Echinococcus Granulosus Hydatid Cyst Location Is Modified by Fasciola Hepatica Infection in Cattle
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Stoore et al. Parasites & Vectors (2018) 11:542 https://doi.org/10.1186/s13071-018-3128-6 RESEARCH Open Access Echinococcus granulosus hydatid cyst location is modified by Fasciola hepatica infection in cattle Caroll Stoore1, Constanza Andrade1, Christian Hidalgo1, Felipe Corrêa1, Mauricio Jiménez1, Marcela Hernandez2,3 and Rodolfo Paredes1* Abstract Background: Natural parasite infection occurs in wild and domestics animals with more than one parasite species at the same time, generating an infection called polyparasitism. Cystic echinococcosis reports are usually based only on infection with Echinoccocus granulosus leaving aside other internal parasitoses that could modulate both the immune response and pathogenesis of the natural infection. Fasciola hepatica is another cosmopolitan parasite in ruminants with a similar distribution to E. granulosus in different parts of the world, but no information of the effect of co-infection with E. granulosus has been described. The aims of this report were to establish E. granulosus prevalence and explore the association of F. hepatica co-infection and natural E. granulosus infections in cattle. Results: From 1725 animals, the prevalence of E. granulosus and F. hepatica was 21.16 and 51.3%, respectively. Considering both infections, older cattle (> 4 years) presented higher prevalence compared to younger animals. In E. granulosus-infected cattle, 5.21% had fertile cysts, 71.78% infertile cysts, and in 23.01% cysts were smaller than 1 cm in diameter. Considering cyst location, 39.72% had lungs cysts, 24.72% had liver cysts and 36.94% had cysts in both organs. Cyst location significantly differed between age groups: 44.68% of younger animals had cysts only in the lungs, while older animals presented hydatid cyst in the lungs and liver simultaneously (44.15%). With E. granulosus infection alone, 30.26% of cysts were found in the lungs, 31.79% in the liver and 37.95% in both organs. Regarding the co-infection of E. granulosus with F. hepatica, the proportion was significantly different (P < 0.05) with most animals having cysts only in the lungs (49.41%) and a lower level of liver infection (15.88%). Analyzing organ cyst distribution and F. hepatica absence/presence ratio within each cyst type, small cysts showed the highest difference in ratio. Conclusions: To the best of our knowledge, this is the first report indicating that F. hepatica co-infection in cattle could be affecting the instate of hydatid cysts in the liver, displacing toward lung localization, suggesting an antagonistic relationship. Keywords: Echinococcus granulosus, Fasciola hepatica, Polyparasitism, Hydatid cyst localization Background the synergistic or antagonic relationship that different para- Co-infection with different parasite species in the same host site species can have within the same host remains poorly (also known as polyparasitism) is a well-documented fact in studied [2]. Helminth parasites are a very diverse group of medical, veterinary and zoological literature. Most of the animals that are classified in four taxonomic groups: nema- animals that live in the wild and humans of rural areas can todes, trematodes, cestodes and acanthocephalans [3]. In be hosts of many concurrent parasite species [1]. However, cattle, two parasites usually represent a frequent infection: the cestode E. granulosus (sensu lato) and the trematode F. hepatica. Echinococcus granulosus (s.l.)hasanindirect * Correspondence: [email protected] life-cycle, with ruminants as intermediate hosts, dogs and 1Escuela de Medicina Veterinaria, Facultad de Ciencias de la Vida, Universidad Andres Bello, Santiago, Chile other canids as definitive hosts, and humans as dead-end Full list of author information is available at the end of the article © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Stoore et al. Parasites & Vectors (2018) 11:542 Page 2 of 7 hosts [4]. The metacestode stage called hydatid cysts de- In routine slaughtering, animals were individually identi- velops in the viscera (mainly lungs and liver) of the inter- fied, age and sex was recorded, and visceral organs of each mediate hosts [5], causing a disease known as cystic animal, mainly the lungs and liver, were visually examined, echinococcosis. Fasciola hepatica also has an indirect palpated and incised along with official veterinarian in- life-cycle; however, herbivores act as the definitive hosts, spectors for the presence of hydatid cysts and F. hepatica. withtheadultwormslocatedinthebileducts[6]. Fasciola hepatica diagnosis was made either by direct Echinococcus granulosus (s.l.) is composed of E. granulo- visualization of adult parasites in bile ducts, or by F. hepatica sus (sensu stricto) (genotypes G1-3), E. equinus (genotype compatible lesions such as enlarged and thickened bile ducts, G4), E. ortleppi (genotype G5), E. canadensis (genotypes calcification of bile ducts, black parasitic material and black G6-8/G10) and E. felidis (“lion strain”), with E. granulosus lymph nodes in the liver: signs of chronic F. hepatica (s.s.) being the most commonly distributed worldwide [7]. infection. Although E. granulosus (s.l.) is able to infect a wide Suspected cystic samples were removed from the in- range of mammalian hosts, the metacestode stage has a fected organ, placed in separate polythene bags and different capacity to produce protoscoleces, the stage in- transported in an isothermal container within 3 h to fective to the definitive host. For unknown reasons, the Universidad Andres Bello Veterinary School for further parasite can, in some animals, produce protoscoleces in- examination. For hydatid cysts confirmation and fertility side the cyst, generating a fertile hydatid cyst, but other determination, cysts were microscopically examined as animals with cystic echonococcosis possess cysts without previously described [25]. Echinococcus granulosus (s.l.) protoscoleces called infertile hydatid cysts [8, 9]. Since genotyping in hydatid cyst samples was determined as cyst fertility is associated with the size of the hydatid previously reported [26]. Briefly, DNA was extracted cyst [4], there is a subset of hydatid cysts that are too from fertile, infertile and small hydatid cysts from both small to be classified as either fertile or infertile. The cel- livers and lungs. The cox1 mtDNA was amplified and se- lular and molecular mechanisms involved in the process quenced, and a 345-nucleotide consensus sequence was of cyst fertility remain unknown [10]. In cattle, hydatid used for comparison analysis. cyst fertility status ranges from 0 to 96% in different parts of the world [11–22] and infection with F. hepatica Study groups is common in many parts of the world. However, there Animals were classified according to their age, hydatid are no studies on relationships between E. granulosus cyst type and location. By age range, individuals were di- (s.l.)andF. hepatica in co-infections. vided into two groups: 4 years-old or younger (≤ 4 years) Although belonging to different higher-level flatworm and over 4 years of age (> 4 years). Cysts were classified taxa, both parasites exhibit common traits regarding their into 3 types: small cysts (< 1 cm in diameter); fertile interaction with the mammalian host; as such, serum of an- cysts (with protoscoleces); and infertile cysts (> 1 cm in imals infected experimentally with E. granulosus (s.l.) can diameter and without protoscoleces). Animals were also recognize F. gigantica antigens in immunoassays [23]but separated into 3 groups according to the location of the there are no specific data on cross-reaction with F. hepat- hydatid cysts: in lungs only; in liver only; and in both or- ica. However, both parasites have the ability to uptake host gans simultaneously. All groups were also separated by glycolipids [6] which could explain the latter. Reports of their F. hepatica co-infection status. polyparasitism involving E. granulosus (s.l.) are scarce. There is only one report that includes the interaction with Data analysis Schistosoma mansoni (a trematode), where in murine Data were recorded in an Excel 2010 datasheet and ana- models simultaneous concomitant infection lead to higher lyzed with RStudio IDE version 1.0.136 and R version IFN-γ profiles, displaying a TH1 response; however, adding 3.3.3 for statistical associations among variables using a E. granulosus infection seven weeks after S. mansoni infec- Chi-square test. Logistic regressions were performed tion led to significant lower IL-10 production, changing the using STATA v.12 software (StataCorp, College Station, immune profile to a TH2response[24]. Here, we provide TX, USA). Statistical significance was considered when the first report that in bovines infected with E. granulosus P-values were below the 0.05 threshold. (s.l.), the presence of co-infection with F. hepatica is associ- ated with changes in the hydatid cysts localization. Results Prevalence of E. granulosus and F. hepatica Methods Of the animals examined, 1217 were