Angelo Belloli

Postresuscitation treatment with argon improves early neurological recovery in a porcine model of cardiac arrest.

Introduction Effects of postresuscitation treatment with argon on neurologic recovery were investigated in a porcine model of cardiac arrest (CA) with an underlying acute myocardial infarction. Methods The left anterior descending coronary artery was occluded in 12 pigs, and CA was induced. After 8 min of untreated CA, cardiopulmonary resuscitation was performed for 5 min before defibrillation. Following resuscitation, animals were subjected to 4-h ventilation with 70% argon/30% oxygen or 70% nitrogen/30% oxygen. Myocardial function was echocardiographically assessed, and serum neuron-specific enolase was measured. Animals were observed up to 72 h for assessment of survival and neurologic recovery. Results All the animals were resuscitated and survived for 72 h, except for a control pig. Ventilation with argon did not have any detrimental effects on hemodynamics and respiratory gas exchange. All the six argon-treated animals had a fast and complete 72-h neurologic recovery, in contrast to only two of the six controls (P < 0.05). Seventy-two-hour neurologic alertness score and neurologic deficit score were, respectively, 100 and 0 in the argon group and 79 and 29 in the control one (P < 0.01 and P < 0.05). Significantly lower increases in serum neuron-specific enolase (12% vs. 234%) and minimal histological brain injury (neuronal degeneration: 0 vs. 1) were also observed in argon-treated animals, in comparison to controls. Conclusions In this model, postresuscitation treatment with argon allowed for a faster and complete neurologic recovery, without detrimental effects on hemodynamics and respiratory gas exchanges.

Spinal epidural abscess in two calves.

OBJECTIVE To report clinical signs, diagnostic and surgical or necropsy findings, and outcome in 2 calves with spinal epidural abscess (SEA). STUDY DESIGN Clinical report. ANIMALS Calves (n=2). METHODS Calves had neurologic examination, analysis and antimicrobial culture of cerebrospinal fluid (CSF), vertebral column radiographs, myelography, and in 1 calf, magnetic resonance imaging (MRI). A definitive diagnosis of SEA was confirmed by necropsy in 1 calf and during surgery and histologic examination of vertebral canal tissue in 1 calf. RESULTS Clinical signs were difficulty in rising, ataxia, fever, apparent spinal pain, hypoesthesia, and paresis/plegia which appeared 15 days before admission. Calf 1 had pelvic limb weakness and difficulty standing and calf 2 had severe ataxia involving both thoracic and pelvic limbs. Extradural spinal cord compression was identified by myelography. SEA suspected in calf 1 with discospondylitis was confirmed at necropsy whereas calf 2 had MRI identification of the lesion and was successfully decompressed by laminectomy and SEA excision. Both calves had peripheral neutrophilia and calf 2 had neutrophilic pleocytosis in CSF. Bacteria were not isolated from CSF, from the surgical site or during necropsy. Calf 2 improved neurologically and had a good long-term outcome. CONCLUSION Good outcome in a calf with SEA was obtained after adequate surgical decompression and antibiotic administration. CLINICAL RELEVANCE SEA should be included in the list of possible causes of fever, apparent spinal pain, and signs of myelopathy in calves.

Variations of serum cortisol in Argentine horses subjected to ship transport and adaptation stress

Abstract The response to stress after transport by ship and adaptation to a new environment was evaluated by variations of serum cortisol (RIA method) in 40 horses imported in Italy from Argentina. The study started at arrival (April 2000) and lasted 6 months (October 2000). As a control group, 42 horses living in Italy have been evaluated. In all subjects, hematocrit, total protein, total bilirubin, urea, creatinine, and creatinine kinase were determined. In control horses, cortisol values ranged between 103 and 278 nM/L (mean 190.24 ± SE 13.50, SD 43.82, CV 23.03), and no significant differences among sex or age groups were observed (P > .05). In subjects coming from Argentina, cortisol values observed at landing were within the normal range (193.73 nM/L ± SE 7.95, SD 50.29, CV 25.95), increased substantially in May (366.06 nM/L ± SE 10.25, SD 64.81, CV 17.70) and in June (337.19 nM/L ± SE 12.16, SD 76.89, CV 22.80) and returned to normal in October (187.87 nM/L ± SE 12.54, SD 43.50, CV 23.15). The differences observed in May and in June were significant in comparison with the values observed in April (P

Preliminary evaluation of total protein concentration and electrophoretic protein fractions in fresh and frozen serum from wild Horned Vipers (Vipera ammodytes ammodytes)

BACKGROUND Determination of the health status of reptiles is based on physical examination and evaluation of hematologic and biochemical values. Evaluation of serum total protein (TP) concentration and protein fractions plays an important role in health assessment; however, little is known about references value for these analytes in wild viperoid snakes. In addition, studies evaluating the stability of proteins in frozen viperoid serum are lacking. OBJECTIVE The aims of this study were to establish preliminary reference values for concentrations of TP and protein fractions in serum from wild vipers and to evaluate the stability of serum proteins in frozen serum samples from viperoid snakes. METHODS Blood samples were collected from wild Horned Vipers (Vipera ammodytes ammodytes). Using fresh serum, TP concentrations were determined using the biuret method and protein fractions were analyzed using agarose gel electrophoresis (AGE); albumin/globulin ratios were calculated. Analyses were also performed on serum frozen at -20°C for 70 days and then thawed. Pre- and post-storage results were compared using the Mann-Whitney U-test. RESULTS Five adult wild Horned Vipers were sampled and comprised 4 males and 1 female. The female snake had higher TP concentrations than the male snakes. The electrophoretic patterns demonstrated 6 protein fractions that were similar for all 5 snakes. There were no significant changes in the concentrations of the 6 protein fractions post-storage; the percentage of the alpha-1 fraction was increased in frozen/thawed serum. CONCLUSION Total protein concentrations in serum from Vipera ammodytes ammodytes were in agreement with published reference intervals for healthy reptiles and viperoid snakes. Serum protein fractions were easy to identify using AGE electrophoresis.

Intravenous immunoglobulin transfusion in colostrum-deprived dairy calves

Immunoglobulin transfusion is employed in the management of the failure of passive transfer (FPT). The aim of this study was to investigate the dose of immunoglobulin G (IgG) needed to reach a protective concentration (>10 g/L) in colostrum-deprived dairy calves. Twenty-eight Holstein Friesian newborn male calves were randomly assigned to either a control group (CG) or a treatment group (PG). Calves in the CG received 4 L of high quality colostrum within 12 h of birth. Calves in the PG received 62.7 ± 3.1 g of IgG IV in 2.6 ± 0.3 L of plasma within 6 h after birth. Serum immunoglobulin G (sIgG) and serum total protein (sTP) concentrations were assayed before and after (24 h, 72 h and 1 week after birth) plasma transfusion or colostrum ingestion. Serum (s) IgG and sTP concentrations increased in both groups throughout the period of observation. Mean sIgG and sTP concentrations after colostrum ingestion or plasma transfusion were higher in the CG than in the PG (P <0.01). Nine treated calves developed diarrhoea during the study and four were humanely euthanased due to progressive clinical deterioration. None of the calves in the CG showed signs of disease or died during the study. The dose of IgG used in this trial effectively provided an adequate sIgG concentration in colostrum-deprived calves (>10 g/L). Calves in the CG had significantly lower morbidity and mortality rates compared to those in the PG, suggesting that plasma transfusion alone is ineffective in providing complete protection against neonatal disease.

Effects of Storage Time on Total Protein and Globulin Concentrations in Bovine Fresh Frozen Plasma Obtained for Transfusion

To evaluate the effects of storage conditions on total protein (TP) and globulin fractions in fresh frozen bovine plasma units prepared and stored for transfusion, TP and globulin fractions were evaluated in fresh plasma and at 1 month and 6 and 12 months after blood collection in plasma stored at −20°C. Significant differences in concentrations were found in the median concentration of total protein (P = 0.0336), between 0 months and 1 month (P = 0.0108), 0 and 6 months (P = 0.0023), and 0 and 12 months (P = 0.0027), in mean concentration (g/dL) of albumin (P = 0.0394), between 0 months and 1 month (P = 0.0131), 0 and 6 months (P = 0.0035), and 0 and 12 months (P = 0.0038), and beta-2 fraction (P = 0.0401), between 0 and 6 months (P = 0.0401) and 0 and 12 months (P = 0.0230). This study suggests that total gamma globulin concentration in bovine frozen plasma is stable for 12 months at −20°C. Total protein, ALB, and beta-2 fraction have significantly different concentrations (g/dL) when compared to prestorage. This study has shown IgG protein fraction stability in bovine fresh frozen plasma collected for transfusion; therefore, bovine fresh frozen plasma seems to be suitable for the treatment of hypogammaglobulinemia (failure of passive transfer) in calves when stored for 12 months at −20°C.

Peculiarity of Porcine Amniotic Membrane and Its Derived Cells: A Contribution to the Study of Cell Therapy from a Large Animal Model

The aim of this work was to provide, for the first time, a protocol for isolation and characterization of stem cells from porcine amniotic membrane in view of their potential uses in regenerative medicine. From three samples of allanto-amnion recovered at delivery, the amniotic membrane was stripped from overlying allantois and digested with trypsin and collagenase to isolate epithelial (amniotic epithelial cells [AECs]) and mesenchymal cells, respectively. Proliferation, differentiation, and characterization studies by molecular biology and flow cytometry were performed. Histological examination revealed very few mesenchymal cells in the stromal layer, and a cellular yield of AECs of 10 × 10(6)/gram of digested tissue was achieved. AECs readily attached to plastic culture dishes displaying typical cuboidal morphology and, although their proliferative capacity decreased to the fifth passage, AECs showed a mean doubling time of 24.77 ± 6 h and a mean frequency of one fibroblast colony-forming unit (CFU-F) for every 116.75 plated cells. AECs expressed mesenchymal stem cell (MSC) mRNA markers (CD29, CD166, CD90, CD73, CD117) and pluripotent markers (Nanog and Oct 4), whereas they were negative for CD34 and MHCII. Mesodermic, ectodermic, and endodermic differentiation was confirmed by staining and expression of specific markers. We conclude that porcine amniotic membrane can provide an attractive source of stem cells that may be a useful tool for biomedical research.

Comparison of VIDAS and radioimmunoassay methods for measurement of cortisol concentration in bovine serum.

Radioimmunoassay (RIA) is the “gold standard” method for evaluation of serum cortisol concentration. The VIDAS cortisol test is an enzyme-linked fluorescent assay designed for the MiniVidas system. The aim of this study was to compare the VIDAS method with RIA for measurement of bovine serum cortisol concentration. Cortisol concentrations were evaluated in 40 cows using both VIDAS and RIA methods, the latter as the reference method. A paired Students t-test, Pearsons correlation analysis, Bland-Altman plot, and Deming regression analysis were used to compare the two methods. There was no statistically significant difference between mean serum cortisol concentrations measured by VIDAS or RIA methods (P = 0.6570). Both methods were able to detect significant differences in mean low and high cortisol concentrations (P < 0.00014 RIA and P < 0.0016 VIDAS). The correlation coefficient was low, but a Bland-Altman plot and Deming regression analysis show neither constant nor proportional error. The VIDAS method produced slightly higher values than RIA, but the difference was small and in no case did the mean value move the normal range. Results suggest that VIDAS method is suitable for the determination of bovine serum cortisol concentration in studies of large numbers of animals.

Aneurysm of the ductus arteriosus in a heifer.

A DUCTUS arteriosus aneurysm was first described in human beings by Martin Saint-Ange (1827), and has since been found in infants, children and adults (Falcone and others 1972). To the authors’ knowledge it has not been reported in cattle. In human beings, a delayed closure of the ductus arteriosus can be caused by neonatal hypoxaemia in premature babies, and by genetics that determine a lack of sufficient smooth muscle cells and an increased content of elastic tissue in full-term babies (Gersony 1986). It has been speculated that the cause of a ductus arteriosus aneurysm is a closure of the pulmonary end of the ductus arteriosus with a patent aortic end, which results in exposure of the vessel to systemic pressure (Bosman and Leoncini 1967, Falcone and others 1972, Lund and others 1992, Zilinskas and others 2000), but the pathogenesis remains unclear (Hornberger 2002). In adults, another hypothesis for a ductus arteriosus aneurysm is that focal degeneration and weakening of the aortic wall adjacent to the ductus arteriosus promotes the formation of an aneurysm (Falcone and others 1972, Lund and others 1992, Hirose and others 1999). Although the lesion can be asymptomatic, patients may have a history of dyspnoea or tachypnoea (Falcone and others 1972, Laurin and others 1992, Lund and others 1992). The aneurysm is often discovered accidentally on chest radiographs or after death from other causes (Lund and others 1991, Zilinskas and others 2000). A ductus arteriosus aneurysm may get bigger or rupture, causing death; progressive enlargement may cause heart failure, tachypnoea or dyspnoea (especially when drinking), sinus tachycardia and hoarseness from recurrent laryngeal nerve compression (Laurin and others 1992, Slovis and others 1992, Sattar and others 1996, d’Udekem and others 1997, Hirose and others 1999, Zilinskas and others 2000, Day and Walesby 2001). This short communication describes a case of ductus arteriosus aneurysm in a heifer. An eight-month-old female Brown Swiss heifer belonging to a dairy herd was admitted to the authors’ clinic for evaluation of a severe swelling of the head and anterior regions of the body that had appeared suddenly five days previously. On clinical examination, the heifer was in a poor physical condition; its body temperature was 39°C, its breathing was abdominal (36 breaths per minute), its pulse rate was regular (94 bpm), the mucous membranes were pale and the lymph nodes were normal in size. The animal presented with dyspnoea, severe ruminal tympany, and was affected by oedema that involved the head, neck, brisket, anterior limbs and extended as far as the umbilical region. Acoustic percussion of the thorax revealed enlargement of the area of cardiac dullness and caudoventral displacement of the lung limits. Auscultation of the thoracic wall revealed amplified heart sounds, increased bronchial tones and pathological lung sounds (wheezes and crackles). Radiographs were taken with the animal positioned in right lateral recumbency, and they showed a loss of the radiolucent space cranial to the heart. The cardiac silhouette could not be seen because an extensive zone of dense soft tissue was superimposed in the cranioventral field of the thorax. The dorsal displacement of the trachea strongly suggested a mediastinal mass (Fig 1). Perihilar distribution of an increased pulmonary radiodensity due to a mixed interstitial-alveolar pattern was also observed, and was consistent with cardiogenic alveolar oedema. Compensatory hyperinflation of the caudal lung lobes was also noticed. Ultrasound examination of the thorax, performed using a 3·5 MHz probe, identified a small degree of pericardial effusion and the presence of a mass adhered to the left thoracic wall; the mass had a wall 2·5 cm thick and an echogenic content. The clinical, radiological and ultrasonographic findings permitted the generic diagnosis of a major neoformation in the cranioventral mediastinum that extended to the left thoracic wall and involved the base of the heart, the trunk of the great arteries and the mediastinum. Because the wall of the neoformation was so thick, a mediastinal abscess or tumour was suspected. Considering the poor prognosis and the extent of the mass, the heifer was euthanased in agreement with the owner. At postmortem examination, the animal’s jugular veins were turgid and there was severe oedema of the cranioventral regions of its body, particularly in the subcutis of the head, neck and brisket. A large whitish mass was detected in the thorax; it measured 20 x 20 x 34 cm, and originated from a cranially displaced aortic arch and ended in the pulmonary artery at the normal insertion point of the ligamentum arteriosus (Fig 2). The mass adhered to the parietal pleura of the left side of the chest and to the apical and medial compressed Veterinary Record (2005) 156, 783-785

Effect of mild hypercapnia on outcome and histological injury in a porcine post cardiac arrest model

AIM OF THE STUDY To evaluate in an established porcine post cardiac arrest model the effect of a mild hypercapnic ventilatory strategy on outcome. METHODS The left anterior descending coronary artery was occluded in 14 pigs and ventricular fibrillation induced and left untreated for 12 min. Cardiopulmonary resuscitation was performed for 5 min prior to defibrillation. After resuscitation, pigs were assigned to either normocapnic (end-tidal carbon dioxide (EtCO2) target: 35-40 mmHg) or hypercapnic ventilation (EtCO2 45-50 mmHg). Hemodynamics was invasively measured and EtCO2 was monitored with an infrared capnometer. Blood gas analysis, serum neuron-specific enolase (NSE) and high sensitive cardiac troponin T (hs-cTnT) were assessed. Survival and functional recovery were evaluated up to 96 h. RESULTS Twelve pigs were successfully resuscitated and eight survived up to 96 h, with animals in the hypercapnic group showing trend towards a longer survival. EtCO2 and arterial partial pressure of CO2 were higher in the hypercapnic group compared to the normocapnic one (p < 0.01), during the 4-hour intervention. Hypercapnia was associated with higher mean arterial pressure compared to normocapnia (p < 0.05). No significant differences were observed in hs-cTnT and in NSE between groups, although the values tended to be lower in the hypercapnic one. Neuronal degeneration was lesser in the frontal cortex of hypercapnic animals compared to the normocapnic ones (p < 0.05). Neurological recovery was equivalent in the two groups. CONCLUSION Mild hypercapnia after resuscitation was associated with better arterial pressure and lesser neuronal degeneration in this model. Nevertheless, no corresponding improvements in neurological recovery were observed.