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Anthelmintic resistance of intestinal nematodes to ivermectin and pyrantel

in Estonian horses

Article  in  Journal of Helminthology · July 2014

DOI: 10.1017/S0022149X14000510 · Source: PubMed

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Journal of Helminthology (2015) 89, 760–763 doi:10.1017/S0022149X14000510
q Cambridge University Press 2014

Anthelmintic resistance of intestinal

nematodes to ivermectin and pyrantel in
Estonian horses
B. Lassen* and S.-M. Peltola
Estonian University of Life Sciences, Institute of Veterinary Medicine
and Animal Sciences, Kreutzwaldi 62, 51014 Tartu, Estonia
(Received 22 January 2014; Accepted 4 June 2014; First Published Online 9 July 2014)

Abstract

There is evidence of resistance in horses to anthelmintic treatment using

ivermectin and pyrantel. However, little information is available about the
parasites, treatment practices or anthelmintic resistance in the horse population
in Estonia. In the present study, we examined 41 trotting and riding horses aged
, 3 years from four stables in Estonia. Faecal samples were collected, and horses
were selected for treatment if the nematode egg count per gram faeces exceeded
200. Horses (n ¼ 32) that shed strongyle-type eggs were treated with pyrantel,
whereas Parascaris equorum-positive animals received ivermectin. Up to 78% of
horses required anthelmintic treatment and the efficiency of the anthelmintics
was evaluated using a faecal egg count reduction test. Resistance of P. equorum
was observed in 50% of horses treated with ivermectin and of strongyles in
27% of horses treated with pyrantel. Ivermectin treatment resulted in a mean
reduction of 100% for strongyle eggs and an 89% reduction in P. equorum,
and pyrantel-treated horses exhibited an 88% reduction in strongyle eggs.
These results are considered to be the first indication of resistance to pyrantel,
but further studies of ivermectin resistance are required. According to
questionnaires completed by the owners of horses, resistance might be
explained by a lack of evidence-based strategies, a strong preference for using
ivermectin and possibly a subjective evaluation of the body weight of horses.

Introduction available from the Baltic region about anthelmintic

The increased resistance of horse nematodes to
anthelmintic drugs (Kaplan, 2002) is a worldwide
concern. The use of uncritical anthelmintic treatment
strategies rather than evidence-based treatments may
partly explain why reports of resistance appear to be
increasing (Kaplan & Nielsen, 2010). Recent studies from
Finland and Sweden indicate that horse nematodes may
possess resistance to ivermectin and pyrantel (Lindgren
et al., 2008; Näreaho et al., 2011). Estonia is estimated to
have a horse population of 10,000 individuals (Kollom,
Estonian Animal Recording Centre, pers. comm.) and
these horses travel widely, but little information is
*Fax: þ 372 7313706
E-mail: brian.lassen@gmail.com
resistance. Thus, we designed a study to investigate
the possible resistance to ivermectin and pyrantel of
Parascaris equorum and strongyles in Estonian trotting and
riding horses.
Materials and methods
Parasitological procedures
For inclusion in this study stables had to have a
minimum of five horses aged ,3 years. The subjects
included trotting horses from two stables in Tallinn county
and riding horses from two stables in Viljandi country.
In total, there were 41 horses, i.e. 23 females and 18 males,
and their owners consented to their participation in
the study. The mean age of horses was 1.9 ^ 1.0 years.
 Anthelmintic resistance of nematodes in Estonian horses
The study design was based on methods described by
Näreaho et al. (2011), except licensed anthelmintic products
available in Estonia were used. Three visits to the stables
were conducted during September and October 2012 to
collect faecal samples for quantitative flotation analysis
using the modified McMaster method (Saari & Nikander,
2009): first, to establish the presence of nematodes in
the faeces of included horses; second to treat animals with
egg counts per gram faeces (EPG) .200; and, third, to
evaluate the efficacy of the drugs using a faecal egg count
reduction test (FECRT). Anthelmintic drugs were given to
13 trotting horses and 19 riding horses. Oral administration
of 19 mg/kg pyrantel embonate (Embotape, Bimeda
Chemicals Export, Ireland) was provided if strongyle-
type eggs were detected and 200 mg/kg ivermectin
(Bimectin, Bimeda Chemicals Export) was provided if the
sample was positive for P. equorum eggs. The weight of
each horse was estimated with a girth tape (Equimax
girth tape, Virbac, Orion Pharma, Espoo, Finland) using
the formula: weight ¼ (girth (cm)2 £ length (cm))/11,877.
Resistance to ivermectin was defined as FECRT ,95%, and
resistance to pyrantel as FECRT ,90% (Kaplan & Nielsen,
2010). Larval cultures were not attempted.
Horse owners in Estonia were invited to answer an
online questionnaire about the use of anthelmintic drugs
in their horses. The questions addressed the decision-
making process, treatment frequency, drug preferences,
drug rotation, evaluation of the weight of horses,
administration practices and use of faecal egg counts.
Data analysis
OpenEpi 2.3.1 (http://www.openepi.com/) was used
to perform the statistical analysis. The mid-P exact test
method was used to estimate the proportions, and chi-
squared tests were employed to detect significant
differences between proportions and sample means.
Results and discussion
Up to 32 horses with an EPG .200 received
anthelmintic treatments and 19% were found to be
infected with P. equorum with a mean EPG of 2766 ^ 1933.
These horses were orally given ivermectin paste during
their second visit. Strongyle-type eggs were detected in
81% of horses with a median EPG of 3904 ^ 3479.
Pyrantel paste was administered to these horses per os.
Table 1. Efficacy (%), with 95% confidence intervals (CI), of anthelmintic treatments of intestinal
nematodes in 32 Estonian horses.
Parascaris/
ivermectin
% CI
Horses treated 19 8–35
Egg count reduction 89 77–100
Anthelmintic resistance 50 15–85
Trotting horses 50 15–85
Riding horses 0 0–39
a
Co-infection with Parascaris equorum.
761
All horses that shed P. equorum eggs and were treated
with ivermectin also shed strongyle-type eggs. The mean
body weight of horses treated with ivermectin was
373.5 kg ^ 121.4, and that of horses treated with pyrantel
was 375.0 kg ^ 132.9.
The results should be interpreted with caution because
a small number of horses participated in the study.
Traversa et al. (2009) established criteria for actual
resistance as the group mean FECRT . 90% and a lower
confidence limit (LCL) , 90%, with suspected resistance
as FECRT $ 90% and/or LCL ,90%. According to these
criteria and the criteria proposed by Kaplan & Nielsen
(2010), P. equorum exhibited resistance to ivermectin and
strongyles were resistant to pyrantel despite the low
sample number. The resistance to ivermectin in P. equorum
was observed in three 2-year-old trotting horses from the
same stable (table 1). The low number of horses treated
with ivermectin means that further studies are required
with more horses to confirm resistance, but our results
should be considered as an early indication of a potential
problem. These results are not significantly different
compared with observations of trotting foals in Finland
by Näreaho et al. (2011), where the low number of subjects
that satisfied the treatment protocol resulted in a high
level of uncertainty. Strongyles exhibited resistance to
pyrantel in both trotting and riding horses, whereas no
resistance to ivermectin was detected (table 1). In the
Finnish study, anthelmintic resistance occurred in 79% of
trotting foals, whereas resistance affected significantly
less (P , 0.001) of the strongyles that parasitized Estonian
horses in the present study. In contrast, other European
studies have reported that resistance to pyrantel is
uncommon in strongyles (Vyšniauskas et al., 2004;
Osterman Lind et al., 2007; Lindgren et al., 2008; Traversa
et al., 2009; Larsen et al., 2011).
We detected ivermectin-resistant parasites in trotting
horses from one stable. Pyrantel resistance was observed
mainly in trotting horses aged , 1 year (n ¼ 1), 1 year
(n ¼ 3) and 3 years (n ¼ 3) in three of the four stables
visited. The difference in resistance between trotting and
riding horses (x2 ¼ 14.7, P , 0.001) may be attributed to
the different roles of the horse as racehorse or for hobby
riding, where the treatments were administered to suit
specific purposes. The value of a fit trotting horse may
promote more intensive anthelmintic treatment practice,
thereby resulting in greater resistance if the effective-
ness is not continuously verified. Previous studies have
examined the differences in anthelmintic practices
Strongyles/ Strongyles/
ivermectina pyrantel
% CI % CI
19 8–35 81 65–92
100 100 –100 88 78–99
0 0–39 27 13–46
0 0–39 23 10–42
0 0–39 4 0–18
762 B. Lassen and S.-M. Peltola
between different types of horse farms and owners. For
example, Lloyd et al. (2000) found that private owners
were not affected by the price of anthelmintics when
considering their treatment options, whereas Fritzen et al.
(2010) reported that different treatment strategies and
other factors promoted resistance to treatments in studs,
riding farms and smallholdings.
In the present study, larval cultures were not attempted
to differentiate the parasites at the species level. At
present, pathogenic Strongylus spp. are considered to be
a minor threat to horses in Europe compared with
cyathostomins, which exhibit increasing resistance to
benzimidazoles, pyrantel/morantel and ivermectin
(Traversa et al., 2009). It is reasonable to assume that the
situation in Estonia is similar to that in other countries in
the area, such as Finland and Sweden, because
anthelmintic treatments are used widely in Estonia. In
these countries, cyathostomins are considered to be of
clinical importance, whereas Strongylus spp. are rare due
to a long tradition of periodic anthelmintic treatments
(Osterman Lind et al., 2007; Näreaho et al., 2011).
Higher EPG levels were detected in the Estonian horses
before treatment (median: 1688, maximum: 7219) compared
with those reported in the Finnish study by Näreaho et al.
(2011), who found up to 2500 EPG faeces. Two horses that
were treated with pyrantel shed higher EPG levels after
treatment compared with their levels before treatment,
which indicated that the drug had no effect on reducing
the egg-producing worm population in these horses. Both
horses had EPG .5000 during the first visit, and the
negative FERCT is likely to represent actual resistance to
pyrantel. This indicates high environmental contamination
and the need for evidence-based treatments to demonstrate
the efficacy of the treatment strategy selected.
In the present study, the weight measurements, dose
calculations and administration of anthelmintics were
performed by the same person, who was a veterinary
student. The protocol was verified by several licensed
veterinarians as correct and ethically sound. Efforts were
made to ensure that accurate evaluations of weights were
obtained in the absence of portable electronic scales,
to facilitate the provision of the correct doses specified
by the manufacturers. According to the 67 owners who
answered our questionnaire about the use of anthelmin-
tics, the treatment plan was usually designed by
themselves (40%), veterinarians (34%) or others (25%).
The owners usually administered the drugs themselves
(64%), but treatments were provided less often by the
stable owners (15%), stable workers (8%), grooms (3%),
veterinarians (3%) or other people (8%). The weights of
horses were evaluated using scales (46%), by eye (28%),
measuring with a girth tape (19%) or by asking another
person for their opinion (6%). Scales are uncommon in
Estonian stables. Thus, we consider that some, or even
the majority, of the owners may have misunderstood the
question where they interpreted it as meaning ‘scaling’
the horse without additional equipment. Given this
possible bias and other responses about the methods used
to estimate body weights, it appears that this practice may
be a cause of the incorrect dosage of anthelmintics.
Rotation between different anthelmintics was practised
throughout the year by 69% of owners, occasionally by
28% and never by 3%. The owners reported that the
anthelmintics they used included ivermectin (39%),
ivermectin and praziquantel (39%), pyrantel (31%),
moxidectin (18%) and fenbendazole (15%). The rotation
of frequently used drugs appeared to favour the selection
of ivermectin, ivermectic/praziquantel and pyrantel. The
use of anthelmintics mainly occurred twice a year (47%),
followed by three (20%), four (20%) and more than four
times a year (14%). The use of faecal egg counts for
diagnosis was known by 79%, but was only used rarely
(54%), occasionally (40%) or not at all (6%). Thus, it
appears that anthelmintic treatments are used widely
by horse owners, but evidence-based methods, such as
those based on faecal egg counts, appear to be employed
rarely in Estonia. Faecal examinations mentioned in the
questionnaires are likely to comprise simple flotation in
most cases because quantitative flotation is not used
widely in Estonia.
This study provides the first evidence of resistance to
pyrantel in Estonia. Thus, there is a need for the stronger
promotion of evidence-based treatment plans, the use of
FECRTs and further studies to investigate the helminth
species that infect horses. Future studies should include
more horses to confirm resistance to ivermectin, which
was suggested in the present study.
Acknowledgements
We thank Reet Herm, Seppo Saari, Toomas Orro, Anu
Näreaho, Pikka Jokelainen and Triin Lepik for their
assistance, as well as the horse owners for allowing their
horses to be involved in the study and for completing the
questionnaire.
Financial support
Funding was provided by Base Financing Science
Theme grant 8-2/T9001VLVL and ETF9433 from the
Estonian Science Foundation.
Conflict of interest
None.
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