nature of microbiome changes in parasitic infections, we found unique microbial taxa associating with hosts infected by each parasite species, as well as taxa only shared between a parasite species and their infected hosts. However, there were some bacterial taxa detected in all parasitised amphipods (regardless of the parasite species), but not in uninfected amphipods or the environment. We propose that shared bacteria associated with all hosts parasitised by distantly related helminths may be important either in helping host defences or parasites' success, and could thus interact with host-parasite evolution.</p>","PeriodicalId":13725,"journal":{"name":"International journal for parasitology","volume":" ","pages":"733-742"},"PeriodicalIF":3.7,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142107086","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}