Canine serum alkaline phosphatase isoenzymes detected by polyacrylamide gel disk electrophoresis

Serum alkaline phosphatase (ALP) isoenzymes were studied in normal dogs using a commercially available polyacrylamide gel disk electrophoresis kit (PAG/disk kit). Serum samples taken from the dogs were incubated with neuraminidase, after which most showed ALP isoenzymes as two characteristic stained bands. To determine the origin of each band, ALP isoenzymes of serum and tissue extracts (liver, intestine and bone) were characterized by heating, wheat germ agglutinin (WGA) and levamisole treatments. The results suggested that the band detected on the anode was liver ALP (LALP) and that the band detected on the cathode represented bone ALP (BALP), and both were corticosteroid-induced ALP (CALP). The percentage of each ALP isoenzyme to total ALP activity was estimated by densitometry. The percentage of BALP was the highest in young dogs (age<1 year, 64.7% ), and this value decreased with age. In contrast, the percentage of LALP in young dogs (22.2%) was much lower than that in middle-aged dogs (ages 1 year to 7 years, 59.3%) and old dogs (ages>7 years, 50.4%). The present results suggested that a commercially available PAG/disk kit is capable of detecting three serum ALP isoenzymes in dogs, and further that it may have clinical applications in the evaluation of ALP isoenzymes in veterinary medicine.     

Whole-cell protein profiles of Haemophilus paragallinarum as detected by polyacrylamide gel electrophoresis

The soluble whole-cell protein profiles of 15 isolates of Haemophilus paragallinarum were examined using standardized sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The patterns were reproducible and the isolates were similar overall. Despite this similarity, two protein profile types were recognized.     

Comparison of electron microscopy and polyacrylamide gel electrophoresis in the diagnosis of avian reovirus and rotavirus infections.

Electron microscopy (EM) and genome electropherotyping by polyacrylamide gel electrophoresis (PAGE) for the detection of avian rotaviruses and reoviruses in intestinal specimens and cell cultures were compared. Fifty-eight field samples of intestine with intestinal contents, referred to as direct specimens, from turkey and chicken flocks located in different regions of California and submitted during 1989 for virus isolation were randomly selected as test samples. Also, 38 field intestinal specimens with suspected viral infection that had been passaged three times in primary chicken embryo kidney (CEK) cell cultures were used in their third passage. The percentage of agreement and the Kappa statistic of positive and negative results between these two tests were calculated. In the comparison, EM was considered the standard test. By statistical analysis, an agreement of 87% was observed in cell-culture samples analyzed by the two virus-detection methods, as contrasted with an agreement of 72% for direct specimens. The analysis of the number of segments and band migration profiles of reference and field virus strains indicated that only reoviruses replicated in CEK cell cultures and mainly rotaviruses were detected by both tests in direct specimens. The Kappa statistic analysis indicated substantial agreement (0.69) between the two tests for CEK samples, with moderate agreement (0.45) for the direct specimens examined.     

Mycoplasma gallisepticum strain variations detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Various strains of Mycoplasma gallisepticum (MG) were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Minor but distinct and reproducible differences in protein banding patterns were detected between strains, which included the vaccine F strain from various sources, an atypical (variant) strain, and the standard (A5969, S6) strains.     

Protein analysis of Babesia caballi merozoites by two-dimensional polyacrylamide gel electrophoresis and western blotting

Babesia caballi merozoites were prepared by combining two improved methods of cultivation and purification of merozoites using Percoll-gradiation, and the protein compositions of merozoites were analyzed by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. The relative molecular masses of the major proteins and protein masses separated by electrophoresis were >94, 80-70, 50-45, 34-30, 30-28 and 18 kDa. By Western blotting, twelve proteins or protein groups were recognized by pooled sera from two horses experimentally infected with B. caballi. Among twelve proteins, five new proteins (54, 30-26, 24, and two 18 kDa) were identified, and the 48 kDa protein was revealed to consist of 2 components in the B. caballi merozoite. One protein (54 kDa) of B. caballi was also recognized by the pooled sera from two horses experimentally infected with B. equi.     

Characterization of eugonic fermenters group EF-4 by polyacrylamide gel electrophoresis and protein immunoblot analysis

Whole-cell lysates and proteinase K-extracted lipopolysaccharide (LPS) of 19 strains of the group eugonic fermenter-4 (EF-4) were analyzed by electrophoresis and protein immunoblotting. These strains were isolated from dog- and cat-bite abscesses in human beings, ferret and human gastric lesions, and cat-lung infections. These strains represent 2 biovar groupings; EF-4a biovars ferment glucose and possess arginine dihydrolase activity, whereas EF-4b biovars do not. Electrophoresis of whole-cell lysates could distinguish between these biovars groups. Electrophoresis of LPS extracts revealed that all strains of EF-4 possess smooth chemotypes. Two strains of EF-4a reacted weekly in protein immunoblots and revealed distinct LPS profiles. These studies suggests that subgroups of EF-4 biovars may exist.     

Evaluation of total protein content in tears of dogs by polyacrylamide gel disk electrophoresis.

Concentration of total proteins was measured and sodium dodecyl sulfate-polyacrylamide gel disk electrophoresis was performed on tear and plasma samples obtained from 26 healthy dogs, and the results were compared. Mean +/- SEM concentration of total proteins in tears was 0.63 +/- 0.04 g/dl, and significant effects of age or gender were not found. The protein composition of tears in dogs was complex, and bands from light and heavy chains of immunoglobulins were identified by electrophoresis.     

聚丙烯酰胺电泳检测犊牛轮状病毒

从疑似轮状病毒腹泻犊牛粪便样品中提取病毒RNA。将提取的核酸进行聚丙烯酰胺凝胶电泳,6个样品中有4个样品的电泳条带具有轮状病毒特征条带．证明该牛群有轮状病毒感染。     

Setaria digitata in cattle of Thailand identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis.

Adult Thai Setaria worms collected from cattle which were bred, housed and slaughtered in Thailand were morphologically identified as Setaria digitata. Furthermore, in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) adult Thai S. digitata had the same protein profiles as adult Japanese S. digitata, but did not possess the protein with a molecular size of 69 kDa which was confirmed in adult S. marshalli. In addition, there were no differences in the protein profiles between male and female S. digitata. In point of the distribution pattern of the proteins ranging from 73 to 64 kDa revealed by 2D-PAGE, there were no differences between Thai and Japanese S. digitata, and between male and female worms of the species.     

Genetic diversity of Mycobacterium avium subspecies paratuberculosis isolates from goats detected by pulsed-field gel electrophoresis

Paratuberculosis in goats occurs worldwide causing considerable economic losses mainly due to reduced milk production. Nowadays, there is still relatively little knowledge about the epidemiology of this disease in goats, and only a few epidemiological studies have been carried out in goats naturally infected with Mycobacterium avium subspecies paratuberculosis (M. a. paratuberculosis). The objective of this study was to characterize forty four clinical caprine isolates of M. a. paratuberculosis by different molecular techniques (pulsed-field gel electrophoresis [PFGE], restriction fragment length polymorphism analysis coupled with hybridization to IS900, and IS1311 polymerase chain reaction-restriction enzyme analysis) to determine the most useful technique for molecular typing of caprine isolates, as well as to disclose the genetic variation amongst caprine isolates and the relationship with strains isolated from other animal species. PFGE was found to be the most discriminative technique identifying a total of 13 'multiplex' PFGE profiles, ten of which were novel profiles found only in caprine isolates to date. All isolates were genotyped as Type II strains, except two isolates that resembled the intermediate group referred as Type III.     

A study of antigenic variants of foot-and-mouth disease virus by polyacrylamide gel electrophoresis of their structural polypeptides

Twenty-nine foot-and-mouth disease (FMD) type A virus strains, previously classified serologically as distinct subtypes were analysed by polyacrylamide gel electrophoresis (PAGE) to determine the extent of variation in the pattern of the structural polypeptides and to evaluate the technique as an aid to existing subtyping techniques. The majority of the subtypes examined had distinct polypeptide patterns, however, some variation also occurred between strains within a subtype. The position of VP2(1B) and VP3(1C) was often unchanged in different strains within a subtype and between geographically related subtypes over long periods of time. Changes in the position of VP1(1D) were also observed within a subtype. The technique was considered to be of value for the screening of isolates prior to conventional serological subtyping procedures and in the tracing of the possible origin of FMD outbreaks.     

Demonstration of polymorphism among Brucella ovis field isolates by pulsed-field gel electrophoresis

Brucella ovis is recognized worldwide as an important pathogen of sheep, and has also been identified in farmed deer in New Zealand. Previously, only one strain type of B. ovis has been identified. The objective of this paper was to perform pulsed-field gel electrophoresis (PFGE) on field isolates of B. ovis to determine whether strain variations exist, whether sheep and deer are affected by the same strains, and to compare the performance of the rare-cutting restriction enzymes XbaI and SwaI. Ten B. ovis isolates from sheep and two from deer in New Zealand, as well as the type strain, were subjected to PFGE analysis using both XbaI and SwaI. PFGE of XbaI restriction fragments produced two banding patterns consisting of 27-28 bands, which were found to be 98% similar by cluster analysis, and were named X1 and X1a. PFGE of SwaI restriction fragments resulted in three banding patterns consisting of 13-15 bands each. Ten of the isolates had identical banding patterns and were named S1. One isolate differed by one band, representing a subtype named S1a. Two isolates differed by six bands, representing a different strain type of B. ovis and this was named S2. Cluster analysis showed S2 to be 78% similar to the S1/S1a cluster. Both strain types were isolated from both sheep and deer. Thus, two distinct strain types of B. ovis were identified in New Zealand, which is the first report of more than one strain type being identified worldwide. Neither strain was species-specific for sheep or deer. The restriction endonuclease SwaI was found to be more discriminatory than the enzyme XbaI, which has been used in previous studies.     

双向凝胶电泳分析感染禽脑脊髓炎病毒鸡脑组织蛋白表达图谱

本研究采用双向凝胶电泳分离接毒组（感染AEV）和对照组（不接毒）的鸡脑组织全蛋白,经过银染后,用Image ScannerⅡ光密度扫描仪获取凝胶图像,并用Image Master 2D Platinum软件进行差异蛋白分析。对蛋白斑点编辑后,接毒组中检测到793个蛋白点,对照组中检测到827个蛋白点,两张图谱的匹配蛋白点数为614个,凝胶匹配率为75．8％。差异蛋白分析后,发现在接毒组中有16个蛋白点表达量显著下调,这些蛋白质成份可能与组织细胞病变有关,它们将为认识禽脑脊髓炎的发病机理提供线索。