Mesocestoides Sp Research Articles

Optimization of CRISPR/Cas12a detection assay and its application in the detection of Echinococcus granulosus

Cystic echinococcosis, resulting from infection with Echinococcus granulosus, poses a significant challenge as a neglected tropical disease owing to the lack of any known effective treatment. Primarily affecting under-resourced, remote, and conflict-ridden regions, the disease is compounded by the limitations of current detection techniques, such as microscopy, physical imaging, ELISA, and qPCR, which are unsuitable for application in these areas. The emergence of CRISPR/Cas12a as a promising tool for nucleic acid detection, characterized by its unparalleled specificity, heightened sensitivity, and rapid detection time, offers a potential solution. In this study, we present a one-pot CRISPR/Cas12a detection method for E. granulosus (genotype G1, sheep strain) integrating recombinase polymerase amplification (RPA) with suboptimal protospacer adjacent motif (PAM) and structured CRISPR RNA (crRNA) to enhance reaction efficiency. The evaluation of the assay's performance using hydatid cyst spiked dog feces and the examination of 62 dog fecal samples collected from various regions of Western China demonstrate its efficacy. The assay permits visual observation of test results about 15 minutes under blue light and displays superior portability and reaction speed relative to qPCR, achieving a sensitivity level of 10 copies of standard plasmids of the target gene. Analytic specificity was verified against four tapeworm species (E. multilocularis, H. taeniaeformis, M. benedeni, and D. caninum) and two other helminths (T. canis and F. hepatica), with negative results also noted for Mesocestoides sp. This study presents a rapid, sensitive, and time-efficient DNA detection method for E. granulosus of hydatid cyst spiked and clinical dog feces, potential serving as an alternative tool for field detection. This novel assay is primarily used to diagnose the definitive host of E. granulosus. Further validation using a larger set of clinical fecal samples is warranted, along with additional exploration of more effective approaches for nucleic acid release.

Comparing copromicroscopy to intestinal scraping to monitor red fox intestinal helminths with zoonotic and veterinary importance.

The red fox acts as reservoir for several helminthic infections which are of interest for both public and animal health. Huge efforts have been made for the assessment of the sensitivity of coprological tests for the detection of Echinococcus multilocularis, while less attention has been paid to other helminthic species. This study aimed at assessing the performance of two copromicroscopic techniques in the detection and prevalence estimation of gastrointestinal helminths in the red fox. Helminths were isolated from the small intestines of 150 red foxes from Bolzano province, Italy, with a scraping, filtration and counting technique (SFCT) and morphologically identified. Rectal contents were collected and submitted to simple flotation (FT) and, only for Taenids, a method based on the concentration of eggs and identification with multiplex PCR (CMPCR). Using SFCT as a reference standard, we estimated the sensitivity of the copromicroscopic tests. Three species of nematodes (namely, Toxocara canis, Uncinaria stenocephala and Pterygodermatites sp.) and five species of cestodes (E. multilocularis, Taenia crassiceps, T. polycantha, Hydatigera taeniaeformis, Mesocestoides sp.) were identified with SFCT, whereas eggs referable to the same taxa were detected with fecal diagnostics, except for Pterygodermatites sp. and Mesocestoides sp. The sensitivity of FT was low for all taxa, ranging from 9.8 to 36.3%, with lower values for Taeniidae. CMPCR was confirmed to perform better for the detection of Taeniidae eggs (23.5%) and the multiplex PCR on retrieved eggs was effective in the identification of the species. A meta-analysis of literature also suggested that our results are consistent with existing data, indicating that copromicroscopy tends to underestimate the prevalence of helminthic infections. The extent of such underestimation varies with taxon, being higher at high prevalence levels, in particular for cestodes. Irregular dynamics of egg shedding, and routine deep freezing of red fox feces may explain the frequency of false negatives with copromicroscopy. Low sensitivity of copromicroscopic tests should be accounted for when estimating prevalence and when defining the correct sample size for the detection of the parasites.

Epidemiological Survey on Gastrointestinal and Pulmonary Parasites in Cats Around Toulouse (France).

Various feline parasites are potentially zoonotic thus establishing factors associated with parasitism is of animal and public health interest. The aim of this study was to determine the prevalence of endoparasites in client-owned cats, living in the area of Toulouse, France, over the period 2015 - 2017, and to investigate possible risk factors. A total of 498 faecal samples from cats of the University Animal Hospital of Ecole Nationale Vétérinaire de Toulouse were analysed, 448 from cats presented for consultation and 50 at post-mortem. Analysis was performed using a commercial flotation enrichment method with a hypersaturated sodium chloride solution and Baermann technique. Further examination of the gastrointestinal tract contents was conducted on necropsied cats. Overall, 11.6 % of cats were positive for endoparasites; 50 (11.2 %) consultation cases and 8 (16 %) post-mortem cases, with no significant difference in prevalence between the groups. Amongst infected cats, most were infected by a single species of parasite and 10.3 % (n=6) were infected with two or more. The most common parasite was Toxocara cati with a prevalence of 9.4 % (n=47). Other endoparasites encountered were: Cystoisospora sp 1.0 % (n=5), Aelurostrongylus abstrusus 1.0 % (n=5), Strongyloides sp 0.6 % (n=3), Dipylidium caninum 0.4 % (n=2), Aonchotheca putorii 0.2 % (n=1), Ancylostomatidae 0.2 % (n=1) and Toxascaris leonina 0.2 % (n=1). The examination of the gastronintestinal tract contents of the necropsied cats revealed Mesocestoides sp 0.4 % (n=2) and Tænia (Hydatigera) tæniaeformis sensu lato 0.2 % (n=1) which are seldomly diagnosed by flotation methods. In this study, increasing age and neutered status were statistically associated with reduced odds of infection by endoparasites (helminth and coccidian). Predictors of significantly increased risk included being male, intact, and not receiving regular anthelmintic treatment. The same risk factors were highlighted specifically for Toxocara cati infections, with rural location being an additional risk factor for infection.

The prevalence of gastrointestinal parasites in Panthera pardus of Katepurna wildlife sanctuary, Akola India

To study the prevalence of gastrointestinal parasites of Panthera pardus, 26 scat samples were collected from Katepurna Sanctuary during the census period conducted by forest department during January 2018. Out of these 21 samples had the highest overall infection rate of Paragonimus sp. Echinostomatidae, Dicrocoeliidae, Pseudophyllidea, Taeniidae, Mesocestoides sp. Hymenolepididae Acanthocephala, Spiruroidea, Gnathostoma sp. Molineus sp. Ancylostomoidea, Mammomonogamus sp. Toxocara-like, Toxascaris sp., Aelurostrongylus sp., Capillaria sp. and Mixed type of infections of Ascaris sp. was found faecal samples. This study provided a first overview on parasites in Panthera pardus in the Katepurna Sanctuary.

Morphological and Molecular Characterization of Post-Larval Pre-Tetrathyridia of Mesocestoides sp. (Cestoda: Cyclophyllidea) from Ground Skink, Scincella lateralis (SAURIA: SCINCIDAE), FROM SOUTHEASTERN OKLAHOMA.

Free pre-tetrathyridia of Mesocestoides sp. are described, for the first time, from samples obtained from the coelomic cavity of a ground skink, Scincella lateralis, from Oklahoma. Closer examination of these early-stage tapeworms revealed they were transitional metamorphosis stages between a post-hexacanth procercoid form to the full metacestode of Mesocestoides. A series of transitional stages was found that span the full period of sucker and apical organ development. However, we did not see any fully developed tetrathyridia, i.e., having classic Mesocestoides morphology but with the apical sucker absent following developmental atrophy. This is the first time that metamorphic pre-tetrathyridial stages of a Mesocestoides sp. have been reported in vivo from a natural infection. These observations corroborate earlier reports of such stages of Mesocestoides vogae developed in vitro, though the previously reported isolate of M. vogae is asexually proliferative, and the species from the present study showed no sign of asexual proliferation. The fact that these immediately post-hexacanth stages can occur in a single lizard intermediate host may suggest that Mesocestoides spp. might develop through a simple 2-host life cycle rather than an obligate 3-host cycle that has been speculated to occur by most previous authors. DNA sequence comparisons and phylogenetic analyses based on mitochondrial 12S, cox1, and nad1 genes have demonstrated that our specimens from S. lateralis represent a species clearly distinct from all previously sequenced Mesocestoides and closely related to 2 forms from domestic dogs and Channel Island fox in California previously published as Mesocestoides sp. C.

Gastrointestinal helminths of raccoons (Procyon lotor) in western Poland (Lubuskie province) - with particular regard to Baylisascaris procyonis

Abstract The aim of the study was to estimate the prevalence of gastrointestinal parasites in raccoons with particular regard to zoonotic parasites. Fifty-five raccoons, hunted or found dead on roads, were examined. The small and large intestines were collected from all raccoons and, additionally, the stomach was collected from 43 animals. The samples were examined with the use of sedimentation and counting technique. The intestines and stomach were examined separately. Samples of raccoon faeces were collected from their environment localised in Słubice district, Lubuskie province (Poland). The samples were collected once a month in 2012. In total, 154 faecal samples were obtained and examined with the use of McMaster flotation technique. The following parasites were detected in the intestinal and stomach contents: tapeworms Mesocestoides sp. (67.3%), Echinostomatidae flukes (34.5%), and nematodes Capillaria sp. (25.5%). Moreover, Acanthocephala were found in the intestines of three raccoons. The highest intensity of infection were observed in case of Mesocestoides sp. Mesocestoides sp. and Echinostomatidae were found statistically more often in the intestines than in the stomach. In the case of these two parasites, there was positive correlation between the intensity of infection in the intestines and the presence of the same parasites in the stomach. Moreover, significantly higher prevalence and intensity of Mesocestoides sp. in males than in females were also observed. Faecal samples contained Baylisascaris procyonis eggs (mean 60 epg). These eggs were found in three samples collected in November and December. Furthermore, in some faecal samples eggs of flukes, Capillaria sp., Mesocystoides sp., and coccidian oocysts were found. It is one of rare reports concerning Baylisascaris procyonis in Poland confirming the presence of this dangerous parasite in Polish raccoon population.

The first report of peritoneal tetrathyridiosis in squirrel monkey (Saimiri sciureus)

This report describes a case of peritoneal larval cestodiasis caused by tetrathyridia of Mesocestoides sp. in an adult female squirrel monkey. The monkey had lived in a zoological garden in Japan and had a clinical history of wasting. At necropsy, numerous whitish oval masses were found in the liver and peritoneal cavity. These masses contained larval cestodes. Morphological observation and molecular analyses of the mitochondrial 12S ribosomal RNA gene and cytochrome c oxidase subunit 1 gene sequences allowed us to identify the larva as the tetrathyridium of Mesocestoides sp. This is the first report of Mesocestoides larvae in a squirrel monkey in Japan.

New Host Records for Mesocestoides sp. Tetrathyridia (Cestoidea: Cyclophyllidea) in Anurans (Bufonidae, Ranidae) from Arkansas, with a Summary of North American Amphibian Hosts

New Host Records for Mesocestoides sp. Tetrathyridia (Cestoidea: Cyclophyllidea) in Anurans (Bufonidae, Ranidae) from Arkansas, with a Summary of North American Amphibian Hosts

NUEVOS HALLAZGOS DE HELMINTOS PARÁSITOS EN CHRYSOCYON BRACHYURUS (CARNIVORA: CANIDAE) EN ARGENTINA

Los estudios sobre las parasitosis de cánidos y en particular de Chrysocyon brachyurus Illiger, 1815 son escasos y existen pocos registros en el norte argentino. El objetivo del presente estudio fue ampliar el conocimiento acerca de los helmintos parásitos de C. brachyurus. Se llevaron a cabo las necropsias de dos ejemplares adultos de C. brachyurus de dos áreas protegidas de Corrientes (Argentina). El material biológico consistió en vísceras de un macho proveniente del Parque Nacional Mburucuyá y de una hembra de la Reserva Provincial Rincón de Santa María. A partir de la necropsia del ejemplar proveniente del Parque Nacional Mburucuyá, se hallaron helmintos localizados en el intestino (Centrorhynchus guira Lunaschi & Drago, 2010 y Ancylostoma caninum Ercolani, 1859) y en el riñón derecho (Dioctophyme renale Goeze, 1782). El ejemplar proveniente de la Reserva Rincón de Santa María presentó únicamente Mesocestoides sp. Vaillant, 1863, localizado en el intestino delgado. Se registra por primera vez las asociaciones C. guira– C. brachyurusy Mesocestoides sp. – C. brachyurus.

A New Host Record for Mesocestoides sp. (Cestoidea: Cyclophyllidea: Mesocestoididae) from a Rough Green Snake Opheodrys aestivus (Ophidia: Colubridae) in Arkansas, U.S.A

ABSTRACT: Twenty-one adult rough green snakes Opheodrys aestivus (Ophidia: Colubridae) were collected during 2010 and 2012 from Montgomery (n = 1) and White (n = 20) counties, Arkansas, U.S.A., and examined for helminths. A single O. aestivus (5%) harbored a massive infection of Mesocestoides sp. This represents a new host record for Mesocestoides sp., and 1 of the rare instances that O. aestivus has been reported to harbor any parasite.

Cestode infection in 2 dogs: cytologic findings in liver and a mesenteric lymph node

Mesocestoides cestode infections in dogs are well known for causing severe peritonitis with larvae or larval fragments (metacestodes, tetrathyridia, or calcareous corpuscles) frequently observed cytologically in peritoneal fluid samples. This case report describes the cytologic and clinical features of 2 dogs infected with cestode larvae, with one case confirmed and the other presumed to be Mesocestoides sp. In these 2 unusual cases, cestode larvae or larval fragments were found in fine-needle aspirates of the liver and a mesenteric lymph node, but no organisms were found in peritoneal fluid samples. The data presented in this report indicate that clinical pathologists should not rule out Mesocestoides sp cestodiasis based on the absence of larvae in peritoneal fluid samples from dogs.

Mesocestoides sp. (Eucestoda, Mesocestoididae) parasitizing four species of wild felines in Southern Brazil

Leopardus colocolo, Leopardus geoffroyi, Leopardus tigrinus and Puma yagouaroundi are wild feline species endangered mainly due to habitat destruction and vehicle run overs. Seventeen felines hit on the roads were collected in Southern Brazil and examined for parasites. Cestodes were identified as Mesocestoides sp. The parasites were found in the small intestine of the hosts with a prevalence of 66.7% (L. colocolo and L. tigrinus), 60% (P. yagouaroundi) and 50% (L. geoffroyi). Rodents and lizards were found in the stomach contents and they possibly were intermediate hosts of Mesocestoides sp. This is the first report of Mesocestoides sp. in wild felines in Brazil.

Wild life surveillance on Echinococcus multilocularis in Sweden

Background Echinococcus multilocularis is a tapeworm whose adult stages parasitize the intestine of canids such as foxes and wolves. Also domestic dogs and cats can act as definitive hosts. The sylvatic life cycle includes small rodents as intermediate hosts but humans may become accidentally infected by ingestion of eggs. Sweden, Finland, UK, Ireland, and Malta are considered to be free of this parasite and therefore have maintained their national rules as regards deworming of pets at movement into the countries. According to the EC regulation, these national rules can be applied during a transitional period to 2010. In order to confirm the absence of E. multilocularis in Sweden, monitoring of foxes is being carried out continuously. These investigations are financed by the Swedish government. Materials and methods In 2007, 245 red foxes were shot and sent to SVA by local hunters in different parts of Sweden. To kill potential tapeworm eggs, the carcasses were placed in –80° C for at least one week before sampling. Faecal samples were then collected from the rectum and sent to Switzerland for testing by coproantigen ELISA (Deplazes et al., 1999). Forty-eight foxes that were positive in the ELISA and additional 28 randomly selected individuals were also examined by a sedimentation technique according to the OIE guidelines. Results and discussion Forty-eight foxes out of 245 were positive for Echinococcus sp. by the coproantigen ELISA. With the sedimentation technique however, Echinococcus sp was not detected in any of the examined animals, including those who had been positive in the ELISA. One possible explanation for obtaining false positive ELISA results was that some kind of cross-reaction had taken place. The majority of the foxes were infected with other parasites, for example Taenia sp, Mesocestoides sp, Alaria alata, Toxocara canis, Toxascaris leonina. Conclusions There is a need for good screening methods with high sensitivities and specificities. The results obtained by the sedimentation technique indicate that Sweden was still free from the fox tapeworm in 2007.

Clinical, Cytological and Molecular Evidence ofMesocestoidessp. Infection in a Dog from Italy

A 12-year-old, 13 kg, mixed-breed male dog was referred for anorexia and depression. The dog showed discomfort on abdominal palpation. Abdominal ultrasound examination revealed multiple, small, round anechoic cystic structures. Cystic fluid obtained with fine needle aspiration contained several 2-4 mm white motile flecks. Microscopic examination of the fluid revealed numerous irregularly shaped organisms measuring several hundred microns to 3 mm, the morphology of which was suggestive of intact and fragmented acephalic metacestodes of the genus Mesocestoides sp. Molecular analysis confirmed that the peritoneal infection was caused by Mesocestoides sp.

Asexually proliferous tetrathyridia of Mesocestoides sp. in the hepatic portal system of the prairie rattlesnake (Crotalus viridis viridis).

A female prairie rattlesnake (Crotalus viridis viridis) was gastric intubated with 250 tetrathyridia of Mesocestoides sp. The snake was killed 12 wk postinfection; a portion of the liver was examined histologically for evidence of tetrathyridia. Five tetrathyridia were seen in two hepatic portal triad vessels. We propose that a blood-borne metastasis of tetrathyridia in reptiles and rodents may occur.

The cestodes of foxhounds and foxes in Powys, mid-Wales.

Cestodes were found in 129 of 875 foxhounds from 20 packs in Powys, mid-Wales, examined by purging in 1983-1988, in the following order of prevalence: Taenia hydatigena in 57 hounds (6.51%), Echinococcus granulosus in 27 (3.09%), Dipylidium caninum in 25 (2.86%), T. ovis in 14 (1.60%), T. pisiformis in 10 (1.14%), T. serialis in five (0.57%) and T. multiceps in four (0.46%). Of 197 foxes examined by autopsy, 22 harboured cestodes: two (1.02%) had E. granulosus, nine (4.57%) T. polyacantha, eight (4.06%) T. pisiformis, two (1.02%) T. serialis, one (0.51%) D. caninum and one (0.51%) Mesocestoides sp. This is the first definite report of T. polyacantha in British foxes. The results are compared with those of previous surveys in Powys and in other parts of Wales, and the reasons for changes in prevalence are discussed.

A New Host and Locality Record for Mesocestoides sp. Tetrathyridia (Cestoidea: Cyclophyllidea), with a Summary of the Genus from Snakes of the World

A new host and distribution record is reported for tetrathyridia of Mesocestoides sp. One of 5 (20%) Namib tiger snakes, Telescopus beetzi, from South Africa was infected. Numerous tetrathyridia were found encapsulated in mesentery attached to the small intestine. Morphological examination of tetrathyridia revealed absence of buds, multiple scoleces, or any other evidence of asexual proliferation. A summary of the snakes of the world reported as hosts of tetrathyridia of Mesocestoides sp. is presented.

Asynchronous Capsule Formation in the Gastrointestinal Tract of the Prairie Rattlesnake (Crotalus viridis viridis) Induced by Mesocestoides sp. Tetrathyridia

The prairie rattlesnake (Crotalus viridis viridis) was experimentally infected with tetrathyridia of Mesocestoides sp. Individual snakes were killed at 4 wk increments, and sections of the stomach, small intestine, large intestine and attached mesenteries were examined for nonencapsulated and encapsulated tetrathyridia. Capsule formation was asynchronous with 9 to 80% encapsulated metacestodes. The distribution of tetrathyridia in the wall of all segments of the gastrointestinal tract is presented as evidence that this metacestode is principally a tissue dwelling parasite.