Echinococcus granulosus sensu stricto and E. canadensis are distributed in livestock of highly endemic area in the Peruvian highlands

doi:10.1016/j.actatropica.2021.106178

Acta Tropica

Volume 225, January 2022, 106178

https://doi.org/10.1016/j.actatropica.2021.106178 Get rights and content

Highlights

•
Echinococcus granulosus sensu lato in livestock hosts in the South highlands of Peru are restricted to E. granulosus s.s. and E. canadensis.
•
Echinococcus granulosus sensu stricto and E. canadensis are reported for this first time in llamas and alpacas.
•
Protoscolices viability of Echinococcus granulosus sensu stricto was significantly higher than E. canadensis.

Abstract

The prevalence of cystic echinococcosis is high in many livestock areas of Peru, where intermediate hosts such as sheep, cattle, and South American camelids can be infected. Several species of E. granulosus have been described in relation to its genetic diversity and distribution. The aim of this study was to determine the species of E. granulosus sensu lato (s.l.) metacestodes collected from sheep, cattle, swine and camelids at different localities in the department of Puno, in the southern highlands of Peru. One hundred and fifty-two echinococcal cysts were collected from 10 different locations. E. granulosus s.l. species were determined by amplification of the Internal transcribed spacer 1 of the ribosomal DNA using a Nested PCR-RFLP technique. The cytochrome C oxidase 1 gene (450 bp) was also amplified and sequenced in samples with different RFLP patterns. Cysts samples were collected from sheep (39.5%), cattle (32.9%), pigs (15.8%) and alpacas/llamas (11.8%). E. granulosus sensu stricto (G1 genotype) was mainly identified in all animal hosts, while, the E. canadensis (G7) was only identified in cysts from pigs and alpacas. This is the first report of E. granulosus sensu stricto and E. canadensis in llamas and alpacas, respectively. Knowledge of species and molecular epidemiology of E. granulosus s.l. in endemic areas in Peru may help to evaluate preventive programs, understand disease transmission, as well as improve vaccine and chemotherapy effectiveness.

Graphical abstract

Introduction

Cystic echinococcosis (CE) is a parasitic zoonosis produced by the larval stage of the taeniid cestode Echinococcus granulosus sensu lato (s.l.). This parasite fulfills its biological cycle among domestic dogs or some wild canids as definitive hosts and livestock animals as intermediate hosts (Bowles and McManus, 1993b; Eckert et al., 2001). Humans may also act as intermediate hosts after accidental ingestion of parasite eggs. The main factors for the transmission and persistence of the infection include the close coexistence of animals or humans with dogs, deficient hygienic-sanitary conditions, cultural customs and low socio-economic status (Apt et al., 2000).

Human CE has a worldwide distribution, occurring in many parts of South America, including Argentina, Peru, Bolivia, Uruguay, Chile and the southern part of Brazil (Cucher et al., 2016; Pavletic et al., 2017). The Andean locations of the central and southern highlands of Peru have the conditions to maintain the biological cycle of the parasite; these areas are endemic for animal (sheep and cattle) and human CE (Moro et al., 2004; Almeida et al., 2007; Gavidia et al., 2008). The highest incidences of human CE are distributed to the south and center regions of Peru, with 14 to 43 CE cases/100000 inhabitants reported in Huancavelica, Junín, Pasco and Puno (Cabrera, 2007). Similarly, 73% of CE cases among livestock have been described in the same regions. The intermediate hosts of E. granulosus are mainly sheep, cattle or pigs (Núñez et al., 2003). Puno is considered a mixed-breeding livestock area and is endemic for CE in the Andean region of Peru (Leo-Velarde and Quiroz, 2004).

The use of molecular techniques allows the discrimination of recognized species within the Echinococcus genus. E. granulosus is genetically diverse, based on the analysis of nuclear or mitochondrial DNA. This diversity is reflected in the phenotypic characteristics of the parasite in relation to its life cycle, development, control and treatment (Bowles and McManus, 1993b). Based on these analyzes, genotypes (G1-10) and species: E. granulosus sensu stricto (s.s.), E. canadensis, E. ortleppi (causative agents of human CE) and E. equinus, E. felidis have been described (McManus, 2013; Chaâbane-Banaoues et al., 2016; Cucher et al., 2016). In South America the presence of different species of E. granulosus has been described; E. granulosus s.s. (G1) in sheep, E. ortleppi (G5) in cattle, E. canadensis (G6) in goats and pigs (G7) (Bowles and McManus, 1993a; Cucher et al., 2016). In Peru, only two species have been reported in a few studies and samples; mainly E. granulosus s.s. (G1) in sheep, cattle and humans, E. canadensis (G7) in pigs and goats (G6) (Moro et al., 2009; Sánchez et al., 2010).

Currently there is a paucity of information about the distribution of circulating E. granulosus s.l. in all livestock species within CE endemic regions in Peru. The aim of this study was to determine the species of E. granulosus s.l. present in echinococcal cysts from sheep, cattle, pigs, alpacas and llamas at different locations in the department of Puno (Southern Highlands), using PCR-RFLP analysis targeting the Internal transcribed spacer (ITS) 1 segment of ribosomal (r) DNA and sequencing of the cytochrome C oxidase subunit 1 (cox1) gene.

Section snippets

Study area

Cyst samples were collected from 10 provinces in the department of Puno (Fig. 1) Puno is located in the southeast of Peru at 3800 m.a.s.l, with a density of 17.5 inhabitants/km². The livestock economic activity is based on mixed breeding of sheep, cattle, pigs, alpacas and llamas.

Sampling collection

One hundred fifty-two samples of fertile echinococcal cysts were collected from the lung and liver of sheep, cattle, pigs, alpacas and llamas. Samples were collected from animals slaughtered in non-authorized centers

Results

Cyst samples from sheep, cattle, alpacas and pigs were collected from different provinces, with the exception of the two cysts from llama that were found only in Melgar and Puno (Fig. 1). A total of 39.5% (60/152) cyst samples were obtained from sheep, 32.9% (50/152) from cattle, and the remaining 27.6% (42/152) were obtained from pigs, llamas and alpacas. The majority of the cysts were identified as E. granulosus s.s (92.1%, 140/152) and were isolated from all provinces, while only 7.9%

Discussion

In this study, the E. granulosus s.s (G1) was the most frequently identified in different livestock hosts with high viability. E. canadensis (G7) was the other species from only pigs and alpacas with less viability. To our knowledge, this is the first report of E. granulosus s.s in llamas and E. canadensis in alpacas worldwide. In Peru, both species of South American camelids number more than 4 million and are used as pack animals, for fiber trading and meat consumption. Camelid rearing is

Conclusion

E. granulosus s.l. in livestock hosts in the South highlands of Peru are restricted to E. granulosus s.s. and E. canadensis. E. granulosus s.s. and E. canadensis are reported for this first time in llamas and alpacas, respectively. Further studies are needed to evaluate the role of camelids in CE transmission and to determine the genetic variability of these genotypes in their definitive host in this endemic area.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

CRediT authorship contribution statement

Leny Sanchez: Investigation, Writing – original draft, Project administration. Holger Mayta: Investigation, Writing – original draft, Project administration. Luis M. Jara: Writing – original draft, Writing – review & editing, Formal analysis, Data curation. Manuela Verástegui: Conceptualization, Resources, Supervision. Robert H. Gilman: Conceptualization, Resources. Luis A. Gómez-Puerta: Resources, Writing – review & editing. Cesar M. Gavidia: Conceptualization, Resources, Writing – review &

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

The authors would like to thank the helpful of PhD. Louisa A. Messenger, who reviewed and enhanced the redaction of the manuscript.

References (36)

F.B. Almeida et al.
Intraspecific variation of Echinococcus granulosus in livestock from Peru
Vet. Parasitol.
(2007)
C.A. Alvarez Rojas et al.
Echinococcus granulosus sensu lato genotypes infecting humans–review of current knowledge
Int. J. Parasitol.
(2014)
J. Bowles et al.
Rapid discrimination of Echinococcus species and strains using a polymerase chain reaction-based RFLP method
Mol. Biochem. Parasitol.
(1993)
J. Bowles et al.
Molecular variation in Echinococcus
Acta Trop
(1993)
G.A. Cardona et al.
A review of the global prevalence, molecular epidemiology and economics of cystic echinococcosis in production animals
Vet. Parasitol.
(2013)
D. Carmena et al.
Canine echinococcosis: global epidemiology and genotypic diversity
Acta Trop.
(2013)
T. Laurimäe et al.
The benefits of analysing complete mitochondrial genomes: Deep insights into the phylogeny and population structure of Echinococcus granulosus sensu lato genotypes G6 and G7
Infect. Genet. Evol.
(2018)
G. Leguia
The epidemiology and economic impact of llama parasites
Parasitol. Today
(1991)
P.L. Moro et al.
Molecular identification of Echinococcus isolates from Peru
Parasitol. Int.
(2009)
W. Pan et al.
Genetic diversity and phylogenetic analysis of EG95 sequences of Echinococcus granulosus: Implications for EG95 vaccine application
Asian Pac. J. Trop. Med.
(2017)

M. Sharbatkhori et al.

Echinococcus granulosus genotypes in livestock of Iran indicating high frequency of G1 genotype in camels

Experimental Parasitol.

(2010)

M. Siles-Lucas et al.

Laboratory diagnosis of Echinococcus spp. in human patients and infected animals

Adv. Parasitol.

(2017)

R.C. Thompson

Biology and Systematics of Echinococcus

Adv. Parasitol.

(2017)

V. Ali et al.

Echinococcus granulosus sensu stricto, Echinococcus ortleppi; and E. intermedius (G7) are present in Bolivia

Parasitology

(2020)

W. Apt et al.

Equinococosis/hidatidosis en la VII Región de Chile: Diagnóstico e intervención educativa

Rev. Panam. Salud Publica

(2000)

J. Bowles et al.

Molecular genetic characterization of the cervid strain ('northern form’) of Echinococcus granulosus

Parasitology

(1994)

G.E. Bradford et al.

Evaluation of production and reproduction of sheep, goat and alpaca genotypes in the Small Ruminant Collaborative Research Support Program

J. Anim. Sci.

(1989)

R. Cabrera

Equinococosis quística en el Perú: necesidades de prevención y control de una enfermedad postergada

Bol. Epidemiol. (Lima)

(2007)

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View full text

Echinococcus granulosus sensu stricto and E. canadensis are distributed in livestock of highly endemic area in the Peruvian highlands

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Study area

Sampling collection

Results

Discussion

Conclusion

Funding

CRediT authorship contribution statement

Declaration of Competing Interest

Acknowledgments

Vet. Parasitol.

Int. J. Parasitol.

Mol. Biochem. Parasitol.

Acta Trop

Vet. Parasitol.

Acta Trop.

Infect. Genet. Evol.

Parasitol. Today

Parasitol. Int.

Asian Pac. J. Trop. Med.

Experimental Parasitol.

Adv. Parasitol.

Adv. Parasitol.

Echinococcus granulosus sensu stricto, Echinococcus ortleppi; and E. intermedius (G7) are present in Bolivia

Parasitology

Equinococosis/hidatidosis en la VII Región de Chile: Diagnóstico e intervención educativa

Rev. Panam. Salud Publica

Molecular genetic characterization of the cervid strain ('northern form’) of Echinococcus granulosus

Parasitology

Evaluation of production and reproduction of sheep, goat and alpaca genotypes in the Small Ruminant Collaborative Research Support Program

J. Anim. Sci.

Equinococosis quística en el Perú: necesidades de prevención y control de una enfermedad postergada

Bol. Epidemiol. (Lima)