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Multi-locus typing of Histomonas meleagridis isolates demonstrates the existence of two different genotypes.

ABSTRACT: Histomonas meleagridis is the aetiological agent of histomonosis or "blackhead disease". Histomonosis is of special importance today, because there is no effective treatment to prevent its occurrence with considerable losses for the poultry industry. Despite its importance only a few molecular studies have yet been performed to investigate the degree of genetic diversity between different isolates of this parasite. In the present study a collection of well defined samples, previously shown positive for the DNA of H. meleagridis, was used to investigate genetic relatedness of the parasite. Samples originated from 25 turkey flocks collected in France between 2007 and 2010. Additionally, diagnostic samples, collected at our Clinic in Vienna, from different European countries and Azerbaijan, during 2010 to 2013 were included in the analyses. For the analysis three different genetic loci were analyzed: 18S rRNA, ?-actinin1 and rpb1 genes. To amplify partial sequences of ?-actinin1 and rpb1 genes, primers specifically targeting H. meleagridis were designed. Following PCR, the sequences of 18S rRNA, ?-actinin1 and rpb1 loci were analyzed. Phylogenetic analyses demonstrated separation of H. meleagridis isolates in two different clusters. The majority of isolates grouped within the cluster 1 and originated from different European countries. The cluster 2 was rare and predominantly found in samples originating from France. Considering that the genetic variability of clusters can be seen as two distinct genetic types we propose the term genotype instead of cluster.

SUBMITTER: Bilic I

PROVIDER: S-EPMC3962415 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Multi-locus typing of Histomonas meleagridis isolates demonstrates the existence of two different genotypes.

Bilic Ivana I Jaskulska Barbara B Souillard Rozenn R Liebhart Dieter D Hess Michael M

PloS one 20140321 3

Histomonas meleagridis is the aetiological agent of histomonosis or "blackhead disease". Histomonosis is of special importance today, because there is no effective treatment to prevent its occurrence with considerable losses for the poultry industry. Despite its importance only a few molecular studies have yet been performed to investigate the degree of genetic diversity between different isolates of this parasite. In the present study a collection of well defined samples, previously shown posit ...[more]

PMID: 24658534

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Project description:BackgroundMelioidosis is a neglected tropical disease with rising global public health and clinical importance. Melioidosis is endemic in Southeast Asia and Northern Australia and is of increasing concern in Malaysia. Despite a number of reported studies from Malaysia, these reports are limited to certain parts of the country and do not provide a cohesive link between epidemiology of melioidosis cases and the nation-wide distribution of the causative agent Burkholderia pseudomallei.Methodology/principle findingsHere we report on the distribution of B. pseudomallei sequence types (STs) in Malaysia and how the STs are related to STs globally. We obtained 84 culture-confirmed B. pseudomallei from confirmed septicaemic melioidosis patients from all over Malaysia. Prior to performing Multi Locus Sequence Typing, the isolates were subjected to antimicrobial susceptibility testing and detection of the YLF/BTFC genes and BimA allele. Up to 90.5% of the isolates were sensitive to all antimicrobials tested while resistance was observed for antimicrobials typically administered during the eradication stage of treatment. YLF gene cluster and bimABp allele variant were detected in all the isolates. The epidemiological distribution patterns of the Malaysian B. pseudomallei isolates were analysed in silico using phylogenetic tools and compared to Southeast Asian and world-wide isolates. Genotyping of the 84 Malaysian B. pseudomallei isolates revealed 29 different STs of which 6 (7.1%) were novel. ST50 was identified as the group founder followed by subgroup founders ST376, ST211 and ST84. A low-level diversity is noted for the B. pseudomallei isolates described in this study while phylogenetic analysis associated the Malaysian STs to Southeast Asian isolates especially isolates from Thailand. Further analysis also showed a strong association that implicates agriculture and domestication activities as high-risk routes of infection.Conclusions/significanceIn conclusion, MLST analysis of B. pseudomallei clinical isolates from all states in Malaysia revealed low diversity and a close association to Southeast Asian isolates.

Project description:Background: Histomonas meleagridis is an anaerobic, intercellular parasite that infects the Galliformes such as turkeys and chickens. In recent years, the reemergence of Histomoniasis has caused serious economic losses as drugs to treat the disease have been banned. At present, studies on H. meleagridis mainly focus on virulence, gene expression analysis, and the innate immunity of the host. However, there are no studies on differential expression of miRNAs (DEMs) in host immune and inflammatory response induced by H. meleagridis infection in chickens. In this study, the expression profile of cecum miRNA at 10 and 15 days post-infection (DPI) with Chinese JSYZ-F strain H. meleagridis was studied by high-throughput sequencing. Results: Compared with the control group, 94 and 127 DEMs were found in the cecum of infected chickens at 10 DPI (CE vs CC) and 15 DPI (CEH vs CCH), respectively, of which 60 DEMs were shared at two-time points. Gene Ontology (GO) enrichment analysis of the target genes of DEMs showed that 881 and 1027 GO terms were significantly enriched at 10 and 15 DPI. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of the target genes of DEMs showed that only 5 and 3 pathways were significantly enriched at 10 and 15 DPI, respectively. The integrated analysis of miRNA–gene network revealed that the DEMs played important roles in the host immune and inflammatory responses to H. meleagridis infection by dynamically regulating the expression of immune and inflammation-related cytokines. Conclusion: Our results not only suggested that host miRNA expression was dynamically altered by H. meleagridis and host, but also revealed that more miRNAs and genes were involved in the later stage of the disease. In addition, host and H. meleagridis regulated the expression of immune and inflammation-related cytokines to respond to H. meleagridis infection. Our results will contribute to future research on miRNA-target interaction during H. meleagridis infection in chickens and provide new ideas for H. meleagridis control.

Project description:Background: Histomonas meleagridis is an anaerobic, intercellular parasite, which infects gallinaceous birds such as turkeys and chickens. In recent years, the reemergence of Histomoniasis has caused serious economic losses as drugs to treat the disease have been banned. At present, research of H. meleagridis focuses on virulence, gene expression analysis, and the innate immunity of the host. However, there are no study on the differentially expressed miRNAs (DEMs) associated with the liver induced by H. meleagridis. In this research, high-throughput sequencing was used to analyze the expression profile of liver miRNA at 10 and 15 days post-infection (DPI) in chickens infected with Chinese JSYZ-F strain H. meleagridis. Results: Compared with the uninfected control, 120 and 118 DEMs were found in the liver of infected chickens at 10 DPI and 15 DPI, respectively, of which 74 DEMs were shared at two-time points. Differentially expressed miRNAs were classified into three types according to the time of infection: L1, 45 miRNAs differentially expressed only at 10 DPI, were predicted to target 1646 genes; L2, 43 miRNAs differentially expressed only at 15 DPI, were predicted to target 2257 genes; L3, 75 miRNAs differentially expressed at both 10 DPI and 15 DPI, were predicted to target 1623 genes. A total of 89, 87 and 41 significantly enriched GO terms (p<0.05) were identified at L1, L2 and L3, respectively. The KEGG pathway analysis of differentially expressed miRNA target genes were shown that 3, 4 and 5 significantly enriched KEGG pathway (p<0.05) were identified at L1, L2 and L3, respectively. Conclusion: This article suggested that liver miRNA expression was dynamically altered by H. meleagridis and host and showed that the expression pattern of the L1 class DEMs were more favourable for controlling the development of the inflammatory response, whereas the L2 class DEMs were more favourable for enhancing the inflammatory response. Inflammation-associated miRNA expression patterns were consistent with the liver inflammatory process after artificial infection. The results of the study laid the foundation for an in-depth analysis of the pathogenic mechanism of H. meleagridis from the perspective of host miRNAs.

Dataset Information

Multi-locus typing of Histomonas meleagridis isolates demonstrates the existence of two different genotypes.

Publications

Multi-locus typing of Histomonas meleagridis isolates demonstrates the existence of two different genotypes.

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