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Animal Science Papers and Reports vol. 36 (2018), no. 1, 33-44
Institute of Genetics and Animal Breeding, Jastrzębiec, Poland
New insights of canine mastitis ‒ a review
Ilona Kaszak1*, Anna Ruszczak1, Szymon Kanafa1,
Olga Witkowska Piłaszewicz2, Mariusz Sacharczuk3, Piotr Jurka1
1 Department of Small Animal Diseases with Clinic, Warsaw University of Life Sciences,
Nowoursynowska 166, 02-787 Warsaw, Poland
2 Department of Pathology and Veterinary Diagnostics, Warsaw University of Life Sciences,
Nowoursynowska 166, 02-787 Warsaw, Poland
3 Laboratory of Neurogenomics, Institute of Genetics and Animal Breeding,
Jastrzebiec, Postępu 36A, 05-552 Magdalenka, Poland
(Accepted January 25, 2018)
Disorders of the mammary gland are frequently diagnosed in bitches of all breeds and of all ages.
They usually appear in the postnatal period, during lactation peak or during false pregnancy. They
may also be related to dierent pathologies of the mammary gland, such as mammary neoplasia.
It is frequently underestimated by clinicians or misdiagnosed. A reliable diagnosis is the key to
success. Conventional diagnosis consists of a clinical diagnosis together with a standard blood test,
while in some cases cytological examination of the mammary gland is performed. In some cases
of mastitis, especially its subclinical presentation, those methods may prove unreliable. For this
reason, determination of specic inammatory biomarkers may enable clinicians to produce a
precise diagnosis, while it is very useful in treatment monitoring. A wide variety of inammatory
biomarkers have been extensively studied both in human and veterinary medicine. In this article,
we will describe the most commonly evaluated inammatory biomarkers - acute phase proteins,
which seem to be promising tools in the diagnostics of canine mastitis. We hope that this paper
will provide clinicians with new ideas for precise diagnosis and more specic treatment of canine
mastitis.
KEY WORDS: biomarkers / bitch / dog / inammation / mammary gland
*Correspoding author: ilonakaszak@gmail.com
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The mammary gland is a transformed skin gland. It is functionally connected with
the secretion of ovarian hormones secretions; therefore, ovarian pathologies may have
an inuence on the mammary glands. Bitches usually have 5 pairs of mammary glands
located symmetrically on both sides of the chest and abdomen, but abnormalities in
the position and number of glands may occur. The mammary gland is basically an
exocrine gland, which basic function is to produce milk to feed the ospring. It is
composed of epithelial cells, which form the lining of alveoli and ducts, surrounded
by myoepithelial cells. The stroma of the gland consists of blood and lymphatic
vessels, brous and fat tissue and nerve endings. The contraction of myoepithelial
cells produces milk secretion into the lumen of secretory ducts which drain in the
opening in the nipple. During the puberty, mammary gland development is related to
cyclic ovarian activity and hormonal changes, with the growth continued until sexual
maturity is reached [Marti et al. 2010, Momont et al. 2002].
Mastitis is a medical term, which refers to an inammation with or without an
infection of the mammary gland. Therefore, we can distinguish septic and aseptic
mastitis. It may involve one or more glands. Inammation of the mammary gland
is a common problem in bitches of all breeds at various ages. Despite that fact,
mastitis is frequently underestimated or incorrectly diagnosed. The pathogens are
usually bacteria, but some cases of fungal mastitis in endemic areas or in dogs with
immunodeciency were observed as well [Ditmyer et al. 2011, Murai et al. 2013].
Prevalence
It is more often observed in non-spayed bitches, as it frequently appears in
the postnatal period, during lactation peak or during false pregnancy. However, it
may also be related to dierent pathological states of the mammary gland, such as
galactostasis, mammary hyperplasia or mammary neoplasia. Sometimes, mastitis is
indistinguishable from a mammary tumor with present inammation. There are four
clinical presentations of mastitis: acute mastitis, gangrenous mastitis, chronic mastitis
and subclinical mastitis. Risk factors include: poor hygienic conditions, trauma and
systemic infections. The most common route of infection is the ascending route from
the nipple, while trauma and haematogenous route are less frequent. Trauma may
be produced by sucking puppies during lactation. In some cases, injuries involving
foreign bodies or insect bites may cause similar lesions. In extreme cases the bitch
may present no signs of the disease or may be already in a critical condition. The
mammary gland is usually changed, but in the case of a subclinical presentation it may
remain normal [Marti et al. 2010, Momont et al. 2002].
I. Kaszak et al.
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Causes of mastitis
Lactation
The term lactation refers to the beginning of milk production and secretion. It
is strongly related to hormonal changes during pregnancy and parturition. Due to
a decrease of progesterone secretion at the end of pregnancy, the concentration of
prolactin increases. Prolactin together with other factors is responsible for the initiation
of lactation. There are several factors that either stimulate (e.g. oxytocin, serotonin,
oestrogens) or inhibit (e.g. dopamine, somatostatin, progestogens) prolactin release
[Concannon et al. 1989]. Mastitis during lactation develops as a consequence of
mammary gland trauma caused by sucking puppies or due to poor hygienic conditions.
Trauma and formed skin lesions facilitate bacteria penetrations into the mammary
tissue. Post-partum metritis through the haematogenous route may also cause mastitis.
Mastitis during lactation can be life – threatening both for the bitch and the puppies
[Schafer-Somi et al. 2003].
New insights of canine mastitis
Fig. 1. Acute mastitis during lactation in a 3 years old Yorkshire Terrier.
False pregnancy
False pregnancy, also called pseudopregnancy is a physiological state of a bitch.
It is connected with hormonal changes (progesterone concentration) after oestrus that
are similar both in pregnant and non-pregnant bitches. The clinical signs are present
usually in 6-8 weeks after oestrus and include mammary gland enlargement, beginning
of milk production, change in behaviour to typical of pregnancy and lactation such
as nesting behaviour, anorexia or agitation. Secretion from mammary glands may be
physiological but sometimes it may even be brown and mastitis may appear. Usually
treatment is necessary, especially in cases of severe lactation, galactostasis, complicated
additionally by infection and inammation [Marti et al. 2010, Momont et al. 2002].
Galactostasis
Galactostasis is the overload of the mammary gland with milk that is seen before
the parturition or shortly afterwards. The enlargement of mammary gland is observed,
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which leads to failure of milk let-down. Animals usually show pain and discomfort,
with the progression of the disease aseptic or sceptic mastitis may develop. It is
observed in highly lactating bitches, especially at very early weaning of puppies.
The treatment consists in limited feeding of the animal for several days (fasting may
be recommended during rst day), cold packs or compresses with baking soda are
recommended, use of diuretics and glucocorticoids may be necessary. A prolactin
antagonist should only be administered in cases of galactostasis due to false pregnancy
or when the ospring was weaned.
I. Kaszak et al.
Mammary gland tumors
Tumors of the mammary gland represent the most frequent cases of neoplasia in
female dogs. Around half of them are malignant neoplasia. They occur in bitches at an
older age, usually between 8 and 10 years old [Benavente et al. 2016, Sorenmo 2003].
It is more common in intact bitches. It is known that ovariohysterectomy before rst
oestrus greatly reduces the risk of mammary tumor [Beauvais et al. 2012]. Still,
the aetiology of canine mammary tumor is unknown. Some risk factors have been
identied, among which the most frequent are: hormonal, nutritional and genetic.
Tumors are usually discovered when macroscopic changes of the mammary gland
are visible. Inammation is very common, especially at a prolonged process of tumor
development is long. The treatment of choice is mastectomy with resection of local
lymph nodes, however, because of mastitis, in some cases antibiotics together with
anti-inammatory drugs have to be administered prior to the surgery.
Fig. 2. Galactostatis after weaning in a young female dog.
Fig. 3. Enormous mammary tumor with gangrenous mastitis.
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Mammary gland hyperplasia
Mammary gland hyperplasia also called mammary hypertrophy or mammary
broadenomatosis is a non-neoplastic proliferative process. It is more typical in
queens than in bitches. It concerns young females, usually after the rst oestrus,
although it may also appear during pregnancy or false pregnancy. It is related to the
progesterone concentration [Martin et al. 2000, Momont et al. 2002]. It is thought that
the administration of progestogens plays an important role in its pathogenesis [Loretti
et al. 2009, Marti et al. 2010]. Mammary glands are usually rm and enlarged, it may
aect one, several or all glands. In severe cases, mastitis with ulcerative necrosis of
the skin may appear. Treatment should always be undertaken.
Clinical presentation of mastitis
Acute mastitis
In the case of acute mastitis mammary glands are usually hot and painful during
palpation, enlarged, swollen and erythematous [Marti et al. 2010]. If acute mastitis is
septic, clinical signs such as fever, apathy, depression, lethargy, anorexia and neglect
of puppies may appear. The puppies may present the toxic milk syndrome, which is
basically a bacterial infection of puppies due to maternal milk contaminated with
bacteria and their toxins. The secretion of the gland is brown, may contain small
amounts of pus and blood, its smell may be changed [Marti et al. 2010]. In cases of
acute mastitis urgent treatment is needed.
Gangrenous mastitis
Gangrenous mastitis is usually a consequence of untreated acute mastitis [Marti
et al. 2010]. It is characterised by pus production and abscess formation may appear.
Secondly, mammary glands may become ulcerated and necrosis may form. Altered
glands become darker, colder and have an unpleasant putrid odour. Signs of sepsis are
usually present at that time.
New insights of canine mastitis
Fig. 4. Severe gangrenous mastitis together with mammary gland neoplasia.
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Chronic mastitis
Little is known about chronic mastitis in dogs. It is typically a consequence of less
severe acute cases of mastitis or are related to mammary neoplasia. During clinical
examination, the gland might by slightly inamed or swollen and its consistency
during palpation may be heterogeneous. Chronic mastitis should be suspected in cases
of an increased rate of falls in new-borns and when the litter is not gaining weight
properly.
Subclinical mastitis
Subclinical mastitis is very dicult to diagnose, it is a challenge for clinicians
as no clinical signs are seen. Moreover, this form of mastitis occurs quite often
[Marti et al. 2010, Vasiu et al. 2015]. Sometimes the only presenting complaint is
slow progress of the ospring growth and their limited weight gain. Additionally,
puppies may also show signs of the toxic milk syndrome. It may develop and lead to
systemic inammation or sepsis. Therefore, subclinical mastitis cannot be neglected
and an early and precise diagnosis must be provided. The mammary gland remains
unchanged and the secretion may also be normal, so clinical diagnosis is usually
unhelpful. Diagnostic imaging may appear to be useful, but not always changes in
the mammary gland structure will be visible. Evaluation of specic inammatory
biomarkers in serum as well as in milk samples may represent the best non- invasive,
fast and very sensitive method of mastitis diagnosis [Vasiu et al. 2015]. In the next
section selected inammatory biomarkers used in the diagnosis of mastitis will be
described.
Diagnosis
If during a veterinary consultation we suspect mammary gland inammation,
rst of all we need to obtain relevant information from the owner. We need to ask:
how long the animal has been presenting this symptom, when was the parturition,
whether it was its rst parturition, how many puppies were born and/ or whether
it is the rst time the bitch presents a false pregnancy. To diagnose mastitis in a
bitch full clinical examination is required [Marti et al. 2010]. All mammary glands
should be evaluated, checking their symmetry, temperature, size, consistency and
skin colour. Secondly, a blood test together with sampling of secretion/ milk from the
abnormal mammary gland should be performed. In the case of secretion, we need to
evaluate its colour, consistency and smell. Cytological examination should always
be performed. In cases of chronic mastitis a microbiological culture of the secretion
together with drug susceptibility testing may be indicated. The most common isolated
pathogens are staphylococci, streptococci and E. coli [Murai et al. 2013, Seweryn
et al. 2009]. Apart from that, X-ray should always be considered, especially if the
I. Kaszak et al.
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mammary gland neoplasia is suspected, in order to exclude metastases to the lungs.
Also, ultrasonography of the mammary gland appears to be a very useful diagnostic
method for mastitis as well as mammary tumor detection. It can precisely estimate
the degree of inammation as well as detect abnormalities inside the gland [Trasch
et al. 2007]. Fine needle biopsy is usually recommended if mammary neoplasia is
suspected, though the obtained results may not always be very precise. Last, but not
the least, determination of specic inammatory biomarkers in serum might prove to
be highly informative.
Selected inammatory biomarkers used in the diagnosis of mastitis
Inammatory biomarkers are usually proteins, which can be measured in blood or
other tissues (e.g. milk) and provide information on the presence of the inammation,
results of treatment or further prognosis for the patient. They are also detectable in
the serum and tissues of healthy dogs, but their levels are signicantly lower. During
local or systemic inammation processes, inammatory cells secrete proinammatory
mediators such as cytokines and chemokines, which are released into systemic
circulation. A wide range of compounds are classied as inammatory biomarkers,
among which acute phase proteins, T-lymphocytes, macrophages, interleukins,
receptor tyrosine kinases, as well as cyclooxygenases play a particularly important
role. In the case of canine mastitis only the determination of acute phase proteins
has been briey described [Vasiu et al. 2017]. The results of this research proved
that evaluation of inammatory biomarkers of inammation may be very useful in
the diagnosis of mastitis, especially its subclinical presentations, which remain a
challenge for the clinicians. Additionally, their assay in cases of clinical mastitis may
be useful in monitoring the treatment outcome.
Acute phase proteins
Acute phase proteins (APP) are serum proteins, which concentrations change as
a part of innate host defence systemic response to infection, inammation or trauma
[Ceron et al. 2005, Eckersall et al. 2010, Paltrinieri et al. 2007]. APP can be divided
into positive, moderate and negative APP, with the concentration of the former
increasing after inammation, the levels of moderate increasing slightly, whereas the
concentration of negative APP is reduced after inammation. The most commonly
evaluated positive APP are C-reactive protein (CRP) and serum amyloid A (SAA),
whereas, albumin is the main negative APP. An APP assay may be helpful in the
diagnostics of the inammation, in treatment monitoring of inammation as well as
in diagnostics of some neoplastic diseases [Ceron et al. 2005, Eckersall et al. 2010,
Planellas et al. 2009, Tecles et al. 2009]. APP production is rapid (being detectable
within several hours after the beginning of the inammatory process), intense, but
also unspecic for the disease. They can be induced by any inammatory stimulus
New insights of canine mastitis
40
or even by pathophysiological conditions (tumors, stress, pregnancy) [Eckersall
et al. 2010]. For these reasons, due to its low specicity its clinical importance is
limited. On the other hand, it gives us a clear sign, that the organism is suering from
some pathological process, which can possibly be life-threatening. Therefore, APP
may be very useful in the detection of particularly subclinical mastitis as well as for
monitoring patients during recovery.
C-reactive protein
C-reactive protein (CRP) is a very well-studied inammatory biomarker, it has
been described in cases of canine pyometra, systemic inammatory diseases and
mammary tumors etc. [Christensen et al. 2015, Dąbrowski et al. 2007, Dąbrowski
et al. 2015, Planellas et al. 2009, Tecles et al. 2009]. Its biggest advantage is that its
concentration increases rapidly after the onset of the tissue damage and decreases
with its recovery [Vasiu et al. 2017]. As it was mentioned before, CRP is a non-
specic acute phase protein and its concertation may be increased due to any disease.
However, an elevated CRP concentration in milk is evidence of mastitis. A recent
study conrmed that CRP concentrations in both serum and milk samples from bitches
presenting mastitis were higher than in healthy bitches [Vasiu et al. 2017]. A positive
correlation was observed between serum and milk CRP concentrations. Interestingly,
no dierences were noticed between subclinical and clinical presentations of mastitis.
The serum values in bitches suering from mastitis ranged from 0.3 to 162.3 µg/
ml, while in healthy bitches it was 2-8.6 µg/ml, thus in diseased bitches the CRP
concentration was as high as 20-fold higher in comparison to healthy bitches. Milk
CRP levels in bitches suering from mastitis ranged from 0.3 to 40.0 µg/ml, while in
healthy females it was 0.1-4.9 µg/ml, so in diseased bitches the concentration was
even 8 times higher then in healthy bitches. Still the range of recorded concentration
values was wide, moreover, other on-going pathologies were not excluded and nally
the number of patients was relatively low. Nevertheless, it seems to be a very good
biomarker for early mastitis diagnosis, especially in cases of subclinical mastitis,
which is dicult to diagnose clinically.
Serum amyloid A
Serum amyloid A is another very important positive APP, assayed in many
systemic diseases in dogs [Christensen et al. 2014, Dąbrowski et al. 2007, Jiptean
et al. 2014]. Some studies indicated that SAA may be a more sensitive maker of
systemic inammation than CRP, as its concentration may be increased, while CRP
concentration would remain normal [Christensen et al. 2014, Jiptean et al. 2014].
Also, the recorded concentration ranges in patients presenting the disease turned out to
be narrower in the case of SAA in comparison to CRP [Jiptean et al. 2014], which was
in contrast to the data from a previous study [Christensen et al. 2014]. Therefore, it
I. Kaszak et al.
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seems to be a more sensitive and more precise inammatory biomarker. Nevertheless,
a simultaneous assay of both SAA and CRP is usually recommended. Still, to the best
of the authors’ knowledge no studies have been conducted on the determination of
SAA in the case of canine mastitis.
Cyclooxygeneses
Cyclooxygenases (COX) are inammatory biomarkers of inammation basically
assayed in neoplastic diseases [Benavente et al. 2016, Carvahlo et al. 2016]. The
cyclooxygenese enzyme catalyses the prostaglandin biosynthesis from arachnoid
acid. There are two isoforms: Cox-1 and Cox-2, but they have dierent biological
functions. Cox-1 is expressed in normal tissues and is responsible for the control of
renal function, reproduction and cytoprotection of stomach, among other things. Cox-
2 is undetectable in normal tissues, as it is expressed in tissue due to inammatory
reactions, growth factor, tumor promoters or oncogenes. Many studies conrmed that
prostaglandins play an important role in tumor’s development. COX-2 expression
was found to be increased in cases of malignant mammary tumors [Benavente et al.
2016, Carvahlo et al. 2016]. Determination of COX-2 in the cases of mastitis may be
interesting, especially in order to distinguish between mastitis and mammary tumors.
However, no such studies have been conducted to date.
Treatment
All the above-mentioned presentations of mastitis require treatment: rstly,
stabilisation of patients by uid administration targeted at correcting metabolic
disturbances. This is followed by therapy involving broad-spectrum antibiotics,
selected based on drug susceptibility testing, while milk pH should also be considered.
If the milk pH is lower than serum pH (<7,3), trimethoprim/ sulfathiazine 15-30 mg/
kg orally BID during 21 days), erythromycin (10 mg/kg orally TID during 21 days),
lincomycin (15 mg/kg orally TID during 21 days) should be used. When pH is >7.4,
ampicillin (20 mg/kg intramuscularly TID during 21 days) or cephalexin (30 mg/kg
orally BID during 21 days) are administered [Jiptean et al. 2014]. Some antibiotics
may reach therapeutic concentrations in milk regardless its pH, but they are usually
not recommended due to their negative eect on the puppies’ growth. In the case of
acute mastitis, the selected antibiotic is not that important as the integrity of the milk/
serum barrier has already been broken [Marti et al. 2010].
Moreover, additional therapy consisting in manual emptying of abnormal
mammary glands to avoid the accumulation of secretion is very important. Keeping
the area clean by washing and disinfecting is also necessary. Application of warm
compresses at least twice a day may help to reduce the inammation of the gland.
In some cases, local application of antibacterial and anti-inammatory tubes used in
cows for intramammary treatment of mastitis (e.g. Tetra Delta, Zoetis) may be helpful.
New insights of canine mastitis
42
Whether to separate puppies from their mother or not remain disputable. In
cases of severe mastitis with the presence of abscesses and gangrenous inammation
it is strongly advised against to continue nursing of puppies, as ingestion of toxic
or antibiotic-containing milk may produce health problems among the ospring.
The owner needs to hand-raise the neonates. In the case of slight mastitis, natural
feeding can be maintained, but it is necessary to remember that during mastitis milk
composition will deteriorate and weight gains among the puppies will be lower.
It is recommended to perform a surgical drainage in cases of gangrenous
inammation; sometimes drains have to be left in the tissue for several days. In some
very severe cases mastectomy is the method of choice.
Prognosis
If sepsis has not been developed, mastitis can be treated without any complications.
Mastitis is usually resolved with appropriate antibiotic therapy, although it takes some
time. The function of the mammary gland remains unaected, unless it was seriously
damaged due to abscesses or necrosis. Bitches may suer from mastitis during every
lactation and every false pregnancy, probably because the mammary gland anatomy
facilitates introduction of pathogens through the nipples. This phenomenon must be
mentioned to the owner and pros and cons of bitch castration should be discussed.
Conclusions
Disorders of the mammary gland are frequent health problems in bitches of
various ages. Among them mastitis is relatively common, especially in lactating
bitches or in false pregnancy. When left untreated it can be life-threatening, so proper
diagnosis and specic treatment are of great importance. Standard diagnostic methods,
involving clinical examination, blood tests and cytological examination of mammary
gland secretion may be insucient, especially in the case of subclinical mastitis. In
order to provide a rapid and more precise diagnosis inammatory biomarkers need to
be assayed. Acute phase proteins (CRP or SAA) detected in serum and milk samples
seem to be the most accurate inammatory biomarkers to diagnose canine mastitis, as
their concentrations are signicantly higher in bitches suering from mastitis than in
healthy bitches. However, more studies need to be conducted to evaluate dierences
in concentrations between dierent mastitis presentations (clinical vs. subclinical).
Identication of dierences between clinical and subclinical presentations may
promote a more adequate treatment. Other inammatory biomarkers such as
cyclooxygenases or interleukins seems to be promising both in diagnosis of mastitis
and dierentiation between mastitis and mammary tumours. However, before it is
clinically feasible, further studies on these biomarkers have to be carried out to obtain
more specic data.
I. Kaszak et al.
43
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