High frequency of benzimidazole resistance alleles in trichostrongyloids from Austrian sheep flocks in an alpine transhumance management system

Abstract Background: Infections of small ruminants with trichostrongyloid nematodes often result in reduced productivity and may be detrimental to the host. Anthelmintic resistance (AR) against most anthelmintic drug classes is now widespread amongst the trichostrongyloids. Baseline establishment, followed by regular monitoring of the level of AR, is necessary for farmers and veterinarians to make informed decisions about parasite management. The detection of single nucleotide polymorphisms (SNPs) is a sensitive method to detect AR against benzimidazoles (BZs), one of the most widely used anthelmintic classes. Alpine transhumance constitutes a special type of pasturing of sheep from many different farms, the aim of this study was to investigate the prevalence of benzimidazole resistance alleles in this particular management system. Results: Sixteen sheep flocks in Styria and Salzburg in Austria were examined by pyrosequencing for SNPs at codons 167, 198 and 200 of the isotype-1 β-tubulin gene. The frequency of the resistance-associated exchange F200Y was 87–100% for H. contortus, 77–100% for T. colubriformis and <5–66% for T. circumcincta. Additionally, the F167Y polymorphism was detected in T. colubriformis from two farms at a frequency of 19% and 23% respectively. Conclusions: The high resistance allele frequency in H. contortus and T. colubriformis in the examined sheep population urgently calls for the development of new treatment strategies to sustainably control trichostrongyloid infections for this kind of pasturing, since the frequent mixing of flocks during the alpine summer grazing must be considered an important risk factor for the spread of resistant nematodes to a large number of farms.

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High frequency of benzimidazole resistance alleles in Haemonchus contortus, Teladorsagia circumcincta and Trichostrongylus colubriformis from Austrian sheep flocks in an alpine transhumance management system

10.21203/rs.2.18014/v1 ◽

2019 ◽

Author(s):

Barbara Hinney ◽

Julia Schoiswohl ◽

Lynsey Melville ◽

Vahel J. Ameen ◽

Walpurga Wille-Piazzai ◽

...

Keyword(s):

Management System ◽

Anthelmintic Resistance ◽

Treatment Strategies ◽

Small Ruminants ◽

Nucleotide Polymorphisms ◽

Sheep Population ◽

Teladorsagia Circumcincta ◽

Benzimidazole Resistance ◽

Drug Classes ◽

New Treatment

Abstract Infections of small ruminants with trichostrongyloid nematodes often result in reduced productivity and may be detrimental to the host. Anthelmintic resistance (AR) against most anthelmintic drug classes is now widespread amongst the trichostrongyloids. Baseline establishment, followed by regular monitoring of the level of AR, is necessary for farmers and veterinarians to make informed decisions about parasite management. The detection of single nucleotide polymorphisms (SNPs) is a sensitive method to detect AR against benzimidazoles (BZs), one of the most widely used anthelmintic classes. Alpine transhumance constitutes a special type of pasturing of sheep from many different farms, the aim of this study was to investigate the prevalence of benzimidazole resistance alleles in this particular management system. Sixteen sheep flocks in Styria and Salzburg in Austria were examined by pyrosequencing for SNPs at codons 167, 198 and 200 of the isotype-1 β-tubulin gene. The frequency of the resistance-associated mutation F200Y was 87–100% for H. contortus, 77–100% for T. colubriformis and <5–66% for T. circumcincta. Additionally, the F167Y polymorphism was detected in T. colubriformis from two farms at a frequency of 19% and 23% respectively. The high resistance allele frequency in H. contortus and T. colubriformis in the examined sheep population urgently calls for the development of new treatment strategies to sustainably control trichostrongyloid infections for this kind of pasturing, since the frequent mixing of flocks during the alpine summer grazing must be considered an important risk factor for the spread of resistant nematodes to a large number of farms.

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Distribution of QPY and RAH haplotypes of granzyme B gene in distinct Brazilian populations

Revista da Sociedade Brasileira de Medicina Tropical ◽

10.1590/s0037-86822012005000001 ◽

2012 ◽

Vol 45 (4) ◽

pp. 496-499

Author(s):

Fernanda Bernadelli Garcia ◽

Simone Kashima ◽

Evandra Strazza Rodrigues ◽

Israel Tojal Silva ◽

Tathiane Maistro Malta ◽

...

Keyword(s):

Granzyme B ◽

Treatment Strategies ◽

Cytotoxic Lymphocytes ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

Exon 2 ◽

Significant Difference ◽

Novel Treatment Strategies ◽

Brazilian Populations ◽

Afro Brazilian

INTRODUCTION: The cytolysis mediated by granules is one of the most important effector functions of cytotoxic T lymphocytes and natural killer cells. Recently, three single nucleotide polymorphisms (SNPs) were identified at exons 2, 3, and 5 of the granzyme B gene, resulting in a haplotype in which three amino acids of mature protein Q48P88Y245 are changed to R48A88H245, which leads to loss of cytotoxic activity of the protein. In this study, we evaluated the frequency of these polymorphisms in Brazilian populations. METHODS: We evaluated the frequency of these polymorphisms in Brazilian ethnic groups (white, Afro-Brazilian, and Asian) by sequencing these regions. RESULTS: The allelic and genotypic frequencies of SNP 2364A/G at exon 2 in Afro-Brazilian individuals (42.3% and 17.3%) were significantly higher when compared with those in whites and Asians (p < 0.0001 and p = 0.0007, respectively). The polymorphisms 2933C/G and 4243C/T also were more frequent in Afro-Brazilians but without any significant difference regarding the other groups. The Afro-Brazilian group presented greater diversity of haplotypes, and the RAH haplotype seemed to be more frequent in this group (25%), followed by the whites (20.7%) and by the Asians (11.9%), similar to the frequency presented in the literature. CONCLUSIONS: There is a higher frequency of polymorphisms in Afro-Brazilians, and the RAH haplotype was more frequent in these individuals. We believe that further studies should aim to investigate the correlation of this haplotype with diseases related to immunity mediated by cytotoxic lymphocytes, and if this correlation is confirmed, novel treatment strategies might be elaborated.

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Large-scale PRNP genotyping of small ruminants using an automated high-throughput MALDI-TOF MS assay

Journal of Veterinary Diagnostic Investigation ◽

10.1177/1040638719859043 ◽

2019 ◽

Vol 31 (4) ◽

pp. 629-633

Author(s):

Sergio Migliore ◽

Maurizio Bivona ◽

Enrico Gagliostro ◽

Onofrio Buttitta ◽

Francesca Lo Mascolo ◽

...

Keyword(s):

High Throughput ◽

Large Scale ◽

Genetic Resistance ◽

Small Ruminants ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Automated Processing ◽

Tof Ms

Scrapie resistance or susceptibility in sheep is associated with single nucleotide polymorphisms (SNPs) at codons 136, 154, and 171 of the prion protein gene ( PRNP). In addition, phenylalanine mutation at codon 141 has been recognized as a risk factor for atypical scrapie. In contrast, K222, D146, and S146 alleles confer genetic resistance to classical scrapie in goats. High-throughput genotyping technologies would provide significant benefits in scrapie eradication plans. The ability to resolve oligonucleotides varying in mass by less than a single nucleotide makes MALDI-TOF mass spectrometry (MS) a suitable platform for PRNP genotyping. We evaluated the commercial Myriapod scrapie kit (Diatech Pharmacogenetics), associated with a highly automated processing platform incorporating MALDI-TOF MS technology, to detect SNPs at codons 136, 154, 171, 141, and 222 of small ruminant PRNP. The Myriapod scrapie kit was accredited according to UNI CEI EN ISO/IEC 17025. We present the genotyping results of 10,960 sheep in Sicily and 1,822 goats in Sicily and Calabria (southern Italy) tested during 2017. We found a high frequency (43.9%) of the protective ARR allele in sheep and a promising 12.3% of the resistant K222 variant in goats. This efficient and high-throughput method is suitable for extensive PRNP genotyping, as requested in the European scrapie eradication plan.

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ABCB1andABCC3Gene Polymorphisms Are Associated with First-year Response to Methotrexate in Juvenile Idiopathic Arthritis

The Journal of Rheumatology ◽

10.3899/jrheum.111593 ◽

2012 ◽

Vol 39 (10) ◽

pp. 2032-2040 ◽

Cited By ~ 50

Author(s):

MAURITS C.F.J. de ROTTE ◽

MAJA BULATOVIC ◽

MARLOES W. HEIJSTEK ◽

GERRIT JANSEN ◽

SANDRA G. HEIL ◽

...

Keyword(s):

Juvenile Idiopathic Arthritis ◽

Gene Polymorphism ◽

Treatment Strategies ◽

First Year ◽

Nucleotide Polymorphisms ◽

Genetic Determinants ◽

Single Nucleotide ◽

American College Of Rheumatology ◽

Individualize Treatment ◽

Rheumatology Pediatric

Objective.Although methotrexate (MTX) is the most widely prescribed drug in juvenile idiopathic arthritis (JIA), 30% of patients fail to respond to it. To individualize treatment strategies, the genetic determinants of response to MTX should be identified.Methods.A cohort of 287 patients with JIA treated with MTX was studied longitudinally over the first year of treatment. MTX response was defined as the American College of Rheumatology pediatric 70 criteria (ACRped70). We genotyped 21 single-nucleotide polymorphisms in 13 genes related to MTX polyglutamylation and to cellular MTX uptake and efflux. Potential associations between ACRped70 and genotypes were analyzed in a multivariate model and corrected for these 3 covariates: disease duration prior to MTX treatment, physician’s global assessment of disease activity at baseline, and MTX dose at all study visits.Results.MTX response was more often achieved by patients variant for the adenosine triphosphate-binding cassette transporter B1 (ABCB1) gene polymorphism rs1045642 (OR 3.80, 95% CI 1.70−8.47, p = 0.001) and patients variant for theABCC3gene polymorphism rs4793665 (OR 3.10, 95% CI 1.49−6.41, p = 0.002) than by patients with other genotypes. Patients variant for the solute carrier 19A1 (SLC19A1) gene polymorphism rs1051266 were less likely to respond to MTX (OR 0.25, 95% CI 0.09−0.72, p = 0.011).Conclusion.ABCB1rs1045642,ABCC3rs4793665, andSLC19A1rs1051266 polymorphisms were associated with response to MTX in 287 patients with JIA studied longitudinally. Upon validation of our results in other JIA cohorts, these genetic determinants may help to individualize treatment strategies by predicting clinical response to MTX.

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Phenotypic and genotypic approaches for detection of anthelmintic resistant sheep gastrointestinal nematodes from Brazilian northeast

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612021048 ◽

2021 ◽

Vol 30 (2) ◽

Author(s):

José Vilemar de Araújo-Filho ◽

Wesley Lyeverton Correia Ribeiro ◽

Weibson Paz Pinheiro André ◽

Géssica Soares Cavalcante ◽

Jéssica Maria Leite dos Santos ◽

...

Keyword(s):

Anthelmintic Resistance ◽

Gastrointestinal Nematode ◽

Gastrointestinal Nematodes ◽

Northeastern Brazil ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

Egg Hatch ◽

Fecal Egg Count ◽

Phenotypic Tests ◽

Brazilian Northeast

Abstract The aim of this study was to characterize the anthelmintic resistance (AR) of a sheep gastrointestinal nematode population, named Caucaia, from northeastern Brazil. Phenotypic tests performed were: egg hatch (EHT), larval development (LDT) and fecal egg count reduction (FECRT). Benzimidazoles (BZs) genotypic evaluation was by frequency of single nucleotide polymorphisms (SNPs) F200Y, F167Y and E198A, and for levamisole (LEV), by frequency of resistance alleles of Hco-acr-8 gene. The primers were designed specifically for Haemonchus contortus. Effective concentrations 50% (EC50) for BZs (EHT), and for macrocyclic lactones (MLs) and LEV (LDT) were 1.02 µg/mL, 1.81 ng/mL and 0.04 µg/mL, respectively. Resistance ratios for MLs and LEV were 0.91 and 3.07, respectively. FECRT efficacies of BZs, MLs, monepantel (MPTL) and LEV were 52.4; 87.0; 94.5 and 99.6%, respectively. qPCR for BZs demonstrated resistance allele frequencies of 0%, 26.24% and 69.08% for SNPs E198A, F200Y and F167Y, respectively. For LEV, 54.37% of resistance alleles were found. There was agreement between EHT, FECRT and qPCR for BZs, and agreement between LDT and qPCR for LEV. Thus, based on higher sensitivity of qPCR, and phenotypic evaluation, the Caucaia population was considered resistant to BZs, MLs, LEV and suspect for MPTL.

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Model-based integration of genomics and metabolomics reveals SNP functionality in Mycobacterium tuberculosis

10.1101/555763 ◽

2019 ◽

Cited By ~ 1

Author(s):

Ove Øyås ◽

Sonia Borrell ◽

Andrej Trauner ◽

Michael Zimmermann ◽

Julia Feldmann ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Treatment Strategies ◽

Nucleotide Polymorphisms ◽

Aminosalicylic Acid ◽

Selective Treatment ◽

Single Nucleotide ◽

Microbial Life ◽

Model Based ◽

Encoding Genes

AbstractHuman tuberculosis is caused by members of the Mycobacterium tuberculosis complex (MTBC) and presents variable disease outcomes. The variation has primarily been attributed to host and environmental factors, but recent evidence indicates an additional role of genetic diversity among MTBC clinical strains. Here, we used metabolomics to unravel the potential role of genetic variations in conferring strain-specific adaptive capacity and vulnerability. To systematically identify functionality of single nucleotide polymorphisms (SNPs), we developed a constraint-based approach that integrates metabolomic and genomic data. Model-based predictions were systematically tested against independent metabolome data; they correctly classified SNP effects in pyruvate kinase and suggested a genetic basis for strain-specific sensitivity to the antibiotic para-aminosalicylic acid. Our method is broadly applicable to mutations in enzyme-encoding genes across microbial life, opening new possibilities for identifying strain-specific metabolic vulnerabilities that could lead to more selective treatment strategies.

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Genetic Polymorphisms and Susceptibility to Sudden Sensorineural Hearing Loss: A Systematic Review

Audiology and Neurotology ◽

10.1159/000497032 ◽

2019 ◽

Vol 24 (1) ◽

pp. 8-19 ◽

Cited By ~ 1

Author(s):

Zaizai Cao ◽

Jinjian Gao ◽

Saiyu Huang ◽

Haijie Xiang ◽

Chuqin Zhang ◽

...

Keyword(s):

Systematic Review ◽

Hearing Loss ◽

Sensorineural Hearing Loss ◽

Treatment Strategies ◽

Sudden Sensorineural Hearing Loss ◽

Sensorineural Hearing ◽

Nucleotide Polymorphisms ◽

Multicenter Studies ◽

Single Nucleotide ◽

Effective Prevention

Background: Recently, genetic factors have been considered as an important risk factor for sudden sensorineural hearing loss (SSNHL). Many studies analyzed the association between SSNHL and polymorphisms. However, most of them gave inconclusive results. Key Message: We performed a systematic review to find out the association between polymorphisms and susceptibility to SSNHL. Finally, 47 studies involving 5,230 SSNHL patients and 68 genes were included for analysis and discussion of results. Polymorphisms in 26 genes have been suggested to be correlated with the susceptibility to SSNHL. Summary: Although a great number of studies support that polymorphisms in genes are associated with susceptibility to SSNHL, we need large multicenter studies, which evaluate multiple single nucleotide polymorphisms in SSNHL patients, to find real genetic risk factors for susceptibility to SSNHL. This is very helpful in designing more effective prevention and treatment strategies for patients with SSNHL.

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Polymorphisms of MTHFR Associated with Higher Relapse/Death Ratio and Delayed Weekly MTX Administration in Pediatric Lymphoid Malignancies

Leukemia Research and Treatment ◽

10.1155/2013/238528 ◽

2013 ◽

Vol 2013 ◽

pp. 1-9 ◽

Cited By ~ 3

Author(s):

Hiroko Fukushima ◽

Takashi Fukushima ◽

Aiko Sakai ◽

Ryoko Suzuki ◽

Ryoko Nakajima-Yamaguchi ◽

...

Keyword(s):

Direct Sequencing ◽

Lymphoblastic Leukemia ◽

Treatment Strategies ◽

Diagnostic Techniques ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

Lymphoid Malignancies ◽

Childhood Malignancies ◽

Taqman Pcr ◽

Pcr Method

Backgrounds. Outcome of childhood malignancy has been improved mostly due to the advances in diagnostic techniques and treatment strategies. While methotrexate (MTX) related polymorphisms have been under investigation in childhood malignancies, many controversial results have been offered. Objectives. To evaluate associations of polymorphisms related MTX metabolisms and clinical course in childhood lymphoid malignancies. Method. Eighty-two acute lymphoblastic leukemia and 21 non-Hodgkin’s lymphoma children were enrolled in this study. Four single nucleotide polymorphisms in 2 genes (MTHFR (rs1801133/c.677C>T/p.Ala222Val and rs1801131/c.1298A>C/p.Glu429Ala) and SLCO1B1 (rs4149056/c.521T>C/p.V174A and rs11045879/c.1865+4846T>C)) were genotyped by Taqman PCR method or direct sequencing. Clinical courses were reviewed retrospectively. Results. No patient who had the AC/CC genotype of rs1801131 (MTHFR) had relapsed or died, in which distribution was statistically different among the AA genotype of rs1801131 (P=0.004). Polymorphisms of SLCO1B1 (rs11045879 and rs4149056) were not correlated with MTX concentrations, adverse events, or disease outcome. Conclusions. Polymorphisms of MTHFR (rs1801131) could be the plausive candidate for prognostic predictor in childhood lymphoid malignancies.

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Multiple Drug Resistance in the canine hookworm Ancylostoma caninum: an Emerging Threat

10.1101/676007 ◽

2019 ◽

Author(s):

Pablo D. Jimenez Castro ◽

Sue Howell ◽

John. J. Schaefer ◽

Russell. W. Avramenko ◽

John. S. Gilleard ◽

...

Keyword(s):

Anthelmintic Resistance ◽

Multiple Drug Resistance ◽

Scale Up ◽

Amplicon Sequencing ◽

The United States ◽

Multiple Drug ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

Ancylostoma Caninum ◽

Health Community

AbstractIn the past few years, diagnoses by veterinarians of recurrent canine hookworm infections have dramatically increased, suggesting that anthelmintic resistance (AR) may have evolved in the parasite Ancylostoma caninum. To investigate this, we established three “suspected-resistant” and two susceptible A. caninum isolates in research dogs for further study. The egg hatch assay (EHA) and the larval development assay (LDA) were used for detecting resistance to benzimidazoles, and macrocyclic lactones, respectively. Resistance ratios ranged from 6.0 to >100 and 5.5-69.8 for the EHA and LDA, respectively. Following treatments with fenbendazole, pyrantel and milbemycin oxime, reduction in faecal egg counts ranged from 64–86%, 0–72% and 58–92%, respectively. Deep amplicon sequencing of the isotype-1 β tubulin gene identified a high frequency of resistance-associated single nucleotide polymorphisms at codon 167 in the resistant isolates and clinical cases.. These data conclusively demonstrate multiple anthelmintic resistance in A. caninum, and provide pivotal evidence that this is an emerging problem in the United States. Consequently, these findings should provide some concern to the global health community, as the scale-up of mass drug administration for soil-transmitted helminths (STH) is now placing similar selection pressures for benzimidazole resistance in human hookworms.

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Discovery of unique loci that underlie nematode responses to benzimidazoles

10.1101/116970 ◽

2017 ◽

Cited By ~ 4

Author(s):

Mostafa Zamanian ◽

Daniel E. Cook ◽

Stefan Zdraljevic ◽

Shannon C. Brady ◽

Daehan Lee ◽

...

Keyword(s):

Drug Resistance ◽

Resistance Genes ◽

Anthelmintic Resistance ◽

Benzimidazole Derivatives ◽

Parasitic Nematodes ◽

C Elegans ◽

Benzimidazole Resistance ◽

Drug Classes ◽

Parasite Populations ◽

Parasite Drug Resistance

Parasitic nematodes impose a debilitating health and economic burden across much of the world. Nematode resistance to anthelmintic drugs threatens parasite control efforts in both human and veterinary medicine. Despite this threat, the genetic landscape of potential resistance mechanisms to these critical drugs remains largely unexplored. Here, we exploit natural variation in the model nematodes Caenorhabditis elegans and Caenorhabditis briggsae to discover quantitative trait loci (QTL) that control sensitivity to benzimidazoles widely used in human and animal medicine. High-throughput phenotyping of albendazole, fenbendazole, mebendazole, and thiabendazole responses in panels of recombinant lines led to the discovery of over 15 QTL in C. elegans and four QTL in C. briggsae associated with divergent responses to these anthelmintics. Many of these QTL are conserved across benzimidazole derivatives, but others show drug and dose specificity. We used near-isogenic lines to recapitulate and narrow the C. elegans albendazole QTL of largest effect and identified candidate variants correlated with the resistance phenotype. These QTL do not overlap with known benzimidazole resistance genes from parasitic nematodes and present specific new leads for the discovery of novel mechanisms of nematode benzimidazole resistance. Analyses of orthologous genes reveal significant conservation of candidate benzimidazole resistance genes in medically important parasitic nematodes. These data provide a basis for extending these approaches to other anthelmintic drug classes and a pathway towards validating new markers for anthelmintic resistance that can be deployed to improve parasite disease control.Author SummaryThe treatment of roundworm (nematode) infections in both humans and animals relies on a small number of anti-parasitic drugs. Resistance to these drugs has appeared in veterinary parasite populations and is a growing concern in human medicine. A better understanding of the genetic basis for parasite drug resistance can be used to help maintain the effectiveness of anti-parasitic drugs and to slow or to prevent the spread of drug resistance in parasite populations. This goal is hampered by the experimental intractability of nematode parasites. Here, we use non-parasitic model nematodes to systematically explore responses to the critical benzimidazole class of anti-parasitic compounds. Using a quantitative genetics approach, we discovered unique genomic intervals that control drug effects, and we identified differences in the genetic architectures of drug responses across compounds and doses. We were able to narrow a major-effect genomic region associated with albendazole resistance and to establish that candidate genes discovered in our genetic mappings are largely conserved in important human and animal parasites. This work provides new leads for understanding parasite drug resistance and contributes a powerful template that can be extended to other anti-parasitic drug classes.

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