Dataset Information

First microscopic and molecular parasitological survey of Strongylus vulgaris in Brazilian ponies.

ABSTRACT: The frequency of gastrointestinal parasites with an emphasis on Strongylus vulgaris was investigated among the Brazilian Pony breed kept on farms in the municipality of Teresópolis, state of Rio de Janeiro. Fecal samples were collected in three stud farms: A (n= 22 animals), B (n= 3), and C (n= 2). Fecal samples were subjected to the quantitative Mini-FLOTAC technique, using three different solutions, and to qualitative techniques. The parasite prevalence was found to be 81.4%. Eggs from strongylids were identified in 74% of the ponies. Eggs of Parascaris spp. were detected in 22.7% of the animals, which were all females of farm A. At this locality, mares were kept with their foals in fenced paddocks all the time. The NaCl solution of d = 1.200 g/ml was generally the one that presented the highest frequency of diagnosis of nematode eggs and the highest mean of fecal eggs per gram. The fecal samples were also subjected to the polymerase chain reaction for amplification of DNA from the ITS2 region for Strongylus vulgaris. Twelve samples presented nucleotide sequences for S. vulgaris. In the end, this study revealed the high frequency (96.3%) of S. vulgaris among ponies on farms in Teresópolis, Rio de Janeiro, Brazil.

SUBMITTER: Martins AV

PROVIDER: S-EPMC10321716 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature

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First microscopic and molecular parasitological survey of Strongylus vulgaris in Brazilian ponies.

Martins André Vianna AV Coelho Aline de Lima AL Corrêa Laís Lisboa LL Ribeiro Mariana Santos MS Lobão Lucas Fernandes LF Palmer João Pedro Siqueira JPS Moura Lucas Cavalcante de LC Molento Marcelo Beltrão MB Barbosa Alynne da Silva ADS

Revista brasileira de parasitologia veterinaria = Brazilian journal of veterinary parasitology : Orgao Oficial do Colegio Brasileiro de Parasitologia Veterinaria 20230623 3

The frequency of gastrointestinal parasites with an emphasis on Strongylus vulgaris was investigated among the Brazilian Pony breed kept on farms in the municipality of Teresópolis, state of Rio de Janeiro. Fecal samples were collected in three stud farms: A (n= 22 animals), B (n= 3), and C (n= 2). Fecal samples were subjected to the quantitative Mini-FLOTAC technique, using three different solutions, and to qualitative techniques. The parasite prevalence was found to be 81.4%. Eggs from strongy ...[more]

PMID: 37377279

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Project description:BackgroundStrongyle parasites are ubiquitous in grazing horses. Strongylus vulgaris, the most pathogenic of the large strongyles, is known for its extensive migration in the mesenteric arterial system. The lifecycle of S. vulgaris is characterised by a long prepatent period where the migrating larvae are virtually undetectable as there currently is no test available for diagnosing prepatent S. vulgaris infection. Presence of S. vulgaris larvae in the arterial system causes endarteritis and thrombosis with a risk of non-strangulating intestinal infarctions. Emergence of anthelmintic resistance among cyathostomins has led to recommendations of reduced treatment intensity by targeting horses that exceed a predetermined strongyle faecal egg count threshold. One study suggests an apparent increase in prevalence of S. vulgaris on farms where reduced anthelmintic treatment intensity has been implemented. These issues highlight the need for an accurate and reliable assay for diagnosing prepatent S. vulgaris infection.MethodsImmunoscreening of a larval S. vulgaris cDNA library using hyperimmune serum raised against S. vulgaris excretory/secretory antigens was performed to identify potential diagnostic antigens. Immunoreactive clones were sequenced, one potential antigen was characterised, expressed as a recombinant protein, initially evaluated by western blot (WB) analysis, the diagnostic potential of the IgG subclasses was evaluated by ELISA, and the diagnostic accuracy evaluated using serum from 102 horses with known S. vulgaris infection status.ResultsThe clone expressing the potential antigen encoded a S. vulgaris SXP/RAL2 homologue. The recombinant protein, rSvSXP, was shown to be a potential diagnostic antigen by WB analysis, and a target of serum IgGa, IgG(T) and total IgG in naturally infected horses, with IgG(T) antibodies being the most reliable indicator of S. vulgaris infection in horses. Evaluation of diagnostic accuracy of the ELISA resulted in a sensitivity of 73.3%, a specificity of 81.0%, a diagnostic odds ratio of 11.69; a positive likelihood ratio (LR) of 3.85 and a negative LR was 0.33. The area under the ROC curve was 0.820.ConclusionIgG(T) antibodies to recombinant SvSXP show potential for use as an antigen for prepatent diagnosis of migrating stages of S. vulgaris with moderate to good diagnostic accuracy.

Project description:BackgroundSurveillance of Strongylus vulgaris and other Strongylus spp. in equids is important for targeted intervention in parasite control, requiring reliable routine diagnostic methods.ObjectivesComparing morphological examination and PCR analyses of larval cultures to identify Strongylus spp. species based on German diagnostic samples from 2018.Study designMethod comparison.MethodsDuring the routine diagnostic investigations, in total 712 strongyle-egg positive equine faecal samples were cultured. Third-stage larvae (L3) were morphologically differentiated. For molecular validation, samples were examined using S. vulgaris real-time PCR and Strongylus edentatus/Strongylus equinus/Strongylus asini high-resolution melting PCRs.ResultsBased on 28S rRNA PCR, 594 samples positive for nematode DNA were included in the study. The inter-rater reliability to compare morphological and molecular species identification was fair for Strongylus spp. without species identification and for S. edentatus, slight for S. equinus and poor for S. vulgaris. The frequency based on morphological and molecular data in this study were for S. vulgaris 0% and 0.8%, respectively, for S. edentatus 0.3% and 1.5%, respectively, and for S. equinus 2.0% and 0.2%, respectively. Based on molecular analyses, one sample obtained from a domestic horse contained S. asini DNA, which was confirmed by sequencing.Main limitationsFor many samples, no or only incomplete data regarding clinical history, the exact geographical location and whether samples were obtained on individual or farm level, were available.ConclusionsResults of morphological and molecular examination methods of strongyle L3 from equine samples can differ substantially. Further evaluation of these methods is required to provide reliable and cost-effective methods of screening equine parasites. Further studies using approaches suitable to detect S. asini are needed to evaluate its clinical and epidemiological relevance.

Project description:Strongylus vulgaris is considered the most pathogenic nematode parasite of the horse. Frequent deworming programs since the 1970s have reduced the prevalence of S. vulgaris to low levels, but to the price of widespread benzimidazole resistance in the small strongyles (cyathostominae) and ivermectin resistance especially in the equine roundworm Parascaris spp. To slow down the progression of anthelmintic resistance in Sweden, horse anthelmintics were made prescription only medicine in 2007 and selective therapy principles were introduced. This means that only individuals with high egg excretion or clinical signs of helminth infection were treated instead of blanket treatment of all horses on a farm. The aims of this study were to investigate the prevalence and risk factors associated with S. vulgaris infection in Sweden ten years after the introduction of a selective therapy regime. A total of 529 faecal samples from 106 farms were collected during March to June in 2016 and 2017. A web-based questionnaire was used to collect information about deworming routines. Strongyle faecal egg counts (FEC) were performed by McMaster and presence of S. vulgaris was demonstrated with a specific PCR on individual larval cultures. Results were analysed by mixed logistic (S. vulgaris prevalence) or linear (FEC) regression models, where farm was included as random factor. The overall prevalence of S. vulgaris at individual and farm levels was 28% and 61%, respectively. We observed a 2.9 increased odds risk of S. vulgaris infection on farms that based their treatment on strongyle FEC only as compared to farms that complemented strongyle FECs with larval cultures or dewormed regularly, 1-4 times per year, without prior diagnosis. We found no association between the prevalence of S. vulgaris and strongyle FEC level, horse age, geographical region or signs of colic. The prevalence of S. vulgaris was 25% in horses shedding ≤150 eggs per gram. Thus horses with low strongyle FECs that are left untreated could be an important source of S. vulgaris infection. This may be an important reason for the approximately three-fold increase in S. vulgaris prevalence since 1999 in Sweden. However, our combined results indicate that selective therapy based on a combination of strongyle FECs and larval cultivation was not associated with an increased risk of S. vulgaris infection. Still, S. vulgaris needs to be monitored continuously and should be taken into careful consideration when the treatment frequency is reduced.

Dataset Information

First microscopic and molecular parasitological survey of Strongylus vulgaris in Brazilian ponies.

Publications

First microscopic and molecular parasitological survey of Strongylus vulgaris in Brazilian ponies.

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