Shin Oikawa

Detection of serum haptoglobin by enzyme-linked immunosorbent assay in cows with fatty liver.

Haptoglobin cannot be detected in the serum of healthy cattle by the haemoglobin-binding assay or single radial immunodiffusion. The present study was designed to examine whether an enzyme-linked immunosorbent assay could be applied to measure serum haptoglobin concentrations in healthy cows and in cows with fatty liver. When either purified cow haptoglobin or haptoglobin-positive serum were used as the antigen in the assay, the standard curves obtained were distinctly different, and haptoglobin in serum was detected more sensitively than the purified protein. The addition of bovine serum albumin to purified haptoglobin shifted the curve towards that obtained with haptoglobin-positive serum, suggesting that an interaction with serum albumin was partly responsible for the different standard curves. By use of the standard curve for haptoglobin in serum, the mean (SEM) concentration of haptoglobin in samples from four cows with fatty liver was 466 (147) micrograms ml-1, but the sera from four apparently healthy cows contained less than 0.01 microgram ml-1 haptoglobin.

Ultrasonographic evaluation of cattle and buffaloes with respiratory disorders

The purpose of the present study was to evaluate cattle and buffaloes with respiratory disorders, determine the extent of the lesions, and assess prognosis. The results were compared with the findings determined following physical examination and at slaughter. Animals were referred to Veterinary Teaching Hospitals because of inappetance, loss of body condition, cough, dyspnea, and nasal discharges. Ultrasonographically, it was possible to detect bronchopneumonia, consolidation, pleural effusion, pulmonary emphysema, and pleuritis. It was not possible to visualize lesions located deeper within the lungs where peripheral tissue was not affected. Laboratory findings included a neutrophilic leukocytosis, γ-globulinemia, and increased activity of aspartate aminotransferase. A diagnosis of respiratory disease was made on the basis of clinical and ultrasonographic findings and confirmed in 25 cases at slaughter. Bovine ultrasonography appears to be suitable as a screening tool for detection of pathologic lung processes near the pleura. Thoracic ultrasonography allows assessment of the extent and severity of pulmonary changes so that further evaluations can be considered.

Ultrasonographic imaging of experimentally induced pancreatitis in cattle.

This study was conducted to determine the ultrasonographic patterns of pancreatitis evoked in cattle, with reference to laboratory and pathological findings. Using ultrasonographic guidance, acute necrotizing pancreatitis was induced in six cows by injecting chloroform into the pancreatic tissue. Ultrasonographic examination was then performed once daily for nine days. Pancreatic lesions were visible 24h after induction of pancreatitis, as represented by a uniform increase in echogenicity and by intralobular and subcapsular fluid accumulation. As the experiment progressed, patchy hypoechogenic foci appeared within the gland parenchyma. Amylase and lipase activities showed rapid increases. Post mortem examination revealed gross and microscopic necrotic and haemorrhagic lesions in the body and right lobe of the pancreas, accompanied by oedema and fibrosis. Ultrasonography was found to be extremely useful for the detection and characterization of experimentally induced pancreatitis and to monitor its progression in the cow. These findings are of potential value as a reference for the diagnostic workup of bovine pancreatitis, and ultrasonography is seen as a promising non-invasive technique for the diagnosis of suspected pancreatitis in cattle.

Acute Escherichia coli Mastitis in Dairy Cattle: Diagnostic Parameters Associated with Poor Prognosis

ABSTRACT This study aimed to identify the diagnostic characteristics associated with poor prognosis and mortality in dairy cows with acute clinical Escherichia coli mastitis. On 17 dairy farms, 24 dairy cows with acute E. coli mastitis that had received therapeutic treatment were categorized into 2 groups by outcome: 17 cows that recovered (survivors) and 7 cows that died or were euthanized (non-survivors). Two days after onset of acute E. coli mastitis, dysstasia was observed in non-survivors, but not in survivors. Compared with survivors, significantly increased hematocrit (HCT) values and non-esterified fatty acid (NEFA) concentrations, and significantly decreased antithrombin activity and platelet counts were found in non-survivors on days 2 and 3 after therapy. Dysstasia, associated with decreased antithrombin activity and platelet counts, and with increased HCT and NEFA concentrations, was considered to be the major prognostic indicator associated with high mortality after therapeutic treatment in acute E. coli mastitis.

Comparison of serum lipid compositions, lipid peroxide, α-tocopherol and lipoproteins in captive marine mammals (bottlenose dolphins, spotted seals and West Indian manatees) and terrestrial mammals

Concentrations of serum lipid components, lipid peroxide (LPO) and alpha-tocopherol and electrophoretic patterns of lipoproteins in serum samples obtained from captive marine mammals and terrestrial mammals were compared. Serum concentrations of total cholesterol, free fatty acid, and phospholipid in fish-eating animals were significantly higher than those in manatees and cows. Serum LPO and alpha-tocopherol concentrations in the fish-eating animals were also significantly higher than those in manatees, cows and dogs. Different patterns of densitometric scans of low density lipoprotein (LDL) and a significantly lower percentage of LDL were demonstrated in the dolphins compared with the seals, cow and dogs. The concentration of LPO was significantly correlated with triglyceride and phospholipid concentrations in serum from the dolphins. These results suggest that triglyceride and phospholipid are susceptible to oxidative reaction in fish-eating animals. Evaluation of serum lipids, LPO and alpha-tocopherol concentrations is needed for nutritional husbandry for fish-eating animals.

Ultrasonographic Prediction of Hepatic Fat Content in Dairy Cows during the Transition Period

The objective of the present study was to emphasize the value of ultrasonography in predicting hepatic fat content in dairy cows during the transition period. For this purpose, five dairy cows during the transition period were examined by ultrasonography 3 times during this phase, 3wk before expected partition (wk -3), at parturition (wk 0) and 3wk after parturition (wk +3). Parallel, blood samples were collected and livers were biopsied. None of the cows showed clinical disease around calving. Compared with precalving values, the hepatic triglyceride and glycogen concentrations were higher at parturition (wk 0) and thereafter (wk +3). Histopathologically, there were no cellular abnormalities in hepatocytes 3 weeks before parturition. However, at parturition the hepatocytes contained some lipid drops in cytosol, and the cells after parturition (wk +3) turn opacificated severely and cellular membranes are indistinct. With increased liver TG content, hepatic ultrasonography showed increased overall echogenicity with vascular blurring. In the field, the visual evaluation of the brightness of hepatic tissue in dairy cows during the transition period can give a rough, but quick estimate of the liver fat content. To the best of the authors’ knowledge, this study is the first that shows hepatic changes by ultrasonography in dairy cows during the transition period.

Ultrasonographic diagnosis of omasal leiomyoma in a cow

GASTROINTESTINAL (GI) tumours in cattle are rare. In 1·3 million animals examined at slaughter in the UK, only three tumours occurred specifically in the GI tract (Anderson and Snadison 1969). Among 1400 cattle examined postmortem in New York, neoplasms were found in the forestomachs of eight animals (0·6 per cent), the only forms being fibroma and lymphosarcoma (Bertone and others 1985). Other reports of GI tumours in cattle include papilloma and squamous cell carcinoma in Kenya (Plowright 1955, Plowright and others 1971) and northern England (Jarrett 1973). Smooth muscle tumours of the GI tract have been found to develop mostly in the small intestine in dogs and cats (Engle and Brodey 1969, ter Haar and others 1998) and cows (Bertone 1990); none has been reported in the forestomachs of cows. Ultrasonography has proved to be the most effective and least invasive diagnostic method of detecting GI tumours in dogs and cats (Paoloni and others 2002), and it has also been used increasingly as part of the diagnostic evaluation of cattle with a variety of GI disorders, such as traumatic reticuloperitonitis (Braun and others 1993). However, the ultrasonographic features of GI tumours in cattle have not yet been delineated. This short communication describes the ultrasonographic, laboratory and clinicopathological findings of an omasal leiomyoma in a cow. A six-year-old Holstein cow, weighing 534 kg, presented with intermittent fever and a one-month history of inappetence. Clinical signs included anorexia, depression and ataxia. The cow remained thin and frequently ran a rectal temperature of 38·8 to 39·5°C; but heart and respiratory rates were within normal ranges (55 bpm and 16 breaths per minute, respectively). Rumen movements were inaudible. The back grip test, pain percussion test on the reticulum and pole test on the sternum elicited only a mild reaction. The left and right sides of the thorax and the right side of the abdomen were clipped and shaved. Ultrasonic coupling gel was liberally applied to the wet skin to ensure good contact and a 3·5 MHz linear transducer was firmly held against the body wall with the cow in a standing position. With the transducer placed over the left ventral thorax, the reticular contractions were monitored for three minutes, as described by Braun and Götz (1994). The entire ventral thorax right and left of the sternum and the right and left thoracic walls were then scanned up to the elbow joints. The outer contours of the reticulum were visualised and the echogenic changes recorded. The omasum was scanned medially and to the right of the ventral midline. The reticular biphasic contractions were not detectable, the reticular contour was uneven and a loss of normal layering was observed. Deposits of echogenic material appeared on the reticular wall, cavitated by hypoechogenic fluid, and a space-occupying mass was identified within the lumen of the omasum. The echotexture of the lesion was inconsistent and echogenic, and the margins were asymmetric (Fig 1). Blood gas analysis showed no abnormalities. Echocardiographic tracings demonstrated atrial fibrillation, a deep QRS complex and irregular T waves. Haematology showed leucocytosis (21,300 leucocytes/μl, reference range 4000 to 12,000/μl) and neutrophilia (18,851 neutrophils/μl, reference range 600 to 4000/μl). Serum chemistry revealed hypoalbuminaemia (8·6 g/litre, reference range 30·3 to 35·3 g/litre) and hyperproteinaemia (100 g/litre, reference range 21 to 36 g/litre), attributed to hypergammaglobulinaemia (73·4 g/litre, reference range 16·9 to 22·5 g/litre), which suggested a chronic inflammatory disease or neoplasia. These findings supported differential diagnoses of traumatic reticuloperitonitis, reticular abscess, reticular adhesions, pleuritis, and pulmonary and hepatic diseases. Ultrasonography of the forestomachs and liver narrowed the diagnosis to either traumatic reticuloperitonitis or a space-occupying mass in the omasum. The results also revealed a high activity of gamma glutamyl transferase (62 U/litre, reference value 6·1 to 17·4 U/litre), indicating a degree of liver impairment, which may have also accounted for the hypoalbuminaemia. All laboratory values were referenced from Kaneko and others (1997). The cow had a poor prognosis and was euthanased. Postmortem examination revealed fibrinous, thick-walled inflammatory deposits interspersed with jelly-like serous fluid adhering to the reticulum, omasum and abomasum, as well as to the rumen. A pedunculated, grape-like mass (18 x 14 cm) was attached to the ventral omasal wall (Fig 2). No ulceration or abrasions were observed on the surface of the mass and the cut surface had a lobulated appearance. Neither a diaphragmatic reticular adhesion nor any metallic foreign body or trace was observed. A specimen from the omasal mass was fixed in buffered 10 per cent formalin, embedded in paraffin, stained with haematoxylin and eosin and examined histologically. There was a proliferation of smooth muscle cells in the omasal muscularis in intertwisting fascicles of well differentiated cells; the fascicles arranged in an exceedingly irregular pattern, transversing at intersecting and obtuse angles to one another. The smooth muscle cells had abundant cytoplasm and irregular fusiform nuclei with vesiculated chromatin. There was marked submucosal oedema with diffuse infiltration of lymphocytes, macrophages and a few neutrophils. Mitotic figures were scarce. The histological picture was compatible with a diagnosis of leiomyoma (Fig 3). Leiomyomas, benign tumours of smooth muscle, are the most common gynaecological tumours in domestic animals FIG 1: Longitudinal sonogram of an omasal leiomyoma imaged at the seventh intercostal space. Note the asymmetric, inhomogenous and the echogenic patterns of the tumour. The mass appeared to project into the omasal lumen. Lt Lateral, Md Medial, 1 Right ventral thoracic wall, 2 Sternal part of the diaphragm and peritoneum, 3 Liver, 4 Omasal wall, 5 Omasal lumen

Transcutaneous ultrasound-guided pancreatic biopsy in cattle and its safety: a preliminary report

This study describes a free-hand technique for percutaneous pancreatic biopsy in cattle with ultrasound-guidance using a 14G spinal biopsy needle. Its safety was evaluated based on 36 consecutive procedures. To assess the immediate effects of pancreatic biopsy, 31 cows were necropsied shortly after the procedure and examined. The remaining five cows were examined daily for eight days and then necropsied and examined. No life-threatening complications nor clinically detectable abnormalities were observed. Changes indicative of inflammation were not apparent in total and differential WBC counts or in total protein and fibrinogen concentrations. A small increase of amylase activity was detected in only one cow kept for the eight-day observation period. Serum lipase activity increased significantly on day four after biopsy. Urea nitrogen, creatinine, glucose and the activities of aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyltransferase remained within reference ranges. Changes in the peritoneum and pancreas observed at necropsy were negligible. We conclude that percutaneous ultrasound-guided pancreatic biopsy did not appear to influence the cows condition adversely and the procedure provided an excellent method of obtaining a pancreatic specimen for histological examination. The procedure was considered safe, fast, cost-effective, and practical when performed properly. We believe that the technique can be used in cows with suspected pancreatic disease for making an ante mortem diagnosis.

Relevance of serum concentrations of non-esterified fatty acids and very low-density lipoproteins in nulli/primiparous and multiparous cows in the close-up period

Serum concentrations of non-esterified fatty acids (NEFA) and very low-density lipoprotein (VLDL) in close-up dairy cattle were compared in relation to parity. Data were obtained from 37 nulli/primiparous (NP) and 24 multiparous (MU, parity: 2–7) cows between 14 days and 1 day prepartum. A positive correlation (r=0.684, P<0.01) was found between serum NEFA and VLDL concentrations in NP cows. Among the VLDL constituents, the NEFA concentration was particularly correlated with the triglyceride (TG) concentration (r=0.658, P<0.01). However, no significant correlation was found between the concentrations of NEFA and VLDL or VLDL-TG in MU cows (r=−0.028 and 0.307). These results suggest the presence of higher hepatic secretion of NEFA-derived VLDL in NP cows.

Effectiveness of Isoprothiolane in Preventing Fatty Liver in Dairy Cows

In this study, we evaluated the effectiveness of isoprothiolane in preventing fatty liver in dairy cows. Eight pregnant Holstein cows with high body condition scores were studied. Four cows were administered isoprothiolane via feed at 10 g/day/head from 21 days before delivery (treated group), while the remaining four cows received no treatment (non-treated group). Liver samples from both groups were obtained 21 days before delivery and 2 days following delivery. No accumulation of lipid droplets in hepatocytes was observed in any cows prior to delivery. Following delivery, the accumulation of lipid droplets in hepatocytes was significantly reduced in the treated group compared to the non-treated group. The number of mitochondria per unit area was also significantly smaller in the treated group than in the non-treated group. Blood and biochemical test results from cows in the treated group showed an increase in mGOT levels and stable intrahepatic ATP production. These results suggest that isoprothiolane treatment intensified beta-oxidation in liver mitochondria, thereby activating the TCA cycle and maintaining intrahepatic ATP production.