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Abstract

Canine distemper virus (CDV) infects wild and domestic Canidae worldwide. The hemag- glutinin (H) gene has the highest genetic variation in the genome of this virus. Thus, the H gene is commonly used for lineage identification and genetic analyses. In order to study the genetic characteristics and pathogenicity of CDV strains prevalent in China, 132 samples were collected from domestic dogs with suspected CDV infection, 58 samples were confirmed to be positive, and the H gene was successfully amplified from 15 samples. The epidemic strain was identified as type Asia-1 and the novel mutations, A51T, V58I, R179K and D262N, were detected in this strain. Isolated strains, BJ16B53, BJ16B14, and BJ17B8, were used for an animal infection experiment in raccoon dogs. BJ16B53 and BJ16B14 were found to cause clinical symptoms, death, and exten- sive lesions in various organs. These results are expected to facilitate the development of effective strategies to monitor and control CDV infection in China.

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Authors and Affiliations

M. Chen
T. Xin
S. Hou
W. Lin
W. Song
H. Zhu
K. Huang
H. Jia
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Abstract

In order to understand infection of avian influenza A virus (AIV) and canine distemper virus (CDV) in the Siberian Tiger in Northeast China, 75 Siberian Tiger serum samples from three cap- tive facilities in northeastern China were collected. AIV and CDV antibody surveillance was test- ed by using hemagglutination inhibition and serum neutralization methods. The results showed that the seroprevalence of H5 AIV, H9 AIV and CDV was respectively 9.33% (7/75), 61.33% (46/75) and 16% (12/75). In the 1<years <2 and > 5 year-old group, the seroprevalence of the H9 AIV was 24% and 80% (P < 0.01), and the CDV seroprevalence was 6% and 36% (P < 0.01), respectively. It was demonstrated that 3 (4%) out of 75 serum samples were AIV+CDV seropos- itive, with 2.67% (2/75) in H9+AIV and 1.33% (1/75) in H5+H9+AIV. To our knowledge, this is the first report of AIV and CDV seroprevalence in Siberian Tigers in China, which will provide base-line data for the control of AIV and CDV infection in Siberian Tigers in China.

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Authors and Affiliations

K. Wang
H. Wang
N. Feng
Y. Zhao
Y. Gao
G. Hu
X. Xia
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Abstract

Serum concentration of thyroid hormones in healthy dogs varies according to age, sex, breed or professional activity. The aim of this study was to determine the influence of both age and dogs’ work involvement on TSH and thyroid hormones values. Thyroid-stimulating hormone (TSH), total thyroxine (tT4) and free thyroxine (fT4) were tested in the serum of 57 healthy, German Shepherd dogs. The dogs were divided into study groups according to age: dogs aged 3 to 6 years (A), dogs over the age of 6 years (B) and involvement: police-working dogs (C) and accompanying animals (D). Mean values of TSH, tT4 and fT4 ranged from 0.19 to 0.31 ng/ml, 15.58 to 17.25 nmol/L and 11.83 to 17.89 pmol/L, respectively. The highest values of TSH were in group B and the lowest were in group A, while there was an inverse dependence in case of fT4 concentration. The highest mean values of tT4 were in dogs in group C and the lowest in group B. There were statistically significant differences in TSH (p=0.007) and fT4 (p=0.003) concentrations between the age groups. The results indicate that a dog’s age is an important factor in the case of thyroid profile results interpretation.
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Authors and Affiliations

I. Taszkun
1
A. Milczak
2
G. Kalisz
3
P. Wilkołek
1
J. Zwolska
4
M. Szczepanik
1

  1. Subdepartment of Clinical Diagnostics and Veterinary Dermatology, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, Głeboka 30, 20-612 Lublin, Poland
  2. Department and Clinic of Animal Internal Diseases, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, Głeboka 30, 20-612 Lublin, Poland
  3. Department of Biopharmacy, Faculty of Pharmacy, Medical University of Lublin, ul. W. Chodzki 4a, 20-093 Lublin, Poland
  4. Department and Clinic of Animal Surgery, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, Głeboka 30, 20-612 Lublin, Poland
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Abstract

Gallbladder mucocele (GM) is a common extrahepatic biliary disease recognized in dogs and is defined as the expansion and extension of the gallbladder by an accumulation of semi-solid bile or bile acid. Histopathological diagnosis of necrotizing cholecystitis and transmural coagulative necrosis of the gallbladder wall shows poor prognosis. Conversely, histopathological diagnosis with partial necrotic findings is often achieved. We hypothesized that histopathological partial necrosis of the gallbladder wall is the primary lesion of necrotic cholecystitis or transmural ischemic necrosis. Therefore, we investigated the relationship between histopathological necrosis/ partial necrosis findings and their clinical conditions. We retrospectively analyzed 55 dogs diagnosed with GM that had undergone cholecystectomy at the Yamaguchi University Animal Medical Center. The group with histopathological necrosis/partial necrosis of the gallbladder wall showed elevated levels of preoperative white blood cells, alanine transaminase, alkaline phosphatase, γ-glutamyltransferase, total bilirubin, and C-reactive protein compared to the non-necrotic group. Partial necrosis of the gallbladder wall may affect the progression of the disease and hematological abnormalities. Additionally, all death cases until 2 weeks were included in the histopathological necrosis/partial necrosis group. In this study, we found that poor prognosis factors were associated with partial necrosis of the gallbladder wall. Furthermore, these cases of partial necrosis showed elevated levels of blood test parameters. These results suggest that necrosis of the gallbladder wall is associated with poor prognosis and poor pathophysiological conditions.
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Bibliography


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Xenoulis PG (2014) The association between hyperlipidemia and gall bladder mucoceles in dogs. Vet J 200: 353-354.
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Authors and Affiliations

H. Itoh
1
K. Igari
2
K. Tani
3
H. Sunahara
3
Y. Nemoto
3
M. Nakaichi
4
T. Iseri
4
H. Horikirizono
4
K. Itamoto
1

  1. Department of Small Animal Clinical Science, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi City, Yamaguchi 753-8515, Japan
  2. Pet Clinic Hallelujah, 2544-1 Nakabaru, Kasuya, Kasuya-gun, Fukuoka 811-2304, Japan
  3. Department of Veterinary Surgery, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi City, Yamaguchi 753-8515, Japan
  4. Laboratory of Veterinary Radiology Yamaguchi University, 1677-1 Yoshida, Yamaguchi City, Yamaguchi, 753-8511, Japan
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Abstract

Many viruses are involved in concomitant infections, which are prevalent in nature. In mixed infections, one or both infectious agents may be increased, reduced, or both may be increased while the other is suppressed. Canine distemper virus (CDV) and Canine parvovirus- 2 (CPV-2) are important causes of gastroenteritis in dogs. Detection of these viruses is challenging since the symptoms are very similar. CDV is a member of the morbillivirus genus in the Paramyxoviridae family, and CPV-2 is a member of the Protoparvovirus genus in the Parvoviridae family; and both predominantly affect puppies and induce gastrointestinal symptoms in dogs. The purpose of this study was to contribute to the differential diagnosis of dogs with gastrointestinal symptoms. A PCR technique with specific primers was used to identify CDV and CPV-2 infections in gastroenteric dogs, and clinical changes in the infected dogs were monitored. The VP2 structural gene of CPV and the nucleocapsid gene of CDV were partially amplified in the study. PCR amplified the partial fragments of the CDV nucleocapsid (287 bp) and CPV-2 VP2 proteins (583 bp) from feces. In total, 3 out of 36 stool samples were positive for CDV and CPV-2 in the same dogs. Gasterointestinal symptoms also supported the diagnosis of concomitant infection with CDV and CPV-2 in these dogs. Dehydration and diarrhea in dogs can be signs of various diseases, such as viral, bacterial, and parasitic infections. After the elimination of non-viral pathogens, CDV and CPV-2 should also be simultaneously investigated to establish what is causing these symptoms. This study demonstrates the potential utility of correct diagnosis for the control of viral infection in dogs, but more research with a broader use of PCR-based detections is needed to assess its impact on differential diagnosis for concomitant infections.
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Authors and Affiliations

H.S. Saltık
1

  1. Department of Virology, Faculty of Veterinary Medicine, University of Burdur Mehmet Akif, 15030, Burdur, Turkey
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Abstract

Canine parvovirus (CPV) is a single-stranded DNA virus that causes severe and fatal gastrointestinal diseases in dogs. CPV has developed several strategies to evade innate immune response mediated by type I interferons (IFN-I) to achieve a successful infection. The aim of this work was to evaluate the capability of CVP-2c to evade the IFN-I mediated response in infected cells. To establish the role of this response, the gene expression of interferon β (IFNβ), IFIT1, IFIT3, MAVS, and STING were estimated in MDCK cells infected with CPV-2c. Viral replication and gene expression was evaluated by quantitative PCR, also, a treatment with IFN-I (interferon omega) was included to confirm the role of IFN-I during CPV infection. The results revealed that CPV-2c infection stimulates the expression of IFNβ moderately, in these cells. Due to low IFNβ induction, the IFIT1 and IFIT3 expression were also low, and therefore CPV-2c was able to replicate in these cells. However, when the cells were treated with exogenous IFN-I, the IFNβ expression was higher, leading to an increased gene expression of IFIT1 and IFIT3, responsible for antiviral control. The overexpression of these proteins reduced the expression of NS1 and VP2 viral genes and hence viral replication. MAVS and STING expression on infected cells showed a mild increase compared to IFNβ, suggesting that the viral infection could partially modify its expression. All results obtained in this study showed that during CPV-2c infection in MDCK cells, the IFNβ expression was altered since this cytokine is one of the most critical factors for the control and inhibition of viral replication.
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Authors and Affiliations

T. Reyes-Cruz
1
D. Martínez-Gómez
2
A. Verdugo-Rodríguez
3
J. Bustos-Martínez
4
J. López-Islas
2
E.T. Méndez-Olvera
2

  1. Doctorado en Ciencias Biológicas y de la Salud, Autonomous Metropolitan University (UAM), Calzada del Hueso 1100, Villa Quietud, C.P. 04960, Coyoacán, México City, México
  2. Department of Agricultural and Animal Production, Autonomous Metropolitan University, campus Xochimilco (UAM-X), Calzada del Hueso 1100, Villa Quietud, C.P. 04960, Coyoacán, México City, México
  3. Department of Microbiology and Immunology, Faculty of Veterinary Medicine, National Autonomous University of Mexico, Av. Universidad 3000, C.P. 04510, Coyoacán, México City, México
  4. Department of Health Care, Autonomous Metropolitan University, campus Xochimilco (UAM-X), Calzada del Hueso 1100, Villa Quietud, C.P. 04960, Coyoacán, México City, México
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Abstract

Canine parvovirus (CPV) causes acute gastroenteritis in domestic dogs, cats, and several wild carnivore species. In this study, the full-length VP2 gene of 36 CPV isolates from dogs and cats infected between 2016 and 2017 in Beijing was sequenced and analyzed. The results showed that, in dogs, the new CPV-2a strain was the predominant variant (n = 18; 50%), followed by the new CPV-2b (n = 6; 16.7%) and CPV-2c (n = 3; 8.3%) strains, whereas, among cats, the predominant strain was still CPV-2 (n = 9; 25%). One new CPV-2a strain, 20170320-BJ-11, and two CPV-2c strains, 20160810-BJ-81 and 20170322-BJ-26, were isolated and used to perform experimental infections. Multiple organs of beagles that died tested PCR positive for CPV, and characteristic histopathological lesions were observed in organs, including the liver, spleen, lungs, kidneys, small intestines, and lymph nodes. Experimental infections showed that the isolates from the epidemic caused high morbidity in beagles, indicating their virulence in animals and suggesting the need to further monitor evolution of CPV in China.

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Authors and Affiliations

M.R. Chen
X.Y. Guo
Z.Y. Wang
Y.T. Jiang
W.F. Yuan
T. Xin
S.H. Hou
T.Q. Song
W.D. Lin
H.F. Zhu
H. Jia
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Abstract

Abnormal DNA methylation is involved in the initiation and progression of lymphoid tumors. Hence, DNA demethylating agents are promising candidate drugs for chemotherapy against these tumors. The salicylic acid derived anti-inflammatory agent, olsalazine, reportedly suppresses DNA methyltransferase in human cells and has the potential to be clinically applied as a DNA demethylating agent. In this study, we investigated the effects of olsalazine on cell proliferation and DNA methylation using canine lymphoid tumor cell lines (CLBL-1, GL-1, and UL-1). Treatment with olsalazine led to significant cell growth inhibition and increased the apoptotic rate in all three cell lines. Treatment with olsalazine reduced the total amount of 5-methylcytosine in genomic DNA, as assessed by enzyme-linked immunosorbent assay. Genome-wide analysis of DNA methylation revealed that 1,801 to 5,626 CpG sites showed decreased DNA methylation levels in three cell lines, including the promoter regions of ADAM23, FES, and CREB3L1 genes. The outcomes of the present study demonstrate that a DNA demethylating agent olsalazine, inhibits cell proliferation and DNA methylation in canine lymphoid tumor cells, suggesting that it can be a candidate drug for the treatment of lymphoid tumors in dogs.
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Authors and Affiliations

S. Itoh
1 2
J. Yamazaki
3 4
M. Iwahana
2
A. Tsukamoto
2

  1. Laboratory of Biology, Azabu University, School of Veterinary Medicine, 1-17-71 Fuchinobe, Chuou-ku, Sagamihara, Kanagawa 252-5201, Japan
  2. Laboratory of Laboratory Animal Science, Azabu University, School of Veterinary Medicine, 1-17-71 Fuchinobe, Chuou-ku, Sagamihara, Kanagawa 252-5201, Japan
  3. Veterinary Teaching Hospital, Graduate School of Veterinary Medicine, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo, Hokkaido, 060-0808, Japan
  4. One Health Research Center, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo, Hokkaido, 060-0808, Japan
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Abstract

Canine status epilepticus (CSE) is characterized by epileptic seizures that are longer than 5 min or more than one seizure with incomplete recovery. Currently, diazepam suppositories are generally prescribed for CSE. Levetiracetam (LEV) is one of the newest antiepileptic drugs currently available. This study compared the pharmacokinetics of intragastric and intrarectal administration in oral formula of LEV in four healthy beagles as a reference data when the owner administers levetiracetam to dogs by himself at home. Blood for measuring plasma LEV concen- trations was collected 0, 30, 60, 90, 120, 240, 360, and 540 min after LEV administration. The time to reach the maximum plasma concentration (Tmax) was markedly shorter with intra- rectal administration (45±26 min) than with intragastric administration (270±99 min). Intrarectal administration of LEV tablets could be an effective option for treating canine seizures although it might be a limit for treating CSE because the absorption rate is not fast enough.

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Authors and Affiliations

H. Takamura
P. Chongphaibulpatana
Y. Uzuka
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Abstract

Seminal plasma (SP) proteins are responsible for sperm functional quality. Developing a reliable method to determine the degree of oxidative damage of these proteins is important for establishing semen fertilizing ability. The main aim of the study was to verify the applicability of protein carbonyl derivatives measurement in the SP of canine and stallion, using a method with 2,4-dinitrophenylhydrazine (DNPH). The research material consisted of ejaculates obtained from eight English Springer Spaniels, and from seven half-blood stallions during the breeding and non-breeding season. The content of carbonyl groups in the SP was measured on the basis of the reactions with DNPH. The following reagent variants were used to dissolve protein precipitates: Variant 1 (V1) – 6M Guanidine solution and Variant 2 (V2) – 0.1M NaOH solution. It has been shown that to obtain reliable results for the measurement of protein carbonylated groups in the dog and horse SP, both 6M Guanidine and 0.1M NaOH may be used. A correlation was also found between the number of carbonyl groups and the total protein content in the canine (V1: r = -0.724; V2: r = -0.847) and stallion (V1: r = -0.336; V2: r = -0.334) SP. Additionally, the study showed a higher content (p≤0.05) of protein carbonyl groups in the stallion SP in the non-breeding season compared to the breeding season. The method based on the reaction with DNPH, due to its simplicity and cost effectiveness, appears to be suitable for large-scale application in the determination of the SP proteins oxidative damage in dog and horse semen.
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Authors and Affiliations

M. Mogielnicka-Brzozowska
1
M.J. Woźniak
1
A.W. Cichowska
1
L. Fraser
1
B. Kraziński
2
R. Strzezek
1
D. Zielińska
3

  1. Department of Animal Biochemistry and Biotechnology, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-957 Olsztyn, Poland
  2. Department of Human Histology and Embryology, School of Medicine, Collegium Medicum, University of Warmia and Mazury in Olsztyn, Warszawska 30, 10-082 Olsztyn, Poland
  3. Department of Chemistry, Faculty of Environmental Management and Agriculture, University of Warmia and Mazury in Olsztyn, Oczapowskiego 8, 10-719 Olsztyn, Poland
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Abstract

Probiotics and prebiotics are viable bacteria with beneficial effects on the host and components that selectively act on the beneficial commensal bacteria, respectively. The combined use of probiotics and prebiotics is termed synbiotics. Probiotic intake improves dysbiosis in the intestinal microbiota and can positively affect canine atopic dermatitis (CAD). However, clinical studies on improvements in CAD using synbiotics remain limited.
In this study, 15 dogs with CAD who received prednisolone, a synthetic glucocorticoid (GC) used in the treatment of CAD, for more than 90 days were continuously treated with Lactobacillus paracasei M-1 from fermented food as a probiotic, and trisaccharide kestose as a prebiotic, for 90 days to determine their synbiotic effects on CAD. The CAD symptoms were evaluated using the canine atopic dermatitis lesion index (CADLI) and pruritus visual analog scores (PVAS) at 30, 60 and 90 days after synbiotic administration. The total prednisolone use for 90 days pre- and post-administration was also evaluated.
Synbiotic administration significantly reduced the CADLI (pre: median, 28.0 [22.0-32.0]; 30 days: median, 20.0 [20.0−28.0]; 60 days: median, 20.0 [10.0−21.0]; 90 days: median, 12.0 [10.0-19.0]) and PVAS (pre: median, 6.0 [5.0-7.0]; 30 days: median, 3.0 [3.0-3.5]; 60 days: median, 3.0 [3.0-3.5]; 90 days: median, 2.0 [2.0-3.5]) scores, and reduced the total prednisone use over 90 days (pre: 112.0 [25-450] mg; post: 80.0 [18.-300.0] mg; p<0.001) in the 15 dogs. Thus, the synbiotic activity of L. paracasei M-1 and trisaccharide kestose can improve CAD.
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Authors and Affiliations

K. Kawano
1 2
K. Iyori
3
N. Kondo
2 4 5
S. Yamakawa
2 4 5
T. Fujii
6
K. Funasaka
2
Y. Hirooka
2 6
T. Tochio
6

  1. Tokyo Animal Allergy Center, Adachi-ku, Tokyo 123-0842, Japan
  2. Department of Gastroenterology and Hepatology, Fujita Health University, Toyoake, Aichi 470-1192, Japan
  3. Vet Derm Tokyo, Dermatological and Laboratory Service for Animals, Fujisawa, Kanagawa 252-0823, Japan
  4. Research and Development Division, Itochu Sugar Co., Ltd., Hekinan, Aichi 447-8506, Japan
  5. WELLNEO SUGAR Co., Ltd., Chuo-ku, Tokyo 103-8536, Japan
  6. Department of Medical Research on Prebiotics and Probiotics, Fujita Health University, Toyoake, Aichi 470-1192, Japan
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Abstract

The aim of this study was to evaluate the plasma levels of chemokines CCL2 and CXCL12 in female dogs with malignant mammary gland tumours without and with metastases. The concentrations of CCL2 and CXCL12 were determined in 25 female dogs with malignant mammary gland tumours (15 without metastases and 10 with metastases) and 10 healthy control animals using a specific canine ELISA assay. The mean plasma concentrations of CCL2 and CXCL12 were significantly higher (p<0.05) in the metastatic group compared to the control group. Moreover, the concentrations of these chemokines were markedly higher in the dogs with metastases than in those without metastases; however, a statistically significant difference was not found. The concentrations of both tested chemokines were numerically increased in the dogs with grade 2 and grade 3 carcinomas compared to the dogs with grade 1 carcinomas but the differences did not reach statistical significance. In conclusion, the results of our study demonstrate that plasma concentrations of chemokines CCL2 and CXCL12 are significantly increased in the dogs with metastatic malignant mammary gland tumours compared to the healthy dogs and show an upward trend compared to those without metastases. However, clarifying whether the increase of these chemokines is a cause or an effect of metastasis in female dogs with malignant mammary gland tumours as well as their potential role in metastatic process requires further research.
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Authors and Affiliations

M. Szczubiał
1
W. Łopuszyński
2
R. Dąbrowski
1
M. Jamioł
3
M. Bochniarz
1
P. Brodzki
1

  1. Department and Clinic of Animal Reproduction, Faculty of Veterinary Medicine, University of Life Sciences, Gleboka 30, 20-612 Lublin, Poland
  2. Department of Pathomorphology and Forensic Veterinary Medicine, Faculty of Veterinary Medicine, University of Life Sciences, Gleboka 30, 20-612 Lublin, Poland
  3. Department of Animal Biochemistry, Faculty of Veterinary Medicine, University of Life Sciences, Akademicka 12, 20-033 Lublin, Poland
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Abstract

The purpose of this study was to evaluate in detail both the in vivo and in vitro efficacy of the enzyme agents, ZYMOX® Plus Otic (ZYMOX-P), in the treatment of canine otitis externa (OE). Eight dogs with a diagnosis of non-seasonal severe chronic OE were recruited for the study. ZYMOX-P was administered for 2-4 weeks. The Otitis Index Score (OTIS3) and bacteria or yeast colony growth were measured. Also, minimum biofilm (BF) formation inhibition concentration (MBIC) and BF bactericidal concentration (BBC) were measured in vitro. OTIS3 showed a statistically significant reduction after treatment (88.2%, p<0.001; pre-treatment = 11.0 ± 0.9; post-treatment = 1.3 ± 0.4, mean ± SEM). The individual OTIS scores, erythema, edema, erosions/ ulcerations, exudate and pruritus showed significant reduction (85.7%, 95.7%, 83.3%, 80.0%, and 89.3%, respectively). Microscopic examination revealed the presence of BF exopolysaccharide in all 8 ear samples when stained with alcian blue. Seven of the 8 dogs (87.5%) showed a reduction in colony growth. ZYMOX-P was effective at 34-fold and 16-fold dilutions on MBIC and BBC, respectively. These findings indicate that ZYMOX-P has efficacy against BF-related infection and is beneficial when used for the management of canine OE.
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Authors and Affiliations

M. Fujimura
1

  1. Fujimura Animal Allergy Hospital, Aomatanihigashi 5-10-26, Minou-city, Osaka 562-0022, Japan
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Abstract

In this study plasma concentrations of IL-1α and IL-8 in 29 female dogs with malignant mammary gland tumours (19 without metastasis and 10 with metastasis) and in 10 healthy control animals were determined. Concentrations of IL-1α and IL-8 were analysed using a specific canine ELISA assay. Mean plasma concentrations of IL-1α and IL-8 were significantly higher (p<0.05) in female dogs with both non-metastatic and metastatic malignant tumours compared to the healthy animals. The concentrations of both tested cytokines were significantly increased (p<0.05) in the dogs with metastasis. In female dogs with mammary carcinomas, the plasma concentration of IL-1α was significantly higher (p<0.05) in the animals with grade 3 tumours compared to the dogs with grade 1 tumours. The concentration of IL-8 was significantly higher (p<0.05) in the dogs with grade 3 tumours compared to that found in the animals with grade 1 and grade 2 tumours. A moderate correlation (r=0.433) was found between IL-1α and IL-8 concentrations in the female dogs. These findings suggest that increased malignancy and invasiveness of canine mammary tumours is associated with an increased production of IL-1α and IL-8 in the tumour microenvironment, which, in turn, leads to an increase in their circulating levels. This may indicate that circulating levels of the cytokines investigated could be considered as diagnostic and prognostic markers in canine malignant mammary tumours. However, further studies in this fields are needed.
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Authors and Affiliations

M. Szczubiał
1
W. Łopuszyński
2
R. Dąbrowski
1
M. Bochniarz
1
P. Brodzki
1

  1. Department and Clinic of Animal Reproduction, Faculty of Veterinary Medicine, University of Life Sciences, Gleboka 30, 20-612 Lublin, Poland
  2. Department of Pathomorphology and Forensic Veterinary Medicine, Faculty of Veterinary Medicine, University of Life Sciences, Gleboka 30, 20-612 Lublin, Poland
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Abstract

Suberoylanilide hydroxamic acid (SAHA) is a histone deacetylase inhibitor (HDACi) that suppresses the growth of tumor cells in humans and canines. SAHA reportedly enhances the antitumor activity of human peripheral blood mononuclear cell (PBMC). However, it is unclear whether a similar effect is exerted in canines. The present study focused on the effect of SAHA on the cytotoxicity of IL-2 activated PBMC in three tumor cell lines (CTAC, CIPm, and MCM-N1). The mRNA expression of a ligand for the NKG2D receptor was upregulated in SAHA-treated cell lines. Moreover, the SAHA-treated cell lines, except MCM-N1 demonstrated a significantly higher PBMC cytotoxicity compared to the untreated cell lines. Therefore, the NKG2DL upregulation likely enhanced the interaction of NKG2D-NKG2DL, leading to enhanced cytotoxicity of PBMC. It was also revealed that activated PBMC treated with SAHA significantly attenuated their cytotoxicity toward all the cell lines. Although the NKG2D, NKp46, NKp44, and NKp30 receptors, involved in PBMC cytotoxicity, were presumed to be downregulated, there was no significant reduction in the mRNA expression of these receptors. This study revealed that SAHA not only sensitizes the canine tumor cells to cytotoxicity due to PBMC activation, but also suppresses the cytotoxicity of PBMC themselves. Therefore, our results highlight the necessity of avoiding this inhibitory action to enhance the antitumor effect of SAHA in canines.
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Authors and Affiliations

T. Oyamada
1 2
S. Okano
2

  1. Animal Medical Center, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8, Saiwai, Fuchu, Tokyo, 183-8509, Japan
  2. Department of Small Animal Surgery 2, School of Veterinary Medicine, Kitasato University, Higashi 23-35-1, Towada, Aomori, 034-8628, Japan
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Abstract

αB-crystallin is a member of a small family of thermal shock proteins that protects cells from stress. Because of lack of its expression in peripheral blood leukocytes, it was proposed as a molecular marker of circulating tumor cells in canine mammary gland tumors. The aim of the present study was to determine if αB-crystallin shows stability of expression, what is the requirement for this type of marker. It was also assessed whether there is co-expression of αB-crystallin with the basal marker, cytokeratin 17. For this purpose, samples of various types of canine mammary gland tumors of epithelial origin, were selected. Using RT-qPCR, we have found αB-crystallin and cytokeratin 17 co-expression in benign and malignant canine mammary gland tumors. It has been demonstrated that the expression of αB-crystallin in tested neoplastic samples is not stable in comparison to the control group. Furthermore αB-crystallin overor down- expression was associated witch the same cytokeratin 17 pattern. αB-crystallin can be a marker of circulating tumor cells in the bloodstream, but for cancers in which basal marker expression occurs and thus not universal for all cancers originating from the mammary gland tissue.

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Authors and Affiliations

M. Chmielewska-Krzesińska
A. Jakimiuk
K. Wąsowicz
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Abstract

Emerging researches in humans, pigs and mice, highlighted that estrogen plays a pivotal role in self-renewal and differentiation of bone marrow mesenchymal stem cells (BMSCs). The present study aimed at evaluating effects of 17 beta-estradiol (E2) on proliferation and apoptosis of canine-derived bone marrow mesenchymal stem cells (cBMSCs) in vitro. The results showed that E2 supplementation at the concentration of 10-11 M promoted the proliferation of cBMSCs by CCK-8 assay and RT-qPCR analysis for the proliferation-related genes, with proliferating cell nuclear antigen (PCNA), cyclin-D1 (CCND1) being up-regulated and cyclin-dependent kinase inhibitor 1B (CDKN1B) being down-regulated. Contrarily, analysis of fluorescence-activated cell sorting (FACS) and RT-qPCR demonstrated that E2 supplementation above 10-11 M had inhibitory effects on the proliferation of cBMSCs and induced apoptosis. Intriguingly, cBMSCs still possessed the capability to differentiate into osteoblasts and adipocytes with 10-11 M E2 addition. Taken together, this study determined the optimal culture condition of cBMSCs in vitro, and has important implications for further understanding the regulatory effect of E2 on the self-renewal of cBMSCs, which are helpful for the clinical application of BMSCs.

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Authors and Affiliations

Z.-H. Zhou
C.-W. Gu
J. Li
X.-Y. Huang
J.-Q. Deng
L.-H. Shen
S.-Z. Cao
J.-L. Deng
Z.-C. Zuo
Y. Wang
ORCID: ORCID
X.-P. Ma
Z.-H. Ren
S.-M. Yu
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Abstract

Staphylococcus pseudintermedius (SP) and methicillin-resistant SP (MRSP) is one of the most important veterinary pathogens in the dog. Herein, from a total of 126 S. pseudintermedius strains, 23 MRSP (18%) were identified. Multi-Locus Sequence Typing (MLST) revealed that most of MRSP strains belonged to ST71 (26%), which have been already reported in Italy and other countries. Interestingly, nine new sequence types (39%), from 1053 up to 1061, were described for the first time. Moreover, the isolated MRSP strains showed relevant antibiotic resistance profiles. This report highlights the circulation of new sequence types of MRSP in Italy and underlines the need of a global epidemiological surveillance to limit the increasing spread of multidrug-resistant MRSP strains worldwide, since they may represent a considerable concern for dog’s health.

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Authors and Affiliations

F.P. Nocera
A. Parisi
M. Corrente
L. De Martino
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Abstract

Erythritol helps both prevent and improve periodontal disease and is therefore widely used for dental care in humans. However, only a few studies have investigated the effects of erythritol on periodontal disease in animals. We hypothesized that erythritol could be used to prevent and improve periodontal disease also in canines and investigated the effects of erythritol on canine periodontal disease–related pathogenic bacteria using both in vitro and in vivo methods.
The effect of erythritol on the proliferation of Porphyromonas gulae, which is reportedly associated with canine periodontal disease, was investigated in vitro. In addition, a 4-week intervention trial using an external gel preparation containing 5% erythritol was performed in canines with mild periodontal disease; changes in the microbiota around periodontal lesions were investigated using next-generation sequencing and bioinformatics analysis.
The growth of P. gulae was significantly suppressed by erythritol in vitro. In the intervention study, the Shannon index, an indicator of the species distribution α-diversity, and the occupancy of several canine periodontal disease – related bacteria ( P. gulae, P. cangingivalis) were significantly decreased in periodontal lesions.
Based on the results of in vitro and in vivo studies, we conclude that, as in humans, erythritol has bacteriostatic effects against periodontal disease – related bacteria in canines.
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Authors and Affiliations

T. Tochio
1 2
R. Makida
1
T. Fujii
1
Y. Kadota
1
M. Takahashi
1
A. Watanabe
2
K. Funasaka
2
Y. Hirooka
2
A. Yasukawa
3
K. Kawano
2 4

  1. B Food Science Co., Ltd., 24-12, Kitahama-machi, Chita, Aichi 478-0046, Japan
  2. Department of Gastroenterology and Hepatology, Fujita Health University, Toyoake, Aichi 470-1192, Japan
  3. Kamishakujii Veterinary Medical Hospital, 1-4-13, Sekimachi-Higashi, Nerima-ku, Tokyo 177-0052, Japan
  4. Tokyo Animal Allergy Center, 4-23-15, Kurihara, Adachi-ku, Tokyo 123-0842, Japan

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