16626

2014
VDIXXX10.1177/1040638713516626Lamanema chavezi infection of a llama in the United StatesJarvinen et al.

Case Report

Journal of Veterinary Diagnostic Investigation

Identification of Lamanema chavezi Becklund 2014, Vol. 26(1) 178­–183

© 2014 The Author(s)
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DOI: 10.1177/1040638713516626
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United States

Julie Ann C. Jarvinen,1 Elizabeth M. Whitley, Amanda J. Kreuder,

Jennifer A. Schleining

Abstract. Infection with Lamanema chavezi, a parasitic nematode of New World camelids, was diagnosed by examination
of feces and formalin-fixed liver from a 14-month-old female llama (Lama glama) that died after a 6-week illness. Infection
with L. chavezi was initially suspected when a granuloma containing an unidentified nematode was detected microscopically
in the hepatic parenchyma from a necropsy specimen. The subsequent diagnosis of L. chavezi infection was based on the
morphologic features of 2 immature nematodes dissected from individual hepatic granulomas, characteristics of eggs detected
in feces of the llama by centrifugal flotation in sugar solution (specific gravity: 1.30), development of third-stage larvae
within the eggs after incubation of the llama feces at room temperature for ≥30 days, and the morphology of third-stage larvae
released from the embryonated eggs. Collectively, these findings indicate that the llama, born and raised in Oregon, harbored
an autochthonous L. chavezi infection. Eggs identified as L. chavezi were also detected by centrifugal flotation of pelleted
feces from 3 of 7 herd mates of the llama indicating this parasite is endemic in the Oregon herd. The findings reported herein
serve to alert diagnosticians and veterinary practitioners to the occurrence of L. chavezi in New World camelids in the United
States and describe diagnostic features of this potential pathogen.

Key words: Alpacas; Lamanema chavezi; llamas; nematode; New World camelid.

Lamanema chavezi is a potentially pathogenic nematode of
New World camelids (NWCs) that, to the authors’ knowl-
edge, has not been previously reported to occur in the United
States.2,3 The direct life cycle of this trichostrongyloid is
unusual in that third-stage larvae (L3s) develop within the
eggs, hatching of the eggs is stimulated by exposure to low
temperatures, and L3s undergo an enterohepatic migration
before returning to the small intestine to become adults.11
Based on the morphologic characteristics of L. chavezi and
the unique features of its life cycle, an autochthonous infec-
tion with this nematode is reported in a llama (Lama glama)
born and raised in the State of Oregon. Furthermore, detec-
tion of L. chavezi eggs in feces from herd mates of this llama
indicates that this nematode is endemic in the Oregon herd.
A 14-month-old female llama was transported from Ore-
gon to Iowa on September 9, 2012 to attend a llama event.
The owner had treated the llama with fenbendazole at 18.5–
40 mg/kg body weight orally for 1–3 days every 2–3 months
beginning at 3 months of age. A final dose of fenbendazole
(25.5 mg/kg body weight) was given at 13 months of age at
the time of transport to Iowa. Four weeks later, on October 8,
2012, the llama was admitted to the Iowa State University
(ISU) Lloyd Veterinary Medical Center (Ames, Iowa) with a
3-week history of lethargy, anorexia, and weight loss. Ultra-
sonographic examination of the liver at the time of presenta-
tion to the teaching hospital revealed small, hyperechoic foci
scattered throughout the parenchyma (Fig. 1A). During hos-
pitalization, serum hepatic enzymes were not elevated. The
llama died on October 22, 2012 following 2 weeks of inten-
sive therapy for suspected septic salmonellosis. A routine,
complete necropsy was performed, and systemic salmonel-
losis and septic peritonitis due to jejunal rupture were diag-
nosed. Samples of liver and other organs were fixed in 10%
neutral buffered formalin, processed routinely, sectioned to 5
μm, and stained with hematoxylin and eosin.
Among the gross lesions observed at necropsy were mul-
tiple, small (pinpoint to 2 mm), white-tan foci on the liver
surface that extended into the parenchyma (Fig. 1B). The
lesions were consistent with the hyperechoic foci previously
observed in the ultrasonogram (Fig. 1A). Microscopically,
many of the hepatic lesions noted at necropsy were well-
organized granulomas that contained a coiled nematode
(Fig. 2A, 2B) or were undergoing mineralization (Fig. 2C).
Longitudinal cuticular ridges that comprise the synlophe are
evident in a section of a single nematode (Fig. 2B). In tissue
From the Departments of Veterinary Pathology (Jarvinen, Whitley)
and Veterinary Diagnostic and Production Animal Medicine (Kreuder,
Schleining), College of Veterinary Medicine, Iowa State University,
Ames, IA.
1
Corresponding Author: Julie Ann C. Jarvinen, Department of
Veterinary Pathology, 1786 College of Veterinary Medicine, Iowa State
University, 1600 South 16th Street, Ames, IA 50011. jarvinen@iastate.edu
 Lamanema chavezi infection of a llama in the United States 179

Figure 1.  Llama (Lama glama). A, ultrasonogram of scattered hyperechoic foci (arrows) in the hepatic parenchyma. Hyperechoic foci
represent areas of the parenchyma that have greater density than the surrounding parenchyma, and, in this case, correspond to multiple, small
hepatic granulomas. B, multiple granulomas (arrows) scattered throughout the parenchyma of the liver. Bar = 1 cm.

Figure 2.  Hepatic granulomas in a llama (Lama glama). A, granuloma containing multiple cross-sections of a coiled nematode (arrows).
Bar = 200 µm. B, a cross-section of the nematode in Figure 1A; note the longitudinal cuticular ridges (arrows). Bar = 50 µm. C, hepatic
granuloma with mineralized contents surrounded by mature fibroplasia (arrow). Hematoxylin and eosin. Bar = 200 µm.

sections, the lamina propria of the small intestine and colon
was expanded by moderate numbers of eosinophils, lympho-
cytes, and plasma cells, but encysted larvae were not
observed.
With the aid of a stereoscopic microscope, portions of
formalin-fixed liver containing grossly visible pale foci were
carefully examined and 2 coiled nematode larvae, 1 intact
male and 1 decapitated female, were dissected from individ-
ual granulomas. The larvae were initially placed in 70%
ethanol–glycerol solution and then transferred to 70% etha-
nol. The female larva had a subterminal anus and a spine
approximately 12 mm long that was directed laterally
approximately at a right angle after it emerged from the dis-
tal extremity of the tail (Fig. 3A, 3B), features previously
described for L. chavezi.22 The spicules of the male larva
(Fig. 3C, 3D) were consistent with descriptions of adult
L. chavezi.3,22 Other features of the formalin-fixed larvae
could not be evaluated because they were obscured by
superimposition of larval coils on one another and by tissue
adherent to the larval surfaces that could not be removed
without compromising the integrity of the larvae. The pres-
ence of adult L. chavezi, previously described as 8–18 mm
long, red, and with a corkscrew-like appearance of the ante-
rior two thirds of the body,11 could not be determined
because gross samples of the intestinal tract of the llama
had been discarded and were not available for examination.
The ISU Clinical Pathology Laboratory initially exam-
ined feces collected from the llama on October 8 and 9, 2012,
and reported 300 trichostrongyle and <100 Nematodirus
eggs per gram of feces by the McMaster method, no Fasciola
hepatica eggs by fecal sedimentation, and no Eimeria mac-
usaniensis oocysts by either method. The remaining feces
were refrigerated until examined by centrifugal flotation in
sugar solution (specific gravity: ≥1.3) on November 2, 2012.
Low numbers of yellow-brown, cylindrical eggs with paral-
lel sides and rounded poles measuring 170.2 µm × 73.4 µm
180
Jarvinen et al.

Figure 3.  Lamanema chavezi larvae dissected from hepatic granulomas in a llama (Lama glama). A and B, posterior end of a female L.
chavezi larva. A, lateral view with subterminal anus (arrow) and spine at distal extremity of tail (arrowhead). Bar = 200 µm. B, lateral view:
terminal spine (arrow) curves at right angle to tail. Bar = 50 µm. C and D, spicules of a male L. chavezi larva. C, paired, external shafts
(arrow) with membranous expansions at tips and an internal spur (arrowhead); the second internal spur is not in focus in this image. Bar =
50 µm. D, distal portion of paired, internal, pointed spurs (arrow). Bar = 50 µm.

(n = 2) and containing a highly cellular internal mass were
present in the llama feces that had been refrigerated for 24
days (Fig. 4A). In comparison, Nematodirus eggs present in
the same fecal sample had 8–16 large, round dark cells.
All remaining feces from the samples were thoroughly
mixed with tap water and incubated for 38 days at room tem-
perature. During this time, aliquots of the cultured feces were
periodically removed and examined by centrifugal flotation
to monitor larval development. By 30 days of incubation of
the llama feces, L3s with 2 retained cuticle sheaths were
present within the L. chavezi (Fig. 4B) and Nematodirus
eggs. When eggs containing an L3 were ruptured on glass
microscope slides by applying pressure to the coverslip, lar-
vae actively emerged. The egg shells remaining after L3s
had emerged from the L. chavezi eggs were yellow-brown
with a darker golden-brown outer wall (Fig. 4C). Third-stage
larvae from 2 L. chavezi eggs averaged 697.2 μm × 29.8 µm,
had 8 large intestinal cells, an unnotched larval tail tapering
abruptly to a relatively broad blunt point, and a short (26.2
µm), sharply pointed sheath tail (Fig. 5A–5C).
On November 27, 2012, fecal samples were obtained
from 7 healthy Oregon herd mates of the llama and low num-
bers of eggs consistent with those of L. chavezi were detected
in feces from 3 llamas. The eggs averaged 171.2 µm × 77.3
µm (n = 11) and contained a developing first-stage larva.
In South America, L. chavezi is a common, small intesti-
nal parasite of NWCs with reported prevalence rates of
18.5% in Argentinian llamas,6 48.2% in Peruvian alpacas
 Lamanema chavezi infection of a llama in the United States
Figure 4.  A, Lamanema chavezi egg from llama (Lama glama) feces after 24 days of storage at 4°C. Bar = 200 µm. B, third-stage
larva (L3) in L. chavezi egg; the short sheath tail (arrow) of the L3 can be seen inside the egg. Bar = 50 µm. C, shell of ruptured egg after
emergence of L3. Bar = 200 µm.
Figure 5. Third-stage larvae (L3) from Lamanema chavezi
eggs. A, an iodine-stained larva. Bar = 200 µm. B, anterior end of an
L3 demonstrating 2 retained cuticle sheaths and esophagus (arrow,
junction of esophagus and intestine). Bar = 50 µm. C, posterior end
of the same L3 demonstrating 2 retained cuticle sheaths, short tail
sheath (arrowhead), and short, bluntly pointed larval tail (arrow).
Bar = 50 µm.
(Vicugna pacos),7 13–61.3% in Chilean llamas,1,20 and 64%
in Bolivian llamas (Sporndly E, Nissen AM: 2008, Preva-
lence of parasites in llamas in the Andean Bolivia. Faculty of
181
Life Sciences, University of Copenhagen, Denmark. Mas-
ter’s thesis. Abstract available at http://www.ddrn.dk/filer/
forum/File/Lamas_Bolivia_ES_AMN.pdf. Accessed June 9,
2013). Although L. chavezi is generally considered to be a
host-specific parasite of NWCs, it has also been described in
2 species of South American rodents.9,22 Consistent with the
age of the llama in the present case, the greatest prevalence
of L. chavezi in South America occurs in NWCs under 2–3
years of age.7,11 Unusual eggs detected in feces of alpacas
and llamas in New Zealand provided the first indication that
L. chavezi might parasitize NWCs outside of South Amer-
ica.16 Subsequently, identification of adults expelled in feces
of a llama following anthelmintic treatment confirmed the
occurrence of L. chavezi in New Zealand.17 Further studies
are needed to determine the prevalence of L. chavezi in
NWCs in the United States and New Zealand.
In addition to eggs of L. chavezi, feces of the llama in the
present case also contained eggs of trichostrongyles, Nema-
todirus spp., and Nematodirus battus. Of nematodes infect-
ing NWCs, only Nematodirus spp. and Marshallagia spp.
produce large eggs (>130 µm) that need to be differentiated
from eggs of L. chavezi. The eggs of Nematodirus spp. in the
present case were readily identified by the presence of a mor-
ula composed of ≤16 large dark cells within a clear to color-
less shell, by their greater size compared with L. chavezi,
and/or by their shape (i.e., wide centrally and narrow at the
poles).2,23 In fresh feces, eggs of L. chavezi and Marshallagia
both have flat parallel sides with rounded poles, contain a
morula comprised of >64 cells, and their size ranges overlap,
but the eggs of L. chavezi are yellow-brown while those of
Marshallagia have a colorless shell.2,6,13,16,19 The eggs of N.
battus and L. chavezi are similar in size and shape, but N.
battus eggs are brown or coffee-colored, have a thickened
shell with an irregular pitted internal surface at the poles, and
fewer and larger morula cells in comparison to L. chavezi.2,6,16
Larvae develop to second-stage larvae within the eggs of
Marshallagia spp. and to L3s in Nematodirus spp. and
L. chavezi.19 In the present case, L3s of Nematodirus spp.
were easily identified by their long filamentous sheath tail;
182
Jarvinen et al.
those of N. battus had a long tapering notched larval tail.24 In
contrast, L3s obtained from the L. chavezi eggs had a short
sheath tail, an unnotched larval tail, and were comparable in
size to L3s previously reported.16
Small, mineralized granulomatous or fibrotic lesions are
frequently reported as incidental findings in the livers of
NWCs in the United States and are attributed to parasite
migration, but the specific parasites causing these lesions are
commonly not identified.4 Migration of L. chavezi, Fasciola
hepatica, Fascioloides magna, Dicrocoelium dendriticum,
and tapeworm larvae can produce lesions in the liver of
NWCs, but, to date, only F. hepatica and F. magna are
reported to cycle in NWCs in the United States.2,8,12 In the
present case, no fluke eggs were found when feces from the
llama were examined by sedimentation, and no immature or
adult liver flukes or tapeworm larvae were found in the liver
or bile ducts at necropsy. Although the llama feces contained
N. battus eggs, infection with this nematode would not be
expected to cause hepatic lesions because it remains in the
intestinal tract and does not migrate in the liver.14 The occur-
rence and appearance of trichostrongyloid larvae in hepatic
granulomas of the llama in the present case are consistent
with descriptions of L. chavezi infection.11
Lamanema chavezi is considered the most pathogenic
nematode of NWCs, and pathogenicity is typically greatest
in younger hosts infected with large numbers of larvae. Ane-
mia, anorexia, emaciation, prostration, colic, and pale feces
were noted in a 4-month-old alpaca that died 20 days after it
was inoculated with 200,000 L3s.10 Inoculation with 10,000
L3s resulted in anemia, eosinophilia, focal granulomatous
liver lesions, and parasites in the bile ducts 30 days after
inoculation of a 20-day-old alpaca10 and an increase in serum
aspartate transaminase (serum glutamic oxaloacetic trans-
aminase) levels 14 days after inoculation of two 3-month-old
alpacas.21 In contrast, a 6-month-old alpaca inoculated with
10,000 L3s became anemic but showed no clinical signs
while neither anemia nor clinical signs occurred in a
12-month-old alpaca inoculated with 7,000 L3s.10 A natu-
rally infected, 2-year-old llama in poor body condition that
was shedding large numbers of L. chavezi eggs had small
hepatic abscesses and hemorrhagic enteritis at necropsy,
while a second naturally infected 3-year-old llama herd mate
shed lesser numbers of L. chavezi eggs and exhibited no clin-
ical signs.5 Natural L. chavezi infections of NWCs in Argen-
tina6 and New Zealand16–18 also had no overt clinical signs.
In the present case, detection of L. chavezi eggs in feces indi-
cates that the llama had been infected with this parasite for at
least the length of the pre-patent period, which is 30–32
days, and mineralization of the granulomas suggests a
chronic infection.11
Anthelmintics with demonstrated efficacy against L.
chavezi in NWCs include levamisole11 and ivermectin,15 but
resistance to ivermectin has also been reported (Ciro-Marino
TA: 2011, Determinacion de resistencia anthlemintica frente
a ivermectina de nematodos gastrointestinales en alpacas
(Vicugna pacos) Puno-Peru [Determination of anthelmintic
resistance against ivermectin of gastrointestinal nematodes
in alpacas (Vicugna pacos) in Puno-Peru]. In Spanish.
Abstract in English. Available at: http://www.buscagro.com/
www.buscagro.com/biblioteca/Ciro-Traverso-Arguedas/
nematodos-en-alpacas.pdf. Accessed July 15, 2013). It is of
interest that a patent L. chavezi infection was present in the
llama of the current study even though it had been repeatedly
treated with fenbendazole at 1–3-month intervals beginning
at 3 months of age with the most recent treatment given 6
weeks prior to death. Whether or not infection with L. chavezi
might have predisposed this llama to Salmonella infection or
otherwise contributed significantly to its illness is unknown.
Declaration of conflicting interests
The author(s) declared no potential conflicts of interest with
respect to the research, authorship, and/or publication of this
article.
Funding
The author(s) received no financial support for the research, author-
ship, and/or publication of this article.
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