Phytotherapeutic effects of Echinacea purpurea in gamma-irradiated mice

Echinacea (E.) purpurea herb is commonly known as the purple coneflower, red sunflower and rudbeckia. In this paper, we report the curative efficacy of an Echinacea extract in γ-irradiated mice. E. purpurea was given to male mice that were divided into five groups (control, treated, irradiated, treated before irradiation & treated after irradiation) at a dose of 30 mg/kg body weight for 2 weeks before and after irradiation with 3 Gy of γ-rays. The results reflected the detrimental reduction effects of γ-rays on peripheral blood hemoglobin and the levels of red blood cells, differential white blood cells, and bone marrow cells. The thiobarbituric acid-reactive substances (TBARs) level, Superoxide dismutase (SOD) and glutathione peroxidase (GSPx) activities and DNA fragmentation were also investigated. FT-Raman spectroscopy was used to explore the structural changes in liver tissues. Significant changes were observed in the microenvironment of the major constituents, including tyrosine and protein secondary structures. E. purpurea administration significantly ameliorated all estimated parameters. The radio-protection effectiveness was similar to the radio-recovery curativeness in comparison to the control group in most of the tested parameters. The radio-protection efficiency was greater than the radio-recovery in hemoglobin level during the first two weeks, in lymphoid cell count and TBARs level at the fourth week and in SOD activity during the first two weeks, as compared to the levels of these parameters in the control group.     

Studies of Enteric Pathogens and γ-Globulin Levels of Neonatal Calves in Sweden

Viring, S., S.-O. Olsson, S. Alenius, U. Emanuelsson, S.-O. Jacobsson, B. Larsson, N. Linde and A. Uggla: Studies of enteric pathogens and γ-globulin levels of neonatal calves in Sweden. Acta vet. scand. 1993, 34, 271-279.– Faecal and blood samples were taken from 10-30% of calves, 36 hours to 14 days old, in 47 dairy herds in different regions of Sweden from September 1987 to October 1988 (Olsson et al 1993). Faecal samples from 279 calves were analysed for the presence of Escherichia coli (K99⁺), rotavirus and Cryptosporidium sp. Twenty (7.2%) of these samples were from diarrhoeic calves. An ELISA was developed and used for the rotavirus analysis. E. coli K99⁺ was detected in 11.5%, Cryptosporidium sp. in 6.1% and rotavirus in 5.4% of the faecal samples. The presence of rotavirus alone and the combination rotavirus and E. coli (K99⁺) was found to be associated with diarrhoea (p<0.001 and p<0.01 respectively).Blood samples from 327 calves were analysed for the level of total protein and γ-globulin. In 43 of these samples (13%) γ-globulin did not separate from the ß₂-region by electrophoresis. The mean total protein concentration was 53.6 g/1 in calves free from diarrhoea. The mean γ-globulin concentration, adjusted to 7 days age was 5.9 g/1. The 20 diarrhoeic calves had lower levels of both total protein and γ-globulin, compared with calves without diarrhoea, but the difference was not significant. One litre more of colostrum at the first feed increased the level of total protein of the calves’ sera by 1.4 g/1 (p = 0.05). Calves born between May and September had a 2.0 g/1 higher (p<0.001) serum concentration of γ-globulin than calves born between October and April.     

Agarose gel electrophoretic fractionation of serum proteins in adult cattle. I. A study of clinically healthy cows.

Blood samples were taken from 25 clinically healthy Swedish red-and-white dairy cows on 11 occasions in the course of about one year. The blood serum proteins were studied, chiefly by agarose gel electrophoresis, in respect of relative and absolute concentrations. The following parameters were studied: Total protein, albumin, total globulin, A/G quotient, α₁-, α₂-, inter-α-β-, β₁-, β₂- and γ-globulin. The material was analysed statistically in respect of variation between and within cows, and of age, pregnancy and seasonal variation. The variation between cows was found to be significantly greater than that between different samplings of the same cow. Older cows exhibited significantly higher γ-globulin concentrations. The variation with season and stage of pregnancy was less pronounced for most serum protein fractions.     

Agarose gel electrophoretic fractionation of serum proteins in adult cattle. II. A study of cows with different diseases.

In 283 dairy cows suffering from internal disorders the serum proteins were studied by agarose gel electrophoresis supplemented by total protein and albumin determinations. The clinical diagnoses could be grouped according to protein pattern. Group 1 (abomasal displacement and traumatic muscle injury) did not appreciably affect the serum protein concentrations and represented primarily non-inflammatory diseases or diseases of non-infectious origin. Group 2 (leukosis) occupied an exceptional position, with heavy lowering of albumin unaccompanied by a corresponding rise of total globulin. The γ-globulin concentration was significantly lowered. Group 3 (acute traumatic peritonitis) represented an acute inflammatory process with increase chiefly of the α-globulins, while the γ-globulin concentration was normal. Group 4 (chronic traumatic peritonitis, summer mastitis, chronic subclinical mastitis, chronic laminitis and polyarthritis, urinary tract infection, abscess, and sub-acute-chronic pneumonia) comprised diseases chiefly of chronic inflammatory character and with infectious origin. Especially characteristic was the heavy rise of globulins in general and of γ-globulin in particular. In most cases there was a large increase also of the α- and p-globulin fractions.     

Some effects of protein deficiency in young growing pigs. I. The serum protein system

No significant differences between the serum protein concentrations of the totally starved pigs and control animals given a milk protein supplemented diet were found. Serum lipid levels rose in the totally starved group. During the first 42 days of the experimental period sharp falls in serum protein concentrations were noted in the protein deprived pigs. The losses were greatest in the albumin and two β-globulin fractions, γ-globulin levels rose but at a much slower rate than in the control group. In the pigs which were refed a protein containing diet during the second 42 day period, the concentrations of all the serum protein fractions except for γ-globulin had reached control levels by the end of the investigation. The pigs which continued with the protein free diet showed further losses of albumin, a reduction in some α-globulin fractions and a cessation of net γ-globulin synthesis. The trends observed by measurements of protein bound carbohydrate were similar to those obtained from polypeptide determinations indicating quantitative rather than qualitative changes in the serum proteins. The reduction in the serum lipid concentrations of the pigs undergoing protein starvation was largely accounted for by losses of α- and β-lipoproteins. The protein deprived pigs maintained their initial body weight, while a continuous fall in weight was found in the totally starved group and a continuous increase in the control pigs.     

Escherichia coli strains isolated from the uterus and urinary bladder of bitches suffering from pyometra: comparison by restriction enzyme digestion and pulsed-field gel electrophoresis.

Pyometra (uterine inflammation with accumulation of pus in the uterus) is regarded as one of the most common illnesses in bitches. The ethiology and pathogenesis are complex with both hormonal and bacterial elements. The bacteria most frequently isolated from the uterine content is Escherichia coli.In this study, 84 E. coli strains from the uteri of 70 bitches suffering from the disease were examined and their DNA-profiles compared by restriction enzyme analysis and pulsed-field gel electrophoresis (PFGE). Through variations in DNA-profiles of the E. coli isolates, this study indicates that pyometra is caused by E. coli originating from the normal flora of the dogs and not by certain clones spread between animals.E. coli strains from the urinary bladder and the uterus of six of the bitches suffering from simultaneous urinary tract infection and pyometra were examined and compared as above. The DNA-profiles of the isolates from each of the six bitches were 100% identical. This study supports the theory suggesting that in cases of simultaneous urinary tract infection and E. coli pyometra, the urinary tract and uterus are infected with the same bacterial strain.To evaluate whether the uterus was infected with a single clone of E. coli or if multiple clones were present, eight to 16 colonies of E. coli isolated from pyometra samples from a further 10 bitches were examined. All bacterial colonies from the culture of the same bitch showed identical DNA-profiles.In 14 of the 70 bitches, two macroscopically different but biochemically identical E. coli colony types were isolated. The two colony types from the same bitch proved to have identical DNA-profiles in 13 cases and almost identical in the remaining bitch.     

Serum bile acids as an indicator of liver disease in dogs

Total serum bile acids were determined in 62 dogs with different primary or secondary liver diseases, using 3α-hydroxysteroid dehydrogenase coupled to nitrobluetetrazolium in a centrifugal analyzer. A reaction time of 4 min was sufficient, yielding a within run coefficient of variation of 7% at 6 µmol/1 and 3% at 27 µmol/1. A reference range of 0–4.4 µmol/1 2 h post prandially was observed. The sensitivity of bile acids as a liver function test was superior to that of alanine and aspartate aminotransferase, alkaline phosphatase, γ-glutamyltransferase and combinations of two of these. The bile acids test detected 36 of 39 patients with a morphological or clinical liver diagnosis. For dogs with heart failure the bile acids test was a markedly more sensitive indicator of secondary liver involvement than alanine aminotransferase or alkaline phosphatase. For secondary liver affections associated with pyometra or epilepsy medication the opposite was the case. Bile acid values in the pooled patient material was not correlated to any of the 4 enzymes measured. For cirrhosis there was positive correlation, however, with the amino transferase values.     

Relationships between the age and blood test results or body sizes in Noma horses

The objective of this study is to analyze the relationships between the age and blood test results or body sizes in Noma horses by using the results of periodical health examination. Out of 45 hematological or physical items examined, statistically significant, but loose correlations were observed in 14 items. Red blood cell count, activities of aspartate aminotransferase, alkaline phosphatase, and creatinine kinase, concentrations of calcium and inorganic phosphorus decreased with aging. Conversely, mean corpuscular volume, mean corpuscular hemoglobin, lipase activity, γ-globulin and chloride concentrations, body height, chest circumference and cannon bone circumference increased with aging. The changes in a few items seemed unique to Noma horse. However, most age-related changes found in this study might be considered as a common trend in horse breeds rather than distinctive characteristic in Noma horse.     

Uropathogenic virulence factors in isolates of Escherichia coli from clinical cases of canine pyometra and feces of healthy bitches

Escherichia coli is commonly isolated in canine pyometra, but little is known of the virulence factors that may be involved in the precipitation of this disease. The aim of this study was to compare the prevalence of uropathogenic virulence factor (UVF) genes in E. coli isolates from canine pyometra and from feces of healthy bitches to evaluate their role in the pathogenesis of pyometra. E. coli from 23 cases of canine pyometra and from the feces of 24 healthy bitches were analyzed, by polymerase chain reaction, for UVF genes associated with canine and human urinary tract infections (UTIs). The prevalences of UVFs in E. coli from canine pyometra were similar to that in canine and human uropathogenic E. coli. The prevalence of pap was greater (P=0.036) for E. coli from pyometra (52%) than for fecal isolates (21%), and the papGIII allele was present in all pap-containing isolates. The prevalences of genes for alpha-haemolysin and cytotoxic necrotising factor 1 were not significantly higher (P=0.075) in E. coli from pyometra than from feces. The proportion of pyometra strains with >or=3 UVFs was higher (P=0.039) than that of fecal strains, suggesting that possession of >or=3 UVF genes enhances the pathogenicity of the strain. Our findings demonstrate that E. coli associated with canine pyometra are similar to uropathogenic strains, and that operons that encode P fimbriae, alpha-haemolysin and cytotoxic necrotising factor 1 probably enhance the virulence and pathogenicity of the strain in the canine genital tract.     

Studies of electrophoresis on cellulose acetate membrane of serum proteins from normal horses, sheep and pigs

A method for the rapid electrophoresis on a cellulose acetate membrane of serum proteins from horses, sheep and pigs is discussed.The various main globulin fractions in the serum of these animals were experimentally identified.Normal values for the percentage composition of serum from normal horses, sheep and pigs were calculated.In the horse there was great individual variation in the shape of the β-fraction, assumed to be due to different transferrin types. The mean value for β-globulin of 19.5 % in the horse was higher than for the other two species.The albumin percentage was highest in the sheep and lowest in the pig, 48.5 % and 43.2 % respectively. The sheep had the highest γ-globulin percentage, 22.8 %, while the horse had the lowest with 19.0 %.Finally the values were compared with corresponding figures reported by other authors and the results discussed.     

Agarose gel electrophoretic separation of blood serum proteins in cattle.

The agarose gel electrophoresis described by Johansson (1972) was modified so that a buffer of pH 7.9 was used in the gel, whereas the buffer in the electrode vessels had a pH of 8.6.The cattle blood serum protein picture is described in detail. The β₁-globulin zone shows a very distinct picture of the genetically polymorphic bovine transferrins. The region between the α- and β-globulins shows a number of faint and often very distinct bands. A faint background staining over the whole electrophoretogram may partly be caused by a rather strong lipoprotein in the α₁-region, lipids thus having migrated all over the electrophoretogram.The modified method described is well suited as a “screen electrophoresis” for cattle serum and is also useful e.g. in studying bovine transferrin polymorphism.     

Electrophoretic Studies of Sera from Cattle Vaccinated or Naturally Infected with Brucella Abortus

Examinations of the electrophoretic behaviour in cellulose acetate were made on the proteins of sera from 18 field cases of brucellosis, 15 calves recently vaccinated with strain 19 Brucella abortus and from 28 normal cattle. The total protein and γ-globulin content of the sera of the naturally infected, serologically positive cattle tended to be somewhat higher than that of sera from most normal cattle. A few of the serologically-negative normals also had defenitely elevated serum γ-globulin values. In the vaccinated calves there was an increase both in total serum protein and γ-globulin but its magnitude in different individuals was not directly related to the rise in their antibody titre.     

Changes in serum protein electrophoresis profiles and acute phase proteins in calves with diarrhea

Calf diarrhea leads to substantial economic losses in the livestock industry worldwide due to medical treatment costs, retarded growth performance, and even death. The objective of this study was to investigate changes in serum protein profiles and acute phase proteins in calves with diarrhea and identify the association between these changes and diarrhea. A total of 185 Korean beef calves were used and divided into 3 groups by age: 1 to 10 days (n = 46), 11 to 20 days (n = 65), and 21 to 30 days (n = 74). Blood and fecal samples were collected from each calf. Serum concentrations of total protein, protein fractions (albumin, α1-globulin, α2-globulin, β-globulin, and γ-globulin), haptoglobin (Hp), and serum amyloid A (SAA) were analyzed. Compared to calves without diarrhea, calves with diarrhea had significantly lower albumin concentrations at 11 to 20 days and 21 to 30 days of age (P = 0.017 and P = 0.000, respectively) and significantly higher α1-globulin fractions at 21 to 30 days of age (P = 0.01). Interestingly, α2-globulin fractions were significantly higher in diarrheic calves in all age groups, whereas γ-globulin fractions were significantly lower in calves with diarrhea aged 1 to 10 days, compared with normal animals. In calves with diarrhea, the concentration of Hp was significantly higher, whereas SAA levels were not different between normal and diarrheic calves. In addition, a positive correlation was found between α2-globulin and Hp (P = 0.0004). Taken together, these results provide useful information about the use of serum protein profiles and Hp as prognostic and diagnostic markers for animal health status.     

一例犬子宫蓄脓的诊断及手术治疗

本试验对一例患有子宫蓄脓的西施犬通过询问病史、临床检查、血常规检测、X光及B超检查进行确诊,采用手术法治疗。结果显示,血常规检查发现白细胞、中性粒细胞均增多,X光检测到明显膨大的子宫,B超影像可观察到子宫角为圆腔状的黑色暗区,子宫壁增生变厚产生无回声液性暗区。对于子宫蓄脓的诊断,血常规检查显示体内有炎症变化,X光和B超确诊病犬为开放性子宫蓄脓,对病犬采取手术疗法,彻底切除卵巢和子宫。病犬术后恢复良好。     

Measurement of C‐reactive protein and Prostaglandin F2α Metabolite Concentrations in Differentiation of Canine Pyometra and Cystic Endometrial Hyperplasia/Mucometra

Canine pyometra is a dioestrus period disease in which systemic inflammatory response syndrome (SIRS) is a common outcome due to the response of the body to the bacterial infection. The purpose of this study was i) to differentiate canine pyometra and cystic endometrial hyperplasia (CEH)/mucometra by measuring serum C‐reactive protein (CRP) and prostaglandin F_2α metabolite (PGFM) concentrations in blood and ii) to compare serum concentrations of CRP and PGFM in bitches with a pathological uterus (pyometra or CEH/mucometra) to concentrations in bitches with a healthy uterus. Mean CRP concentrations were found significantly higher (p < 0.001) in dogs with pyometra compared to those with CEH/mucometra or healthy uterus. However, no statistical difference could be detected between the groups for mean PGFM concentrations. Mean white blood cell count (WBC), alkaline phosphatase (ALP) and total protein concentrations were found significantly higher (p < 0.001) in dogs with pyometra. Escherichia coli was the most frequently isolated microorganism from dogs with pyometra (64.3%). Edwardsiella spp. was detected in a single case of pyometra for the first time. In conclusion, our results demonstrate that serum CRP concentrations were increased in dogs with pyometra and thus we conclude that serum CRP concentration but not PGFM might be useful as a marker to differentiate a case of CEH/mucometra from pyometra in female dogs. To the authors' knowledge, this is the first report in which Edwardsiella spp. has been isolated in the canine uterus.     

Peptococcus (S. Micrococcus) indolicus. The demonstration of two varieties of hemolysin forming strains.

On the basis of biochemical and serological criteria 2 hemolysin forming varieties of peptococci were identified as Peptococcus indolicus.Of a total of 16 hemolytic strains examined 9 originated from the vagina of clinically healthy cows, 4 from mastitis secretions from dry cows, 2 from the interdigital skin of clinically healthy sows, and 1 from a subcutaneous abscess in a pig.Two strains were designated a-hemolytic and 14 β-hemolytic.On blood agar plates colonies of the α-hemolytic variety were surrounded by narrow zones of almost complete hemolysis, while colonies of the β-hemolytic variety were surrounded by broad zones of incomplete hemolysis. The hemolysins were termed α- and β-hemolysin, respectively.The β-hemolysin, but not the α-hemolysin, could be demonstrated in cultures grown in liquid media. The β-hemolysin was found to be filtrable, relatively thermoresistant, and non-dermonecrotic.By gel diffusion analyses the 2 α-hemolytic strains were referred to Serotype C. Ten of the β-hemolytic strains belonged to Serotype C, 2 to Type B, 1 to Type D, and 1 to Type E.     

Hysterectomy leads to fast improvement of haematological and immunological parameters in bitches with pyometra

OBJECTIVES: To investigate whether a combination of hysterectomy and antibiotic treatment leads to an improvement of altered haematological and immunological parameters in bitches affected by pyometra. METHODS: Blood samples obtained from 13 bitches affected by pyometra were investigated before hysterectomy and seven days later for a total and differential leucocyte count, activity of neutrophils and lymphocytes, and quantification of total serum immunoglobulins, lysozyme and circulating immune complexes. RESULTS: The parameters most affected included changes in blood profile (leucocytosis because of neutrophilia or leucopenia) and inhibition of lymphocyte activity. Seven days after hysterectomy, all affected parameters returned to normal levels comparable to clinically healthy dogs. CLINICAL SIGNIFICANCE: Application of immunostimulating therapy is not necessary to manage changes in blood cell counts and suppression of lymphocyte activity in bitches with pyometra. Removal of the infected uterus, being the source of infection, leads to improvement.     

Enhanced immune response with foot and mouth disease virus VP1 and interleukin-1 fusion genes

The capsid of the foot and mouth disease (FMD) virus carries the epitopes that are critical for inducing the immune response. In an attempt to enhance the specific immune response, plasmid DNA was constructed to express VP1/interleukin-1α (IL-1α) and precursor capsid (P1) in combination with 2A (P1-2A)/IL-1α under the control of the human cytomegalovirus (HCMV) immediateearly promoter and intron. After DNA transfection into MA104 (monkey kidney) cells, Western blotting and an immunofluorescence assay were used to confirm the expression of VP1 or P1-2A and IL-1α. Mice were inoculated with the encoding plasmids via the intradermal route, and the IgG1 and IgG2a levels were used to determine the immune responses. These results show that although the immunized groups did not carry a high level of neutralizing antibodies, the plasmids encoding the VP1/IL-1α, and P1-2A/IL-1α fused genes were effective in inducing an enhanced immune response.     

Milk plasmin, antitrypsin, N-acetyl-beta-D-glucosaminidase and bacterial growth in lactoserum during the early post partum period

The activities of plasmin, N-acetyl-β-d-glucosaminidase (NAGase) and antitrypsin in milk were determined during 1 week post partum. The concentration of NAGase and antitrypsin in milk decreased significantly during this period. A slight decrease in plasmin activity was also seen. Replication rates of E. coli and S. aureus in lactoserum were also determined. Both test bacteria showed a tendency for increasing growth rates towards the end of the period. Growth of E. coli was significantly (p < 0.01) enhanced in day 2 samples as compared with samples collected during the first day post partum.