V.H. Jarquín-Díaz, et al.

IJP: Parasites and Wildlife 10 (2019) 29–40

infections were caught at farms at which infections with the both

Eimeria species was found in other mice independently.

(1.00–1.36)

(1.02–1.56)

(0.99–1.45)

Levine and Ivens (1965); Ankrom et al. (1975)

(1.1–1.2)

(1.0–1.6)

(1.1–1.4)
We used the number of mice caught per farm as a proxy for popu-
lation density, assuming roughly equal trapping effort at all localities.
L/W ratio

1.17 We then question whether population density affects prevalence by

1.17
1.23
1.22
1.29
1.25
testing differences in the likelihood of a mouse individual to be infected

Eimer (1870); Haberkorn (1970)

dependent on that population density. We detect that the likelihood of
infection is significantly increased for both E. ferrisi and for E. falciformis
(logistic regression, p < 0.05; Table 3). Infection with E. falciformis got
15.57 (12.37–18.44)

more likely by 19%, infection with E. ferrisi by 14% with each mouse
15.92 (13.89–17.6)

Ernst et al. (1971)

14 (10.97–17.44)

caught at the same locality. We also included the detection of other

Eimeria species in the model for each species and did not find a sig-
18.4 (15–21)
Width (μm)

18 (13–24)

14 (11–18)

This work

This work

This work
Reference

nificant influence (p > 0.05) on likelihood of infection.

4. Discussion

In this study we identify Eimeria species in wild commensal popu-

Jejunum, ileum and cecum lations of house mice (Mus musculus). We show that detection and
Ileum, cecum and colon

identification of this group of rodent coccidia can be challenging and

18.62 (15.29–20.76)

17.32 (13.59–21.47)

20.02 (16.22–22.8)

propose to complement classical coprological assessment with mole-

Cecum and colon
Tissue localization

Cecum and ileum

cular tools: a highly sensitive detection PCR, genotyping PCRs for

23.1 (18–26)
Lenght (μm)

species identification and qPCR for localization and detection of double

21 (15–26)

17 (12–22)

infections. Based on this we identified three different Eimeria species in

Cecum

Cecum

the house mouse: E. ferrisi, E. falciformis and E. vermiformis.

Morphological characteristics and preferential occurrence were con-
gruent with the assignment of isolates to the above species. We use our
results to show a positive effect on host density on prevalence of E.
5.52 (4.16–6.93)

5.03 (2.88–7.73)

5.43 (4.19–7.00)

ferrisi and E. falciformis.

Absent/Present
Morphological and morphometrical characteristics from Eimeria wild-derived isolates and reference morphotypesa.

Few studies report prevalences of Eimeria in wild populations of Mus

Width (μm)

7.9 (6–10)
Residuum

5.5 (5–6)

musculus. Prevalences range from 3% to 40% for isolates classified ei-

7 (6–8)
Absent

Absent
Absent
Absent
Absent

ther as E. falciformis, E. ferrisi or E. vermiformis (Ball and Lewis, 1984;

Ernst et al., 1971; Golemanski, 1979; Levine and Ivens, 1965; Tattersall
et al., 1994). Other studies make no assessment at the species level
(detection as Eimeria spp) (Moro et al., 2003; Owen, 1976; Parker et al.,
8.88 (7.03–10.30)

7.75 (5.17–11.47)

8.35 (6.18–11.29)

2009; Yakimoff and Gousseff, 1938).

Polar granule

A recent study in rodents (other then Mus musculus) in central

10.5 (10–11)

12.8 (11–14)
Lenght (μm)

11 (10–12)

Europe reported an Eimeria spp. prevalence of 32.7% based on copro-

Presentb

Presentb
Present
Present

Present

Present

logical observations (Mácová et al., 2018). At 37.6% the overall pre-

valence for all Eimeria species in our study in house could be considered
high in comparison to all these studies.
While flotation is the most commonly used for detection and
Stieda body

quantification of coccidia (Hobbs et al., 1999; Hu et al., 2016; Rinaldi

Micropyle

Presentc

et al., 2011), we here used a complementary approach of flotations and

Present
Present
Present
Present
Present
Absent
Absent
Absent
Absent
Absent
Absent

diagnostic PCR. We observed relatively large discrepancies between

Measurements are means in micrometers with ranges in parenthesis.

both methods. Flotation and counting of oocysts has a relatively high

limit of detection (Ballweber et al., 2014; Webster et al., 1996), ex-
plaining negative findings in oocyst flotations positive for PCR. Nega-
Refractile body
Lightly pitted

tive PCR results for samples with visible oocyst in flotations could be a
result of a failure to break oocyst walls during DNA extractions and/or
Smooth
Smooth
Smooth
Smooth
Smooth

Present
Present
Present
Present
Present
Present

faecal PCR inhibition (Raj et al., 2013). Importantly, tested but couldn't
Wall

find any species-specific bias in both methods making e.g. relative

Observed in more than 80% of the oocysts.

species prevalences reliable.

Traditional identification of Eimeria, depends on the expertise to
Ellipsoidal/Subspherical

recognise the morphology of sporulated oocyst (Levine and Ivens,

Spherical/Ellipsoidal

Spherical/Ellipsoidal

Spherical/Ellipsoidal
Spherical/Ellipsoidal

1965). We show that interpretation of morphometrical data is complex

Ovoid/Spherical

Absent/Present

due overlap between species while measurement means agree with

literature (Table 1). Considering the challenges of identification and
Sporocysts
Residuum

characterisation of Eimeria isolates from field samples, we conclude that

Present but not evident.
Oocysts

Present
Present
Present

Present
Present
Shape

characterisation of Eimeria species requires molecular markers and

phylogenetic analysis.
Sequence identity of our isolates to reference sequences from
Eimeria species previously described in M. musculus was above 98% for
vermiformis
vermiformis

vermiformis
vermiformis
falciformis
falciformis

falciformis
falciformis

COI, which is sometimes assumed sufficient correspond differences

ferrisi
ferrisi

ferrisi
ferrisi

within species of Eimeria (Yang et al., 2015). We confirm taxonomic

Species

Species
Table 1

assignment based on highly supported maximum likelihood and Baye-

E.
E.
E.
E.
E.
E.

E.
E.
E.
E.
E.
E.

b
a

sian phylogenetic clustering of 18S and COI sequences. Moreover, the

35