Morphologic evaluation of acute endometritis in mares with differing resistance to uterine infections

The study was designed to determine differences between normal mares and mares with endometrial pathology in the inflammatory response after bacterial challenge. Six normal mares (biopsy category I) and 4 mares with pathological endometrial changes (biopsy category II) were given an intrauterine infusion of β-hemolytic streptococci on the second day of estrus. All mares had a similar kind of inflammatory response after the bacterial inoculation as assessed by rectal and vaginal examinations. There were no significant differences in the amount of discharge, uterine tone, uterine size and cervical relaxation between the groups. Leukocytic response, as determined by endometrial smears and biopsies, was of the same magnitude in both groups. Two mares from the pathological group were not able to eliminate the infection, but had vaginal discharge and bacteriologically positive uterine swabs until the end of the experiment. It is concluded that the inability of some mares to clear uterine infections cannot be explained by a deficient inflammatory response.     

Simultaneous Flow Cytometric Analysis of Phagocytosis and Oxidative Burst Activity in Equine Leukocytes

This paper describes a method for simultaneously measuring phagocytosis and oxidative burst activity in equine peripheral blood leukocytes by flow cytometry. Opsonized propidium iodide-labelled Staphylococcus aureus (PI-Sa) was used to measure the uptake of bacteria by equine phacocytes and the oxidative burst activity by oxidation of dihydrorhodamine 123. The requirements to achieve optimal activity of phagocytosis and oxidative burst are described. The advantage of the simultaneous technique is that it provides both independent and comparative values for phagocytosis and the oxidative burst, for the detection of impaired mechanisms of microbial destruction. Furthermore, the technique allows evaluation of opsonization activity in this context.     

犬瘟热患犬中性粒细胞cAMP PDE活性变化及中药作用研究

研究犬瘟热患犬中性粒细胞(PMN)中环磷酸腺苷-磷酸二酯酶（cAMP-PDE）的活性变化及中药对其活性的体外调节作用，为抗炎药研究和病毒性炎症的中药治疗积累资料。提取犬瘟热患犬（n=9）和正常犬(n=6)的中性粒细胞制备cAMP-PDE样品，通过高效液相色谱(HPLC)法测定其活性及中药对其活性的影响。结果显示，犬瘟热患犬组中性粒细〖JP2〗胞cAMP-PDE活性（18.55%±2.66%）显著（P<0.05）高于正常组（11.54%±5.16%）。解表药等5类114味中药中芦根（77.27%）、胡黄连（70.41%）、女贞子（65.50%）、香薷（65.16%）、贝母（61.43%）、黄芩（60.67%）、淫羊藿（60.34%）、黄芪（59.13%）、青蒿（56.93%）、秦皮（55.55%）、辛夷（55.03%）、甘草（54.67%）、牛蒡子（53.40%）、麻黄（53.32%）14味对cAMP-PDE活性具有明显抑制作用。结果表明，中性粒细胞cAMP-PDE活性在犬瘟热患犬炎症过程中具有重要作用，对cAMP-PDE活性有明显抑制作用的中药对其炎症可能具有疗效，同时，中药抑制中性粒细胞中cAMP-PDE活性可能是其抗炎作用机理之一。     

A Preliminary Comparison of Plasma Fibrinogen Concentrations,Leukocyte Numbers and Erythrocyte Sedimentation Rate as Non-Specific Indicators of Inflammatory Conditions in Buffalo (Bubalis Bubalis)

Zargham Khan, M., Muhammad, G., Umar, A. and Ali Khan, S., 1997. A preliminary comparison of plasma fibrinogen concentrations, leukocyte numbers and erythrocyte sedimentation rate as non-specific indicators of inflammatory conditions in buffalo (Bubalis bubalis). Veterinary Research Communications, 21 (4), 265-271The plasma fibrinogen concentration (Fib), total leukocyte count (TLC), neutrophil, lymphocyte and monocyte numbers, and the erythrocyte sedimentation rate (ESR) were determined in 153 buffaloes suffering from different clinical conditions. Fib increased significantly (p < 0.05) in chronic mastitis, pyrexia, pyometra, cutaneous abscesses, tail gangrene and acute indigestion, whereas in most of the other conditions studied it varied non-significantly. TLC increased significantly in chronic mastitis, pyrexia, endometritis, cutaneous abscesses and infected skin wounds. An increase in neutrophils was associated with an increased TLC. Numbers of lymphocytes varied non-significantly in most of the conditions. Monocytes decreased significantly in most of the acute conditions. ESR was significantly elevated in all clinical conditions. Significantly increased mean Fib values in the different conditions varied from 703 ± 119 to 725 ± 140 mg/dl, while the maximum individual value was 1510 mg/dl in a case of cutaneous myiasis. The significantly increased mean TLC ranged from 9.48 ± 2.91 to 11.1 ± 3.5 × 103/µl, while it was 21.7 × 103/µl in a case of meningitis. ESR values in sick buffaloes varied from 57 to 111 mm in the first hour.     

Italian University System has to Match Educational Needs in Adherence to European Standards and Professional Scenario

The effect of a water-soluble fraction (WSF) of a non-pathogenic strain of Mycobacterium phlei was studied in bovine subclinical mastitis (SCM) by measuring the myeloperoxidase and acid phosphatase enzyme levels in the milk leukocytes. Forty-five cows were divided into three equal groups. Group I, consisting of 15 healthy cows, served as the control, whereas groups II and III each contained 15 cows with subclinical mastitis on the basis of a positive reaction in the California mastitis test (CMT). The cows in group II received 100 microg of WSF in 5 ml sterile phosphate-buffered saline, pH 7.4 (PBS) once only, while those in group III received 5 ml sterile PBS daily for 7 days, both treatments being given by the intramammary route. Observations were made up to 30 days after treatment (AT). The CMT of the healthy milk was negative (0), whereas it ranged between 1 and 2 points in SCM. The somatic cell count (SCC) increased significantly (p < 0.05) on day 3, then fell steeply from day 7 up to day 30 AT in the cows in group II. A steady decrease in the total bacterial count (TBC) was observed in the group treated with WSF but the bacterial counts remained high in the groups treated with PBS. The mean acid phosphatase level was enhanced by 119% on day 3 AT in group II but only by 18.7% in the cows in group III. The mean myeloperoxidase level was enhanced by 100% in the cows in group II but only by 18% in those in group III on day 3 AT. This significant reduction in the bacterial load in infected cows caused by intramammary infusion of WSF may be due to activation of the microbicidal activity of the neutrophils, but this requires confirmation.     

In vitro effect of recombinant human tumor necrosis factor-alpha on canine neutrophil apoptosis

Neutrophils (polymorphonuclear leukocytes: PMNs) are essential for the host defense against various infections and are often injurious to the host, causing inflammatory diseases where tumor necrosis factor-alpha (TNF-alpha) is suggested to play an important role. Since an effect of TNF-alpha on canine PMN apoptosis has not been studied, canine PMNs were stimulated with recombinant human (rh)TNF-alpha in the present study to investigate the effect of TNF-alpha on canine PMN apoptosis. PMN apoptosis and function to produce ROS were assessed by flow cytometry. Delayed apoptosis was observed in the PMNs treated with rhTNF-alpha at 100 ng/ml, accompanied by retention of capability to produce ROS. However, PMN apoptosis was accelerated by rhTNF-alpha combined with cycloheximide. Therefore, it is indicated that TNF-alpha is able to activate anti- and pro-apoptotic pathways in PMNs and that the inhibition of PMN apoptosis by TNF-alpha requires protein synthesis in the PMNs.     

Clenbuterol Affects the Expression of Messenger RNA for Interleukin 10 in Peripheral Leukocytes from Horses Challenged Intrabronchially with Lipopolysaccharides

On four occasions, four horses with heaves and four horses with small airway inflammatory diseases inhaled 0.9% saline based aerosol mixtures with or without lipopolysaccharides (LPS). Prior to the first saline and LPS inhalation, horses were untreated, while three and a half days prior to the third and forth inhalation horses had received 0.8 μ g/kg clenbuterol intravenously twice daily. The messenger RNA (mRNA) expression of tumour necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-10 and interferon- γ (IFN- γ) was investigated by RT-PCR, all of which were expressed in the white blood cells of samples collected. Inhalation of LPS only changed the cytokine expression profile of IL-10, IL-4 and TNF-α mRNA which were higher after challenge with LPS. However in those horses that were treated with clenbuterol the LPS-induced IL-10 mRNA expression was shown to be suppressed. Further changes in IL-4 and TNF-α were not significant. Thus the results of this study indicated that clenbuterol can modulate the expression of IL-10 mRNA in peripheral white blood cells in those horses with small airway diseases that have been exposed to LPS. van den Hoven, R., Duvigneau, J.C., Hartl, R.T. and Gemeiner, M., 2006. Clenbuterol affects the expression of messenger RNA for interleukin 10 in peripheral leukocytes from horses challenged intrabronchially with lipopolysaccharides. Veterinary Research Communications, 30(8), 921–928     

Studies on Bovine Leukosis

Summary

An enzyme linked immunosorbent assay (ELISA) and the agar gel immunodiffusion test with bovine leukosis virus glycoprotein as antigen (AGIDT‐BLV gp) were further used to test 633 bovine sera for antibodies to BL V. Both tests detected the same number of sera positive (149) or negative (464) for antibodies. Nine sera were negative in the ELISA but found to be weakly positive (2 sera) or bending the control line (7) in the AGIDT‐BLV gp. On the other hand 11 sera were scored negative in the AGIDT‐BLV gp but were weakly positive (9 sera), positive (1), and strongly positive (I) in the ELISA. Both tests are used routinely in this Institute as they complement each other, specially if sera with low antibody titers are under investigation. It is concluded that ELISA can fully replace radioimmunoassays in the serodiagnosis of enzootic bovine leukosis.     

Diurnal variation of canine blood leukocyte counts

A diurnal variation was detected in the numbers of total leukocytes, neutrophils, lymphocytes and eosinophils in the blood of 19 dogs (10 beagles and nine German shepherds). Blood samples were collected every fourth hour for 24 hours and once a day for 7 days. The neutrophil count increased during the day and reached its maximum in the late afternoon. The lymphocyte count had its maximum values in the late evening and its minimum values in the early morning. The eosinophil numbers were low around noon, increased during the afternoon and reached their maximum numbers in the late evening. Leukocyte numbers had statistically significant diurnal variation, so in designing research protocols with repeated blood sampling and closely controlled factors it may be important to take blood samples at the same time every day. The mild normal leukocyte changes during the day are not likely to confuse interpretation of clinical cases where patient results are compared with wide reference ranges.     

IN VITRO EVALUATION OF FELINE LEUKOCYTES RADIOLABELED IN WHOLE BLOOD WITH 99MTC STANNOUS COLLOID

Technetium-99m stannous colloid (^99mTcSnC) has been used to radiolabel human leukocytes to investigate various inflammatory disorders. We investigated the in vitro behavior of feline leukocytes labeled in whole blood with ^99mTcSnC. Heparinized blood samples were collected from healthy cats and divided into control and test aliquots. The latter were labeled with ^99mTcSnC using a standard procedure. Leukocyte viability was determined for each sample using a trypan blue exclusion test. Labeling efficiency was determined for test aliquots. Test aliquots were layered onto Histopaque-1077^® and centrifuged before measurement of radioactivity of the blood components. Leukocytes from radiolabeled and control samples were washed and incubated with opsonized zymosan particles to allow assessment of phagocytic function. Aliquots were taken from radiolabeled feline leukocyte samples at 1, 3, 4, and 7 h postlabelling. After centrifugation of each aliquot, radioactivity of the supernatant and pellet was measured and the labeling retention determined. Leukocyte viability in both radiolabeled and control samples was >98%. The labeling efficiency was 95.2±0.14%. The distribution of radioactivity in feline blood was found to be 3.4±0.18% in plasma, 39.0±0.37% in erythrocytes, and 57.6±0.38% in leukocytes. Labeled feline leukocytes had phagocytic activity of 90.9±0.18% (control 91.3±0.15%). The radiolabeled leukocytes retained 93.4±0.19% of the radioactivity up to 7 h postlabeling. ^99mTcSnC efficiently labeled feline leukocytes with no effect on viability and minimal effect on phagocytic function. The percentage retention of radioactivity by the leukocytes was still high at 7 h postlabeling.