Distribution of 5'-nucleotidase and gamma glutamyl transferase activities in the tissues of the horse

In the horse, 5'-nucleotidase (5'-NT) activity is found mainly in homogenates of lung, kidney, small intestine, mammary gland, liver and pancreas. Lower activities are present in brain and muscle. Activity can be demonstrated histochemically in the glomeruli and tubules of the kidney, in the sinusoidal borders of the hepatocytes and the bile duct epithelium as well as in the blood vessels of all organs. There is no significant difference between the 5'-NT activity in serum and plasma of normal horses and of horses suffering from a range of orthopaedic conditions. Previous findings that gamma glutamyl transferase activity is greatest in kidney and high in liver and pancreas have been confirmed.     

Further studies on the diagnostic value of gamma-glutamyl transpeptidase and 5'-nucleotidase in cattle, sheep and horses

The distribution of 5'-nucleotidase (5'-NT) and gamma-glutamyl transpeptidase (gamma-GT) is similar in the tissues of the sheep, calf and horse, except that there is relatively less gamma-GT in calf liver than in the liver of the other two species. The liver lesion produced by the oral administration of chloroform is similar in the three species and is accompanied by the release of 5'-NT into the plasma of the sheep and calf but not of the horse. Conversely, gamma-GT is released into plasma of the horse but not of the sheep or calf. This difference is not related to the tissue distribution of the two enzymes. The kidney lesion in sheep produced by the intravenous administration of mercuric chloride is accompanied by a reduction in the rate of excretion of an injected dose of inulin and by an increase in the concentration of urea in plasma and in the activity of gamma-GT in plasma and urine. There was no increase in 5'-NT activity in plasma or urine.     

Observations on gamma-glutamyl transferase, 5'-nucleotidase and leucine aminopeptidase activities in the plasma of the horse

In 18 horses there was no effect of age or sex on plasma activities of gamma-glutamyl transferase (gamma-GT), 5'-nucleotidase (5'-NT) and leucine aminopeptidase (LAP). All the enzymes were equally stable after storage for one month at -20 degrees C and there was no significant difference between their activities in serum and plasma in clinically normal horses. The pattern of release of gamma-GT, 5'-NT and LAP into plasma was studied in 114 horses which had a variety of orthopaedic, gastrointestinal, cardiovascular and hepatic (necrosis, lipidosis, neoplasia and cirrhosis) conditions. A definitive diagnosis of hepatic disease was established by histological examination of the liver. gamma-GT and 5'-NT were leaked into plasma in hepatic disease and gamma-GT was the more sensitive indicator of liver damage. There was some evidence that gamma-GT and 5'-NT plasma activities may increase in hepatic necrosis as well as in biliary obstruction. LAP was insensitive and not hepatic specific in the horse.     

PRRSV在仔猪体内的分布规律及其对组织的病理性损伤

为了研究猪繁殖与呼吸综合征病毒在仔猪体内的分布规律及其对组织的病理性损伤,试验将3头60日龄的健康仔猪随机分为2组,A组2头,B组l头.A组滴鼻接种PRRSV LC毒株(TC:ID50为1×10-5.25/0.1 mL)3 mL/头,B组为对照组,观察临床症状.攻毒25 d后剖杀,采集淋巴结、脾脏、肺脏和肝脏各2份,其中一份进行中性福尔马林固定,制作石蜡切片,观察组织病理学变化;另一份在-70℃保存,利用RT-PCR检测PRRSV.病理学检查结果表明:肺脏出现明显的间质性肺炎,肺泡渗出物增多,可见局灶性出血;脾脏和淋巴结可见出血、增生;肝脏中央静脉及门管区静脉可见明显的淤血.利用RT-PCR方法在淋巴结、脾脏、肺脏和肝脏中都检测到了PRRSV.     

PRRSV在仔猪体内的分布规律及其对组织的病理性损伤

为了研究猪繁殖与呼吸综合征病毒在仔猪体内的分布规律及其对组织的病理性损伤,试验将3头60日龄的健康仔猪随机分为2组,A组2头,B组1头.A组滴鼻接种PRRSV LC毒株(TCID50为1×10-5.25/0.1 mL)3 mL/头,B组为对照组,观察临床症状.攻毒25 d后剖杀,采集淋巴结、脾脏、肺脏和肝脏各2份,其中一份进行中性福尔马林固定,制作石蜡切片,观察组织病理学变化;另一份在-70℃保存,利用RT-PcR检测PRRSV.病理学检查结果表明:肺脏出现明显的间质性肺炎,肺泡渗出物增多,可见局灶性出血;脾脏和淋巴结可见出血、增生;肝脏中央静脉及门管区静脉可见明显的淤血.利用RT-PCR方法在淋巴结、脾脏、肺脏和肝脏中都检测到了PRRSV.     

The influence of feeding conditions on urine pH and plasma bicarbonate levels in healthy sheep

In healthy sheep the reaction of the urine has generally been accepted to be on the alkaline side (Bentinck-Smith 1963). Thus Healy et al. (1928) only found 2 of 40 healthy sheep with an acid urine. Bentinck-Smith reported that nursing animals in contrast to adults will produce an acid urine. However, the influence of feeding on urine pH and plasma bicarbonate levels in sheep does not seem to have been systematically investigated. During recent years it has often been observed in this laboratory that healthy animals, examined as a matter of routine, may produce an acid urine. Based on these observations healthy sheep under different feeding conditions were examined in order to elucidate the influence of the diet on the pH of the urine and the plasma bicarbonate content. Since an excessive lactic acid production and increased fat metabolism may also influence the acid-base balance, the lactic acid concentration of rumen fluid was determined, and the urine was tested for ketone bodies.     

Kidney-derived proteins in urine as biomarkers of induced acute kidney injury in sheep

Acute kidney injury (AKI) is a life-threatening condition for which an early diagnosis is problematic. The aim of the present study was to identify kidney-derived urinary proteins specific to AKI in sheep. AKI was induced in six sheep by an overdose of ketoprofen. Six untreated sheep served as controls. Urine samples were collected for up to 24 h after drug administration and pooled according to time and treatment. Tissue samples from kidney were taken immediately after euthanasia. Urinary proteins were separated by two-dimensional gel electrophoresis (2DE) and the proteins of interest were identified by mass spectrometry. Calbindin-D28k, retinol-binding protein 4 and CD1d were identified in ketoprofen-treated sheep, but not in controls. In addition, calbindin-D28k and CD1d were localized in kidney tissues by immunohistochemical staining. These preliminary results suggest that urinary calbindin-D28k and CD1d represent potential useful biomarkers of AKI, at least in sheep.     

Characterization of an ecto-5'-nucleotidase activity present on the cell surface of Tritrichomonas foetus

Tritrichomonas foetus is the causative agent of sexually transmitted trichomoniasis in cattle. In females, the infection can be associated with infertility, vaginitis, endometritis, abortion or pyometra, leading to significant economic losses in cattle raising. T. foetus is devoid of the ability to synthesize purine nucleotides de novo, depending instead on salvaging purines from the host environment. Ecto-5'-nucleotidase catalyzes the final step of extracellular nucleotide degradation, the hydrolysis of nucleoside 5'-monophosphates to the corresponding nucleosides and Pi. In this work we show that living, intact cells of T. foetus were able to hydrolyze 5'AMP at a rate of 12.57 ± 1.23 nmol Pi × h(-1) × 10(-7) cells at pH 7.2 and the 5'AMP hydrolysis is due to a plasma membrane-bound ecto-enzyme activity. The apparent K(m) for 5'AMP was 0.49 ± 0.06 mM. In addition to 5'AMP, the enzyme hydrolyzed all substrate monophosphates tested except 3'AMP. No divalent metals or metal chelators were able to modulate enzyme activity. Phosphatase inhibitors did not have an effect on ecto-5'-nucleotidase activity while ammonium molybdate did inhibit the activity in a dose dependent manner. The presence of adenosine in the culture medium negatively modulated the enzyme. These results indicate the existence of an ecto-5'-nucleotidase that may play a role in the salvage of purines.     

Identification of Chlamydophila abortus and the development of lesions in placental tissues of experimentally infected sheep

Chlamydophila (C.) abortus is a major cause of infectious abortion in sheep in many countries. Twenty-one pregnant sheep were experimentally infected intranasally with C. abortus at 70 days of gestation (dg). Thereafter, a number of animals were killed at weekly intervals and a post-mortem examination was carried out. Evidence of chlamydial infection in the placenta was determined by isolation of the bacterium by tissue culture and detection of C. abortus DNA by real-time polymerase chain reaction (real-time PCR). In addition, histopathological changes in the placenta were assessed, as was the detection of chlamydial antigen by immunohistochemistry (IHC). Evidence of placental infection was observed as early as 2 weeks after inoculation, and while only relatively low numbers of bacteria were isolated by culture and/or detected by real-time PCR prior to 113-114dg, at 119-121dg, it was more numerous. This study, using the four criteria for assessment of infection, showed that while C. abortus gained access to the placenta as early as 85dg, characteristic histopathological changes were not apparent until 119/121dg. While the chronology of when the bacterium arrived in the placenta and subsequent lesion development is remarkable for its consistency this paper provides more reliable data on the former which in turn now allows study of the factors that permit its access to this tissue and govern its multiplication and the ensuing triggering of damage.     

Cervical scoliosis and kidney lesions in sheep following dosage with vitamin D.

A case of cervical deformity of lambs following dosage with vitamin D was investigated. The lesions of a scoliosis are described together with the kidney lesions. It is suggested that doses of vitamin D normally considered to be non-toxic to sheep may prove to be so under certain conditions.     

The activity of enzymes in serum and tissues of clinically normal sheep

Extract

Feline infectious peritonitis was first described as a distinct disease entity in 1966 in the United States (Wolfe and Griesemer, 1966), although it had been observed prior to that date (Holzworth, 1963). The disease is widespread in that country (Disque et al., 1968: Hardy and O'Reilly, 1969; Ward and Pederson, 1969; Colby and Low, 1970; Colgrove and Parker, 1971) and has been recorded in Canada (Stephenson et al., 1971), England (Ingram, 1970), Ireland (Hartigan and Wilson, 1972), Japan (Konishi et at., 1971), the Netherlands (Mieog and Richter, 1971), Switzerland (Stunzi and Grevel, 1973) and most recently in Australia (Watson et al., 1974; Jones and Hogg, 1974). Two cases of feline infectious .peritonitis have, been seen in New Zealand (R. C. Gumbrell, pers. comm.). One experimental cat inoculated with peritoneal fluid from this case developed clinical signs and lesions said to be consistent with feline infectious peritonitis.     

Diurnal and seasonal variations of serum enzyme activity in cattle and sheep

In order to test the variation of enzyme activity in serum of cattle and sheep during the day, blood samples were taken at three hrs. interval from 6 a.m. to 9 p.m. The following enzymes were assayed: Aspartate aminotransferase (AspAT = GOT), alanine aminotransferase (AlAT = GPT), total lactate dehydrogenase (LDH), and a-hydroxybutyrate dehydrogenase (HBD). The variation between animals contributed by far to the greatest part of the total variation in clinical healthy animals. The time-of-day-dependant variation was less than 3 %, except for alanine aminotransferase.During the first two weeks of spring pasture serum aspartate and alanine aminotransferase levels were significantly raised in both cows and ewes, compared with serum levels of the same animals on indoor feeding. No such increase occurred in total lactate dehydrogenase.     

N-acetyl-beta-D-glucosaminidase activity in urine of cows with renal parenchymal lesions

OBJECTIVE: To measure N-acetyl-beta-D-glucosaminidase (NAG) activity in urine of cows with renal diseases and to correlate values for NAG activity with renal lesions. ANIMALS: 8 lactating Holstein cows and a Japanese Shorthorn cow, all of which had renal disease. PROCEDURE: Urine samples were collected, and urinary NAG activity and creatinine concentration were measured. The NAG activity was expressed as units per gram of creatinine (NAG index). Cows were euthanatized, necropsy was performed, and correlations between results for urinary NAG index and histopathologic findings for the kidneys were evaluated. RESULTS: The NAG activity and NAG index in urine samples obtained from cows with interstitial nephritis were high, ranging from 4.2 to 13.6 U/L and 3.5 to 23.0 U/g, respectively. A cow with renal amyloidosis also had high values for urinary NAG activity and NAG index. Histologic examination of the kidneys revealed various kinds of parenchymal lesions. However, urinary NAG index in cows with enzootic bovine leukosis was low. CONCLUSIONS AND CLINICAL RELEVANCE: Cows with renal diseases had high urinary NAG indexes that correlated well with their renal lesions, except for cows with enzootic bovine leukosis. Therefore, measurement of NAG index in urine samples has the potential to provide new perspectives on clinical diagnosis of renal disease in cattle.