Detection of Cyclospora cayetanensis, Echinococcus multilocularis, Toxocara spp. and microsporidia in fresh produce using molecular methods: – A review

Cyclospora cayetanensis is a coccidian protozoan that causes cyclosporiasis, a severe gastroenteric disease, especially for immunocompromised patients, children, and the elderly. The parasite is considered as an emerging organism and a major contributor of gastroenteritis worldwide. Although the global prevalence of cyclosporiasis morbidity and mortality has not been assessed, global concern has arisen since diarrheal illness and gastroenteritis significantly affect both developing countries and industrialized nations. In the last two decades, an increasing number of foodborne outbreaks has been associated with the consumption of fresh produce that is difficult to clean thoroughly and is consumed without processing. Investigations of these outbreaks have revealed the necessity to increase the awareness in clinicians of this infection, since this protozoan is often ignored by surveillance systems, and to establish control measures to reduce contamination of fresh produce. In this review, the major cyclosporiasis outbreaks linked to the consumption of ready to eat fresh fruits and vegetables are presented.

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Evaluation of the U.S. Food and Drug Administration validated method for detection of Cyclospora cayetanensis in high-risk fresh produce matrices and a method modification for a prepared dish

Food Microbiology ◽

10.1016/j.fm.2018.07.013 ◽

2018 ◽

Vol 76 ◽

pp. 497-503 ◽

Cited By ~ 9

Author(s):

Sonia Almeria ◽

Alexandre J. da Silva ◽

Tyann Blessington ◽

Tami Craig Cloyd ◽

Hediye Nese Cinar ◽

...

Keyword(s):

High Risk ◽

Drug Administration ◽

Food And Drug Administration ◽

Fresh Produce ◽

Cyclospora Cayetanensis ◽

The U.S

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Cyclospora cayetanensis and Cyclosporiasis: An Update

Microorganisms ◽

10.3390/microorganisms7090317 ◽

2019 ◽

Vol 7 (9) ◽

pp. 317 ◽

Cited By ~ 13

Author(s):

Sonia Almeria ◽

Hediye N. Cinar ◽

Jitender P. Dubey

Keyword(s):

Clinical Symptoms ◽

Fresh Produce ◽

Developed Countries ◽

Future Research ◽

Immunocompromised Patients ◽

Fecal Matter ◽

Cyclospora Cayetanensis ◽

Transmission Cycle ◽

Geographical Regions ◽

Susceptible Populations

Cyclospora cayetanensis is a coccidian parasite of humans, with a direct fecal–oral transmission cycle. It is globally distributed and an important cause of foodborne outbreaks of enteric disease in many developed countries, mostly associated with the consumption of contaminated fresh produce. Because oocysts are excreted unsporulated and need to sporulate in the environment, direct person-to-person transmission is unlikely. Infection by C. cayetanensis is remarkably seasonal worldwide, although it varies by geographical regions. Most susceptible populations are children, foreigners, and immunocompromised patients in endemic countries, while in industrialized countries, C. cayetanensis affects people of any age. The risk of infection in developed countries is associated with travel to endemic areas and the domestic consumption of contaminated food, mainly fresh produce imported from endemic regions. Water and soil contaminated with fecal matter may act as a vehicle of transmission for C. cayetanensis infection. The disease is self-limiting in most immunocompetent patients, but it may present as a severe, protracted or chronic diarrhea in some cases, and may colonize extra-intestinal organs in immunocompromised patients. Trimetoprim-sulfamethoxazole is the antibiotic of choice for the treatment of cyclosporiasis, but relapses may occur. Further research is needed to understand many unknown epidemiological aspects of this parasitic disease. Here, we summarize the biology, epidemiology, outbreaks, clinical symptoms, diagnosis, treatment, control and prevention of C. cayetanensis; additionally, we outline future research needs for this parasite.

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A novel multiplex real-time PCR for the detection of Echinococcus multilocularis, Toxoplasma gondii, and Cyclospora cayetanensis on berries

Food Research International ◽

10.1016/j.foodres.2019.108636 ◽

2019 ◽

Vol 125 ◽

pp. 108636 ◽

Cited By ~ 5

Author(s):

Tamirat Tefera Temesgen ◽

Lucy Jane Robertson ◽

Kristoffer Relling Tysnes

Keyword(s):

Toxoplasma Gondii ◽

Real Time ◽

Real Time Pcr ◽

Echinococcus Multilocularis ◽

Cyclospora Cayetanensis

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Efficacy of Wash Solutions in Recovering Cyclospora cayetanensis, Cryptosporidium parvum, and Toxoplasma gondii from Basil

Journal of Food Protection ◽

10.4315/0362-028x.jfp-13-381 ◽

2014 ◽

Vol 77 (8) ◽

pp. 1348-1354 ◽

Cited By ~ 19

Author(s):

VENESSA CHANDRA ◽

MARIA TORRES ◽

YNÉS R. ORTEGA

Keyword(s):

Toxoplasma Gondii ◽

Cryptosporidium Parvum ◽

Fresh Produce ◽

Pure Water ◽

Sodium Dodecyl ◽

Recovery Process ◽

Optimal Recovery ◽

Cyclospora Cayetanensis ◽

Wash Solution ◽

Phosphate Buffered Saline

Parasitic diseases can be acquired by ingestion of contaminated raw or minimally processed fresh produce (herbs and fruits). The sensitivity of methods used to detect parasites on fresh produce depends in part on the efficacy of wash solutions in removing them from suspect samples. In this study, six wash solutions (sterile E-Pure water, 3% levulinic acid–3% sodium dodecyl sulfate, 1 M glycine, 0.1 M phosphate-buffered saline, 0.1% Alconox, and 1% HCl–pepsin) were evaluated for their effectiveness in removing Cyclospora cayetanensis, Cryptosporidium parvum, and Toxoplasma gondii from basil. One hundred or 1,000 oocysts of these parasites were inoculated onto the adaxial surfaces of 25 g of basil leaves, placed in stomacher bags, and stored for 1 h at 21°C or 24 h at 4°C. Leaves were hand washed in each wash solution for 1 min. DNA was extracted from the wash solutions and amplified using PCR for the detection of all parasites. Oocysts inoculated at a concentration of 1,000 oocysts per 25 g of basil were detected in all wash solutions. At an inoculum concentration of 100 oocysts per 25 g, oocysts were detected in 18.5 to 92.6% of the wash solutions. The lowest variability in recovering oocysts from basil inoculated with 100 oocysts was observed in 1% HCl–pepsin wash solution. Oocyst recovery rates were higher at 1 h than at 24 h postinoculation. Unlike most bacteria, parasites cannot be enriched; therefore, an optimal recovery process for oocysts from suspected foods is critical. The observations in this study provide guidance concerning the selection of wash solutions giving the highest retrieval of parasite oocysts.

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Echinococcus multilocularis and Echinococcus granulosus in canines in North-Khorasan Province, northeastern Iran, identified using morphology and genetic characterization of mitochondrial DNA

Parasites & Vectors ◽

10.1186/s13071-019-3859-z ◽

2019 ◽

Vol 12 (1) ◽

Cited By ~ 2

Author(s):

Zahra Heidari ◽

Mitra Sharbatkhori ◽

Iraj Mobedi ◽

Seyed Hossein Mirhendi ◽

Bahram Nikmanesh ◽

...

Keyword(s):

Mitochondrial Dna ◽

Echinococcus Granulosus ◽

Echinococcus Multilocularis ◽

Genetic Characterization ◽

Comparative Sequence Analysis ◽

Molecular Methods ◽

Morphological Description ◽

Intraspecific Divergence ◽

Morphological Criteria

Abstract Background Canids are definitive hosts of Echinococcus multilocularis and Echinococcus granulosus. This study aimed to survey these two Echinococcus species in canids of North-Khorasan Province, northeastern Iran, using morphological criteria and genetic characterization of mitochondrial DNA. Methods The carcasses of 106 canids, namely 61 jackals (Canis aureus), 23 foxes (Vulpes vulpes), 19 dogs (Canis familiaris) and three wolves (Canis lupus) were collected from the study area in 2013–2014 and examined for Echinococcus species. Morphological features were assessed by microscopy of adult worms. For molecular characterization, DNA was extracted, mostly from the adult worms but also from eggs. DNA fragments of the cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) mitochondrial genes were amplified and sequenced. Sequences were aligned and compared with reference sequences. Intraspecific and interspecific diversity were calculated and phylogenetic analysis was performed. Results Overall, 9.4% of the canids (eight jackals and two foxes) were found infected with E. multilocularis by molecular methods, of which seven cases were also confirmed using morphological description of the adult worms. Echinococcus granulosus was found in 6.6% of the canines (four dogs, two jackals and one wolf) as determined by both molecular methods and adult cestode morphology. All E. granulosus isolates were identified as the G1 genotype. Comparative sequence analysis indicated 0–0.7% and 0% intraspecific divergence within E. granulosus isolates and 0% and 0–0.2% within E. multilocularis isolates for cox1 and nad1, respectively. Conclusions This study revealed the presence of E. multilocularis and E. granulosus in canids of North-Khorasan Province of Iran. Jackals were found infected with both E. multilocularis and E. granulosus, but infection with the former species was higher.

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Evaluation of an Improved U.S. Food and Drug Administration Method for the Detection of Cyclospora cayetanensis in Produce Using Real-Time PCR

Journal of Food Protection ◽

10.4315/0362-028x.jfp-16-492 ◽

2017 ◽

Vol 80 (7) ◽

pp. 1133-1144 ◽

Cited By ~ 18

Author(s):

Helen R. Murphy ◽

Seulgi Lee ◽

Alexandre J. da Silva

Keyword(s):

Real Time ◽

Drug Administration ◽

Real Time Pcr ◽

Nested Pcr ◽

Food And Drug Administration ◽

Fresh Produce ◽

Pcr Assay ◽

Detection Rates ◽

Cyclospora Cayetanensis ◽

The Real

ABSTRACTCyclospora cayetanensis is a protozoan parasite that causes human diarrheal disease associated with the consumption of fresh produce or water contaminated with C. cayetanensis oocysts. In the United States, foodborne outbreaks of cyclosporiasis have been linked to various types of imported fresh produce, including cilantro and raspberries. An improved method was developed for identification of C. cayetanensis in produce at the U.S. Food and Drug Administration. The method relies on a 0.1% Alconox produce wash solution for efficient recovery of oocysts, a commercial kit for DNA template preparation, and an optimized TaqMan real-time PCR assay with an internal amplification control for molecular detection of the parasite. A single laboratory validation study was performed to assess the method's performance and compare the optimized TaqMan real-time PCR assay and a reference nested PCR assay by examining 128 samples. The samples consisted of 25 g of cilantro or 50 g of raspberries seeded with 0, 5, 10, or 200 C. cayetanensis oocysts. Detection rates for cilantro seeded with 5 and 10 oocysts were 50.0 and 87.5%, respectively, with the real-time PCR assay and 43.7 and 94.8%, respectively, with the nested PCR assay. Detection rates for raspberries seeded with 5 and 10 oocysts were 25.0 and 75.0%, respectively, with the real-time PCR assay and 18.8 and 68.8%, respectively, with the nested PCR assay. All unseeded samples were negative, and all samples seeded with 200 oocysts were positive. Detection rates using the two PCR methods were statistically similar, but the real-time PCR assay is less laborious and less prone to amplicon contamination and allows monitoring of amplification and analysis of results, making it more attractive to diagnostic testing laboratories. The improved sample preparation steps and the TaqMan real-time PCR assay provide a robust, streamlined, and rapid analytical procedure for surveillance, outbreak response, and regulatory testing of foods for detection of C. cayetanensis.

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The expression of TGF-β 1, TβR and Smad2/3 in liver infected with Echinococcus multilocularis

Cell Biology International ◽

10.1042/cbi034s057c ◽

2010 ◽

Vol 34 (8) ◽

pp. S57-S57

Author(s):

Jianbing Ding ◽

Yao Li ◽

Xu Qi ◽

Xiumin Ma ◽

Renyong Lin

Keyword(s):

Echinococcus Multilocularis

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Surgical treatment of alveolar echinococcosis: a single centre experience and systematic review of the literature

Perspectives in Surgery ◽

10.33699/pis.2019.98.4.167-173 ◽

2019 ◽

Vol 98 (4) ◽

pp. 167-173

Keyword(s):

Case Report ◽

Echinococcus Multilocularis ◽

Alveolar Echinococcosis ◽

Lymphatic System ◽

Surgical Intervention ◽

Clinical Symptoms ◽

Safety Margin ◽

Surgical Removal ◽

Single Centre Experience ◽

Therapeutic Procedures

Introduction: Alveolar echinococcosis (AE) is a zoonosis caused by Echinococcus multilocularis. AE is primarily localised in the liver. Echinococcus multilocularis imitates tumour-like behaviour. It can metastasise through blood or lymphatic system to distant organs. Echinococcosis often remains asymptomatic due to its long incubation period and indistinct symptoms. Clinical symptoms are determined by the parasite’s location. Diagnosis of echinococcosis is based on medical history, clinical symptoms, laboratory tests, serology results, imaging methods and final histology findings. Surgical removal of the cyst with a safety margin, followed by chemotherapy is the therapeutic method of choice. Case report: We present a case report of alveolar echinococcosis in a thirty-year-old female patient in whom we surgically removed multiple liver foci of alveolar echinococcosis. The disease recurred after two years and required another surgical intervention. Conclusions: Alveolar echinococcosis is a disease with a high potential for a complete cure provided that it is diagnosed early and that the recommended therapeutic procedures are strictly adhered to.

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