The use of loop-mediated isothermal amplification improves <i>Toxoplasma gondii</i> detection in wildlife

Trisciuoglio, Andrea; Zanet, Stefania; Marello, Giuseppina; Chiesa, Francesco; Nucera, Daniele; Bergallo, Massimiliano; Gennero, Maria Silvia; Ferroglio, E.

doi:10.1177/1040638715611170

Cited by 11 publications

(9 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A number of studies have reported the presence of this parasite in wild rodents and members of Carnivora, Didelphimorphia, and Xenarthra (18) (19) (20) . These findings confirm the worldwide distribution of T. gondii, and highlight the wide variety of intermediate hosts that form part of the epidemiological chain responsible for transmission of this infection and the associated disease.…”

Section: Discussionmentioning

confidence: 99%

Genotyping of Toxoplasma gondii and Sarcocystis spp. in road-killed wild mammals from the Central Western Region of the State of São Paulo, Brazil

Richini-Pereira

Marson

Silva

et al. 2016

Rev. Soc. Bras. Med. Trop.

View full text Add to dashboard Cite

Introduction: Road-killed wild animals host zoonotic pathogens such as Toxoplasma gondii, offering a new opportunity for the epidemiological study of these infectious organisms. Methods: This investigation aimed to determine the presence of T. gondii and other apicomplexan parasites in tissue samples of 64 road-killed wild animals, using polymerase chain reaction (PCR). Positive samples were then typed by PCR-restriction fragment length polymorphism (RFLP) using 7 markers: SAG1, 5′-3′SAG2, SAG3, BTUB, c29-6, PK1, and Apico. PCR-RFLP targeting 18S ribosomal RNA (rRNA) genes was also performed on all samples to detect other apicomplexan parasites. Results: T. gondii DNA was detected in 16 tissue samples from 8 individual animals, as follows: 1 Cerdocyon thous (crab-eating fox), 1 Didelphis albiventris (white-eared opossum), 1 Lutreolina crassicaudata (lutrine opossum), 2 Myrmecophaga tridactyla (giant anteater), 1 Procyon cancrivorus (crab-eating raccoon), and 2 Sphiggurus spinosus (Paraguay hairy dwarf porcupine). Seven different T. gondii genotypes were identified, 6 of which were novel. Typing by 18S rRNA verified these 16 T. gondii-infected samples, and identified 1 Sarcocystis spp.-infected animal [Dasypus novemcinctus (nine-banded armadillo)]. The amplified T. gondii (GenBank accession No. L37415.1) and Sarcocystis spp. 18S rRNA products were confirmed by sequencing. Conclusions: Our results indicate that T. gondii is commonly present in wild mammals, which act as sources of infection for humans and animals, including other wild species. The approach employed herein proved useful for detecting T. gondii and Sarcocystis spp. in the environment and identifying their natural reservoirs, contributing to our understanding of host-parasite interactions.

show abstract

Section: Discussionmentioning

confidence: 99%

Genotyping of Toxoplasma gondii and Sarcocystis spp. in road-killed wild mammals from the Central Western Region of the State of São Paulo, Brazil

Richini-Pereira

Marson

Silva

et al. 2016

Rev. Soc. Bras. Med. Trop.

View full text Add to dashboard Cite

show abstract

“…Epidemiological data for each game species included in the review are provided below. Serological and molecular studies showed a level of infection of hunted wildlife ranging from 2% to 68% [81]. Comparison of the studies should be performed carefully, considering different methodologies, lack of standardized research methods, different types, number and collection methods of tested samples and geographical and climatic differences [10].…”

Section: Epidemiology In Mammalian Game Species In the European Unionmentioning

confidence: 99%

“…Tissues samples can be examined by using immunohistochemical staining or applying molecular techniques for the detection of parasite DNA [76], by polymerase chain reaction (PCR), quantitative real-time PCR (qPCR) and loop-mediated isothermal amplification (LAMP). Quantitative real-time PCR is the most sensitive diagnostic technique; however, due to extreme simplicity and speed, LAMP provides a valuable alternative for fast and effective diagnosis in contexts where economic resources are limited and no valid alternative are available (e.g., game meat screening in rural areas) [81]. For genotyping, the PCR-RFLP method, together with the microsatellite analysis, is considered particularly useful due to its simplicity and inexpensiveness [74].…”

Section: Parasite Detection At Slaughtering and Identificationmentioning

confidence: 99%

A Review on Alaria alata, Toxoplasma gondii and Sarcocystis spp. in Mammalian Game Meat Consumed in Europe: Epidemiology, Risk Management and Future Directions

Guardone

Armani

Mancianti

et al. 2022

Animals

Self Cite

View full text Add to dashboard Cite

Game meat is increasingly appreciated and consumed in Europe, also due to the growing population of wild ungulates. In addition to interesting nutritional properties and market opportunities, game meat is characterized by some specific public health issues. This review focuses on the etiology, epidemiology, public health aspects and risk management along the supply chain, including parasite detection at slaughtering and inactivation in meat, of three selected foodborne parasitic hazards (Alaria alata, Toxoplasma gondii and Sarcocystis spp.) in the main mammalian game meat species in the EU: wild boar (Sus scrofa), red deer (Cervus elaphus), roe deer (Capreolus capreolus), fallow deer (Dama dama), Alpine chamois (Rupicapra rupicapra), moose (Alces alces), hare (Lepus europaeus) and wild rabbit (Oryctolagus cuniculus). The presented data point out the main issues, and knowledge gaps as well as the potential for improved control in order to contribute to the risk analysis process. To pursue an effective management of these parasitic zoonoses, awareness raising should involve all figures in the supply chain, including hunters, restaurateurs and consumers. Human behaviour and the lack of knowledge regarding meat borne parasitic zoonoses and the health risks they pose seem to be the most important factors responsible for human infections. However, detection methods, starting from the sampling procedure, should be further developed and standardized in order to improve the collection of accurate and up-to-date epidemiological data.

show abstract

“…Sterile water and T. gondii genomic DNA extracted from parasite cultures and identified by species-specific PCR and sequencing were used in each set of reactions as negative and positive controls, respectively. Sensitivity and specificity of the LAMP assay were assessed [21].…”

Section: Loop Mediated Isothermal Amplification (Lamp)mentioning

confidence: 99%

Absence of Viable Toxoplasma gondii in Artisanal Raw-Milk Ewe Cheese Derived from Naturally Infected Animals

et al. 2020

Self Cite

View full text Add to dashboard Cite

The presence of viable Toxoplasma gondii was investigated in artisanal cheeses made from milk of naturally infected ewes. Ewe milk was analyzed beforehand for the presence and vitality of T. gondii by loop-mediated isothermal amplification (LAMP) and reverse-transcriptase PCR (RT-PCR), respectively. Cheeses were prepared from raw milk following a traditional cheesemaking process. The cheese obtained from T. gondii-positive milk was analyzed by LAMP to detect Toxoplasma DNA-positive samples. RT-PCR was then carried out to assess the viability of the parasites in T. gondii-positive milk samples and fresh cheese, after 5 and 15 days of ripening. Physical-chemical parameters of cheeses were also investigated. All cheese samples derived from T. gondii-positive milk were positive according to LAMP, at both 5 and 15 days of ripening, while none of the samples were positive according to RT-PCR. Thus, while the presence of the parasite was demonstrated by the detection of specific DNA, the absence of detectable T. gondii RNA supports the hypothesis that changes in the chemical and physical characteristics occurring during the cheesemaking process and ripening period, could be sufficient to inactivate viable T. gondii in milk, minimizing the risk of human infection through consumption of raw sheep milk cheese.

show abstract

The use of loop-mediated isothermal amplification improves Toxoplasma gondii detection in wildlife

Cited by 11 publications

References 12 publications

Genotyping of Toxoplasma gondii and Sarcocystis spp. in road-killed wild mammals from the Central Western Region of the State of São Paulo, Brazil

Genotyping of Toxoplasma gondii and Sarcocystis spp. in road-killed wild mammals from the Central Western Region of the State of São Paulo, Brazil

A Review on Alaria alata, Toxoplasma gondii and Sarcocystis spp. in Mammalian Game Meat Consumed in Europe: Epidemiology, Risk Management and Future Directions

Absence of Viable Toxoplasma gondii in Artisanal Raw-Milk Ewe Cheese Derived from Naturally Infected Animals

Contact Info

Product

Resources

About