Nondomestic, Exotic, Wildlife and Zoo Animals–Original Article

Veterinary Pathology
2017, Vol. 54(2) 298-311
Muscle Pathology in Free-Ranging Stranded ª The Author(s) 2016
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Cetaceans DOI: 10.1177/0300985816660747
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E. Sierra1, A. Espinosa de los Monteros1, A. Fernández1,

J. Dı́az-Delgado1, C. Suárez-Santana1, M. Arbelo1,
M. A. Sierra2, and P. Herráez1

Abstract
Despite the profound impact that skeletal muscle disorders may pose for the daily activities of wild terrestrial and marine
mammals, such conditions have been rarely described in cetaceans. In this study, the authors aimed to determine the nature and
prevalence of skeletal muscle lesions in small and large odontocetes and mysticetes (n ¼ 153) from 19 different species. A
macroscopic evaluation of the epaxial muscle mass and a histologic examination of the longissimus dorsi muscle were performed in
all cases. The only macroscopically evident change was variable degrees of atrophy of the epaxial muscles (longissimus dorsi,
multifidus, spinalis) in emaciated specimens. The histopathological study revealed single or combined morphological changes in
91.5% of the cases. These changes included the following: degenerative lesions (75.2%), muscle atrophy (37.9%), chronic myo-
pathic changes (25.5%), parasitic infestation (9.2%), and myositis (1.9%). The skeletal muscle is easily sampled during a necropsy
and provides essential microscopic information that reflects both local and systemic conditions. Thus, skeletal muscle should be
systematically sampled, processed, and examined in all stranded cetaceans.

Keywords
cetaceans, skeletal muscle, degenerative changes, atrophy, hypertrophy, chronic myopathic changes, Sarcocystis, Toxoplasma,
myositis

Skeletal muscle disorders have been rarely described in ceta-
ceans. These disorders include granulomatous mucormycosis
in a killer whale (Orcinus orca);1 sarcocystosis in several ceta-
cean species (beluga whale, Delphinapterus leucas; Atlantic
white-sided dolphin, Lagenorhynchus acutus; long-finned pilot
whales, Globicephala melaena;9,17,28 striped dolphin, Stenella
coeruleoalba;15 sperm whale, Physeter macrocephalus; and
northern right whale dolphin, Lissodelphis borealis);11 and
bacterial necrotizing fasciitis and myositis in the captive com-
mon bottlenose dolphin (Tursiops truncatus).97
The different cetacean species exhibit a wide range of body
masses.26,55 However, the distribution of the skeletal muscle is
similar in all the species and provides similar tail-stroke power
for swimming. The longissimus dorsi is part of the epaxial
musculature, which lies along both sides of the vertebral col-
umn, and is involved primarily in cetacean locomotion and has
been shown to be 1 of the 2 muscles that power the dolphin’s
upstroke.44,63 The impact of skeletal muscle disorders may
range from subclinical or non-life-threatening situations (eg,
mild intra or interspecific interactions) to grave localized inju-
ries (eg, those caused by ship collision) or disseminated disease
processes (eg, infectious myositis), which could impede loco-
motion and lead to death. The presence of complex polysac-
charide inclusions,78 myopathic changes related to capture
myopathy, and ship-strike-associated muscle lesions have been
previously reported in the populations studied here.42,43,76 Age-
related histological findings, including variations in the number
and the size of different types of myofibers, have also been
described in this group of animals.75 In addition, the presence
of sarcoplasmic masses within the skeletal muscle of a stranded
pygmy sperm whale (Kogia breviceps) have been previously
published.74
Given the scarcity of reports on the muscle pathology of
these species and the wide range of muscle lesions encountered
in the stranded cetaceans of our study, this article aimed to
1
Department of Veterinary Pathology, Institute for Animal Health, Veterinary
School, University of Las Palmas de Gran Canaria, Arucas, Spain
2
Department of Anatomy and Comparative Pathology, Veterinary Faculty,
University of Córdoba, Córdoba, Spain
Supplemental material for this article is available on the Veterinary Pathology
website at http://journals.sagepub.com/doi/suppl/10.1177/0300985816660747.
Corresponding Author:
E. Sierra, Department of Veterinary Pathology, Institute for Animal Health,
Veterinary School, University of Las Palmas de Gran Canaria, Trasmontaña
S/N, 35416, Arucas, Las Palmas, Spain.
Email: esierra@becarios.ulpgc.es
Sierra et al 299

Table 1. Muscle Pathology in Stranded Cetaceans (1996–2013): Number of Cases by Species in the Different Etiological Categories.

Lesion

Degenerative Changes Atrophy Parasites

Species n CM Trauma Sepsis FG FI UE Senile C/M Dnv Dss H CMC S. spp. T.g. Myositis CPI SM

Fin whale (Bph) 3 1 1a N N N 1 N 1 N N 1 N 1 N N 1 N

Short-beaked common 10 3 N N N N 1 1 3 3 N N 4 1 N N 3 N
dolphin (Dde)
Short-finned pilot whale (Gma) 12 4 2b 1 N N 2 3 1 1 1 1 3 N N N 2 N
Risso’s dolphin (Ggr) 3 2 N N N N 1 1 1 N N N 1 1 N N 1 N
North Atlantic bottlenose 1 1 N N N N N N N N N N 1 N N N N N
whale (Ham)
Pygmy sperm whale (Kbr) 12 2 2a, 2b 1 N N 2 2 3 N N N 1 1 N N 3 1
Fraser’s dolphin (Lho) 2 N N N N N N N N N N N N N N N N N
Sowerby’s beaked whale (Mbi) 1 N N 1 N N N N 1 N N 1 N 1 N N 1 N
Blainville’s beaked whale (Mde) 3 2 N 1 N N N N N 1 N N N N N N 1 N
Gervai’s beaked whale (Meu) 6 3 1b 1 N 1b N N N 1 N N N N N N N N
Harbor porpoise (Pph) 1 N N N N N N N N N N N N N N N N N
Sperm whale (Pma) 13 1 9a, 1b N N N 2 N 2 N N N N 1 N N N N
False killer whale (Pcr) 1 1 N N N N N N 1 N N N N N N 1 N N
Striped dolphin (Sco) 25 10 1b 3 2a 2 12 N N N 1 12 3 N 1 6 N
Atlantic spotted dolphin (Sfr) 23 8 N N N 1a, 2b 3 9 2 1 N N 10 2 1 1 3 N
Spinner dolphin (Slo) 2 1 N N N N N 1 N N N N 1 1 N N 1 N
Rough-toothed dolphin (Sbr) 3 N 1c 1 N N N 1 N N N N 1 N N N N N
Bottlenose dolphin (Ttr) 13 5 N 1 N 2a 1 2 1 N N 1 2 1 N N 4 N
Cuvier’s beaked whale (Zca) 19 3 1a N 10 1b 3 1 1 N N N 3 N N N N N
Total 153 47 21 10 10 9 18 33 17 7 1 5 39 13 1 3 26 1
Abbreviations: Species: abbreviated scientific names in alphabetic order: Bph, Balaenoptera physalus; Dde, Delphinus delphis; Ggr, Grampus griseus; Gma, Globicephala
macrorhynchus; Ham, Hyperoodon ampullatus; Kbr, Kogia breviceps; Lho, Lagenodelphis hosei; Mbi, Mesoplodon bidens; Mde, Mesoplodon densirostris; Meu, Mesoplodon
europaeus; Pcr, Pseudorca crassidens; Pma, Physeter macrocephalus; Pph, Phocoena phocoena; Sbr, Steno bredanensis; Sco, Stenella coeruleoalba; Sfr, Stenella frontalis; Slo,
Stenella longirostris; Ttr, Tursiops truncatus; and Zca, Ziphius cavirostris. CM, capture myopathy. Trauma: aShip collision, bInter- or intraspecific interactions;
c
Undetermined. FG, fat and gas embolic syndrome. FI, fish interaction. This is defined as acute death of cetaceans after interaction with any type of fishing activity,
whether by accidental capture (bycatch)a or as a result of severe injuries caused by fishermen or fishing equipmentb. UA, undetermined etiology. C/M, cachexia/
malnutrition. Dnv, denervation. Dss, Desuse. H, hypertrophy. CMC, chronic myopathic changes. S. spp., Sarcocystis spp. T.g., toxoplasma gondii. CPI, complex
polysaccharide inclusions. SM, sarcoplasmic masses. N, none.

determine the nature and prevalence of skeletal muscle lesions
in small and large odontocetes and mysticetes stranded on the
Canary Islands, Spain.
Material and Methods
Stranding Epidemiologic Data and Necropsy
Examination
Animals included in this study were stranded cetaceans on the
Canary Islands, except for 5 animals from other locations. The
specimens included in this study are summarized in Table 1
(individual case details are given in Table S1 in the supple-
mental material).
Skeletal muscle samples from stranded odontocetes and
mysticetes (n ¼ 153), collected over a 14-year period (1996
to 2013), were examined. The animals were of both sexes
and ranged in age from neonatal to mature adults, as deter-
mined by biological and morphometric parameters. The ske-
letal muscle samples were collected during necropsies,
following standard protocols.53 Necropsies were performed
either on site (the beach or coast) (17.6%), in facilities built
specifically for this purpose (33.3%), or when possible, in
the necropsy room at the Veterinary College of the Univer-
sity of Las Palmas of Gran Canaria (Spain) (29.4%). Large
cetaceans necropsies were also frequently performed at the
rubbish dump of the corresponding island (19.6%). Permis-
sion for the management of the stranded dead cetaceans was
issued by the environmental department of the Canary
Islands government.
The preservation status of the carcasses was established
according to Geraci and Lounsbury.35 The body condition
of each animal was estimated morphologically based on the
prominence of the spinous and transverse vertebral processes,
the mass of the epaxial musculature (longissimus dorsi, multi-
fidus, spinalis), and the absence or limited presence of sub-
cutaneous fat deposits.51 These estimates were made taking
into account the species and age of the animals. Using these
parameters, we classified their body condition as good, mod-
erate, poor or emaciated.
The muscle examination included a macroscopic evaluation
of the epaxial and hypaxial muscle mass based on transverse
and longitudinal sections throughout the entire muscle body
that were approximately 1–2 cm apart.
300
Histopathology and histochemistry. All skeletal muscle samples
were taken from the region of the longissimus dorsi muscle
immediately lateral to the dorsal fin as previously described.62,84
Samples from other skeletal muscle tissue that displayed grossly
visible alterations were also collected for histopathology.
The samples were mounted on a tongue depressor, fixed at
both ends by pins with the myofibers oriented lengthwise, and
immersed in 4% neutral-buffered formalin for 24–48 hours.
Transverse and longitudinal muscle sections were routinely
processed, embedded in paraffin, serially sectioned and stained
with hematoxylin and eosin (HE), periodic acid-Schiff (PAS)
with and without amylase digestion, phosphotungstic acid-
hematoxylin (PTAH), osmium tetroxide (OsO 4), and von
Kossa staining as previously described.2,4,6,34,52
Histological assessments of the longissimus dorsi muscle
samples were obtained from all animals and were performed
in a blind manner by 3 veterinary pathologists (ES, AEM, PH).
Diagnostic criteria for degenerative changes included seg-
mental hypercontraction (characterized by a hyperacidophilic
sarcoplasm and endomysial edema) and/or degeneration/necro-
sis (with a hyaline, floccular, granular, and/or discoid appear-
ance), contraction band necrosis, presence of globules in the
sarcolemmal tube and/or under the basal lamina, an inflamma-
tory response, and fiber regeneration. Lesions were confirmed as
ante mortem when accompanied by histochemical demonstra-
tion of a loss of myofibril striations (affected fibers failed to
show the normal dark-blue coloration after PTAH staining and
stain pale blue, yellow, or pink instead), and myoglobin leakage
and intrafibrillar fibrinogen immunolabeling. The degree of ske-
letal myodegeneration was subjectively judged as follows: mild,
when scattered individual fibers distributed randomly through-
out the section were detected; moderate, when multiple foci of
affected fibers were distributed randomly through the section; or
severe, when multiple, large, widespread, often coalescing areas
of myodegeneration were detected.
Evaluation criteria for myofiber atrophy/hypertrophy
included an assessment of muscle fiber size and morphology.
Atrophic fibers usually displayed decreased sizes and angular
shapes, while hypertrophic fibers tended to have increased
sizes and more rounded contours along with internal disorga-
nized myofibrils. Determination of the fiber types involved and
any alterations in the distribution of the fibers required immu-
nohistochemical fiber typing in each case. This analysis
required the previous knowledge of the normal histology of the
specimens in relation to their different diving behaviors and
their different ages. Variations in fiber type composition
among marine mammals have been proposed to result from
differences in routine dive duration and swimming speed, as
well as from age.16,75,77
Intramyofiber inclusions/deposits were identified with the
HE and PAS stains. PAS-positive, diastase-resistant inclusions
within skeletal muscles are consistent with deposits of abnor-
mal glycogen, often referred to as a complex polysaccharide.85
Intramyofiber vacuolations diagnoses were based on the pres-
ence of various, small, clear vacuoles within the sarcoplasm of
the myofibers as shown in HE-stained sections.
Veterinary Pathology 54(2)
Chronic myopathic changes (CMCs) were determined based
on excessive fiber size variation, increases in the number of
internal nuclei, and intramyofiber pigment deposits, which
were identified by the presence of large amounts of juxtanuc-
lear brownish pigmentation, which is a pattern of PAS staining
that has been show to be compatible with lipofuscinosis.
Ring fibers were identified as cytoarchitectural alterations
consisting of peripheral rings around maloriented myofilaments.
Myositis was determined based on the presence of focal/
multifocal areas of inflammatory infiltrates (mainly macro-
phages and polymorphonuclear leucocytes).
Intramuscular parasites were identified by a combination of
HE, PAS, and PTAH stains.
Immunohistochemistry. Fast and slow myosin heavy chain
(MHC) isoforms, indicating type II and type I fibers, respec-
tively, were detected using monoclonal mouse antihuman myo-
sin heavy-chain isoform type I (slow twitch) (skeletal, slow,
M8421) and type II (fast twitch) (skeletal, fast, M4276) anti-
bodies (Sigma Co, St. Louis, MO, USA), and myoglobin and
fibrinogen were detected using polyclonal rabbit antihuman
myoglobin (A0324) and fibrinogen (A0080) (Dako, Glostrup,
Denmark). Antibodies were visualized using the avidin-biotin-
peroxidase method (Vector Laboratories, Burlingame, CA,
USA) as previously described.42 In addition, polyclonal anti–
Toxoplasma gondii (Dako, Glostrup, Denmark) diluted 1:200
with a previous pronase treatment (5 min) was used to confirm
histopathological diagnoses of toxoplasmosis. Tissue sections
in which the primary antibodies were replaced by phosphate
buffered saline or nonimmune serum were used as negative
controls to confirm the specificity of the tests.65 Goat and
human tissues were used as positive controls.
Electron microscopy. For ultrastructural studies, skeletal muscle
samples were fixed in 2.5% glutaraldehyde in 0.1 M sodium
cacodylate buffer (pH 7.2). The specimens were postfixed in
1% osmium tetroxide in 0.2% veronal buffer, gradually dehy-
drated in an alcohol series, and embedded in EMbed 812 epoxy
resin (Electron Microscopic Science, Hatfield, PA, USA). Thin
sections were stained with uranyl acetate and lead citrate. The
sections were examined with a ZEISS EM-912 transmission
electron microscope (Carl Zeiss, Oberkochen, Germany).
Myofiber sizes comparisons. Myofiber size measurements were
performed in a selected group of animals from our study
(n ¼ 39). For morphometric analysis, 10 randomly selected
fields/images per slide (200X magnification) of a muscle
section that was immunohistochemically stained for antifast
myosin were captured using an Altra20 digital camera (2 Mega-
Pixel CMOS color camera for light microscopy, Olympus Soft
Imaging Solutions GmbH, Münster, Germany). The cross-
sectional area (CSA) and the lesser diameter (the greatest dis-
tance between the opposite sides of the narrowest aspect of the
fiber) of all the fibers in a field with red-stained fast-twitch
muscle fibers (type II) and blue-stained slow-twitch muscle
fibers (type I), were measured. The relative number of type I
Sierra et al
and type II fibers was assessed by counting each fiber type from
the 10 randomly selected fields of each animal. The percentage
of each specific fiber type was calculated as the specific fiber
count divided by the total fiber count. The muscle morphology
data (CSA, lesser diameter and number of fiber types) was
calculated using the digital software CellA (Imaging Solu-
tions). The mean values were obtained using the Soft Imaging
System for Life Science Microscopy.
Interpretation/analysis. The combination of the histopathologic
findings and data regarding the age, diving profile, type of
stranding, and the cause of death for each case (hereafter
referred as the necropsy report)5 were used to establish an
etiological diagnosis of muscle pathology in the current study.
In addition, fiber size measurements were taken into consider-
ation for the diagnosis of myofiber atrophy in selected cases,
including 25 animals with a morphological diagnosis of focal/
multifocal or generalized myofiber atrophy and 14 specimens
with no myofiber size changes.
Results
A varied and often compound set of morphophysiological and
pathological changes were microscopically identified in 140 of
the 153 (91.5%) animals (these data are summarized in Table 1,
while full descriptions are available in Table S1 in the supple-
mental material). The percentage of animals with 1, 2, 3, 4, or 5
histopathologic features were 49.3%, 19.3%, 17.9%, 7.1%, and
2.1%, respectively. Some lesions tended to occur together:
chronic myopathic changes were frequently present in animals
with senile or denervation muscle atrophy (82.9%), while
69.2% and 46.15% of the animals with complex polysaccharide
inclusions displayed at least 1 of the muscle atrophies
described in our study or had degenerative changes associated
to stranding stress syndrome, respectively.
The only macroscopically visible muscle change among the
animals evaluated was atrophy, which was more evident along
the epaxial musculature than in the muscles of the thoracic wall.
Several degrees of atrophy, ranging from mild to a profound
reduction in muscle mass with marked prominence of the adja-
cent axial osseous relieves (ribs, transverse and spinous vertebral
apophyses) were observed in 41 of the 153 (Supplemental
Table S2) animals. Generalized, focal, or multifocal myofiber
atrophy was observed in 58 of the 153 (37.9%) animals, although
only cases of emaciation corresponded with generalized
myofiber atrophy when examined by light microscope.
Myofiber Atrophy/Hypertrophy
The histologic features (fiber size and morphology, and dis-
tribution pattern), based on the available epidemiological
data, were combined to classify myofiber atrophies as senile
atrophy, cachexia/inanition atrophy, denervation atrophy, and
disuse atrophy.
Senile atrophy represented 56.9% of the cases of atrophy
and was detected in 33 animals (old adults) belonging to
301
10 different species. Most of the animals presented a
moderate-good overall body condition. However, histologi-
cally, all of the animals in this group were morphologically
diagnosed with generalized muscle atrophy of type II fibers,
while type I fibers were enlarged compared with those of
juvenile and young adult specimens. No remarkable morpho-
logical changes were observed within the atrophic myofi-
bers. Cytoplasmic accumulation of PAS-positive pigment,
indicating lipofuscin, within myofibers alone or in combi-
nation with CMCs was observed in 54.4% and 33.3% of the
cases, respectively.
Cachexia/inanition atrophy represented 29.3% of the cases of
atrophy. Similar to the patterns observed with senile atrophy, all
the animals in this group showed generalized myofiber atrophy
at the microscopic level. However, a severe wasting of the epax-
ial musculature was only determined in this category based on
gross examination, from which 14 of the 17 (82.4%) animals
showed evidence of a consumptive pathological process. In 4
cases, foreign bodies lodged in the mouth, tongue or stomachs in
the affected specimens were responsible for their condition. His-
tologically, generalized muscle atrophy, primarily of type II
fibers, was noted. In addition, atrophied myofibers showed evi-
dence of variable degrees of flattening and angular shape
changes (Fig. 1). Occasional irregular shapes were also noticed.
Denervation atrophy represented 16.7% of the atrophies
diagnosed. The morphologic diagnosis in this group was multi-
focal myofiber atrophy, frequently adjacent to an intramuscular
nerve (Fig. 2). The affected fibers were of both types, and fiber
type grouping (the clustering of a specific fiber type instead a
random distribution) was common (Fig. 2, inset). Focal con-
centration of myofiber nuclei was a specific feature of this type
of muscle atrophy. More than half of the specimens in this
group were considered senile based on biological and morpho-
metrical parameters. Intramuscular nerves in this group showed
an increase in perineurial and epineurial connective tissue and
decreased myelin, as shown by osmium tetroxide postfixation
staining, when compared to other cases.
Disuse atrophy was diagnosed in 1 specimen; an adult female
short-finned pilot whale (Globicephala macrorhynchus) that had
severe lateral column deviation (scoliosis). Histologically, there
were diffuse atrophic fibers of both types, as well as a slight
enlargement of the endomysial fibrous stroma.
Presumed compensatory hypertrophy was observed in 5
specimens with myofiber atrophy regardless of the etiology
(Fig. 3). Hypertrophied myofibers usually showed longitudi-
nal fiber splitting (muscle fiber divisions).
Degenerative Muscle Changes
Degenerative muscle changes (DMCs) were the most fre-
quently identified lesions (115/153; 75.2%).
Capture myopathy in active-stranded animals, which died
shortly after the stranding or after a period of rehabilitation,
was the most commonly identified underlying disease associ-
ated with acute skeletal rhabdomyolysis in our study
(40.9%).42,43
302 Veterinary Pathology 54(2)

Figures 1–3. Muscle atrophy. Figures 1 and 2. Cachexia/inanition atrophy. Figure 1. Active stranded juvenile male of false killer whale.
Generalized muscle atrophy. Atrophied myofibers were variably flattened and angular (arrowheads). Type II fibers (red) were primarily affected.
Fiber type II IHC. Figure 2. Denervation atrophy. Active stranded aged female common dolphin. Myofiber atrophy, with focal concentration of
myofiber nuclei, adjacent to an intramuscular nerve (asterisk). PAS. Multifocal areas of type 1 fiber grouping were observed (inset). Fiber type II
IHC. Figure 3. Stranded adult female fin whale. Generalized muscle atrophy primarily, but not exclusively, involving type II fibers, in which
hypertrophic muscle fibers were occasionally observed (asterisk). Fiber type II IHC. Figures 4–9. Muscle degeneration. Figure 4. Passively
stranded adult female short-finned pilot whale affected by a traumatic process. Hemorrhages and acute monophasic myonecrosis. HE. Acutely
degenerated myofibers showed intense intrafibrillar fibrinogen positivity (inset). Fibrinogen IHC. Figure 5. Passively stranded adult male rough-
toothed dolphin with nonsuppurative leptomeningitis. Flavobacterium spp. were isolated from the skeletal muscle sample. Scattered acutely
degenerate myofibers showed intense intrafibrillar fibrinogen immunolabelling. Inset: magnification of an affected immunstained myofiber.
Fibrinogen IHC. Figure 6. Passively stranded adult male Sowerby’s beaked whale. Polyphasic myocyte degeneration and necrosis, in which
acutely degenerated fibers (asterisk) and regenerating myofibers, characterized by central rows of large nuclei (arrowheads), were observed
within the same field. HE. Figure 7. Passively stranded adult female Cuvier’s beaked whale affected by the fat and gas embolic syndrome. Acute
monophasic myonecrosis and replacement of interstitial connective and adipose tissue (arrowheads) by large amounts of amorphous hyaline
material. HE. Inset upper right: hyaline material and gas bubbles (asterisk) are present within the interstitial connective and adipose tissue. PTAH.
Inset lower right: passively stranded juvenile male Cuvier’s beaked whale affected by the fat and gas embolic syndrome. Acute monophasic
myonecrosis. Fibers in advanced degeneration with hyaline filled sarcolemmal sheaths. HE. Figure 8. By-caught juvenile male Atlantic spotted
dolphin. Altered sarcolemmal permeability, evidenced by large amounts of myoglobin-positive globules between the unaffected segments of the
myofibers. Myoglobin IHC. Figure 9. Degenerative changes of undetermined cause. Passively stranded adult female Atlantic spotted dolphin.
Edema and primary type I fiber degeneration with intercellular globular accumulation of muscle cell components (asterisks). PTAH. Inset:
calcificated degenerated myofiber. von Kossa.

Physical trauma was the second most frequent etiological frequently observed upon gross examination of traumatized
diagnosis (18.3%) associated with active degenerative muscle muscle areas, which correlated with histological diagnoses of
lesions in our study. Hematomas and hemorrhages were hemorrhages and acute monophasic myonecrosis affecting
Sierra et al 303

Table 2. Muscle Degenerative Changes Associated With Systemic

Infection or Septicemia.

Specie n Necropsy Report CA

Globicephala 1 Septicemia NP
macrorhynchus
Kogia breviceps 1 Septicemia NP
Mesoplodon bidens 1 Septicemia NP
Mesoplodon densirostris 1 Herpesvirus PCR
infection
Mesoplodon europaeus 1 Septicemia NP
Stenella coeruleoalba 3 Nonsuppurative NP
meningoencephalitis
(1/3)
Septicemia (2/3)
Steno bredanensis 1 Nonsuppurative Skeletal muscle:
leptomeningitis Flavobacterium
spp.
Tursiops truncatus 1 Septicemia NP
Abbreviations: CA, complementary analysis; NP, not performed; PCR,
polymerase chain reaction.
Figures 10–11. Electron microscopy of degenerative changes in ske-
letal muscle. Figure 10. Actively stranded juvenile male short-finned
both types of myofibers (Fig. 4). Affected myofibers often pilot whale. A hypercontracted myofiber (asterisk) is surrounded by
unaffected larger myofibers. Bar ¼ 2 mm. Figure 11. Actively
displayed a hyaline and/or a floccular appearance, contraction
stranded juvenile female bottlenose dolphin. Contraction band necro-
band necrosis, fragmentation of myofibers and discoid degen- sis corresponds with thin hypercontracted portions of the sarcoplasm
eration. Discoid degeneration was exclusively observed in (arrowheads) within the affected myofibers. Bar ¼ 10 mm.
animals in which sharp trauma from ship strikes was the cause
of death.
Systemic infection or septicemia was the third most com- fishing-related trauma. Moreover, altered sarcolemmal perme-
mon cause of myonecrosis (8.7%) in our study and was found ability, evidenced by large amounts of myoglobin-positive glo-
in 8 different species (Table 2). The related histological muscle bules between the unaffected segments of the myofibers, was a
findings ranged from scattered acute single-myocyte degenera- common finding with both of these causes (Fig. 8).
tion and necrosis (Fig. 5) to, more frequently, diffuse and poly- Degenerative muscle lesions of undetermined etiology rep-
phasic myocyte degeneration and necrosis (Fig. 6). resented 15.6% of the cases and were found in 18 specimens of
Regenerating myofibers were commonly encountered along 10 different species. Despite the heterogeneity of this group,
with acute degenerative changes. An inflammatory reaction, some common features were shared among some animals.
mainly composed of reactive and phagocytizing macrophages, Notably, 3 animals in this group showed acute degenerative
was frequently observed within the affected areas. changes that primarily affected type I fibers, which were sim-
Other causes associated with DMCs in the present study ilar to those observed in capture myopathy (Fig. 9). Calcifica-
included fat and gas embolism syndrome (8.7%) and interac- tion (von Kossa-positive) of a necrotic myofiber was noted in 1
tions with fishing activities (7.8%). of these 3 individuals (the only detected case of calcification in
Those cases with an etiological diagnosis of fat and gas the present study) (Fig. 9, inset). Interestingly, 9 of the 18
embolism as the cause of stranding/death not only displayed muscle samples in this group showed additional histological
degenerative changes in their muscle fibers (acute monophasic features compatible with muscle atrophy. In the remaining
myonecrosis) but also in the interstitial connective tissue and 6 animals, a cause for the DMCs was not apparent even thought
intramuscular nerves. Fibers in advanced stages of degenera- the changes were severe in some cases.
tion frequently appeared as hyaline-filled sarcolemmal sheaths, Degenerative muscle lesions typically showed immunohis-
and the interstitial connective tissue was replaced by similarly tochemical depletion of the sarcoplasmic myoglobin coupled
large amounts of amorphous hyaline material (Fig. 7). Intra- with intra and extracellular globular accumulation of myoglo-
neural and intermyofiber edema was a consistent finding in this bin and sarcoplasmic fibrinogen deposition (all of these
group of animals. strongly support an antemortem degeneration).
An interaction with fishing activities was considered to be Some of these changes were confirmed by the electron
responsible for DMCs in 9 individuals, including 5 different microscopy study. The ultrastructural changes depended on the
species. In these cases, entanglement (55.6%) and/or a lethal stage of the degeneration process, and included the formation
interaction with any other fishing gear (44.4%) was determined of contraction band necrosis, fragmentation of myofibrils,
as the cause of death. Entanglement caused milder and more mitochondrial swelling, and the disappearance of the myofi-
discrete histopathological muscle lesions than any other brillar structure in necrotic fibers (Figs. 10, 11).
304
Chronic Myopathic Changes
Chronic myopathic changes, detected in 39 of the 153
(25.5%) animals, included ring fibers (Fig. 12), increased
fiber size variation (Fig. 13), longitudinal splitting in hyper-
trophied fibers (Fig. 13, inset), and the presence of juxta-
nuclear lipofuscin.
Complex Polysaccharide Inclusions
Complex polysaccharide inclusions, as previously reported,78
were observed in 26 specimens representing 11 different spe-
cies. An association was established with histological skeletal
muscle lesions that was compatible with rhabdomyolysis, and
generalized or multifocal muscular atrophy in 20 and 12 of the
26 animals, respectively.
Parasitic Infestation
Parasitic infestation of skeletal muscle was observed in 14 of
the 153 (9.2%) animals, including Sarcocystis spp. (8.8%) and
Toxoplasma gondii (0.7%).
Sarcocystis spp. were detected in at least 1 specimen of 10
different species: fin whale (Balaenoptera physalus), short-
beaked common dolphin (Delphinus delphis), Risso’s dolphin
(Grampus griseus), pygmy sperm whale, Sowerby’s beaked
whale (Mesoplodon bidens), sperm whale, striped dolphin,
Atlantic spotted dolphin (Stenella frontalis), spinner dolphin
(Stenella longirostris), and bottlenose dolphin. Two species
were represented by more than 1 individual: the striped dolphin
(n ¼ 3) and the Atlantic spotted dolphin (n ¼ 2). Histologically,
the protozoal cysts appeared round to oval and were located
within myofibers. There was no detectable inflammation in any
case (Figs. 14–15). Cysts contained metrocytes or bradyzoites
and were surrounded by a fibrinogen-positive wall (Fig. 15,
inset). Cysts varied in size, ranging from 43.5 mm  28.3 mm
to 113 mm  121.7 mm.
Muscle parasitization by T. gondii was only detected in an
adult Atlantic spotted dolphin. The protozoal cysts were intra-
cellular, affecting the myofibers and intramuscular nerves.
They were typically ovoid-shaped and approximately 8.6 mm
 8.6 mm size (Fig. 16). They stained positive with PAS and
showed positive immunolabeling with a specific monoclonal
antibody. Moderate interstitial lymphohistiocytic myositis was
associated with these zoites.
Myositis
Myositis was detected in 3 of the 153 (1.9%) animals. All of
these cases showed multifocal pyogranulomatous inflamma-
tion associated with a systemic inflammatory process of bac-
terial origin (Fig. 17). Nocardia farcinica were isolated from
the lung tissue of an adult striped dolphin and Staphylococcus
spp from the pulmonary tissue of an Atlantic spotted dolphin.
In the remaining case, a juvenile false killer whale (Pseudorca
crassidens), the etiologic agent was not determined, although a
Veterinary Pathology 54(2)
penetrating wound was observed in the adjacent skin and sub-
cutis suggested a secondary bacterial infection.
Sarcoplasmic Masses
The presence of sarcoplasmic masses within skeletal myocytes
in an adult pygmy sperm whale has been previously reported.74
Other histopathologic muscle changes in that case included a
high number of internal nuclei, variations in fiber size and
shape, and a predominance of type I fibers compared to that
in the control group.
Discussion
Very little information regarding muscle pathology in marine
mammals was available at the time of this study. Our results
indicate that muscle lesions are more common in cetaceans
(92.2%) than in other domestic and wild species based on the
small amount of published data in terrestrial mammals.81,86 In a
postmortem study in equids, 65% of the analyzed specimens
showed muscle lesions of 1 or more type at necropsy,86 exclud-
ing the polysaccharide storage myopathy, which was previ-
ously reported on the same population (44.1%).88 In equids,
myonecrosis was the most common finding (17%) followed by
chronic myopathic changes (15.7%), generalized muscle atro-
phy (13.1%), intramyofiber protozoa (8.3%), denervation atro-
phy (6.1%), and neoplasia (1.3%), as well as other conditions.86
In wild animals, a high incidence of muscle lesions have been
previously described in the pichi (Zaedyus pichiy) (76.5%).
These lesions were mainly due to rhabdomyolysis (66.9%),
Sarcocystis parasitization (10.3%), and inflammation (6.2%).81
Determining the cause of muscle injury based exclusively
on gross and/or microscopic findings is seldom possible. Addi-
tional tests and life histories are often required. However, this is
particularly difficult in free-ranging or stranded cetaceans,
where their life history is frequently lacking and only necropsy
data are available. In our study, the muscle condition classifi-
cation and establishment of a correlation with an etiological
diagnosis for the stranding/death were deemed necessary in
each case. This allowed us to better understand the underlying
pathogenic mechanisms of muscle pathology, as well as the
pathological significance and the repercussions of muscle
lesions in these marine mammals.
Degenerative Muscle Changes
Degenerative muscle change was the most common finding in
our study (75.2%), as was the case in the 2 previous terrestrial
species.81,86 However, rhabdomyolysis is multifactorial in ori-
gin, and possible causes include sepsis, muscle compression,
seizures, trauma, hypothermia, and severe exertion.33
In our study, degenerative muscle lesions were observed in
association with a variety of disorders, including capture myo-
pathy syndrome, 42,43 trauma, septicemia, gas- and fat-
embolism syndrome and interactions with fishery activities.
Sierra et al 305

Figures 12–13. Chronic myopathic changes. Figure 12. Passively stranded adult female Cuvier’s beaked whale. Ring fiber characterized by
peripheral rings around maloriented myofilaments. PTAH. Inset: actively stranded male Atlantic spotted dolphin. Internal nuclear migration
(asterisks) and a ring fiber (arrowhead). HE. Figure 13. Actively stranded adult female bottlenose dolphin. Numerous hypertrophic myofibers
with myofibrillar disorganization (arrowheads). HE. Inset: hypertrophic fiber with longitudinal splitting. PAS. Figures 14–15. Sarcocystis spp.
Figure 14. Passively stranded adult female pygmy sperm whale. A round protozoal cyst composed of compartmentalized arrangement of zoites
with septal partitions. PTAH. Figure 15. Passively stranded adult male common dolphin. A enlarged oval type I myofiber containing a single
306
Active muscle degeneration was mainly associated with a
stressful situation in 47 of the active-stranded cetaceans
(40.9%), as has been previously reported.42 In addition to the
degenerative lesions observed in the skeletal muscles, this
group of animals also displayed hemodynamic lesions indica-
tive of multiorgan vascular shock, degenerative muscle lesions
affecting cardiac muscles and myoglobinuric nephrosis typical
of capture myopathy (CM).42 Terrestrial and marine wildlife
experience CM, often with fatal consequences, after human
capture/rescue interactions, as described previously,42,80 and,
in the case of active-stranded cetaceans, stress-associated myo-
pathy contributes to the comparatively low success rate of
rehabilitation.68,96
Degenerative muscle changes related to traumatic processes
(18.3%) were mainly attributed to inter/intraspecific interac-
tions and to ship collisions,76 representing 33.3% and 61.9%
of the cases, respectively. The first category refers to animals
with gross lesions that varied from superficial parallel skin
marks, due to inter/intraspecific interaction, to severe traumatic
lesions represented by fractured ribs and subcutaneous and
pulmonary hemorrhages.5 The second category includes ani-
mals that suffered severe trauma that resulted in peracute or
acute death associated with a collision with a ship. The cause of
trauma was not determined in the remaining 4.7% of the trau-
matized cetaceans with skeletal muscle degeneration in our
study. Interstitial edema, hemorrhages and severe, diffuse,
acute monophasic myonecrosis were the main histopathologi-
cal findings in this group of animals, although the major and
more distinguishable features were discoid degeneration or
fragmentation of the myofibers. This type of myodegeneration
was observed predominantly in the cetaceans in which the
sharp trauma was deep enough to cause evisceration or bone
fractures.76 An acute stress response was mainly, although not
exclusively, implicated in the pathogenesis of DMCs in this
physically traumatized group of animals.71,72 The typical stress
response is activated after a wounding or painful trauma.18,37
Although physiological stress has some benefits, an extreme or
prolonged response with the continued release of catechola-
mines and sustained, elevated blood cortisol and aldosterone
levels are potentially damaging to the heart and the skeletal
muscle.10,20 In addition to stress, rhabdomyolysis is a recog-
nized complication of traumatic injury,25,45 in which ischemia,
reperfusion and/or vascular damage are the main pathophysio-
logical mechanisms underlying muscle damage.92
Degenerative muscle changes related to infectious processes
(8.7%) were characterized by scattered single-cell necrosis in
our study. Myodegenerative changes associated with a sys-
temic infection or septicemia have been previously reported
in humans and animals33,41 and have been attributed to direct
muscle invasion by the pathogens, direct damage to muscle
fibers by endotoxins, and/or the increased levels of
Figure 15. (continued) protozoal cyst. Fiber type II HC. Inset: the cyst was surrounded by a fibrinogen-positive wall. Fibrinogen IHC.
Figure 16. Passively stranded adult male Atlantic spotted dolphin. Moderate interstitial lymphohistiocytic myositis was associated with T.
gondii. HE. Inset: An intracellular protozoal cyst. HE. Figure 17. Myositis. Passive stranded adult male striped dolphin. Multifocal pyogranulo-
matous inflammation associated with Nocardia farcinica. PAS. Inset: Nocardia (detected as gram-positive filamentous bacteria). Gram stain.
Veterinary Pathology 54(2)
inflammatory mediators, such as tumor necrosis factor alpha
(TNF-a).54 Acute, multifocal degenerative changes were also
present in muscle samples from animals where gas- and fat-
embolism syndrome was the cause of stranding/death in our
study. Animals with gas- and fat-embolism syndrome included
10 Cuvier’s beaked whales (Ziphius cavirostris) that had evi-
dence of lesions consistent with a severe decompression pro-
cess associated with the use of sonar during military
maneuvers.29,30 Diffuse, mild to moderate acute monophasic
myonecrosis (hyaline degeneration) was a common feature in
this group of animals, although additional pathological findings
included the ‘‘disintegration’’ of interstitial connective tissue
and related structures (intramuscular nerves, adipose tissue,
and/or intrafusal muscle fibers) and their replacement by large
amount of amorphous hyaline material, which was assumed to
be degraded material. A previous association between arterial
gas embolism, decompression sickness, and rhabdomyolysis
(as suggested by elevated levels of plasma creatine kinase) has
been reported.64,73,79 Although the mechanisms leading to the
release of muscle enzymes after arterial gas embolism remain
unclear, it has been suggested that gas emboli entering the
systemic circulation and reaching the skeletal muscle is the
most plausible explanation for the muscle hypoxic and
ischemic damage that occurs in the muscle. Decompression
sickness is the result of the supersaturation of body tissues with
nitrogen gas and the subsequent release of nitrogen bubbles to
the bloodstream. The amount of gas dissolved in specific tis-
sues depends on the dive depth and duration, the descent and
ascent rates, the lipid content of the tissue, and time at the
surface between successive dives.31 Tissues with a high lipid
content can take up a larger amount of nitrogen because of its
higher solubility in lipids than in any other biological tissue.36
A variable fatty component in the interstitial connective tissue
has been observed in the deep-diving cetaceans that were
examined in this study (mainly pygmy sperm whales, short-
finned pilot whales, sperm whales, and the Ziphiidae family)
compared with that of other species.77 The disruption of inter-
stitial fat depots may result in the release of nitrogen gas bub-
bles from supersaturated tissues and could explain the acute
muscle degenerative lesions observed in this group of animals
when suffering from decompression sickness.
An interaction with fishing activities (ie, accidental capture
[bycatch], and/or severe injuries directly caused by fishermen
or fishing equipment) was the etiological diagnosis in 9 ceta-
ceans with a morphological diagnosis of skeletal muscle
degeneration (7.8%). In our study, although no distinctive
degenerative findings were detected in this group of animals,
the leakage of myoglobin strongly supports myofiber damage.
Rhabdomyolysis can be defined as the disruption of skeletal
muscle leading to leakage of intracellular muscle constituents,
especially myoglobin, into the extracellular fluid.45,50 It has
Sierra et al
been proposed that tuna purse-seine and net fishery operations
could elicit short-term responses in dolphins, including severe
muscle damage, resulting in a condition similar to CM, and
hyperthermia.14,94 Capture myopathy is a multifactorial, patho-
physiological syndrome associated with stress, excessive mus-
cular activity, heat, trauma, prolonged muscle compression,
restraint and being transported. In by-caught animals, some
of these mechanisms could contribute to the muscle damage.
Despite the fact that there is currently not full descriptions of
skeletal muscle disorders in net-caught cetaceans, signs rang-
ing from minor temporary effects (eg, muscle stiffness) to mas-
sive muscle damage and death have been sporadically
described in some individual cases.66 In addition, net-caught
cetaceans probably share common behaviors, such as extreme
and desperate struggles to escape, which often causes severe
trauma and eventual death by asphyxiation. Related to this,
near-drowning in humans has been shown to induce rhabdo-
myolysis, and the potential causes include hypothermia, phys-
ical exercise and tissue hypoxia.8
The cause of myonecrosis was not determined in 18 indi-
viduals, although CM syndrome was suspected in 3 cases.
Despite the fact that these cases were found dead-stranded, the
fresh state of the carcasses along with multiple cutaneous ero-
sions, ulcerations and/or abrasions on the head of all or some
of these animals, strongly suggested an active stranding in all
cases. Histological features of muscle atrophy were observed
in 50% of the animals in which the cause of myonecrosis was
not apparent.
One exception for myonecrosis in cetaceans from our study,
when compared with myonecrosis in other species, is that post
necrotic calcification (dystrophic mineralization) was infre-
quent. Myofiber mineralization is considered a nonspecific
sequela of myofiber necrosis, in which diverse circumstances
are involved.91 Since overt mineralization was rarely observed
within the injured skeletal muscle from the stranded cetaceans
in our study, the causes of this difference require further
investigation.
Stress associated with capture and handling and endo-
toxic injury from bacterial infection were the main under-
lying causes of rhabdomyolysis in wild pichis and equids,
respectively.81,86,90 However, although rhabdomyolysis may
be caused by a wide range of mechanisms, it is always
characterized by a loss of muscle integrity and subsequent
leakage of intracellular enzymes and electrolytes into the
bloodstream, which can be life-threatening.83 Therefore,
regardless of the etiology and depending on the severity
of the lesions, rhabdomyolysis can adversely affect the
health status of the affected cetaceans.
Muscular Atrophy/Hypertrophy
Generalized muscle atrophy was related to senescence in
56.9% of the cases with a morphological diagnosis of muscle
atrophy. This group of animals was considered ‘‘senile’’
according to the available epidemiological data. Loss of skele-
tal muscle mass can occur during aging (sarcopenia) in humans
307
and animals, including cetaceans, as we have previously
reported,75 although the many potential cellular and molecular
mechanisms that underlie sarcopenia are unclear. Oxidative
stress, inflammation, endocrine disorders, malnutrition, and
inactivity have all been proposed as responsible cofactors for
muscular senescence in humans and animals.59
Generalized muscle atrophy was detected in 17 specimens in
association with an etiological diagnosis of cachexia/inanition in
our study, which represented 29.3% of the atrophy-related
causes. The term atrophy implies a reduction in either the dia-
meter of the muscle as a whole or in the diameter of a myofi-
ber.91 Both conditions were observed in this group of animals, in
which type II fibers were preferentially affected. Histological
features of the muscles of this group of animals were similar
to previous descriptions in sheep and human beings that were
associated with consumptive disease and inanition.46,70
Cachexia is a metabolic condition associated with an underly-
ing illness and inflammation,27 which cannot be completely
reversed by nutritional interventions. Limited success has been
shown in preserving fat but not muscle mass in these cases.57 In a
similar way, inanition atrophy is due to reduced caloric intake and
results in a general reduction in muscle fiber size, although recov-
ery is typically relatively quickly when nutrition is restored. An
identical histological pattern was detected in all the specimens
in this group regardless of the cause of the atrophy, which was
either inanition (foreign bodies that prevented feeding, such as
a long and thick cable wrapped around the tongue of a fin
whale, or a strand of plastic wrapped around the body of the
mandible in a sperm whale) or chronic systemic diseases.5
Generalized muscle atrophy related to inactivity (disuse)
was recognized in only 1 animal in the present study. This case
displayed external signs of scoliosis, a condition previously
described in cetaceans.7,19 It was not possible to determine the
cause of this malformation, although it could be 1 of the several
causative factors previously associated in documented cases of
vertebral column malformation in cetaceans, including stress or
exertion, spondylodiscitis or spondylo-osteomyelitis, trauma,
or congenital malformations. The etiopathogenic mechanisms
underlying the atrophy of both fiber types in this case might be
related to abnormal swimming patterns and/or starvation due to
impaired feeding.
Although starvation, cachexia, and sarcopenia can be
defined as distinct clinical syndromes, a certain degree of over-
lap might have occurred in animals from the present study, as
has been previously suggested.70 Regardless of the etiology,
muscle atrophy has important implications for the biomecha-
nical performance and health status because a reduction in fiber
size is related to lower force production.93
Scattered fiber necrosis has been described as an additional
histological change in cachexia,95 aggravating the wasting pro-
cess. Multifocal muscle atrophy associated to denervation atro-
phy was detected in 7 specimens, representing the 16.7% of the
atrophy-related etiologies, in which identical histological
features (group atrophy and fiber type grouping with no fiber
type predisposition) to previous reports were observed.23,89
Involvement of both types of fibers was confirmed by IHC
308
(fiber typing) in 6/7 animals. Although determination of fiber
types is necessary to confirm denervation atrophy,24 small
group atrophy and fiber type grouping are virtually pathogno-
monic of denervation atrophy following reinnervation.56,89
More than half of the specimens in this group were consid-
ered senile (based on morpho-pathological findings) and myo-
fiber atrophy might have been partially attributable to aging, as
has been previously proposed.69 Denervation atrophy of skele-
tal muscle is a consequence of damage to lower motor neurons
and results in focal areas characterized by fibers with reduced
size, although selective atrophy of a specific fiber type does not
generally occur.23 Fundamental physiological and metabolic
skeletal muscle changes have been described as occurring after
damage to the nerve supply.32
As found in our study, fiber hypertrophy often accompanies
fiber atrophy, which contributes to increased variation in fiber
size.91 Pathologically hypertrophic fibers have decreased oxy-
gen diffusion from the interstitial capillaries to the internal
regions. This leads to fiber splitting to ensure an adequate
oxygen supply. Hypertrophied fibers could also undergo seg-
mental necrosis if overloading occurs.91 Type II fiber atrophy
together with enlarged type I fibers, resulting in similar sized
fiber types, have been described in the aged animals from our
study. This likely represents an adaptive mechanism related to
the diving abilities of cetaceans.75
Chronic Myopathic Changes
Chronic myopathic changes were the third most common
finding (25.5%) in our study and preferentially involved
specimens with concomitant age-related muscle changes,
such as senile or denervation atrophy.75 Some of the CMCs,
in particular, increased myofiber size variation and the num-
ber of internal nuclei, have been defined as nonspecific
findings that are associated with a variety of neuromuscular
disorders.23 However, they have also been shown to be
related to aging in equids86 and humans,49 an association
that is also apparent in the cetaceans examined here. In
addition, the presence of lipofuscin and lipofuscin-like gran-
ules has been observed in a variety of pathologic and phy-
siologic conditions, including aging.47
Chronic myopathic changes are nonspecific changes
that are mainly related to advanced age, which could vari-
ably affect the fiber integrity and viability. However, the
functional significance of some of these changes are cur-
rently unknown.91
Complex Polysaccharide Inclusions
The presence of complex polysaccharide inclusions, which has
previously been reported,78 was the fourth most common myo-
pathic change identified (22.6%) in our study. An interesting
finding was that 69.2% of the animals with CPI displayed other
muscle atrophies described in our study, as follows: senile
atrophy (50%), denervation atrophy (27.7%), cachexia/inani-
tion (16.6%), and disuse atrophy (5.5%). All of the animals
Veterinary Pathology 54(2)
affected by senile atrophy in our study were older adults, which
explains why some of them also had complex polysaccharide
depositions; a condition previously reported to be associated
with aging in other species.39,87 Muscular atrophy and neuro-
muscular disorders have also been reported in horses with poly-
saccharide storage myopathy.58,91 In addition, 46.15% of the
animals with CPI displayed degenerative changes associated to
the stranding stress syndrome. This association introduces the
intriguing possibility that a polysaccharide storage disease
affecting myocytes might predispose some cetaceans to
capture-related myopathy, which has been reported to occur
in exertional rhabdomyolysis in horses.68,78
Parasitic Infestation
Skeletal muscle parasitism was observed in 9.2% of the ani-
mals in our study. Intrasarcoplasmic protozoan cysts of Sarco-
cystis spp were observed in 13 specimens from 10 different
species. This represents the first description of this type of
infestation in 8 species: Atlantic spotted dolphin, short-
beaked common dolphin, common bottlenose dolphin, spinner
dolphin, pygmy sperm whale, fin whale, Sowerby’s beaked
whale, and Risso’s dolphin. Our results suggest a lower pre-
valence in these species compared to the relatively high pres-
ence of muscle sarcocystosis found in stranded Atlantic white-
sided dolphins (39%). 28 No inflammatory infiltrate was
detected in any of the muscle tissue samples displaying vari-
able Sarcocystis parasitization, suggesting that this protozoan
is largely an incidental finding with non apparent pathogenic
relevance. Although the life cycle of Sarcocystis spp in marine
mammals is unknown, it has been suggested that dolphins are
probably intermediate hosts for this parasite.28
Tissue cysts containing bradyzoites of T. gondii were only
observed in the skeletal muscle of 1 specimen in our study, an
adult male Atlantic spotted dolphin. To our knowledge, this is
the first description of skeletal muscle toxoplasmosis in a ceta-
cean species. Two major presentations for T. gondii infection
have been reported in different cetacean species: localized
encephalitis, and systemic infection,12,21,22,48,60,67 including
in the Atlantic spotted dolphin.5 T. gondii is able to infect a
broad spectrum of cell types, although neurons and muscle
cells are considered the preferred cell type for parasite persis-
tence during infection, which in turn affects their transmission
to new hosts.82 A major route of transmission of T. gondii is the
ingestion of contaminated food from chronically infected inter-
mediate hosts, which in turns relies on the development of a
bradyzoite stage. These can survive within muscle tissue cysts
for at least 31 days.38 The life cycle of T. gondii in marine
mammals remains unsolved. Although it is a matter of specu-
lation at this time, the role of skeletal muscle toxoplasmosis in
Toxoplasma transmission is conceivable.
Myositis
Myositis was uncommon in the current study and was primarily
related to bacterial infections. In 2 of the 3 specimens with
Sierra et al
myositis from this group, bacterial myositis was related to a
systemic infection due to Nocardia farcinica and Staphylococ-
cus spp,5 while no microbiological analysis was performed in
the remaining case. It is possible that several opportunistic
bacteria might have enter through the wound contributing to
the development of the lesions.
Several cases of pyo- and polymyositis have been reported
in humans associated with Nocardia septicemia.3,40,61 How-
ever, muscle involvement in bacterial infections is relatively
uncommon and may result from contiguous sites of infection,
penetrating trauma, vascular insufficiency, or hematogenous
dissemination.13 Associations between the route of infection
and the infecting organism have been recognized. For instance,
in human beings, acute bacterial infections of skeletal muscle
secondary to hematogenous spread are mostly associated with
Staphylococcus aureus, whereas infections as a result of pene-
trating wounds are often polymicrobial.13
Depending on the etiological agents responsible for the
infection and on the severity of the lesions, myositis in ceta-
ceans can have different outcomes, with some fatal cases hav-
ing been reported.97
Sarcoplasmic Masses
The histopathological findings observed in the muscle of a
pygmy sperm whale were comparable to those classically
described as typical of myotonic dystrophy in humans and
other animals.74 The pathogenic relevance of this myopathy
in the present case remains uncertain, although it might have
contributed to the increase in the reported cases of primary
muscle disorders in wild animals.
Conclusion
Our results provide evidence for a wide spectrum of muscle
changes in wild cetaceans, which include lesions associated
with systemic disorders (eg, generalized atrophy in consump-
tive processes, or bacterial myositis), incidental findings (eg,
Sarcocystis spp parasitization), and degenerative processes of
variable severity that might involve other organs (eg, myode-
generation and necrosis with subsequent myoglobinuric tubu-
lonephrosis) and indicate a grave prognosis, aggravate the
underlying disease processes, or have a negative impact on
rehabilitation attempts.42,43
Skeletal muscle is easily sampled during necropsy examina-
tions and provides essential microscopic information on both
local and systemic pathological conditions. Accordingly, ske-
letal muscle should be systematically sampled, processed and
examined in all stranded cetaceans.
In conclusion, muscle lesions have been shown to be com-
mon among free ranging cetaceans. Because myopathic disor-
ders can adversely affect their locomotion,86 these conditions
are highly significant for cetaceans, which strictly depend on
effective skeletal muscle function for survival and the perfor-
mance of specific daily activities in their aquatic environment.
309
Acknowledgements
The authors thank the members of the different stranding networks:
Canary Islands (the SECAC and the Canarias Conservación), Almeria,
and United Kingdom.
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) disclosed receipt of the following financial support for
the research, authorship, and/or publication of this article: This study
was possible thanks to the partial funding by National Project
CGL2012-39681 (Subprograma BOS); Regional Project SolSub
C200801000288, and ProID 20100091; Technical Assistant Contract
by Canary Islands Government delegation (TEC0002955); and pre-
competitive project ULPGC2013-21.
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