Investigations on the Endometrial Response to Intrauterine Administration of N‐Acetylcysteine in Oestrous Mares

In mares, mating‐induced persistent endometritis contributes to low fertility. The condition is in part related to delayed clearance of mucus accumulated within the uterine lumen. The objective of this study was to investigate the endometrial response of healthy mares to intrauterine (i.u.) treatment with N‐acetylcysteine (NAC). Oestrous mares (n = 12) were randomly assigned to a treatment (TM) or control (C) group and received an i.u. infusion of 5% NAC and saline (total volume 140 ml), respectively. Endometrial biopsies were collected in five of the mares 24 h after treatment, in the remaining seven mares 72 h after treatment. Endometrial biopsies were evaluated for integrity of the luminal epithelium, number of polymorphonuclear neutrophils (PMN), staining for cyclooxygenase 2 (COX2), staining with Kiel 67 antigen (Ki‐67), lectins and periodic acid‐Schiff (PAS). The integrity of endometrial epithelial cells was not affected by treatment (no statistical differences between groups or times). At 24 h after treatment, the mean number of PMN in endometrial biopsies from NAC‐ and C‐mares did not differ, but at 72 h after treatment, number of PMN was significantly higher (p < 0.05) in C (3.9 ± 0.6 PMN/field) compared with NAC‐treated mares (2.3 ± 0.2 PMN/field). At 72 h after treatment, the intensity of staining for COX2 was significantly higher after saline than after NAC treatment (p < 0.05). In the epithelium, no differences in staining for the proliferation marker Ki‐67 were seen with respect to time and treatment. Score for the lectin wheat germ agglutinin (WGA) was slightly higher in NAC‐treated mares than in C‐mares 72 h after treatment (p < 0.05). Score for PAS staining of mucus in deep uterine glands differed significantly between groups at 24 h after treatment (p < 0.05). The present study demonstrates that NAC does not adversely affect the endometrial function. Moreover, an anti‐inflammatory effect on the equine endometrium was observed.     

The evaluation of lycopene and cysteamine supplementation effects on sperm and oxidative stress parameters during chilled storage of canine semen

The aim of this study was to evaluate the effects of different concentrations of lycopene and cysteamine on characteristics of sperm, liquid peroxidation and enzymatic activities in seminal plasma of canine semen preserved at 5°C for 72 hr. The semen samples were divided into eight aliquots: control, control sham (dimethyl sulfoxide 5%), lycopene groups (250, 500 and 750 µg/ml) and cysteamine groups (2.5, 5 and 10 mM). Motility, viability, membrane integrity, DNA integrity, total antioxidant capacity (TAC) and malondialdehyde (MDA) levels were evaluated. Progressive motility and total motility were higher with the 500 and 750 µg/ml lycopene concentrations, respectively, compared to the control group and the cysteamine groups following 72 hr of storage in the liquid storage. Motility characteristics, viability and hypo-osmotic swelling test (HOST) percentages were significantly improved in 500 µg/ml lycopene compared to other groups. The 500 and 750 µg/ml lycopene concentrations, respectively, showed significantly reduced percentages of spermatozoa with DNA integrity compared to the control group. The 500 and 750 µg/ml lycopene concentrations, respectively, led to the significant decrease of MDA levels. The 500 µg/ml lycopene enhanced TAC levels after 48 and 72 hr that was not observed in other groups. In conclusion, the findings of this study showed that lycopene supplementation in canine semen extenders improved canine semen parameters and TAC levels and decreased MDA levels in the chilling process.     

Prognostic value of tumour‐associated macrophages in canine mammary tumours

Tumour‐associated macrophages (TAMs) have already been associated in human breast cancer to a poor prognosis. As a part of a tumoural microenvironment, TAMs have an important contribution influencing neoplastic progression. Hitherto, in canine mammary tumours (CMT) the prognostic value of TAMs has not been reported. In this study, MAC387 immunohistochemical expression was evaluated in 59 CMTs (20 benign and 39 malignant). The TAM value was significantly higher in malignant than benign CMT (P = 0.011). In malignant CMT, TAMs were associated with skin ulceration (P = 0.022), histological type (P = 0.044), nuclear grade (P = 0.031) and tubular differentiation (P = 0.042). The survival analysis revealed a significant association between tumours with higher levels of TAMs and the decrease in overall survival (P = 0.030). TAMs have proven to have a prognostic value. These findings suggest the future possibility of using TAMs as a novel therapeutic target in CMT.     

硝化应激在神经型犬瘟热疾病发展中的临床意义

为了研究硝化应激在神经型犬瘟热疾病发展中的作用,并探讨其对该病的诊断价值,选取犬瘟热临床病例113例,其中有神经症状组(NCD)共37例,无神经症状组(NNS)共76例,并设25例无犬瘟热健康犬作为对照组(CG).就诊时分别采取血液和脑脊液,测定一氧化氮(NO)、神经元型一氧化氮合成酶(nNOS)和神经元特异性烯醇化酶...     

Prognostic usefulness of blood leukocyte changes in canine parvoviral enteritis

BACKGROUND: Despite treatment, many dogs still die of complications related to canine parvoviral (CPV) enteritis. Effective prognostication would be beneficial in managing this disease. HYPOTHESIS: We hypothesize that the occurrence of leukocytopenias at admission and at 24 and 48 hours after admission, and changes in absolute leukocyte counts over time, could be used to predict outcome. ANIMALS: Sixty-two puppies with confirmed CPV. METHODS: A prospective study was performed. CBC was performed daily until discharge or death (in which case a postmortem examination was performed). RESULTS: Of the nonsurvivors (10/62; 16%), 9 died because of complications of the disease and 1 was euthanized because of a poor prognosis. There was a statistical significant difference in the occurrence of leukocytopenias between groups at 24 and 48 hours postadmission. The survivors showed a significant increase over time in certain leukocyte types (specifically lymphocytes) compared with values at admission. The positive predictive value for survivors was high. Nonsurvivors had marked thymic and lymphoid atrophy and marked bone marrow hypocellularity. CONCLUSION: An accurate prognosis could be obtained at 24 hours after admission by evaluating the change in total leukocyte, band neutrophil, lymphocyte, monocyte, and eosinophil counts.     

Use of oseltamivir in the treatment of canine parvoviral enteritis

Objective – To determine if oseltamivir with standard therapy for canine parvoviral enteritis ameliorates disease morbidity, mortality, or both; to document significant adverse effects associated with its use.
Design – Prospective, randomized, blinded, placebo-controlled clinical trial.
Setting – University veterinary teaching hospital.
Animals – Thirty-five dogs.
Interventions – Standard therapy was administered to all dogs. Treatment dogs also received oseltamivir, while control dogs received an equivalent volume of placebo.
Measurements and Main Results – Dogs were monitored daily according to a clinical scoring system, physical parameters, and diagnostic evaluations. Dogs in the treatment group gained a significant percentage of weight during hospitalization (mean, +2.6%; SD, 7.1%) versus the control dogs (mean, −4.5%; SD, 6.9%) ( P =0.006). Treatment dogs did not have any significant changes in their white blood cell (WBC) count, while control dogs experienced a significant drop in their WBC counts during their initial stay. In addition, it did not appear that oseltamivir use was associated with any major adverse clinical effects.
Conclusions – While a clear advantage to the use of oseltamivir was not established, a significant weight loss during hospitalization, as well as a significant decrease in WBC count were documented in the control group. No major adverse effects were identified that could be associated with oseltamivir administration. Based on these results, the true role of oseltamivir in the treatment of parvoviral enteritis remains speculative, although it is believed that further investigation is warranted.     

Oxidative stress indices in gastroenteritis in dogs with canine parvoviral infection

Gastroenteritis of viral origin has emerged as a major cause of morbidity and mortality in dogs during the last two decades. Amongst the viral etiologies responsible for gastroenteritis in dogs, canine parvovirus (CPV) is considered as the most pathogenic. The disease is characterized by hemorrhagic enteritis, bloody diarrhoea and myocarditis in young pups. The present study was carried out to examine alterations in oxidative stress indices in the erythrocytes from dogs suffering from gastroenteritis with or without canine parvoviral infection as confirmed by CPV-DNA amplification from faeces using specific primers for CPV-2 as well as CPV-2a and CPV-2b variants by polymerase chain reaction (PCR). The present investigation utilized clinical cases of dogs with signs of acute diarrhea (n = 56), and 14 more apparently healthy dogs of similar age group. Erythrocytic oxidative stress indices such as lipid peroxides level and antioxidant enzymes like superoxide dismutase and catalase activity, and blood micro-mineral (iron, copper, cobalt and zinc) status were analyzed in each dog (n = 70). The acute cases of gastroenteritis in dogs were associated with altered erythrocytic lipid peroxidation as evident by estimation of malonaldehyde (MDA) concentration. The activities of antioxidant enzymes catalase and superoxide dismutase, the first line of antioxidant defense against damaging effects of free radicals, were also altered. The alterations in oxidative stress indices were more pronounced in cases with involvement of canine parvovirus as compared to parvo-negative cases. Our results also revealed decreased blood zinc level in diarrhoea in dogs irrespective of involvement of canine parvovirus.     

Possible application of oxidative stress parameters for the evaluation of animal welfare in sheltered dogs subjected to different environmental and health conditions

The objective of this study was to establish an additional prognostic parameter to assess animal welfare. We explored the validity of using the thiol antioxidant barrier (SHp) and determination of reactive oxygen metabolites (dROMs) as indicators of stress and welfare in sheltered dogs. The shelter is a stressful environment for a dog because of its unpredictable and uncontrollable nature. The dogs enrolled in the present study belonged to 2 types of animal shelters (shelter 1 and shelter 2) that differed for social and spatial restrictions for the dogs. Animals were tested for leishmaniosis and ehrlichiosis and divided into 4 groups on the basis of their negative (group A and B) or positive (group C and D) status and the shelter they belonged to. The Student t test showed significant differences on SHp concentrations between groups A and B (negative titers; different shelters) (P < 0.0001; t₍₁₀₎ = 11.08), groups C and D (positive titers; different shelters) (P < 0.02; t₍₇₎ = 2.998), and groups A and C (different infection status and same shelter) (P < 0.004; t₍₈₎ = 3.975). Levels of dROMs showed significant differences between groups A and C (different infection status and same shelter) (P < 0.03; t₍₈₎ = 2.552) and groups B and D (different infection status and same shelter) (P < 0.02; t₍₉₎ = 2.817). The high dROMs values recorded in all dogs during the study may be because of the stressful environment of animal shelters, in general. The low SHp concentrations that we found in dogs from shelter 1 may suggest a highly stressful condition related to the poor social environment for interaction. SHp and dROMs may provide useful information about the responsiveness of sheltered dogs subjected to different environmental (social and spatial restrictions, management practices, and diet) and health (negative or positive status for leishmaniosis and ehrlichiosis) conditions and may suggest the possibility of establishing an additional prognostic tool for the assessment of welfare and health in sheltered dogs.     

Oxidative burst and phagocytic activity of phagocytes in canine parvoviral enteritis

Canine parvoviral enteritis (CPE) is a severe disease characterized by systemic inflammation and immunosuppression. The function of circulating phagocytes (neutrophils and monocytes) in affected dogs has not been fully investigated. We characterized the functional capacity of canine phagocytes in CPE by determining their oxidative burst and phagocytic activities using flow cytometry. Blood was collected from 28 dogs with CPE and 11 healthy, age-matched, control dogs. Oxidative burst activity was assessed by stimulating phagocytes with opsonized Escherichia coli or phorbol 12-myristate 13-acetate (PMA) and measuring the percentage of phagocytes producing reactive oxygen species and the magnitude of this production. Phagocytosis was measured by incubating phagocytes with opsonized E. coli and measuring the percentage of phagocytes containing E. coli and the number of bacteria per cell. Complete blood counts and serum C-reactive protein (CRP) concentrations were also determined. Serum CRP concentration was negatively and positively correlated with segmented and band neutrophil concentrations, respectively. Overall, no differences in phagocyte function were found between dogs with CPE and healthy control dogs. However, infected dogs with neutropenia or circulating band neutrophils had decreased PMA-stimulated oxidative burst activity compared to healthy controls. Additionally, CPE dogs with neutropenia or circulating band neutrophils had decreased PMA- and E. coli–stimulated oxidative burst activity and decreased phagocytosis of E. coli compared to CPE dogs without neutropenia or band neutrophils. We conclude that phagocytes have decreased oxidative burst and phagocytic activity in neutropenic CPE dogs and in CPE dogs with circulating band neutrophils.     

Investigation of the usability of kisspeptin and oxidative stress parameters in the early diagnosis of asymptomatic cystic endometrial hyperplasia in dogs

This study aimed to investigate the differences in oxidative stress index (OSI) and kisspeptin levels in clinically asymptomatic dogs with cystic endometrial hyperplasia (CEH) compared to healthy and pregnant dogs, and to determine the usability of the obtained results in the diagnosis of asymptomatic CEH. The study comprised three groups; a healthy (n = 8), a pregnant (n = 10) and a CEH (n = 10). All dogs in the three groups were included in the study at the 30 ± 3th day after estrus, and blood samples were collected for analysis of kisspeptin, total antioxidant status (TAS), total oxidant status (TOS), progesterone (P4), estradiol (E2) and some biochemical parameters (TSH; thyroid stimulating hormone, ALT; alanine aminotransferase, AST; aspartate aminotransferase, ALP; alkaline phosphatase, LDH; lactate dehydrogenase, CRE; creatine and BUN; blood urea nitrogen). In addition, OSI value was calculated. P4 and ALT and BUN levels were significantly lower and higher in CEH group than the pregnant group, respectively (p < .05). While kisspeptin and TAS levels were significantly lower in CEH group compared to the healthy and pregnant groups (p < .01), OSI level increased dramatically. In conclusion, it was confirmed that CEH clearly affected kisspeptin and OSI levels, and it is thought that these parameters may be an alternative diagnostic tool for the detection of CEH after further studies.     

犬细小病毒南京分离株基因型分析

采用PCR技术对不同来源的犬细小病毒毒株进行了基因型鉴定,发现所鉴定的1982~2004年的5株犬细小病毒南京分离株和荷兰Intervet疫苗株(CPV-V154)均为CPV-2b亚型,而且相互之间的同源性均高于98.4%。     

Glucose and lipid metabolism disorders in the chickens with dexamethasone‐induced oxidative stress

The purpose of this study was to investigate the effects of long‐term treatment with dexamethasone (DEX) on the antioxidation and nutrition metabolism in broiler chickens. Broilers were placed on a high‐nutrient diet for 41 days, and half were given orally DEX‐supplemented water at 20 mg/L every other day from 19 to 41 days of age. DEX treatment downregulated superoxide dismutase activity as well as the mRNA expression of CuZn‐superoxide dismutase and glutathione peroxidase with a decrease in GSH/GSSG ratio and an increase in malondialdehyde level in the liver of broilers. DEX treatment aggravated oxidative damage in the liver and, therefore, increased the sensitivity of broilers to ascites syndrome with higher mortality and reduced growth performance. Serum metabolomics analysis showed that DEX treatment significantly increased the levels of glucose, intermediates in protein metabolism (valine, proline, serine, threonine and urea) and lipid metabolism‐related products (palmitic acid, stearic acid and cholesterol) while decreasing the levels of β‐hydroxy butyric acid, succinic acid and malic acid, demonstrating that DEX treatment inhibited the Krebs cycle and the oxidation of fatty acids, and promoted the de novo synthesis of fatty acids as well as protein decomposition in the liver of broilers. Additionally, detection of metabolism‐related enzymes revealed that DEX treatment inhibited glycolysis and promoted glycogen decomposition. In summary, DEX treatment resulted in oxidative stress and glucose and lipid metabolism disorders in the broilers.     

COX‐2 and c‐kit expression in canine gliomas

Gliomas are among the most common primary neural tumours of dogs. Cyclooxygenase‐2 (COX‐2) and c‐kit overexpression are associated with increased aggressiveness of gliomas and decreased survival in human beings. COX‐2 is the inducible form of cyclooxygenase, which catalyzes prostaglandin formation and may increase tumour proliferation and angiogenesis. C‐kit is a tyrosine kinase receptor involved in normal cell physiology; c‐kit is upregulated in some canine tumours. In this retrospective study, 20 canine gliomas were identified: 11 (55%) oligodendrogliomas, including 1 anaplastic variant; 1 (5%) oligoastrocytoma; and 8 (40%) astrocytomas, of which 2 were glioblastoma multiforme. None of the gliomas expressed COX‐2. None of the gliomas were immunoreactive for c‐kit, although all three high‐grade tumours had intramural vascular expression. Consequently, COX‐2 inhibitors would likely be ineffective against canine gliomas. C‐kit inhibitors may have an anti‐angiogenic effect in high‐grade gliomas, but would likely be ineffective in low‐ and medium‐grade tumours.     

Protein restriction in pregnant‐ and lactating rats‐induced oxidative stress and hypohomocysteinaemia in their offspring

Hyperhomocyteinaemia has been associated with pathological and stressful conditions and considered as a risk factor for cardiovascular diseases. Maternal protein restriction during late pregnancy and postnatal period is a stressful state to their offspring which is associated with disruption of various physiological processes. The objective of the current study was to evaluate the effects of maternal protein restriction during late pregnancy and early postnatal periods on systemic oxidative stress parameters and on homocysteine metabolism of their offspring. For this purpose, 12 female rats were divided into two groups: a control group (C) with free access to standard diet (20% protein) and a protein restricted group (PR) with free access to a low protein diet (7% protein) from the day 14 of pregnancy until day 14 after delivery. An increase of thiobarbituric acid‐reactive substance levels (TBARS) in plasma (+40%, p < 0.01) and in liver (+37%, p < 0.001) with a concomitant decrease in the activities of catalase (CAT) and superoxide dismutase (SOD) observed in undernourished pups suggest the possibility of oxidative stress. Moreover, no significant changes in total glutathione levels (tGSH) were also observed to be associated with a significant decrease in homocysteine concentrations (?35%; p < 0.001). In their mothers, an alteration of antioxidant enzyme activities (CAT and SOD) and slight increase of TBARS levels were observed. Whereas homocysteine and tGSH levels were unchanged in comparison with those fed ad libitum. These results suggested that PR diet given to mothers during late pregnancy and early postnatal periods induced oxidative stress and hypohomocysteinaemia in their offspring.     

南京地区犬细小病毒基因型鉴定

为调查南京地区犬细小病毒(canine parvovirus,CPV)基因型分布情况,采集南京市某宠物医院2012-2019年犬细小病毒病患犬粪便43份作为检测病料,通过PCR扩增犬细小病毒VP2基因片段,并将扩增产物测序,分析CPV基因型.结果显示,43份病料皆扩增出目的片段,经测序后使用MegAlign软件与Gen...     

Effects of the supplementation with an high‐polyphenols extra‐virgin olive oil on kinetic sperm features and seminal plasma oxidative status in healthy dogs

The aim of this work was to evaluate the effects of the supplementation of two extra‐virgin olive oils (EVOO) having different polyphenols content, on canine spermatozoa kinetic parameters and seminal plasma oxidative status. The study was conducted on 12 clinically healthy dogs of different breeds (2–7 years, 5–48 kg of body weight) divided into two groups: an experimental group supplemented with EVOO (Coratina cultivar) high in polyphenols (H‐P) and a control group fed EVOO (Cima di Bitonto cultivar) low in polyphenols (L‐P). The oil was daily administered per os (1 ml/3 kg BW) before meal. Semen collection was made twice at 15 days distance (D0₁ and D0₂) and then at 30 (D30), 60 (D60) and 90 (D90) days. Semen concentration and kinetic parameters were measured using computer‐assisted sperm analysis (CASA) system to evaluate: sperm total count, sperm motile (MOT%), progressive motility (PROGR%) and its fractions, straight‐line velocity (VSL, μm/s), curvilinear velocity (VCL, μm/s), average path velocity (VAP, μm/s), amplitude of lateral head displacement (ALH, μm), beat cross frequency (BCF, Hz), straightness (STR%) and linearity (LIN%). On seminal plasma, reactive oxygen species (ROS) and biological antioxidant potential (BAP) were tested. From findings, no differences were found for sperm MOT, VSL, VCL, VAP, ALH, BCF, STR, LIN and BAP. A gradual enhancement of PROGR% was observed in H‐P group (p < .01). The ROS levels were higher in dogs H‐P compared to the other group (p < .05). In conclusion, our results highlight the positive effects of EVOO polyphenols on sperm PROGR% in healthy dogs.     

Effects of in vitro hemodilution of canine blood on platelet function analysis using the PFA‐100

Background: The platelet function analyzer (PFA)‐100 is a point‐of‐care instrument previously evaluated in humans and dogs. In both species, artificially prolonged platelet closure time (CT) occurs with anemia. Reliability of the analyzer in dogs becomes a concern when the HCT is between 0.25 and 0.35 L/L. Objective: The objective of this study was to further define the level of HCT at which CT is prolonged, using in vitro diluted canine blood. Methods: Citrated whole blood samples were collected from 22 healthy dogs. Initial HCT was determined and autologous platelet‐rich plasma was added to samples to achieve HCTs of 0.33, 0.30, and 0.27 L/L. CT was determined in duplicate on the PFA‐100 using collagen/adenosine‐5′‐diphosphate cartridges. Results: Compared with the initial CT in samples with HCT 0.39–0.54 L/L (CT mean±SD=57.8±5.75 seconds), significantly prolonged CTs were found in hemodiluted samples with HCT 0.33 L/L (61.1±4.64 seconds), 0.30 L/L (64.3±6.79 seconds), and 0.27 L/L (70.8±7.90 seconds) (P=0.029; repeated measures ANOVA). Conclusion: Although statistical differences were found, further studies are needed to determine the clinical significance of the mild prolongation in CT associated with mild anemia. Until then, dogs with HCTs slightly <0.35 L/L should be evaluated cautiously for platelet dysfunction using the PFA‐100.