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. 2024 Sep;10(5):e1554.
doi: 10.1002/vms3.1554.

Morphometrical and molecular identification of Echinococcus granulosus genotypes in wild canids in north of Iran

Moein Abolhasani Darounkola  1 , Elahe Ebrahimzadeh  1 , Hassan Borji  1 , Mohammadreza Khoshvaght  1
Affiliations

Morphometrical and molecular identification of Echinococcus granulosus genotypes in wild canids in north of Iran

Moein Abolhasani Darounkola et al. Vet Med Sci. 2024 Sep.

Abstract

Background/objective: The cestode Echinococcus granulosus causes cystic echinococcosis, a zoonotic parasitic infection that constitutes a significant public health risk. This parasite has been documented to have potential reservoirs and carriers among wild canids, namely wolves, foxes and jackals. This study aimed to determine the prevalence and molecular characteristics of E. granulosus sensu lato species/genotypes among wild canids in three northern, northeastern and north-western Iran regions.

Methods: From 2019 to 2022, 93 wild canid carcasses (69 jackals), (22 foxes) and (2 wolves) were collected that were killed in car accidents or illnesses. Analyses of morphology and morphometry were performed to verify the presence of E. granulosus. To determine E. granulosus s.l. species/genotypes, polymerase chain reaction (PCR)-RFLP (ITS1) was performed utilizing the Bsh1236I (BstUI) restriction enzyme. COX1, NADH1 and ITS1 gene sequencing were also performed to confirm the PCR-RFLP results.

Results: During this study, 93 wild canids were examined, and 3.2% (95% CI: 0%-7%) of the 93 were infected with Echinococcus. The north-western region of Iran showed two out of 30 jackals (6.6%) infected with adult Echinococcus compared to one out of 35 jackals (2.8%) in the northern region. DNA from Echinococcus was detected in these individuals by PCR. Based on PCR-RFLP analysis of the ITS1 gene and sequencing of COX1, NADH1 and ITS1 gene, E. granulosus sensu stricto genotype was confirmed in the jackals that had been infected.

Conclusion: Evidence shows that E. granulosus occurs in jackals in Iran, with the E. granulosus s.s. genotype being the most common. This parasite has been identified as a zoonotic parasite with a genotype that can be transmitted to livestock and humans. Establishing effective control measures to prevent the spread of echinococcosis and ensure public health is crucial.

Keywords: Echinococcus granulosus; Iran; genotypes; molecular identification; wild canids.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

FIGURE 1
FIGURE 1
Map of Iran, geographical locations of north‐east (Razavi Khorasan and North Khorasan Provinces), north (Mazandaran Province) and north‐west (Ardabil Province).
FIGURE 2
FIGURE 2
Morphological appearance of Echinococcus granulosus adult worm from north of Iran (a) and northwest of Iran (b) the uterus containing the eggs is indicated by the arrow: (a and b).
FIGURE 3
FIGURE 3
(a) Complete view of the morphological appearance of Echinococcus granulosus hooks and (b) large hooks (a) and small hooks (b).
FIGURE 4
FIGURE 4
(a) Agarose gel electrophoresis of ITS1–polymerase chain reaction (PCR) products. M: DNA ladder (100 bp), N: negative control, P: positive control, north of Iran (1,2,3) and northwest of Iran (4,5,6). (b) Agarose gel electrophoresis of the PCR‐ITS1 products of Echinococcus granulosus isolate digested with the restriction enzyme Bsh1236I.
FIGURE 5
FIGURE 5
Agarose gel electrophoresis of Cox1–polymerase chain reaction (PCR) products. M: DNA ladder (100 bp), N: negative control, P: positive control, north of Iran (1,2,3) and northwest of Iran (4,5,6).
FIGURE 6
FIGURE 6
Agarose gel electrophoresis of NAD1–polymerase chain reaction (PCR) products. M: DNA Ladder (A: 100 bp and B: 50 bp), N: negative control, P: positive control, north of Iran (A) and northwest of Iran (B).
FIGURE 7
FIGURE 7
The phylogenetic tree of Echinococcus granulosus COXI derived in this study (showed with ●) and known E. granulosus genotypes and other species of Echinococcus in GenBank. The evolutionary analyses were conducted in MEGA11, and using the neighbour‐Joining method. The percentages of replicate trees in which the associated taxa clustered together in the bootstrap test (500 replicates) are shown next to the branches. The evolutionary distances were computed using the maximum composite likelihood method.
FIGURE 8
FIGURE 8
The phylogenetic tree of Echinococcus granulosus NADH1 derived in this study (showed with ●) and known E. granulosus genotypes and other species of Echinococcus in GenBank. The evolutionary analyses were conducted in MEGA11 using the neighbour‐Joining method. The percentages of replicate trees in which the associated taxa clustered together in the bootstrap test (500 replicates) are shown next to the branches. The evolutionary distances were computed using the maximum composite likelihood method.
FIGURE 9
FIGURE 9
The phylogenetic network of Echinococcus granulosus CoxI and NADH1 derived in this study.
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