Accepted Manuscript Title: Trichinella pseudospiralis in the Iberian peninsula Author: M.J. Zamora M. Alvarez J. Olmedo M.C. Blanco E. Pozio PII: DOI: Reference:
S0304-4017(15)00176-4 http://dx.doi.org/doi:10.1016/j.vetpar.2015.04.004 VETPAR 7596
To appear in:
Veterinary Parasitology
Received date: Revised date: Accepted date:
17-2-2015 31-3-2015 2-4-2015
Please cite this article as: Zamora, M.J., Alvarez, M., Olmedo, J., Blanco, M.C., Pozio, E.,Trichinella pseudospiralis in the Iberian peninsula, Veterinary Parasitology (2015), http://dx.doi.org/10.1016/j.vetpar.2015.04.004 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
Highlights
2
Trichinella pseudospiralis is widespread in most of European countries.
3
This is the first report of T. pseudospiralis in the Iberian peninsula.
4
Artificial digestion test allows to discover this ‘elusive’ Trichinella species.
5
T. pseudospiralis biomass in wild boar is low in Europe.
cr
6
ip t
1
Ac ce p
te
d
M
an
us
7
1
Page 1 of 17
7
SHORT COMMUNICATION
8
10
Trichinella pseudospiralis in the Iberian peninsula Zamora, M.J a, Alvarez, M.a, Olmedo, J.a, Blanco, M.C.a, Pozio, E.b
ip t
9
11 12
a
13
Española de Consumo, Seguridad Alimentaria y Nutrición. Ctra. Pozuelo a Majadahonda,
14
Km 5,100. 28220 Majadahonda. Madrid. Spain.
15
b
16
Sanità, viale Regia Elena, 299, 00161 Rome, Italy.
us
an
Departament of Infectious Parasitic and Immunomediated Diseases, Istituto Superiore di
M
17 18
Corresponding autor:
20
Edoardo Pozio
21
Tel. +390649902304
22
Fax +390649903561
23
Email: [email protected]
Ac ce p
te
d
19
24
cr
Servicio de Microbiología Alimentaria. Centro Nacional de Alimentación. Agencia
2
Page 2 of 17
Abstract
25
Nematode worms of the genus Trichinella are zoonotic parasites circulating in most
26
continents, including Europe. In Spain, Trichinella spiralis and Trichinella britovi are highly
27
prevalent in wildlife but seldom in domestic pigs. In Portugal, only T. britovi was
28
documented in wild carnivores. In the period 2006-2013 in Spain, 384 (0.0001%) pigs and
29
1,399 (0.20%) wild boars (Sus scrofa) were positive for Trichinella spp. larvae, which were
30
identified as T. spiralis or T. britovi. In 2014, Trichinella pseudospiralis larvae were isolated
31
from a wild boar hunted in the Gerona province, Cataluña region, North-East of Spain, near
32
the border to France. This is the first report of T. pseudospiralis in the Iberian peninsula,
33
which suggests a broad distribution area of this zoonotic nematode in Europe. Since larvae
34
of this Trichinella species do not encapsulate in the host muscles, they can be detected only
35
by artificial digestion of muscle samples. T. pseudospiralis is the only Trichinella species
36
infecting both mammals and birds. Birds can spread this pathogen over great distances
37
including islands triggering new foci of infections in areas previously considered at low risk
38
for these pathogen.
cr
us
an
M
d
te
Ac ce p
39
ip t
24
40
Key words: Trichinella pseudospiralis; wild boar, Sus scrofa; Spain; Iberian peninsula;
41
epidemiology
42
3
Page 3 of 17
42
1. Introduction Trichinellosis is a relevant foodborne zoonosis caused by parasite nematodes of the
44
genus Trichinella, that are present in all the continents but Antarctica (Pozio and Zarlenga,
45
2013). Humans acquire the infection by the consumption of raw or semi-raw meat and meat
46
derived products from domestic and wild swine, carnivores and horses (Gottstein et al.,
47
2009). Two clades have been identified in the genus Trichinella, the encapsulated clade
48
infecting only mammals, which includes nine taxa (Trichinella spiralis, T. nativa, T. britovi,
49
T. murrelli, T. nelsoni, T. patagoniensis, Trichinella T6, T8 and T9) and the non-
50
encapsulated clade infecting mammals and birds (T. pseudospiralis) or mammals and
51
reptiles (T. papuae and T. zimbabwensis) (Pozio and Zarlenga, 2013). In Europe, four
52
species have been documented. T. britovi is prevalent among sylvatic carnivores of most
53
European countries but can be detected occasionally in domestic and sylvatic swine (Pozio
54
et al., 2009). T. spiralis was introduced to Europe from Asia since the Middle Ages and it is
55
now scattered in 20 out of 28 EU countries (Pozio et al., 2009; Pozio and Zarlenga, 2013). It
56
is prevalent among sylvatic swine, sometimes in backyard and free-ranging pigs from
57
remote areas, and in carnivores. T. nativa is prevalent in carnivores of the arctic and
58
subarctic regions (Pozio and Zarlenga, 2013). The fourth species, T. pseudospiralis, is
59
detected in swine and carnivores of Europe although rarely (Merialdi et al., 2011; Moskwa
60
et al., 2013; Learmount et al., 2015).
cr
us
an
M
d
te
Ac ce p
61
ip t
43
In the European Union, the implementation of a new legislation on Trichinella
62
control of domestic and wild susceptible animals through detection by digestion methods
63
(European Commission, 2005), strongly increased the number of tested animals and,
64
consequently, the detection of Trichinella spp. larvae, which, in some cases, have been
65
identified as T. pseudospiralis. Due to the lack of a collagen capsule, larvae of this species
66
cannot be identified easily among the muscle fibers by trichinoscopy (Gottstein et al., 2009). 4
Page 4 of 17
The aim of the present work was to describe the first isolation and identification of T.
67 68
pseudospiralis in the Iberian peninsula, where only T. spiralis and T. britovi had been
69
documented so far.
71
2. Materials and Methods
72
2.1
Muscle sampling and testing
cr
73
ip t
70
To follow the Regulation 2075/2005 of the European Union (European Commission, 2005), all Trichinella spp. susceptible domestic and sylvatic animals intended for human
75
consumption were tested in Spain from 2006 to 2014. Five g of diaphragm were collected
76
from pigs slaughtered at the abattoirs and from pigs slaughtered at home; and 60 g of
77
diaphragm, masseter muscles, and/or tongue, were collected from hunted wild boar (Sus
78
scrofa) at game abattoirs by the veterinarian services or directly by hunters in the field.
M
an
us
74
Muscle samples (1 g for pigs, and 10 g for wild boar) were tested by the method of
80
pooled sample digestion (European Commission, 2005). The digestion fluid sediment was
81
observed by a stereo-microscope at 40 x magnification. When a pool tested positive, a
82
further 1 g for pigs, and 50 g for wild boar were tested from single animals. From 2009 to
83
2014, an average of 150g of muscle samples from each infected animal was frozen and send
84
to the National Reference Laboratory (NRL) for Trichinella, National Food Center, Madrid,
85
Spain, to confirm the positivity, to evaluate the number of larvae per g (lpg), and to identify
86
the Trichinella species. The number of lpg was counted as ≤ 12 or >12 lpg under a
87
stereomicroscope at 40 x magnification. Any Trichinella spp. larva was washed three times
88
in distilled water and stored in 96% ethanol at 4 ºC. Then, two-five single larvae from each
89
isolate were identified at the species level by multiplex PCR according to a published
90
protocol (Pozio and La Rosa, 2003). A reference larva for each of the four species
91
circulating in Europe was used as control. Not all the Trichinella spp. infected muscles were
Ac ce p
te
d
79
5
Page 5 of 17
92
forwarded from peripheral laboratories to the NRL for the positivity confirmation, larval
93
count, and larva identification at the species level.
94
96
3. Results
ip t
95
In Spain in the period 2006-2013, Trichinella spp. larvae were detected in 384
(0.0001%) and 1,399 (0.19%) domestic pigs and wild boars, respectively (Table 1). The
98
number of pigs and wild boar tested for Trichinella spp. and the number of animals, which
99
were positive in the whole Spain in 2014, are not available yet. In the period 2009-2014, 405
us
cr
97
swine (374 hunted wild boars and 31 pigs), which had been tested as Trichinella spp.
101
positive at the slaughterhouses and game abattoirs, were forwarded to the NRL, and 49 of
102
them were not confirmed as positive. An average of 5.91 lpg (range 3.77-11.07) was
103
detected in 243 wild boars, and more than 12 lpg were detected in 162 wild boars. Out of
104
356 Trichinella spp. isolates, 223 (62.6%) were identified as T. spiralis, 112 (31.4%) as T.
105
britovi, 1 (0.3%) as T. pseudospiralis, 2 (0.5%) showed a mixed T. spiralis and T. britovi
106
infection, and 18 (5.0%) did not show any amplification suggesting a DNA degradation. T.
107
pseudospiralis larvae (24.8 lpg of diaphragm muscle) were isolated from a male wild boar of
108
54 kg, hunted in Saint Hilary de Sacalm (longitude 2,5167; latitude 41,8833), Gerona
109
province, Cataluña region, North-East of Spain near the border to France, in January 2014.
M
d
te
Ac ce p
110
an
100
111
4. Discussion
112
The circulation of Trichinella spp. in the Iberian peninsula has been documented since 1881,
113
when human outbreaks occurred in Portugal and Spain (Kozar, 1970). T. spiralis and T.
114
britovi were identified in domestic pigs and wildlife of Spain from more than 20 years (La
115
Rosa et al., 1992; Pozio et al., 1992; 1996), and T. britovi in wild carnivores of Portugal
6
Page 6 of 17
116
(Pozio, 2007; Lopes et al., 2015); whereas, this is the first time that T. pseudospiralis is
117
documented in this region. T. pseudospiralis was first described in a raccoon (Procyon lotor) of the Krasnodar
119
region, European Russia (Garkavi, 1972) and, for ten years, this species was considered an
120
enigma for the lack of epidemiological information (Dick, 1983). Whereupon, this parasite
121
species was isolated from mammals and birds of Asia (Shaikenov et al., 1983), marsupials
122
and birds of Tasmania (Obendorf et al., 1990; Obendorf and Clarke, 1992), mammals of
123
North and South America (Gamble et al., 2005; Krivokapich et al., 2015; Reichard et al.,
124
2015), and from a bird of North America (Lyndsay et al., 1995).
us
cr
ip t
118
The finding of T. pseudospiralis larvae in a wild boar hunted in the Iberian peninsula
126
completes the information on the distribution on this ‘elusive’ zoonotic pathogen in Europe.
127
T. pseudospiralis has been detected from the eastern (Krasnodar region of Russia) to western
128
(United Kingdom, Spain) regions, and from northern (Finland and Sweden) to southern
129
(Bulgaria, Italy, Spain) regions of Europe (Garkavi, 1972; Pozio et al., 2004; Airas et al.,
130
2010; Merialdi et al., 2011; Moskwa et al., 2013; Conedera et al., 2014; Learmount et al.,
131
2015). In the European Union, T. pseudospiralis has been documented in animals of 18 out
132
of 28 countries (Table 2). The locality of Saint Hilary de Sacalm in Spain, where the
133
infected wild boar was hunted, is about 230 km as the crow flies from Camargue (France),
134
where a T. pseudospiralis-infected wild boar was the source of a human outbreak of
135
trichinellosis (Ranque et al., 2000).
M
d
te
Ac ce p
136
an
125
In Spain, non-encapsulated nematode larvae resembling those of the genus
137
Trichinella were detected in histological sections of muscles of a common buzzard (Buteo
138
buteo) from the zoological garden of Jerez de la Frontera (southern Spain) by Calero and co-
139
workers (1978). This finding was considered only as a suspected case since the parasite was
7
Page 7 of 17
140
not identified at the species level by a biological, biochemical or molecular method (Pozio et
141
al., 1992).
142
The T. pseudospiralis infected wild boar was detected during routine meat inspection by artificial digestion. The artificial digestion of pooled samples is the reference method
144
according to Commission Regulation (EC) No. 2075/2005 (European Commission, 2005).
145
The theoretical sensitivity of the artificial digestion method of pooled samples for pigs is of
146
1 lpg, since this worm burden is considered sufficient to cause clinical disease in humans
147
(OIE 2012). The implementation of the artificial digestion has allowed the description of the
148
distribution of T. pseudospiralis (Pozio et al., 2013). In fact, in the last 10 years, the reports
149
of this non-encapsulated species in the European Union have increased (Pozio, 2013;
150
International Trichinella Reference Center, ITCR, Rome, Italy).
M
an
us
cr
ip t
143
T. pseudospiralis is an enigmatic parasite species. It shows a cosmopolitan
152
distribution but its prevalence in both carnivores and omnivores is generally much lower
153
than that of the other Trichinella species circulating in the same regions (Pozio and
154
Zarlenga, 2013). According to the ITRC database, T. pseudospiralis accounts only for
155
0.25% (3/1,157) and 1.8% (40/2,252) of the total number of Trichinella spp. isolates from
156
domestic pigs and wild boar, respectively, identified in Europe so far (T. spiralis for 85.56%
157
and 54.44%, T. britovi for 14.17% and 43.33%, in domestic pigs and wild boar, respectively;
158
and T. nativa for 0.44% in wild boar only). If we apply the percentage of each species for
159
the Trichinella spp. isolates from the 1,399 wild boars and 384 domestic pigs detected in
160
Spain in the period 2006-2013, we can speculate that no more than 1 domestic pig and 25
161
wild boars could be infected by T. pseudospiralis in Spain.
162 163
Ac ce p
te
d
151
In Spain, Trichinella spp. infected pigs were detected only in extensive production farms (i.e., backyard and free-ranging pigs); whereas in intensive production farms,
8
Page 8 of 17
164
Trichinella spp. infected pigs were not detected since the 1960s (Ministry of Agriculture,
165
Food and Environment).
166
A different epidemiological picture has been observed in Tasmania and Florida, where a 30% and 20.4% prevalence was detected in Tasmanian devils (Sarchophilus
168
harrisii) and in Florida panthers (Puma concolor coryi), respectively (Obendorf et al., 1990;
169
Richards et al., 2015). Furthermore in Tasmania, T. pseudospiralis was detected in 3.3%
170
(3/91) of raptorial birds (Obendorf and Clarke, 1992). This is still the highest prevalence
171
never detected in birds up to now. At present, we do not have information on the role of the
172
main reservoir hosts of the encapsulated Trichinella species circulating in Europe, as
173
reservoir of T. pseudospiralis. We can suspect that other animal species such as birds and
174
rodents could be reservoir of this non-encapsulated species; alternately, we can assume that
175
the biomass of this species is much lower than that of the other Trichinella species in nature.
176
However, we have to keep in mind that wild animal movements have a relevant role in the
177
spreading of zoonotic pathogens (Gortázar, et al., 2007).
cr
us
an
M
d
te
178
ip t
167
In the last decades, the European wild boar populations have increased possibly due to changes in hunting practices, the expansion of single-crop farming, and climate change
180
(Bieber and Ruf, 2005, Acevedo et al., 2006). The exponential increase of the number of
181
wild boar heads has also increased the biomass of Trichinella spp. (Kirjusina et al., 2015).
182
Ac ce p
179
Meat and meat derived products from wild boar are the main source of infection for
183
humans in Spain (Murrell and Pozio, 2011). Recently, meat of wild boar preserved under
184
vacuum from a supplier of Girona (Cataluña, Spain), was imported to Belgium and
185
consumed in three restaurants by 16 persons who got ill at the beginning of November 2014
186
(Pozio, 2015).
187 188
No Trichinella spp. infection was documented in domestic pigs and wild boar of Portugal since 1966, when the last human case was reported due to the consumption of 9
Page 9 of 17
189
sausages from a domestic pig (Ferreira et al., 2014). In Portugal, only T. britovi was
190
documented in wild carnivores (foxes, wolves and a river otter) (ITRC,
191
www.iss.it/site/trichinella/index.asp). There is the need to educate hunters on the importance of the systematic examination
ip t
192
for Trichinella spp. larvae of game intended for human consumption to prevent human
194
infection. Furthermore, veterinary services should educate hunters not to spread game
195
carcasses or their scraps and offal in the environment, and should organize a system for a
196
proper collection and disposal of these biological samples. The increase of the wild boar
197
population size may have enhanced the common habit of hunters to leave animal carcasses
198
in the field after skinning, or removing and discarding the entrails, which has been
199
demonstrated to strongly increase the probability of Trichinella spp. transmission among
200
wildlife, and to free-ranging and backyard pigs, particularly if the pig owner is a hunter
201
(Pozio, 2014).
202
te
d
M
an
us
cr
193
Acknowledgements
204
We thank the Public Health Agency of Catalonia, which sent us the muscle samples for the
205
diagnosis. This work was supported by the Spanish Agency for Consumer Affairs, Food
206
Safety and Nutrition.
207
Ac ce p
203
10
Page 10 of 17
207
References
208
Airas, N., Saari, S., Mikkonen, T., Virtala, A.M., Pellikka, J., Oksanen, A., Isomursu, M., Kilpelä, S.S., Lim, C.W., Sukura, A., 2010. Sylvatic Trichinella spp. infection in
210
Finland. J. Parasitol. 96, 67-76.
212
Acevedo, P., Escudero, M. A., Munoz, R., Gortázar C., 2006. Factors affecting wild boar
abundance across an environmental gradient in Spain. Acta Theriologica 51, 327-336.
cr
211
ip t
209
Bieber, C., Ruf, T., 2005. Population dynamics in wild boar Sus scrofa: ecology, elasticity
214
of growth rate and implications for the management of pulsed resource consumers. J.
215
Appl. Ecol. 42, 1203–1213.
us
213
Calero, R., Martinez, F., Hernandez, S. and Acosta, I., 1978. Parasitacion de Buteo buteo
217
(Aves: Accipitridae) por Trichinella sp. en el parque zoologico de Jerez de la
218
Frontiera. Revista Iberica Parasitolgia 38, 135-138.
M
Conedera, G., Vio, D., Ustulin, M., Londero, M., Bregoli, M., Perosa, G., Rigo, S.,
d
219
an
216
Simonato, G., Marangon, S., Interisano, M., Gomez Morales, M.A., Pozio, E., Capelli,
221
G., 2014. Trichinella pseudospiralis in wildboars (Sus scrofa) of the Friuli Venezia
222
Giulia region, Italy. Wiener Klinische Wochenschrift 126, S70-S71.
224 225 226 227 228
Ac ce p
223
te
220
Dick, T.A., 1983. Species, and infraspecific variation. In: Campbell, W.C. (Ed.), Trichinella and Trichinosis. Plenum Press, New York and London, pp. 31-73.
European Commission, 2005. Regulation (EC) no. 2075/2005 of the European Parliament and of the Council of 5 December 2005 laying down specific rules on official controls for Trichinella in meat. Off. J. Eur. Comm. 338, 60–82.
Ferreira, I., Martins, S., Reis, T., Vilares, A., Mendes, A., Cardoso, S., do Céu Costa, M.,
229
João Gargaté, M., 2014. Triquinelose humana: estudo observacional em dois grupos
230
populacionais expostos à infeção por Trichinella sp. Observações Boletim
231
Epidemiológico 3, 20-22. 11
Page 11 of 17
234 235 236 237 238
pseudospiralis from a wild pig in Texas. Vet. Parasitol. 132, 147-150. Garkavi, B.L., 1972. Species of Trichinella isolated from wild animals. Veterinariya 10, 9091.
ip t
233
Gamble, H.R., Pozio, E., Lichtenfels, J.R. and Zarlenga, D.S., 2005. Trichinella
Gottstein, B., Pozio, E., Nöckler, K., 2009. Epidemiology, diagnosis, treatment, and control of trichinellosis. Clin. Microbiol. Rev. 22, 127-145.
cr
232
Gortázar, C., Ferroglio, E., Höfle, U., Frölich, K., Vicente, J., 2007. Diseases shared between wildlife and livestock a European perspective. Eur. J. Wildl. Res. 53, 241–
240
256.
243
an
242
International Trichinella Reference Center, www.iss.it/site/trichinella/index/asp, dowloaded on February 9, 2015.
M
241
us
239
Kirjušina, M., Deksne, G., Marucci, G., Bakasejevs, E., Jahundoviča, I., Daukšte, A., Zdankovska, A., Bērziņa, Z., Esīte, Z., Bella, A., Galati, F., Krūmiņa, A., Pozio, E.,
245
2015. A 38-year study on Trichinella spp. in wild boar (Sus scrofa) of Latvia shows a
246
stable incidence with an increased parasite biomass in the last decade. Parasit. Vectors
247
8, 753.
249 250 251 252 253
te
Ac ce p
248
d
244
Krivokapich, S.J., Gonzalez Prous, C.L., Gatti, G.M., Saldía, L., 2015. First finding of Trichinella pseudospiralis in the Neotropical region. Vet. Parasitol. 208, 268-271.
Kozar, Z., 1970. Trichinosis in Europe. In: Gould S.E. (Ed.), Trichinosis in man and animals. Charles Thomas Publisher, Spingfield, pp. 423-436.
La Rosa, G., Pozio, E., Rossi, P., Murrell, K.D., 1992. Allozyme analysis of Trichinella isolates from various host species and geographical regions. J. Parasitol. 78, 641-646.
254
Learmount, J., Boughtflower, V., Allanson, P.C., Hartley, K.M., Barrecheguren Gutierrez,
255
A., Stephens, N.A., Marucci, G., Smith, G.C., 2015. First report of Trichinella
256
pseudospiralis in a red fox in mainland Britain. Vet. Parasitol. 208, 259-262. 12
Page 12 of 17
257
Lindsay, D.S., Zarlenga, D.S., Gamble, H.R., Al-Yaman, F., Smith, P.C., Blagburn, B.L.,
258
1995. Isolation and characterization of Trichinella pseudospiralis Garkavi, 1972 from
259
a black vulture (Coragyps atratus). J. Parasitol. 81, 920-923. Lopes, A.P., Vila-Viçosa, M.J., Cardoso, L., Gottstein, B., Müller, N., Cortes, H.C.E., 2015.
261
Trichinella britovi in a red fox (Vulpes vulpes) from Portugal. Vet. Parasitol. (in
262
press).
cr
263
ip t
260
Merialdi, G., Bardasi, L., Fontana,M.C., Spaggiari,B., Maioli, G., Conedera, G., Vio, D.,Londero, M., Marucci, G., Ludovisie, A., Pozio, E., Capelli, G., 2011. First reports
265
of Trichinella pseudospiralis in wild boars (Sus scrofa) of Italy. Vet. Parasitol. 178,
266
370–373.
an
Ministry of Agriculture, Food and Environment. Report on results of the national program of
M
267
us
264
surveillance in wildlife 2012.
269
http://rasve.magrama.es/Recursos/Ficheros/Historico/83_Resultados%20del%20Progr
270
ama%20Nacional%20de%20Vigilancia%20en%20fauna%20silvestre%202012..pdf,
271
downloaded on March 31, 2015.
te
d
268
Moskwa, B., Goździk, K., Bień, J., Borecka, A., Gawor, J., Cabaj, W., 2013. First report of
273
Trichinella pseudospiralis in Poland, in red foxes (Vulpes vulpes). Acta Parasitol. 58,
274 275 276 277
Ac ce p
272
149-154.
Murrell, K.D., Pozio, E., 2011. Worldwide occurrence and impact of human trichinellosis, 1986-2009. Emerg. Infect. Dis. 17, 2194-2202.
Nöckler, K., Reckinger, S., Pozio, E., 2006. Trichinella spiralis and Trichinella
278
pseudospiralis mixed infection in a wild boar (Sus scrofa) of Germany. Vet. Parasitol.
279
137, 364–368.
280 281
Office International of Epizooties, 2013. Chapter 8.14, Infection with Trichinella spp. Terrestrial Animal Health Code, pp. 1–4. 13
Page 13 of 17
282 283 284
Obendorf, D.L., Clarke, K.P., 1992. Trichinella pseudospiralis infections in free-living Tasmanian birds. J. Helminthol. Soc. Washington 59, 144-147. Obendorf, D.L., Handlinger, J.H., Mason, R.W., Clarke, K.P., Forman, A.J., Hooper, P.T., Smith, S.J., Holdsworth, M., 1990. Trichinella pseudospiralis infection in Tasmanian
286
wildlife. Aust. Vet. J. 67, 108–110.
292 293 294 295
cr
us
291
and habitats. Vet. Parasitol. 194, 128-132.
Pozio, E., 2014. Searching for Trichinella: not all pigs are created equal. Trends Parasitol.
an
290
Pozio, E., 2013. The opportunistic nature of Trichinella-Exploitation of new geographies
30, 4-11.
M
289
Vet. Parasitol. 149, 3-21.
Pozio, E., 2015. Trichinella spp. imported with live animals and meat. Vet. Parasitol. (in press)
d
288
Pozio, E., 2007. World distribution of Trichinella spp. infections in animals and humans.
Pozio, E., La Rosa, G., 2003. PCR-derived methods for the identification of Trichinella
te
287
ip t
285
parasites from animal and human samples. Methods Mol. Biol. 216, 299–309.
297
Pozio, E., Zarlenga,D.S., 2013. New pieces of the Trichinella puzzle. Int. J. Parasitol. 43,
298 299 300 301 302 303 304
Ac ce p
296
983-997.
Pozio, E., Nöckler, K., Reckinger, S., 2006. Trichinella spiralis and Trichinella pseudospiralis mixed infection in a wild boar (Sus scrofa) of Germany. Vet. Parasitol. 137, 364–368.
Pozio, E., La Rosa, G., Murrell, K.D., Lichtenfels, J.R., 1992. Taxonomic revision of the genus Trichinella. J. Parasitol. 78, 654-659. Pozio, E., Shaikenov, B., La Rosa, G., Obendorf, D.L., 1992. Allozymic and biological
305
characters of Trichinella pseudospiralis isolates from free-ranging animals. J.
306
Parasitol. 78, 1087-1090. 14
Page 14 of 17
307
Pozio, E., La Rosa, G., Serrano, F.J., Barrat, J., Rossi, L., 1996. Environmental and human
308
influence on the ecology of Trichinella spiralis and Trichinella britovi in Western
309
Europe. Parasitol. 113, 527-533. Pozio, E., Christensson, D., Stéen, M., Marucci, G., La Rosa, G., Bröjer, C., Mörner, T.,
ip t
310
Uhlhorn, H., Agren, E., Hall, M., 2004. Trichinella pseudospiralis foci in Sweden. Vet
312
Parasitol. 125, 335-342.
313
cr
311
Pozio, E., Rinaldi, L., Marucci, G., Musella, V., Galati, F., Cringoli, G., Boireau, P., La Rosa, G., 2009. Hosts and habitats of Trichinella spiralis and Trichinella britovi in
315
Europe. Int. J. Parasitol. 39, 71–79.
318
an
317
Ranque, S., Faugère, B., Pozio, E., La Rosa, G., Tamburrini, A., Pellissier, J.F., Brouqui, P., 2000. Trichinella pseudospiralis outbreak in France. Emerg. Infect. Dis. 6, 543-547.
M
316
us
314
Reichard, M.V., Criffield, M., Thomas, J.E., Paritte, J.M., Cunningham, M., Onorato, D., Logan, K., Interisano, M., Marucci, G., Pozio, E., 2015. High prevalence of
320
Trichinella pseudospiralis in Florida panthers (Puma concolor coryi). Parasit. Vectors.
321
8, 67.
323 324
te
Shaikenov, B.S., Boev, S.N., 1983. Distribution of Trichinella species in the old world.
Ac ce p
322
d
319
Wiad. Parazytol. 29, 595-608.
15
Page 15 of 17
324
Table 1
325
Pigs and wild boar (Sus scrofa) tested for Trichinella spp. in Spain from 2006 to 2013. Trichinella positive/tested (%) Domestic pigs slaughtered
Hunted
ip t
Year
at homeb
wild boar
2006
3/37,601,623 (8x10-6)
6/132,760 (0.004)
172/70,566 (0.24)
2007
48/41,198,179 (10-4)
4/75,514 (0.005)
2008
75/38,806,715 (10-4)
2/90,889 (0.002)
2009
59/39,929,716 (10-4)
5/60,295 (0.008)
2010
2/41,224,861 (5x10-6)
0/53,485
160/78,571 (0.2)
2011
8/41,597,557 (2x10-5)
1/32,138 (0.003)
176/104,869 (0.16)
2012
115c/40,783,578 (3x10-4)
2013
45/39,190,053 (10-4)
103/51,718 (0.19)
M
an
us
182/81,248 (0.24)
1/32,987 (0.003)
245/123,597 (0.19)
10/44,438 (0.022)
257/117,552 (0.21)
d
355/314,332,282 (10-4)
104/64,557 (0.16)
29/521,667 (0.005)
1,399/692,678 (0.20)
te
Total
cr
at abattoirsa
326
a
327
pigs) have been slaughtered in official abattoirs; b backyard and free-ranging pigs only; c of
328
them 114 originated from the same farm of Salamanca, Castile and León, Spain.
Ac ce p
329
Pigs reared in different containment systems (including some backyard and free-ranging
16
Page 16 of 17
329
Table 2
330
Trichinella pseudospiralis in animals of the European Union countries. Number of animals
331
are reported between brackets. Domestic
Carnivores and other
wild boar
pigs (3)
animals (24)
Italy (4)
Croatia (1),
Bulgaria (2), Denmark (1),
Slovakia (2)
Finland (14), Italy (1),
Czech Rep. (2), Estonia (2),
Italy (2)
Lithuania (1), Poland (1),
us
Finland (1), France (1),
Birds
ip t
Austria (1), Bulgaria (1),
Farmed
cr
Hunted wild boar (38)
Slovakia (2), Sweden (1),
an
Germany (14), Hungary (1), Italy (2), Netherlands (1),
United Kingdom (1)
M
Spain (1), Sweden (11) 332
Ac ce p
te
d
333
17
Page 17 of 17
S0304-4017(15)00176-4 http://dx.doi.org/doi:10.1016/j.vetpar.2015.04.004 VETPAR 7596
To appear in:
Veterinary Parasitology
Received date: Revised date: Accepted date:
17-2-2015 31-3-2015 2-4-2015
Please cite this article as: Zamora, M.J., Alvarez, M., Olmedo, J., Blanco, M.C., Pozio, E.,Trichinella pseudospiralis in the Iberian peninsula, Veterinary Parasitology (2015), http://dx.doi.org/10.1016/j.vetpar.2015.04.004 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
Highlights
2
Trichinella pseudospiralis is widespread in most of European countries.
3
This is the first report of T. pseudospiralis in the Iberian peninsula.
4
Artificial digestion test allows to discover this ‘elusive’ Trichinella species.
5
T. pseudospiralis biomass in wild boar is low in Europe.
cr
6
ip t
1
Ac ce p
te
d
M
an
us
7
1
Page 1 of 17
7
SHORT COMMUNICATION
8
10
Trichinella pseudospiralis in the Iberian peninsula Zamora, M.J a, Alvarez, M.a, Olmedo, J.a, Blanco, M.C.a, Pozio, E.b
ip t
9
11 12
a
13
Española de Consumo, Seguridad Alimentaria y Nutrición. Ctra. Pozuelo a Majadahonda,
14
Km 5,100. 28220 Majadahonda. Madrid. Spain.
15
b
16
Sanità, viale Regia Elena, 299, 00161 Rome, Italy.
us
an
Departament of Infectious Parasitic and Immunomediated Diseases, Istituto Superiore di
M
17 18
Corresponding autor:
20
Edoardo Pozio
21
Tel. +390649902304
22
Fax +390649903561
23
Email: [email protected]
Ac ce p
te
d
19
24
cr
Servicio de Microbiología Alimentaria. Centro Nacional de Alimentación. Agencia
2
Page 2 of 17
Abstract
25
Nematode worms of the genus Trichinella are zoonotic parasites circulating in most
26
continents, including Europe. In Spain, Trichinella spiralis and Trichinella britovi are highly
27
prevalent in wildlife but seldom in domestic pigs. In Portugal, only T. britovi was
28
documented in wild carnivores. In the period 2006-2013 in Spain, 384 (0.0001%) pigs and
29
1,399 (0.20%) wild boars (Sus scrofa) were positive for Trichinella spp. larvae, which were
30
identified as T. spiralis or T. britovi. In 2014, Trichinella pseudospiralis larvae were isolated
31
from a wild boar hunted in the Gerona province, Cataluña region, North-East of Spain, near
32
the border to France. This is the first report of T. pseudospiralis in the Iberian peninsula,
33
which suggests a broad distribution area of this zoonotic nematode in Europe. Since larvae
34
of this Trichinella species do not encapsulate in the host muscles, they can be detected only
35
by artificial digestion of muscle samples. T. pseudospiralis is the only Trichinella species
36
infecting both mammals and birds. Birds can spread this pathogen over great distances
37
including islands triggering new foci of infections in areas previously considered at low risk
38
for these pathogen.
cr
us
an
M
d
te
Ac ce p
39
ip t
24
40
Key words: Trichinella pseudospiralis; wild boar, Sus scrofa; Spain; Iberian peninsula;
41
epidemiology
42
3
Page 3 of 17
42
1. Introduction Trichinellosis is a relevant foodborne zoonosis caused by parasite nematodes of the
44
genus Trichinella, that are present in all the continents but Antarctica (Pozio and Zarlenga,
45
2013). Humans acquire the infection by the consumption of raw or semi-raw meat and meat
46
derived products from domestic and wild swine, carnivores and horses (Gottstein et al.,
47
2009). Two clades have been identified in the genus Trichinella, the encapsulated clade
48
infecting only mammals, which includes nine taxa (Trichinella spiralis, T. nativa, T. britovi,
49
T. murrelli, T. nelsoni, T. patagoniensis, Trichinella T6, T8 and T9) and the non-
50
encapsulated clade infecting mammals and birds (T. pseudospiralis) or mammals and
51
reptiles (T. papuae and T. zimbabwensis) (Pozio and Zarlenga, 2013). In Europe, four
52
species have been documented. T. britovi is prevalent among sylvatic carnivores of most
53
European countries but can be detected occasionally in domestic and sylvatic swine (Pozio
54
et al., 2009). T. spiralis was introduced to Europe from Asia since the Middle Ages and it is
55
now scattered in 20 out of 28 EU countries (Pozio et al., 2009; Pozio and Zarlenga, 2013). It
56
is prevalent among sylvatic swine, sometimes in backyard and free-ranging pigs from
57
remote areas, and in carnivores. T. nativa is prevalent in carnivores of the arctic and
58
subarctic regions (Pozio and Zarlenga, 2013). The fourth species, T. pseudospiralis, is
59
detected in swine and carnivores of Europe although rarely (Merialdi et al., 2011; Moskwa
60
et al., 2013; Learmount et al., 2015).
cr
us
an
M
d
te
Ac ce p
61
ip t
43
In the European Union, the implementation of a new legislation on Trichinella
62
control of domestic and wild susceptible animals through detection by digestion methods
63
(European Commission, 2005), strongly increased the number of tested animals and,
64
consequently, the detection of Trichinella spp. larvae, which, in some cases, have been
65
identified as T. pseudospiralis. Due to the lack of a collagen capsule, larvae of this species
66
cannot be identified easily among the muscle fibers by trichinoscopy (Gottstein et al., 2009). 4
Page 4 of 17
The aim of the present work was to describe the first isolation and identification of T.
67 68
pseudospiralis in the Iberian peninsula, where only T. spiralis and T. britovi had been
69
documented so far.
71
2. Materials and Methods
72
2.1
Muscle sampling and testing
cr
73
ip t
70
To follow the Regulation 2075/2005 of the European Union (European Commission, 2005), all Trichinella spp. susceptible domestic and sylvatic animals intended for human
75
consumption were tested in Spain from 2006 to 2014. Five g of diaphragm were collected
76
from pigs slaughtered at the abattoirs and from pigs slaughtered at home; and 60 g of
77
diaphragm, masseter muscles, and/or tongue, were collected from hunted wild boar (Sus
78
scrofa) at game abattoirs by the veterinarian services or directly by hunters in the field.
M
an
us
74
Muscle samples (1 g for pigs, and 10 g for wild boar) were tested by the method of
80
pooled sample digestion (European Commission, 2005). The digestion fluid sediment was
81
observed by a stereo-microscope at 40 x magnification. When a pool tested positive, a
82
further 1 g for pigs, and 50 g for wild boar were tested from single animals. From 2009 to
83
2014, an average of 150g of muscle samples from each infected animal was frozen and send
84
to the National Reference Laboratory (NRL) for Trichinella, National Food Center, Madrid,
85
Spain, to confirm the positivity, to evaluate the number of larvae per g (lpg), and to identify
86
the Trichinella species. The number of lpg was counted as ≤ 12 or >12 lpg under a
87
stereomicroscope at 40 x magnification. Any Trichinella spp. larva was washed three times
88
in distilled water and stored in 96% ethanol at 4 ºC. Then, two-five single larvae from each
89
isolate were identified at the species level by multiplex PCR according to a published
90
protocol (Pozio and La Rosa, 2003). A reference larva for each of the four species
91
circulating in Europe was used as control. Not all the Trichinella spp. infected muscles were
Ac ce p
te
d
79
5
Page 5 of 17
92
forwarded from peripheral laboratories to the NRL for the positivity confirmation, larval
93
count, and larva identification at the species level.
94
96
3. Results
ip t
95
In Spain in the period 2006-2013, Trichinella spp. larvae were detected in 384
(0.0001%) and 1,399 (0.19%) domestic pigs and wild boars, respectively (Table 1). The
98
number of pigs and wild boar tested for Trichinella spp. and the number of animals, which
99
were positive in the whole Spain in 2014, are not available yet. In the period 2009-2014, 405
us
cr
97
swine (374 hunted wild boars and 31 pigs), which had been tested as Trichinella spp.
101
positive at the slaughterhouses and game abattoirs, were forwarded to the NRL, and 49 of
102
them were not confirmed as positive. An average of 5.91 lpg (range 3.77-11.07) was
103
detected in 243 wild boars, and more than 12 lpg were detected in 162 wild boars. Out of
104
356 Trichinella spp. isolates, 223 (62.6%) were identified as T. spiralis, 112 (31.4%) as T.
105
britovi, 1 (0.3%) as T. pseudospiralis, 2 (0.5%) showed a mixed T. spiralis and T. britovi
106
infection, and 18 (5.0%) did not show any amplification suggesting a DNA degradation. T.
107
pseudospiralis larvae (24.8 lpg of diaphragm muscle) were isolated from a male wild boar of
108
54 kg, hunted in Saint Hilary de Sacalm (longitude 2,5167; latitude 41,8833), Gerona
109
province, Cataluña region, North-East of Spain near the border to France, in January 2014.
M
d
te
Ac ce p
110
an
100
111
4. Discussion
112
The circulation of Trichinella spp. in the Iberian peninsula has been documented since 1881,
113
when human outbreaks occurred in Portugal and Spain (Kozar, 1970). T. spiralis and T.
114
britovi were identified in domestic pigs and wildlife of Spain from more than 20 years (La
115
Rosa et al., 1992; Pozio et al., 1992; 1996), and T. britovi in wild carnivores of Portugal
6
Page 6 of 17
116
(Pozio, 2007; Lopes et al., 2015); whereas, this is the first time that T. pseudospiralis is
117
documented in this region. T. pseudospiralis was first described in a raccoon (Procyon lotor) of the Krasnodar
119
region, European Russia (Garkavi, 1972) and, for ten years, this species was considered an
120
enigma for the lack of epidemiological information (Dick, 1983). Whereupon, this parasite
121
species was isolated from mammals and birds of Asia (Shaikenov et al., 1983), marsupials
122
and birds of Tasmania (Obendorf et al., 1990; Obendorf and Clarke, 1992), mammals of
123
North and South America (Gamble et al., 2005; Krivokapich et al., 2015; Reichard et al.,
124
2015), and from a bird of North America (Lyndsay et al., 1995).
us
cr
ip t
118
The finding of T. pseudospiralis larvae in a wild boar hunted in the Iberian peninsula
126
completes the information on the distribution on this ‘elusive’ zoonotic pathogen in Europe.
127
T. pseudospiralis has been detected from the eastern (Krasnodar region of Russia) to western
128
(United Kingdom, Spain) regions, and from northern (Finland and Sweden) to southern
129
(Bulgaria, Italy, Spain) regions of Europe (Garkavi, 1972; Pozio et al., 2004; Airas et al.,
130
2010; Merialdi et al., 2011; Moskwa et al., 2013; Conedera et al., 2014; Learmount et al.,
131
2015). In the European Union, T. pseudospiralis has been documented in animals of 18 out
132
of 28 countries (Table 2). The locality of Saint Hilary de Sacalm in Spain, where the
133
infected wild boar was hunted, is about 230 km as the crow flies from Camargue (France),
134
where a T. pseudospiralis-infected wild boar was the source of a human outbreak of
135
trichinellosis (Ranque et al., 2000).
M
d
te
Ac ce p
136
an
125
In Spain, non-encapsulated nematode larvae resembling those of the genus
137
Trichinella were detected in histological sections of muscles of a common buzzard (Buteo
138
buteo) from the zoological garden of Jerez de la Frontera (southern Spain) by Calero and co-
139
workers (1978). This finding was considered only as a suspected case since the parasite was
7
Page 7 of 17
140
not identified at the species level by a biological, biochemical or molecular method (Pozio et
141
al., 1992).
142
The T. pseudospiralis infected wild boar was detected during routine meat inspection by artificial digestion. The artificial digestion of pooled samples is the reference method
144
according to Commission Regulation (EC) No. 2075/2005 (European Commission, 2005).
145
The theoretical sensitivity of the artificial digestion method of pooled samples for pigs is of
146
1 lpg, since this worm burden is considered sufficient to cause clinical disease in humans
147
(OIE 2012). The implementation of the artificial digestion has allowed the description of the
148
distribution of T. pseudospiralis (Pozio et al., 2013). In fact, in the last 10 years, the reports
149
of this non-encapsulated species in the European Union have increased (Pozio, 2013;
150
International Trichinella Reference Center, ITCR, Rome, Italy).
M
an
us
cr
ip t
143
T. pseudospiralis is an enigmatic parasite species. It shows a cosmopolitan
152
distribution but its prevalence in both carnivores and omnivores is generally much lower
153
than that of the other Trichinella species circulating in the same regions (Pozio and
154
Zarlenga, 2013). According to the ITRC database, T. pseudospiralis accounts only for
155
0.25% (3/1,157) and 1.8% (40/2,252) of the total number of Trichinella spp. isolates from
156
domestic pigs and wild boar, respectively, identified in Europe so far (T. spiralis for 85.56%
157
and 54.44%, T. britovi for 14.17% and 43.33%, in domestic pigs and wild boar, respectively;
158
and T. nativa for 0.44% in wild boar only). If we apply the percentage of each species for
159
the Trichinella spp. isolates from the 1,399 wild boars and 384 domestic pigs detected in
160
Spain in the period 2006-2013, we can speculate that no more than 1 domestic pig and 25
161
wild boars could be infected by T. pseudospiralis in Spain.
162 163
Ac ce p
te
d
151
In Spain, Trichinella spp. infected pigs were detected only in extensive production farms (i.e., backyard and free-ranging pigs); whereas in intensive production farms,
8
Page 8 of 17
164
Trichinella spp. infected pigs were not detected since the 1960s (Ministry of Agriculture,
165
Food and Environment).
166
A different epidemiological picture has been observed in Tasmania and Florida, where a 30% and 20.4% prevalence was detected in Tasmanian devils (Sarchophilus
168
harrisii) and in Florida panthers (Puma concolor coryi), respectively (Obendorf et al., 1990;
169
Richards et al., 2015). Furthermore in Tasmania, T. pseudospiralis was detected in 3.3%
170
(3/91) of raptorial birds (Obendorf and Clarke, 1992). This is still the highest prevalence
171
never detected in birds up to now. At present, we do not have information on the role of the
172
main reservoir hosts of the encapsulated Trichinella species circulating in Europe, as
173
reservoir of T. pseudospiralis. We can suspect that other animal species such as birds and
174
rodents could be reservoir of this non-encapsulated species; alternately, we can assume that
175
the biomass of this species is much lower than that of the other Trichinella species in nature.
176
However, we have to keep in mind that wild animal movements have a relevant role in the
177
spreading of zoonotic pathogens (Gortázar, et al., 2007).
cr
us
an
M
d
te
178
ip t
167
In the last decades, the European wild boar populations have increased possibly due to changes in hunting practices, the expansion of single-crop farming, and climate change
180
(Bieber and Ruf, 2005, Acevedo et al., 2006). The exponential increase of the number of
181
wild boar heads has also increased the biomass of Trichinella spp. (Kirjusina et al., 2015).
182
Ac ce p
179
Meat and meat derived products from wild boar are the main source of infection for
183
humans in Spain (Murrell and Pozio, 2011). Recently, meat of wild boar preserved under
184
vacuum from a supplier of Girona (Cataluña, Spain), was imported to Belgium and
185
consumed in three restaurants by 16 persons who got ill at the beginning of November 2014
186
(Pozio, 2015).
187 188
No Trichinella spp. infection was documented in domestic pigs and wild boar of Portugal since 1966, when the last human case was reported due to the consumption of 9
Page 9 of 17
189
sausages from a domestic pig (Ferreira et al., 2014). In Portugal, only T. britovi was
190
documented in wild carnivores (foxes, wolves and a river otter) (ITRC,
191
www.iss.it/site/trichinella/index.asp). There is the need to educate hunters on the importance of the systematic examination
ip t
192
for Trichinella spp. larvae of game intended for human consumption to prevent human
194
infection. Furthermore, veterinary services should educate hunters not to spread game
195
carcasses or their scraps and offal in the environment, and should organize a system for a
196
proper collection and disposal of these biological samples. The increase of the wild boar
197
population size may have enhanced the common habit of hunters to leave animal carcasses
198
in the field after skinning, or removing and discarding the entrails, which has been
199
demonstrated to strongly increase the probability of Trichinella spp. transmission among
200
wildlife, and to free-ranging and backyard pigs, particularly if the pig owner is a hunter
201
(Pozio, 2014).
202
te
d
M
an
us
cr
193
Acknowledgements
204
We thank the Public Health Agency of Catalonia, which sent us the muscle samples for the
205
diagnosis. This work was supported by the Spanish Agency for Consumer Affairs, Food
206
Safety and Nutrition.
207
Ac ce p
203
10
Page 10 of 17
207
References
208
Airas, N., Saari, S., Mikkonen, T., Virtala, A.M., Pellikka, J., Oksanen, A., Isomursu, M., Kilpelä, S.S., Lim, C.W., Sukura, A., 2010. Sylvatic Trichinella spp. infection in
210
Finland. J. Parasitol. 96, 67-76.
212
Acevedo, P., Escudero, M. A., Munoz, R., Gortázar C., 2006. Factors affecting wild boar
abundance across an environmental gradient in Spain. Acta Theriologica 51, 327-336.
cr
211
ip t
209
Bieber, C., Ruf, T., 2005. Population dynamics in wild boar Sus scrofa: ecology, elasticity
214
of growth rate and implications for the management of pulsed resource consumers. J.
215
Appl. Ecol. 42, 1203–1213.
us
213
Calero, R., Martinez, F., Hernandez, S. and Acosta, I., 1978. Parasitacion de Buteo buteo
217
(Aves: Accipitridae) por Trichinella sp. en el parque zoologico de Jerez de la
218
Frontiera. Revista Iberica Parasitolgia 38, 135-138.
M
Conedera, G., Vio, D., Ustulin, M., Londero, M., Bregoli, M., Perosa, G., Rigo, S.,
d
219
an
216
Simonato, G., Marangon, S., Interisano, M., Gomez Morales, M.A., Pozio, E., Capelli,
221
G., 2014. Trichinella pseudospiralis in wildboars (Sus scrofa) of the Friuli Venezia
222
Giulia region, Italy. Wiener Klinische Wochenschrift 126, S70-S71.
224 225 226 227 228
Ac ce p
223
te
220
Dick, T.A., 1983. Species, and infraspecific variation. In: Campbell, W.C. (Ed.), Trichinella and Trichinosis. Plenum Press, New York and London, pp. 31-73.
European Commission, 2005. Regulation (EC) no. 2075/2005 of the European Parliament and of the Council of 5 December 2005 laying down specific rules on official controls for Trichinella in meat. Off. J. Eur. Comm. 338, 60–82.
Ferreira, I., Martins, S., Reis, T., Vilares, A., Mendes, A., Cardoso, S., do Céu Costa, M.,
229
João Gargaté, M., 2014. Triquinelose humana: estudo observacional em dois grupos
230
populacionais expostos à infeção por Trichinella sp. Observações Boletim
231
Epidemiológico 3, 20-22. 11
Page 11 of 17
234 235 236 237 238
pseudospiralis from a wild pig in Texas. Vet. Parasitol. 132, 147-150. Garkavi, B.L., 1972. Species of Trichinella isolated from wild animals. Veterinariya 10, 9091.
ip t
233
Gamble, H.R., Pozio, E., Lichtenfels, J.R. and Zarlenga, D.S., 2005. Trichinella
Gottstein, B., Pozio, E., Nöckler, K., 2009. Epidemiology, diagnosis, treatment, and control of trichinellosis. Clin. Microbiol. Rev. 22, 127-145.
cr
232
Gortázar, C., Ferroglio, E., Höfle, U., Frölich, K., Vicente, J., 2007. Diseases shared between wildlife and livestock a European perspective. Eur. J. Wildl. Res. 53, 241–
240
256.
243
an
242
International Trichinella Reference Center, www.iss.it/site/trichinella/index/asp, dowloaded on February 9, 2015.
M
241
us
239
Kirjušina, M., Deksne, G., Marucci, G., Bakasejevs, E., Jahundoviča, I., Daukšte, A., Zdankovska, A., Bērziņa, Z., Esīte, Z., Bella, A., Galati, F., Krūmiņa, A., Pozio, E.,
245
2015. A 38-year study on Trichinella spp. in wild boar (Sus scrofa) of Latvia shows a
246
stable incidence with an increased parasite biomass in the last decade. Parasit. Vectors
247
8, 753.
249 250 251 252 253
te
Ac ce p
248
d
244
Krivokapich, S.J., Gonzalez Prous, C.L., Gatti, G.M., Saldía, L., 2015. First finding of Trichinella pseudospiralis in the Neotropical region. Vet. Parasitol. 208, 268-271.
Kozar, Z., 1970. Trichinosis in Europe. In: Gould S.E. (Ed.), Trichinosis in man and animals. Charles Thomas Publisher, Spingfield, pp. 423-436.
La Rosa, G., Pozio, E., Rossi, P., Murrell, K.D., 1992. Allozyme analysis of Trichinella isolates from various host species and geographical regions. J. Parasitol. 78, 641-646.
254
Learmount, J., Boughtflower, V., Allanson, P.C., Hartley, K.M., Barrecheguren Gutierrez,
255
A., Stephens, N.A., Marucci, G., Smith, G.C., 2015. First report of Trichinella
256
pseudospiralis in a red fox in mainland Britain. Vet. Parasitol. 208, 259-262. 12
Page 12 of 17
257
Lindsay, D.S., Zarlenga, D.S., Gamble, H.R., Al-Yaman, F., Smith, P.C., Blagburn, B.L.,
258
1995. Isolation and characterization of Trichinella pseudospiralis Garkavi, 1972 from
259
a black vulture (Coragyps atratus). J. Parasitol. 81, 920-923. Lopes, A.P., Vila-Viçosa, M.J., Cardoso, L., Gottstein, B., Müller, N., Cortes, H.C.E., 2015.
261
Trichinella britovi in a red fox (Vulpes vulpes) from Portugal. Vet. Parasitol. (in
262
press).
cr
263
ip t
260
Merialdi, G., Bardasi, L., Fontana,M.C., Spaggiari,B., Maioli, G., Conedera, G., Vio, D.,Londero, M., Marucci, G., Ludovisie, A., Pozio, E., Capelli, G., 2011. First reports
265
of Trichinella pseudospiralis in wild boars (Sus scrofa) of Italy. Vet. Parasitol. 178,
266
370–373.
an
Ministry of Agriculture, Food and Environment. Report on results of the national program of
M
267
us
264
surveillance in wildlife 2012.
269
http://rasve.magrama.es/Recursos/Ficheros/Historico/83_Resultados%20del%20Progr
270
ama%20Nacional%20de%20Vigilancia%20en%20fauna%20silvestre%202012..pdf,
271
downloaded on March 31, 2015.
te
d
268
Moskwa, B., Goździk, K., Bień, J., Borecka, A., Gawor, J., Cabaj, W., 2013. First report of
273
Trichinella pseudospiralis in Poland, in red foxes (Vulpes vulpes). Acta Parasitol. 58,
274 275 276 277
Ac ce p
272
149-154.
Murrell, K.D., Pozio, E., 2011. Worldwide occurrence and impact of human trichinellosis, 1986-2009. Emerg. Infect. Dis. 17, 2194-2202.
Nöckler, K., Reckinger, S., Pozio, E., 2006. Trichinella spiralis and Trichinella
278
pseudospiralis mixed infection in a wild boar (Sus scrofa) of Germany. Vet. Parasitol.
279
137, 364–368.
280 281
Office International of Epizooties, 2013. Chapter 8.14, Infection with Trichinella spp. Terrestrial Animal Health Code, pp. 1–4. 13
Page 13 of 17
282 283 284
Obendorf, D.L., Clarke, K.P., 1992. Trichinella pseudospiralis infections in free-living Tasmanian birds. J. Helminthol. Soc. Washington 59, 144-147. Obendorf, D.L., Handlinger, J.H., Mason, R.W., Clarke, K.P., Forman, A.J., Hooper, P.T., Smith, S.J., Holdsworth, M., 1990. Trichinella pseudospiralis infection in Tasmanian
286
wildlife. Aust. Vet. J. 67, 108–110.
292 293 294 295
cr
us
291
and habitats. Vet. Parasitol. 194, 128-132.
Pozio, E., 2014. Searching for Trichinella: not all pigs are created equal. Trends Parasitol.
an
290
Pozio, E., 2013. The opportunistic nature of Trichinella-Exploitation of new geographies
30, 4-11.
M
289
Vet. Parasitol. 149, 3-21.
Pozio, E., 2015. Trichinella spp. imported with live animals and meat. Vet. Parasitol. (in press)
d
288
Pozio, E., 2007. World distribution of Trichinella spp. infections in animals and humans.
Pozio, E., La Rosa, G., 2003. PCR-derived methods for the identification of Trichinella
te
287
ip t
285
parasites from animal and human samples. Methods Mol. Biol. 216, 299–309.
297
Pozio, E., Zarlenga,D.S., 2013. New pieces of the Trichinella puzzle. Int. J. Parasitol. 43,
298 299 300 301 302 303 304
Ac ce p
296
983-997.
Pozio, E., Nöckler, K., Reckinger, S., 2006. Trichinella spiralis and Trichinella pseudospiralis mixed infection in a wild boar (Sus scrofa) of Germany. Vet. Parasitol. 137, 364–368.
Pozio, E., La Rosa, G., Murrell, K.D., Lichtenfels, J.R., 1992. Taxonomic revision of the genus Trichinella. J. Parasitol. 78, 654-659. Pozio, E., Shaikenov, B., La Rosa, G., Obendorf, D.L., 1992. Allozymic and biological
305
characters of Trichinella pseudospiralis isolates from free-ranging animals. J.
306
Parasitol. 78, 1087-1090. 14
Page 14 of 17
307
Pozio, E., La Rosa, G., Serrano, F.J., Barrat, J., Rossi, L., 1996. Environmental and human
308
influence on the ecology of Trichinella spiralis and Trichinella britovi in Western
309
Europe. Parasitol. 113, 527-533. Pozio, E., Christensson, D., Stéen, M., Marucci, G., La Rosa, G., Bröjer, C., Mörner, T.,
ip t
310
Uhlhorn, H., Agren, E., Hall, M., 2004. Trichinella pseudospiralis foci in Sweden. Vet
312
Parasitol. 125, 335-342.
313
cr
311
Pozio, E., Rinaldi, L., Marucci, G., Musella, V., Galati, F., Cringoli, G., Boireau, P., La Rosa, G., 2009. Hosts and habitats of Trichinella spiralis and Trichinella britovi in
315
Europe. Int. J. Parasitol. 39, 71–79.
318
an
317
Ranque, S., Faugère, B., Pozio, E., La Rosa, G., Tamburrini, A., Pellissier, J.F., Brouqui, P., 2000. Trichinella pseudospiralis outbreak in France. Emerg. Infect. Dis. 6, 543-547.
M
316
us
314
Reichard, M.V., Criffield, M., Thomas, J.E., Paritte, J.M., Cunningham, M., Onorato, D., Logan, K., Interisano, M., Marucci, G., Pozio, E., 2015. High prevalence of
320
Trichinella pseudospiralis in Florida panthers (Puma concolor coryi). Parasit. Vectors.
321
8, 67.
323 324
te
Shaikenov, B.S., Boev, S.N., 1983. Distribution of Trichinella species in the old world.
Ac ce p
322
d
319
Wiad. Parazytol. 29, 595-608.
15
Page 15 of 17
324
Table 1
325
Pigs and wild boar (Sus scrofa) tested for Trichinella spp. in Spain from 2006 to 2013. Trichinella positive/tested (%) Domestic pigs slaughtered
Hunted
ip t
Year
at homeb
wild boar
2006
3/37,601,623 (8x10-6)
6/132,760 (0.004)
172/70,566 (0.24)
2007
48/41,198,179 (10-4)
4/75,514 (0.005)
2008
75/38,806,715 (10-4)
2/90,889 (0.002)
2009
59/39,929,716 (10-4)
5/60,295 (0.008)
2010
2/41,224,861 (5x10-6)
0/53,485
160/78,571 (0.2)
2011
8/41,597,557 (2x10-5)
1/32,138 (0.003)
176/104,869 (0.16)
2012
115c/40,783,578 (3x10-4)
2013
45/39,190,053 (10-4)
103/51,718 (0.19)
M
an
us
182/81,248 (0.24)
1/32,987 (0.003)
245/123,597 (0.19)
10/44,438 (0.022)
257/117,552 (0.21)
d
355/314,332,282 (10-4)
104/64,557 (0.16)
29/521,667 (0.005)
1,399/692,678 (0.20)
te
Total
cr
at abattoirsa
326
a
327
pigs) have been slaughtered in official abattoirs; b backyard and free-ranging pigs only; c of
328
them 114 originated from the same farm of Salamanca, Castile and León, Spain.
Ac ce p
329
Pigs reared in different containment systems (including some backyard and free-ranging
16
Page 16 of 17
329
Table 2
330
Trichinella pseudospiralis in animals of the European Union countries. Number of animals
331
are reported between brackets. Domestic
Carnivores and other
wild boar
pigs (3)
animals (24)
Italy (4)
Croatia (1),
Bulgaria (2), Denmark (1),
Slovakia (2)
Finland (14), Italy (1),
Czech Rep. (2), Estonia (2),
Italy (2)
Lithuania (1), Poland (1),
us
Finland (1), France (1),
Birds
ip t
Austria (1), Bulgaria (1),
Farmed
cr
Hunted wild boar (38)
Slovakia (2), Sweden (1),
an
Germany (14), Hungary (1), Italy (2), Netherlands (1),
United Kingdom (1)
M
Spain (1), Sweden (11) 332
Ac ce p
te
d
333
17
Page 17 of 17
