HISTOPATHOLOGY REPORT

DATE: 26/10/18
REPORTING PATHOLOGISTS: Celine Berjot (440456326), Cheryl Chan and Lesca Sofyan
PATIENT: 8yo M Canine
SPECIMEN: Specimen #3: Bronchi (transverse section)

MACROSCOPY: Not provided

MICROSCOPY:

At low magnification, two transverse sections of two bronchi is visible, with cartilage rings that form a
Y between both bronchi, indicating that this section is taken from the site of bifurcation of the trachea
into two bronchi. Both sections appear to be very similar, therefore only one will be reported on. Both
bronchi have abnormal large, highly cellular and vacuolated masses (3.4mm x 4.9mm for the left mass)
attached to the mucosa and protruding into the lumen. One nodule is proximally and centrally situated
on the lumen of the bronchi whilst the other nodule is protruding on the left distal rim of the bronchi.
Both seem to be of a similar size.

The lining of the bronchus seems to be composed of a corrugated mucosa internally, a submucosa,
several overlapping layers of incomplete cartilage ring, and a serosa externally. The masses seem to
have disrupted the mucosa lining and have extend the submucosa into the lumen, resulting in disruption
to the ring of cartilage. A piece of cartilage is present in the tissue around the mass, which is abnormal.
A mass of basophilic tissue is present at the top of the slide.

Upon medium magnification, there is a pseudostratified columnar respiratory epithelium which lines
both bronchi and which lines the mass, indicating that the mass is a proliferation of submucosal tissue.
The epithelium is too damaged to observe live goblet cells, however the shape of degenerate cells
suggests a high degree of goblet cells (10-20%) suggestive that this section if of the proximal bronchus.
The nodules are supported by proliferation of submucosal tissue and cartilage, and consist of a dense
population of cells with a dozen circular structures dispersed through this, containing multiple circular
structures arranged with bilateral symmetry, which resembles typical sections through metazoans such
as worms. The different length of the various transverse sections indicates that the metazoan is
cylindrical and the sections varying angles of oblique.

The right nodule contains areas of strong fibrosis (fibrocytes and collagen fibres) in the shape of worm
cross-sections which seem to be either areas of successful immune response that have cleared the worm
or strongly fibrotic areas immediately adjacent to adult worms. The presence of fibrocytes and collagen
forming elongated shapes of connective tissue could suggest a post-infection where the host’s immune
system was able to eradicate the parasite, leaving behind some scarring after regeneration. The right
nodule predominates with fibrous tissues within the connective tissue matrix which might suggest a
previous lesion that has been resolved by a successful immune response.

At high magnification, the submucosa and the fatty serosa appear normal, while the pseudostratified
columnar epithelium appears very damaged, being thinner than usual, discontinuous, with necrotic cells
with areas having sloughed off. This could be due to the increased effort breathing due to the reduced
diameter of the lumen caused by the nodule, indicative that this condition is chronic.
The mass contains some capillaries and a small amount of irregular, dense connective tissue similar to
the normal submucosa. However, there is a large infiltration of round cells with abundant cytoplasm,
large nuclei with a clockwork chromatin pattern and perinuclear halos, and round cells with scant,
eosinophilic cytoplasm and dense, often bilobed nuclei, appearing to be plasma cells and eosinophils
respectively. Eosinophils are in particularly in high concentration around the worms. Large,
pleomorphic cells with finely granular, eosinophilic cytoplasm and sparse nuclei embedded within

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eosinophilic, fibrous tissue are found particularly around the worms, likely corresponding to be either
epithelioid macrophages or fibroblasts, or both, as these are often difficult to distinguish. The worms
themselves many round and coiled structures within a membrane which are thought to represent eggs
and larvae within uteri of gravid female worms. In the left bronchus, 23 worm cross-sections are present
in the nodule, 3 in the bronchial lumen, while only 5 cross-sections are found in the right bronchus.
There are therefore up to 31 worms visible on this specimen.

Blood vessels in the submucosa and mucosa appear to be hyperaemic, and small areas of haemorrhage
are visible from the mucosa, such as the eosinophilic tissue top left of the left nodule, between the
nodule and the normal mucosa. This eosinophilic tissue seems to contain dense, coiled structures which
could correspond to hatched larvae. Areas of dark red material, which could correspond to necrotic cell
debris and/or blood, are occasionally found in the mucosa. Some areas of the submucosa appear slightly
disorganised, with pale eosinophilic to unobservable nuclei, indicating low level of necrosis, although
tissue architecture is still maintained. A more basophilic area in the centre of the Y cartilage appears to
correspond to either to necrotic cartilage or to a staining artefact. More basophilic tissue at the top of
the slide, external to the serosa of the left bronchus, appears to be a dense population of small round
cells with scant cytoplasm and large, dense nuclei, which are likely small lymphocytes. Some plasma
cells and fibrocytes and collagen fibres, as well as capillaries are also present. This seems unrelated to
the parasite infection and no aetiological agent is visible, therefore the significance of this is unknown.

MORPHOLOGICAL DIAGNOSIS: Bifocal, chronic, nodular, granulomatous tracheobronchitis in

response to a parasite infestation of the bronchus submucosa at the site of bifurcation of the trachea into
the bronchi, which one nodule in each bronchus. The right nodule appears to be older and more
resolving than the left, which appears more actively infected. Although only part of the lumen is visible,
the nodules appear to be occupying about 30% of the lumen of each bronchus, which would likely be
causing severe physiological impairment.

AETIOLOGICAL DIAGNOSIS: Dog lungworm (Filaroides (Oslerus) osleri), causing verminous

nodular tracheobronchitis

PATHOLOGY COMMENTS:

Differential diagnosis: (in order of likelihood) Eucoleus aerophilus, Angiostrongylus vasorum (French
heartworm), Filaroides milksi (canine verminous pneumonia), Paragoniumus spp. (lung fluke),
Aleurostrongylus abstrusus, Dipetalonema reconditium

The nodules and their location at the bifurcation of the trachea into the bronchi, is quite characteristic
of Filaroides (Oslerus) osleri. O. osleri lungworms produce fibrous, granulomatous and firm nodules
that are encapsulated with one small pore for mature female eggs to lay (Taylor et al., 2014). Only E.
aerophilus can form tracheobronchial nodules, other dog lungworms infecting only the lung
parenchyma (Yao, et al., 2010), allowing these to be excluded. To confirm O. osleri infection, a
collection sample of the tracheal and bronchus mucus (by tracheal or bronchial wash) should be used
to identify L1, eggs and adult worms. Adults are 6.5 to 13.5mm in length, with a characteristic S-shaped
kink, and L1 larvae are around 250um in length, have an indentation in their tail and no dorsal spines.
Eggs are distinguished from E. aerophilus by their size (80um by 50um) (Conboy, 2009) and lack of
bipolar plugs (Yao, et al., 2010). Although faecal floats can be performed to identify eggs and larvae,
it less useful and not of gold standard as O. osleri eggs are shed intermittently (Taylor et al., 2009). To
differentiate O. osleri from its closely associated relative, A. vasorum, a sandwich enzyme-linked
immunosorbent assay detection of circulating antigens of A. vasorum is the most sensitive method,
although not having perfect sensitivity at only 92% (Conboy, 2009). Otherwise, performing PCR on
tracheobronchial wash samples can immediately confirm the species of the parasitic worm present.

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Although there is distortion in architecture and two prominent masses, neoplasia can be excluded from
the differentiation as the pathological features constituting a neoplasia such as neoplastic cells are not
present. Cells are regularly shaped, with no mitoses or distortion of architecture.
Evidence of tracheobronchial nodules can be revealed on lateral thoracic radiographs however, this
technique alone is not suitable for definite diagnosis.
Due to rareness of this pathogen, there is insufficient literature to confirm that degenerative changes, as
well as the accumulation of lymphocytes, seen in this specimen are typical of O. osleri infection.

REFERENCES:
Conboy, G., 2009. Helminth Parasites of the Canine and Feline Respiratory Tract. Veterinary Clinics
Small Animals, 39, pp. 1109-1126.
Yao, C., O’Toole, D., Driscoll, M., McFarland, W., Fox, J., Cornish, T., and Jolley, W., 2010.
Filaroides osleri (Oslerus osleri): Two case reports and a review of canid infections in North America.
Veterinary Parasitology, 179, pp.123-129.
Taylor, M.A., Coop, R.L., Wall, R.L., 2007, Veterinary Parasitology, third edition, Blackwell
Publishing, Oxford.