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1.
Streptococcus uberis is an environmental udder pathogen that infects cattle and can cause persistent intramammary infection (IMI), despite the fact that isolates are mainly susceptible to antibiotics. As biofilm growth can cause persistent infection, the ability of ten S. uberis isolates from clinical and subclinical IMIs to form biofilms on the polystyrene surface of a conventional 96-microplates model was examined. Biofilm formation was judged by different staining methods (crystal violet and resazurin) and by atomic force and fluorescence microscopy. These analyses revealed that two out of ten S. uberis strains tested were able to form biofilms. Upon treatment with Proteinase K, biofilms of S. uberis were completely disintegrated, which indicates that biofilm formation is protein-mediated in these strains. Addition of trace amounts of milk, the natural growth medium of S. uberis, significantly increased biofilm formation by most of the strains initially classified as non-biofilm producers. Alpha-casein and β-casein were the primary inducers of biofilm growth, and casein degradation by serine protease activity was required to achieve maximal biofilm production. These results suggest that the extracellular proteolytic activity of S. uberis contributes to an increased biofilm formation. Such a mode of growth induced by host proteins might help to explain the persistence of IMIs caused by this pathogen.  相似文献   

2.
The goal of this work was to develop a protocol for rapid genotyping of A and G variants at the CSN2 locus and genotyping of T and C variants at the CSN3 locus in sheep breeds (Sumava and Valachian) by means of PCR and LightCycler analysis. The LightCycler technique combines rapid and efficient in vitro amplification of DNA in glass capillaries with melting curve analysis based on fluorescence resonance energy transfer for the sensitive detection of point mutation. The A variant had a greater frequency (Sumava, 0.778; Valachian, 0.835) than did variant G (Sumava, 0.222; Valachian, 0.165) in both sheep breeds. The CSN3 locus was found to be monomorphic, with no polymorphism identified in either population.  相似文献   

3.
2-Agonist drugs may be illegally used as growth promoters for feedlot calves, when mixed into milk replacer immediately before feeding. To check for the presence of clenbuterol, salbutamol and terbutaline in such food, an analytical system was established using a screening method based on two commercial qualitative competitive ELISA tests, with antibodies raised against the arylamino group and thet-butyl group. The extraction procedure was based on precipitation of the milk samples with acetonitrile followed by filtration. The absence of any significant interference by other substances in the filtrate allowed detection of 2-agonist drugs in spiked samples at the lowest concentration having a repartitioning effect (50 ppb for clenbuterol, mabuterol and terbutaline, 500 ppb for salbutamol). In view of a false positive response with tetracycline in milk samples and a cross-reaction between clenbuterol and mabuterol, an HPLC-MS technique was developed which, after extraction and purification of the samples with SPE C18 Polar Plus, was able to confirm the presence of these drugs. The good recovery after extraction (ranging from 84% to 90.2%) and the low detection limit with this method (250 ng/ml for clenbuterol, mabuterol and terbutaline, and 2.5 µg/ml for salbutamol) allowed easy confirmation and simultaneous detection of the four 2-agonists at the lowest concentrations at which they are used in adulterated milk for calves.Abbreviations B optical density of the sample - B maximal optical density in total absence of competition - %B/B 0 percentage of inhibition - ELISA enzyme-linked immunosorbent assay - EIA enzyme immunoassay - HPLC-MS high-performance liquid chromatography-mass spectrometry - m/z mass to charge ratio - ppb parts per billion - ppm parts per million - SPE solid-phase extraction  相似文献   

4.
Osteosarcoma (OSA) is the most frequently occurring malignant primary bone tumour in dogs and children and arises from cells of the osteoblast lineage. Inappropriate Wnt signalling activity has been implicated in human OSA. Altered expression of β-catenin, an integral member of the Wnt signalling pathway, has been associated with numerous human cancers, including OSA. In this study, 30 of the 37 primary canine OSA tissues and 2 of the 3 metastatic OSAs were positive for β-catenin expression as determined by immunohistochemistry, whereas 2 normal bones stained negative for β-catenin. No mutations were identified in exon 3 of β-catenin in the three OSA cases in which DNA sequencing was performed. Finally, there was no relationship between β-catenin expression and overall survival time or disease-free interval. Our results indicate β-catenin is frequently expressed within the cytoplasm of neoplastic cells in canine OSA but contains no detectable mutations in exon 3, similar to human OSA.  相似文献   

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6.
Veterinary Research Communications -  相似文献   

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8.
Beta-alanine is an important amino acid involved in several metabolic reactions in the body. The study aimed to investigate the effect of β-alanine supplementation on intestinal development and the immune performance of weaned piglets. Thirty-two 21-day-old healthy weaned piglets(half female and half male; Duroc × Landrace × Yorkshire) with an initial body weight of 8.11 ± 0.21 kg were randomly divided into 4 groups with 8 replicates of 1 pig each. The control group was fed a basal diet and the ...  相似文献   

9.
Defensin is a small cationic peptide widely distributed in animals and plants. It is an important component of the innate immune defense system of mammals. It has a broad spectrum of antibacterial, antiviral and antifungal activities. Porcine β-defensin-2 (PBD-2) is one of the natural defensins expressed in pigs and has good antibacterial activity in vitro. The purpose of this study is to evaluate the therapeutic effect of PBD-2 in the treatment of Streptococcus suis infection in animals and to provide more insights for evaluating the value of PBD-2 as a therapeutic drug. Firstly, the bactericidal activity of PBD-2 against S. suis clinical isolate SC-19 was measured in vitro, and the cytotoxicity of PBD-2 was tested on mouse-derived macrophages (RAW264.7). Subsequently, C57 female mice aged 4-6 weeks and weighing between 18-22 g infected with S. suis SC-19 were treated with PBD-2 or PBS (n ≥ 6). The survival rate of mice, and the bacteria loads, inflammatory cytokine levels, and pathological changes of tissues were determined. The results showed that PBD-2 (25-200 μg·mL-1) could significantly inhibit the growth of S. suis SC-19 in a concentration-dependent manner (P<0.05). At the same time, PBD-2 had no significant toxic effect on RAW cells (P>0.05). The in vivo study revealed that PBD-2 treatment could effectively reduce the mortality of mice infected with S. suis SC-19. At the same time, PBD-2 treatment significantly reduced bacteria loads in brain, lung, spleen and blood of mice, and decreased the levels of inflammatory cytokines IL-6, IL-12 and TNF-α in mouse sera (P<0.05). At the same time, PBD-2 treatment also significantly alleviated the degree of pathological damages in lung and spleen tissues of mice infected with bacteria (P<0.05). These results indicate that PBD-2 confers great resilience to S. suis in vitro without significant cytotoxicity, while it also exhibits a therapeutic effect on S. suis infection in mice, suggesting that the use of PBD-2 as a therapeutic drug and substitutes for antibiotics is of an excellent prospect.  相似文献   

10.
Abstract

AIMS: To assess the occurrence of, and characterise, extended-spectrum β-lactamase (ESBL) and AmpC β-lactamase (AmpC)-producing Enterobacteriaceae isolated by veterinary diagnostic laboratories from infection sites in companion animals in New Zealand.

METHODS: Selected Enterobacteriaceae isolates were submitted by seven New Zealand veterinary diagnostic laboratories. They were isolated from infection sites in companion animals between June 2012 and June 2013, and were resistant to amoxicillin-clavulanic acid, fluoroquinolones, or any combination of two or more antimicrobials. Based on disk diffusion test results, the isolates were phenotypically categorised according to production of ESBL and AmpC. Genes for ESBL and AmpC production were amplified by PCR and sequenced. Escherichia coli isolates were also typed by multilocus sequence typing.

RESULTS: A total of 115 isolates matching the inclusion criteria were obtained from the participating laboratories, of which 74 (64%) originated from dogs and 29 (25%) from cats. Seven bacterial species were identified, of which E. coli was the most common (87/115, 76%). Of the 115 isolates, 10 (9%) expressed the ESBL phenotype, 43 (37%) the AmpC phenotype, and seven (6%) both ESBL and AmpC phenotypes. Of the 60 ESBL and AmpC-producing isolates, 36 (60%) were E. coli. Amongst these isolates, 27/60 (45%) were classified as multidrug resistant, compared with 15/55 (27%) non-ESBL or AmpC-producing isolates (p<0.01). Ninety five isolates were resistant to amoxicillin-clavulanic acid and 58 (61%) of these were ESBL or AmpC-producing. The predominant ESBL genes were blaCTX-M-14 and blaCTX-M-15, and the dominant plasmid-encoded AmpC gene was blaCMY-2. Thirty-eight E. coli multilocus sequence types (ST) were identified, and the most prevalent were ST12 (12/89, 13%), ST131 (6/89, 7%) and ST648 (6/89, 7%). ESBL and AmpC-producing isolates accounted for 35/1,082 (3.2%) of the Enterobacteriaceae isolated by one laboratory network over the study period.

CONCLUSIONS AND CLINICAL RELEVANCE: ESBL and AmpC-producing Enterobacteriaceae were associated with clinical infections in companion animals in New Zealand, and were often multidrug resistant. In this study, these organisms accounted for <5% of all Enterobacteriaceae isolated from infection sites by one laboratory network, but their prevalence among isolates resistant to amoxicillin-clavulanic acid was 61%. Therefore routine secondary testing for ESBL and AmpC production by Enterobacteriaceae that are resistant to amoxicillin-clavulanic acid in primary testing could improve the accuracy of definitive antimicrobial therapy in companion animals in New Zealand.  相似文献   

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13.
Colloidal gold immunochromatographic was uesd for the determination of β-lactams and tetracyclines in milk. Tetracyclines hapten was obtained by chemical synthesis. Immunogen and envelope antigen were prepared by cephalosporins and tetracyclines hapten coupled to carrier protein. Monoclonal antibodies were obtained by immunogen immunizing mice. A rapid and simple colloidal gold immunochromatographic assay was established for the determination of β-lactams and tetracyclines in milk. 12 kinds of β-lactams and 4 kinds of tetracyclines were detected at the same time by the strip. The milk sample could be used to be detected without dilution. The detection time could be 3 to 5 min. The method could be efficiently used for mass samples screening in primary laboratory.  相似文献   

14.
Objectives : To evaluate accuracy and precision of a hand‐held ketone meter measuring β‐hydroxybutyrate and to determine its diagnostic performance to rule out ketoacidaemia in diabetic cats. Methods : The ketone meter was validated by calculating within‐day precision at different β‐hydroxybutyrate concentrations and by comparison with a laboratory method. To determine its diagnostic performance to diagnose ketoacidaemia, 217 sets of data (venous blood gas analysis and β‐hydroxybutyrate measurements) were retrospectively analysed. Sensitivities and specificities were calculated with the help of receiver‐operating characteristic curves. Results : The ketone meter reliably detected β‐hydroxybutyrate at concentrations >0·1 mmol/L and reproducibility was acceptable. Measurements highly correlated with laboratory results (r=0·97; P<0·001), but a significant negative bias was found at high concentrations. A β‐hydroxybutyrate concentration of >2·55 mmol/L had a sensitivity of 94% and a specificity of 68% for diagnosing ketoacidaemia. Many cats with high β‐hydroxybutyrate concentrations and normal blood pH had an elevated chloride gap suggestive of superimposed hypochloraemic metabolic alkalosis. Clinical Significance : The commercially available point‐of‐care ketone meter Precision Xtra is a valid tool to measure β‐hydroxybutyrate in diabetic cats. Concentration <2·55 mmol/L enable ketoacidaemia to be excluded and should lead to redirection of differential diagnoses.  相似文献   

15.
This study assessed the capacity of β-lactam antibiotics to prevent salmonella-mediated encephalopathy in calves given the putative neuroprotective effects of these drugs of increasing glutamate export from the brain. Both ampicillin and ceftiofur prevented the development of encephalopathy despite resistance of the inoculated Salmonella enterica serovar Saint-Paul isolate to both drugs. A glutamate receptor antagonist also prevented this salmonella-mediated encephalopathy. Glutamate exporters were hyper-expressed in the presence of β-lactam antibiotics while a glutamate export inhibitor obviated the effects of these antibiotics, demonstrating a neuroprotective effect through glutamate export from the brain. The findings indicate that β-lactam antibiotics remain an important treatment option for this atypical form of bovine salmonellosis.  相似文献   

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18.
In this study, effects of oral β-carotene supplementation to mares (β-carotene group: 1000 mg/day, n = 15; control group: n = 15) from 2 weeks before foaling until 6 weeks thereafter on concentrations of β-carotene, vitamin A and α-tocopherol in plasma, colostrum and milk and plasma of their foals were determined. In addition, effects on fertility were studied. Beta-carotene concentrations increased in plasma and colostrum of β-carotene-supplemented mares compared to control mares (p < 0.05). In mares of both groups, β-carotene concentrations were higher in colostrum than in milk (p < 0.05). In foals, β-carotene concentrations increased with colostrum uptake and were higher in foals born to supplemented mares (p < 0.05; control group: 0.0003 ± 0.0002 μg/ml on day 0, 0.008 ± 0.0023 μg/ml on day 1; β-carotene group: 0.0005 ± 0.0003 μg/ml on day 0, 0.048 ± 0.018 μg/ml on day 1). Concentrations of vitamin A and α-tocopherol were higher in colostrum than in milk (p < 0.05) but did not differ between groups. Concentration of α-tocopherol in plasma of mares decreased over time and in foals, increased markedly within 4 days after birth. All but one mare (control group) showed oestrus within 2 weeks post-partum. Occurrence of oestrus did not differ between groups. More mares of the control group (7/7 vs. 5/12 in the β-carotene group) became pregnant after being bred in first post-partum oestrus (p < 0.05). In conclusion, β-carotene supplementation to mares increased β-carotene concentrations in plasma, colostrum and milk of mares and plasma of their foals but had no positive effects on fertility.  相似文献   

19.
This study investigated the effects of supplementary β-glucosidase on the carcass composition, meat quality, weight of digestive organ and apparent digestibility in male broilers. Two hundred and forty male, 1-day avine broiler chicks were randomly allocated into four treatment groups and fed with corn-soya bean meal supplemented with 0 (control), 0.6, 1.2 and 1.8 U/g of β-glucosidase respectively. The results showed that there were no significant differences (p > 0.05) among groups in carcass composition (percentages of eviscerated yield, half-eviscerated yield, muscle yield of breast and leg). However, adding 0.6 U/g β-glucosidase to the diet not only altered the meat quality by decreasing the drip loss ratio (p < 0.05) and relative lightness (L*) value (p < 0.01), increasing relative redness (a*) value (p < 0.01), but also significantly decreased the pancreas to body weight ratio (p < 0.05), however, with little effect on liver, proventriculus and gizzard to body weight ratio (p > 0.05). The length and width of duodenum villus were not affected by the addition of β-glucosidase, but the coefficients of total tract apparent digestibility of protein and fat increased by 9.02% (p < 0.05) and 7.40% (p < 0.01) respectively; the parameters of ash were not affected by β-glucosidase addition (p < 0.05). This study provided valuable information for evaluation of the effect of supplementary β-glucosidase on the meat quality and digestibility of broilers.  相似文献   

20.
β-Conglycinin, a major seed-storage protein in soybeans, is one of the primary antigenic proteins responsible for soybean-meal hypersensitivity in weaned piglets. The protein is a heterotrimer composed of subunits α, α′ and β. It is currently unknown which of the β-conglycinin subunits are allergenic for piglets. The aim of this study was to identify potential allergenic subunits of β-conglycinin for soybean sensitive piglets and to characterise these subunits by immunoglobulin (Ig) G and E immunoblotting, ELISA, ‘skin prick’ and whole blood histamine-release testing. The IgG and IgE binding capabilities of the purified α, α′ and β subunits of β-conglycinin were determined by immunoblot analysis and ELISA with sera from β-conglycinin sensitised piglets. Skin prick testing and whole blood histamine release testing were also performed to detect the activated effector cell response to specific allergens. Specific IgG and E antibodies were identified that recognised all three subunits of β-conglycinin in the sera of β-conglycinin sensitised piglets. All three subunits of β-conglycinin elicited positive skin test and specific histamine release responses from the whole blood of β-conglycinin sensitised piglets. These results suggest that all three β-conglycinin subunits are potential allergens for piglets.  相似文献   

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