Haematologic and biochemical changes in bitches with clinical and subclinical mastitis

1* 1 1 1 Iosif VASIU , Orsolya SARPATAKI1 , Ioana BEDECEAN , Raul Alexandru POP , Florinel Gheorghe BRUDAŞCĂ 1 Faculty of Veterinary Medicine,[email protected] University of Agricultural Science and Veterinary Medicine Cluj- Napoca, 3-5 CaleaMănăştur, Cluj-Napoca, Romania * Corresponding author:

Bulletin UASVM Veterinary Medicine 73(2)/2016 Print ISSN 1843-5270; Electronic ISSN 1843-5378 doi:10.15835/buasvmcn-vm:11995

Abstract

Mastitis in bitch is in general superficially researched and evaluated. Data regarding blood tests Lactatioduring bitch sine graviditateglandular tissue inflammation is scarcely documented, therefore this paper aims to assess blood response in such cases. A total of 74 blood samples were obtained from lactating dams. These included females with , with or without overt signs of mammary gland illness and with or without other signs of disease. Routine complete blood count and serum total proteins were evaluated. Complete blood count and total serum protein levels showed great flexibility, thus making it very difficult to establish and describe a clear and common haematological and biochemical pathway during mastitis inflammatory response. Clincians shoulds not relay only on blood tests whenever dealing with such pathology. Complete blood count and total serum protein levels should only be usedKeywords as complementary: bitch, complete tests, alongsideblood count, other mastitis, assays such serum as milktotal microbiology protein or cytology.

INTRODUCTION et al. important changes in total serum protein levels In curent veterinary practice, mammary (Ververidis , 2007). ( gland inflammationet al. diagnostic is based mainly on The aim of this research was antepartumto evaluate history, clinical and microbiological examination postpartumwhether completeLactatio blood count sine graviditateCBC) and serum (Trasch , 2008). There is scarce data regardinget al. total proteins in lactating bitches ( , blood changes during clinical or subclinical and ), with mastitis episodeset al. in this species (Hasegawa , or without overt signs of mastitis and other 1993). However, there is one experimental study concurrent signs of illness, follow the same (Ververidis , 2007). Where type of haematological or biochemical blood was observed immediately after mammary response.MATERIALS AND METHODS gland bacterial challenge. At 12 hours, shortly after inoculation, a drop in the leukocyte count was observed, then followed by an immediate In order to evaluate haematological and increase. On the sixt day, after challenge, biochemical changes during bitch mastitis cases, leukocytes were in normal range. 100 lactating dams were included in a case- was also observedth shortly after challenge, but controlled study, but because of owners reluctance, after the 6 day, this parameter dropped in range we were only able to obtain 74 blood samples. as well. Severe thrombocytopenia was recorded Dams in this study were divided into two in the first hours afterth inoculation, followed by groups. Females with primary mastitis and an increase in the 4 day. However, there were no females with secondary mastitis. During consult, 249

Haematologic and biochemical changes in bitches with clinical and subclinical mastitis females with concurrent affections were included The CBC showed that was present in the second category (i.e. secondary mastitis), in 21% (4/19, CI 95%: 6.05-45.57) of cases whilst whereas clinically healthy females or just with was present in only 5% (1/19, CI 95%: clinical or subclinical mastitis, were included in 0.13-26.03) of the tested females. the first category. with eosinophilia was encountered in 11% (2/19, Haematology and biochemstriy assays were CI 95%: 1.30-33.14) of the samples, followed by processed on venous blood after a thorough local with eosinophilia and by Lymphocytosis disinfection with betadine (Betadine 100 ml, Egis with eosinopenia in 5% (1/19, CI 95%: 0.13-26.03) Pharmaceuticals Ltd., Budapest, Hungary). After of the cases, each (Tab.1) (p>0.05). collection, blood samples were rushed as quickly In 16% (3/19, CI 95%: 3.38-39.58) of as possible to the laboratory at the University of samples, CBC encountered Neutrophilia with left Agricultural Science and Veterinary Medicine Cluj- shift along with eosinophilia, 11% (2/19, CI 95%: Napoca. 1.30-33.14) were with Neutrophilia with left shift Complete blood count was determined with along with monocytosis, showing chronic signs of Abacus Junior Vet analyzer (Diatron Messtechnik, disease (p>0.05). Budapest, Hungary) whilst differential blood Without correlation with clinical features, count was counted on blood film stained with 5% (1/19, CI 95%: 0.13-26.03) of the samples Dia-Quick Panoptic dye kit (Reag-Fix Panoptic, presented, Neutrophilia with a left shift with ReagensKft., Budapest Hungary). Biochemical eosinopenia, lymphocytosis and eosinopenia, results for total serum protein, were obtained with a lymphopenia, monocytopenia and eosinophilia Spectrophotometer analyzer UV-VIS Screen Master and also with with a right shift along Touch (Hospitex Diagnostix, Fiorentino, Italy). with monocytosis, lymphocytosis and eosinophilia, Complete blood count and serum proteinogram each (Tab.1) (p>0.05). changes prevalence andp its 95% confidence In 31% (5/16, CI 95%: 11.02-58.66) of tested intervals were assessed. The differences were samples Hyperproteinemia with hyperglobulinemia­ considered significant if values were lower than was encountered, followed by 25% (4/16, CI 95%: 0.05. Data analysis was performed using EpiInfo 7 7.27-52.38) of dams with Hyperglobulinemia. In softwareRESULTS (CDC, ANDUSA). DISCUSSION 13% (2/16, CI 95%: 1.55-38.35) of cases presence of Hyperproteinemia with hyperalbuminemia and Hyper­ pro­ teinemia­ with hyperalbuminemia In bitches with primary mastitis, 40% and hyperglobulinemia­ was noticed. In just 6% (4/10, CI 95%: 6.67-65.25) of cases were (1/16, CI 95%: 0.16-30.23) of cases Hyperalbumi­ recorded with Normocytic hyperchromic nemia followed by Hyporproteinemia and followed by Normocytic normochromic and Hyperalbuminemia with hyperglobulinemia were Microcytic hypochromic mild anemia in 20% encountered (Tab.1) (p>0.05). (2/10, CI 95%: 2.52-55.61) of tested samples. Usually, elevated levels of total serum proteins, Normocytic normochromic moderate anemia and including, albumins and globulins could be caused Normocytic hypochromic mild anemia were also by dehydration, chronical inflammation or chronic present in 10% (1/10, CI 95%: 0.25-44.50) of the subacute bacterial infection (Kerr, 2002b). These tested cases, each (Tab.1) (p>0.05). Presence of hypothesis can explain the presenceMastitis of gangre high levels­nosa mild anemia could have been caused by the loss of these parameters in primary mastitis bitches. of blood due to inflamed mammary glands and In one documented bitch infections (Kerr, 2002a). case report, where biochemical and haematological Thrombocytopenia was recorded in 60% changes were recorded, results showed presence (3/5, CI 95%: 14.66-94.73) of cases, whilst 40% of severe leukocytosis (36,300/µl) with marked (2/5, CI 95% 5.27-85.34) were recorded with elevated α, β and γ-globuline levels, moderate Thrombocytosis (p>0.05). anemia, a slightly increased level of alkaline Mild Lecukocytosis was encountered in 4 phosphatase and creatine phosphokinase with (4/4, CI 95%: 10.00) of the primary mastitis moderate hipernatremia and hypokalemia. On bitches (p>0.05). the day of admission, the amount of circulating leukocytes decreasedBulletin temporarilyUASVM Veterinary Medicineto 26,000/ml, 73 (2) / 2016 250 et al

VASIU Tab. 1. Hemoleucogram and biochemical results in both types of bitch mastitis

Percen­ Diagnostic – Secondary mastitis Percen­ No. Diagnostic – Primary mastitis No. tage tage

WBC 1 WBC

2 Severe 14% 1 3 Moderate leukocytosis 29% Mild leukocytosis 100% 4 Severe leukocytosis 29%

RBC Mild leukocytosis RBC 29%

2 5

3 Normocytic normochromic moderate anemia 10% 6 Normocytic hypochromic moderate anemia 14%

4 Normocytic hypochromic mild anemia 10% 7 Normocytic hypochromic mild anemia 14%

5 Microcytic hypochromic mild anemia 20% 8 Normocytic normochromic mild anemia 14%

6 Normocytic normochromic mild anemia 20% Normocytic hyperchromic mild anemia 57%

Normocytic hyperchromicPLT mild anemia 40% PLT

8 9

9 Thrombocytosis 40% 10 Thrombocytosis 33%

Thrombocytopenia CBC 60% Thrombocytopenia CBC 67%

10 11

11 Eosinophilia 5% 12 Monocitopenia with eosinophilia 6% Neutrophilia with left shift with 12 Lymphocytosis with eosinopenia 5% 13 6% lymphocytosis and eosinopenia 13 Monocytosis with eosinophilia 5% 14 Neutrophilia with left shift with lymphopenia 6% Neutrophilia with left shift with lymphopenia, 14 Neutrophilia with left shift with eosinopenia 5% 15 6% monocytopenia and eosinopenia Neutrophilia with left shift with 15 5% 16 Neutrophilia with left shift with monocytosis 6% lymphocytosis and eosinopenia 16 Neutrophilia with left shift with lymphopenia 5% 17 Neutropenia with eosinophilia 6% Neutrophilia with left shift with 17 5% 18 Neutrophilia with left shift 12% monocytopenia and eosinophilia Neutropenia with right shift with monocytosis, 18 5% 19 Neutrophilia with left shift with eosinopenia 12% lymphocytosis and eosinophilia Neutrophilia with left shift with 19 Monocytopenia with eosinophilia 11% 20 12% monocytosis and eosinophilia Neutrophilia with left shift with 20 Neutrophilia with left shift with monocytosis 11% 29% lymphopenia and eosinopenia 21 Neutrophilia with left shift with eosinophilia 16%

Eosinopenia 21%

22 Total Proteins 21 Total Proteins

23 Hyperalbuminemia 6% 22 Hyperalbuminemia 13%

24 Hyperalbuminemia with hyperglobulinemia 6% 23 Hyperglobulinemia 13%

25 Hyporproteinemia 6% 24 Hyperproteinemia with hyperalbuminemia 20%

26 Hyperproteinemia cu hyperalbuminemia 13% 25 Hyperproteinemia with hyperglobulinemia 20% Hyperproteinemia with hyperalbuminemia Hyperproteinemia with hyperalbuminemia 27 13% 33% and hyperglobulinemia and hyperglobulinemia 28 Hyperglobulinemia 25%

Hyperproteinemya with hyperglobulinemia 31% Bulletin UASVM Veterinary Medicine 73 (2) / 2016 251

Haematologic and biochemical changes in bitches with clinical and subclinical mastitis but then increased to 48,600/ml. Along with these bu­minemia and hyperglobulinemia were re­ etpara al.­meters, thrombocytopenia (179,000/ml) cor­ded, followed by Hyperproteinemia with with giant platelets were also recorded (Hasegawa hyper­al­bu­minemia and Hyperproteinemia with , 1992).These changes are partially confirmed hyper­globulinemia in 20% (3/15, CI 95%: 4.33- by our study, where mild to severe leukocytosis 48.09) of cases, each and Hyperalbuminemia and were recordedMastitis (Tab. 1)acuta alongside with a left shift Hyperglobulinemia in 13% of tested samples and increase in total protein serum levels (Tab. 1). (2/15, CI 95%: 1.66-40.46), each (Tab.1). In bitch reports,Mastitis leukocytosis acuta In secondary mastitis bitches, blood parame­ with or without a left shiftet al. was encountered ters should always be interpreted according to the (Lopate, 2012). In a recent bitch case primaryCONCLUSION underlying causes. report, the authors (Vasiu , 2015) indicated presence of moderate anemia with leukocytosis with a left shift. Presence of eosinophilia with Haematology and biochemical assays proved dehydration along with elevated liver enzymes and to be modest and limited. The use of these tests in renal biomarkers with electrolyte imbalance, had order to evaluate mammary gland health status also been reported (Lopate, 2012). Eosinophilia in bitch should resume only as complementary and eosinopenia, alongside various combinations assays. However, one should try to evaluate bitch of other types of leukocytes were, as well, recorded mammary gland health status, alongside clinical in our study. and other paraclinical (i.e. microbiological) In secondary mastitis cases, Normocytic hy­ testing. Clinicians should avoid relaying only per­­chromic­ mild anemia was diagnosed in 57% on these test whenever treating or diagnosing (8/14, CI 95%: 28.86-82.34) of cases, alongside mastitis in the bitch. In cases where mastitis is of Normocytic hypochromic moderate anemia, secondary, one should always carefully interpret Normocytic hypochromic mild anemia and blood test and treat the dam, according to the Normocytic normochromic mild anemia in 14% underlyingAcknowledgments. cause. (2/14, CI 95%: 1.78-42.81) of samples, each. Thrombocytopenia was registered in 67% We want to thank DVM PhD (4/6, CI 95%: 22.28-95.67) of cases, whilst 33% student Matei Ioana Adriana for the completion of (2/6, CI 95%: 4.33-77.72) of the samples were the statistical analysis and to Tressa Rebek-Nagy with Thrombocytosis (Tab.1). for reviewingREFERENCES and proofreading this manuscript. Test results showed that in secondary mastitis cases, mild, moderate and severe Leukocytosis were encountered in 29% (2/7, CI 95%: 3.67- 1. Hasegawa T, Fuji M, Fukada T, Tduji C, FujitaStaphylococcus T, Goto Y, aureusShinjo T, Ogawa H (1993). Platelet abnormalities in a dog 70.96) of cases, each. Severe Leukopenia was suffering from gangrenous mastitis by registered only in 14% (1/7, CI 95%: 0.36-57.87) infection. J Vet Med Sci 55:169-171. of the tested bitches. 2. Kerr M (2002a). Part I: Haematology, p. 1-66. In: Kerr Complete blood count showed presence of M (ed.). Veterinary Laboratory Medicine, Clinical Neutrophilia with left shift with lymphopenia and Biochemistry and Haematology, Second edition, Blackwell eosinopenia in 29% (5/17, CI 95%: 10.31-55.69) Science. London UK. of cases, followed closely by Neutrophilia with left 3. Kerr M (2002b). Part II: Clinical Biochemistry, p. 67-196. shift with eosinopenia and with monocytosis and In: Kerr M (ed.). Veterinary Laboratory Medicine, Clinical Biochemistry and Haematology, Second edition, Blackwell eosinophilia in 12% (2/17, CI 95%: 1.46-36.44) of Science. London UK. cases, each. 4. Lopate C (2012). Reproductive physiology of canine Neutrophilia with left shift with monocytosis, pregnancy and parturition and conditions of the peri­ with lymphopenia, monocytopenia and parturient period, p. 25-41. In: Lopate C (ed.). Management eosinopenia and lymphocytosis and eosinopenia of pregnant and neonatal dogs, cats andexotic pets.Wiley- were diagnosed in 6% (1/17, CI 95%: 0.15-28.69) Blackwell. of cases, each. 5. Trasch K, Wehrend A (2008). Klinische, sonographische In 33% (5/15, CI 95%: 11.82-61.62) of eva­ und mikrobiologische Untersuchungenbei Hündinnen mit acuter Mastitis [Clinical, ultrasonographical and lua­­ted samples, Hyperproteinemia with hyperal­ Bulletin UASVM Veterinary Medicine 73 (2) / 2016 252 et al

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Bulletin UASVM Veterinary Medicine 73 (2) / 2016