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- Bagge, Elisabeth, et al.
(författare)
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Detection and identification by PCR of Clostridium chauvoei in clinical isolates, bovine faeces and substrates from biogas plant
- 2009
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Ingår i: Acta Veterinaria Scandinavica. - : Springer Science and Business Media LLC. - 0044-605X .- 1751-0147. ; 51
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Tidskriftsartikel (refereegranskat)abstract
- Background: Clostridium chauvoei causes blackleg, an acute disease associated with high mortality in ruminants. The apparent primary port of entry is oral, during grazing on pasture contaminated by spores. Cases of blackleg can occur year after year on contaminated pastures. A method to determine the prevalence of C. chauvoei spores on pasture would be useful.The standard method for C. chauvoei detection is culture and biochemical identification, which requires a pure culture. In most muscle samples from cattle dead from blackleg the amount of C. chauvoei in samples is high and the bacterium can easily be cultured, although some samples may be contaminated. Detection by PCR would be faster and independent of contaminating flora.Digested residues from biogas plants provide an excellent fertiliser, but it is known that spore-forming baeria such as Clostridium spp. are not reduced by pasteurisation. The use of digested residues as fertiliser may contribute to the spread of C. chauvoei. Soil, manure and substrate from biogas plants are contaminated with other anaerobic bacteria which outgrow C. chauvoei. Therefore, detection by PCR is would be useful. This study applied a PCR-based method to detect of C. chauvoei in 25 muscle and blood samples, 114 manure samples, 84 soil samples and 33 samples from the biogas process.Methods: Muscle tissues from suspected cases of blackleg were analysed both by the standard culture method followed by biochemical identification and by PCR, with and without preculture. To investigate whether muscle tissue samples are necessary, samples taken by swabs were also investigated. Samples from a biogas plant and manure and soil from farms were analysed by culture followed by PCR. The farms had proven cases of blackleg. For detection of C. chauvoei in the samples, a specific PCR primer pair complementary to the spacer region of the 16S-23S rRNA gene was used.Results: Clostridium chauvoei was detected in 32% of muscle samples analysed by culture with identification by biochemical methods and in 56% of cases by culture in combination with PCR. Clostridium chauvoei was detected in 3 (out of 11) samples from the biogas plants collected before pasteurisation, but samples taken after pasteurisation and after digestion all tested negative. Clostridium chauvoei was not detected in any soil or silage samples and only one manure samples tested positive.Conclusion: The diagnostic method used for C. chauvoei was not applicable in estimating the risk of blackleg on particular pastures from manure or soil samples, but found to be highly useful for clinical samples.
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- Bagge, Elisabeth, et al.
(författare)
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Diversity of spore-forming bacteria in cattle manure, slaughterhouse waste and samples from biogas plants
- 2010
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Ingår i: Journal of Applied Microbiology. - : Oxford University Press (OUP). - 1364-5072 .- 1365-2672. ; 109, s. 1549-1565
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Tidskriftsartikel (refereegranskat)abstract
- Aims:As biowaste intended for biogas production can contain pathogenic micro-organisms, the recommended treatment is pasteurization at 70 degrees C for 60 min. This reduces pathogens such as Salmonella spp., whereas spore-forming bacteria (Bacillus spp. and Clostridium spp.) survive. Most spore-forming bacteria are harmless, but some can cause diseases such as blackleg, botulism and anthrax. In this study, the effect of the biogas process on Bacillus spp. and Clostridium spp. was investigated.Methods and Results:We analysed 97 faecal samples, 20 slaughterhouse waste samples and 60 samples collected at different stages in the biogas process. Bacillus spp. and Clostridium spp. were quantified and subcultured. The isolates were identified by biochemical methods and by 16S rRNA gene sequencing. Phylogenetic trees were constructed from the sequences obtained from isolates from the samples. Clostridium botulinum/Clostridium spp. and Clostridium sordellii were found both before and after pasteurization, but not after digestion (AD). Some of the isolated strains probably represented new members of the genera Clostridium and Bacillus.Conclusion:After digestion, the numbers of clostridia decreased, but none of the pathogenic bacteria did, whereas Bacillus spp. remained constant during the process.Significance and Impact of the Study:Biogas is gaining in importance as an energy source and because the residues are used as fertilizers, we needed to study the prevalence of pathogenic bacteria in such material.
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- Bagge, Elisabeth
(författare)
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Hygiene aspects of the biogas process with emphasis on spore-forming bacteria
- 2009
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Biogas is a renewable source of energy which can be obtained from processing of biowaste. The digested residues can be used as fertiliser. Biowaste intended for biogas production contains pathogenic micro-organisms. A pre-pasteurisation step at 70°C for 60 min before anaerobic digestion reduces non spore-forming bacteria such as Salmonella spp. To maintain the standard of the digested residues it must be handled in a strictly hygienic manner to avoid recontamination and re-growth of bacteria. The risk of contamination is particularly high when digested residues are transported in the same vehicles as the raw material. However, heat treatment at 70°C for 60 min will not reduce spore-forming bacteria such as Bacillus spp. and Clostridium spp. Spore-forming bacteria, including those that cause serious diseases, can be present in substrate intended for biogas production. The number of species and the quantity of Bacillus spp. and Clostridium spp. in manure, slaughterhouse waste and in samples from different stages during the biogas process were investigated. The number of species of clostridia seemed to decrease following digestion, likewise the quantity. However, Bacillus spp. seemed to pass unaffected through the biogas process. In laboratory-scale experiments the effects on clostridia during pasteurisation and digestion were investigated. Pathogenic clostridia were inoculated in substrates from homogenisation tanks and digester tanks. The inoculated clostridia remained after pasteurisation, but the impacts of digestion differ between different species. Culture followed by identification of C. chauvoei by PCR in samples from cattle died from blackleg, is faster and safer than culture followed by biochemical identification of C. chauvoei. However, for environmental samples the PCR method is not practically applicable for detection of C. chauvoei. To avoid spreading of diseases via biogas plants when digested residues are spread on arable land, a pasteurisation stage at 70°C for 60 min before anaerobic digestion gives adequate reduction of most non spore-forming bacteria, such as salmonella. However, caution should be exercised before digested residues are spread in areas without endemic problems of pathogenic spore-forming bacteria. In Sweden, official recommendation is that digested residues from biogas plants only should be applied on arable land, and not on grasslands for animal pasture.
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- Bagge, Ebba, et al.
(författare)
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Pattern of endocrine treatment for epithelial ovarian cancer in the Southeast medical region of Sweden: a population-based study
- 2019
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Ingår i: Acta Oncologica. - : TAYLOR & FRANCIS LTD. - 0284-186X .- 1651-226X. ; 58:3, s. 320-325
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Tidskriftsartikel (refereegranskat)abstract
- Aim of the study: Endocrine treatment (ET) is an alternative as salvage therapy in epithelial ovarian cancer (EOC) but the usage in routine care is unknown. We evaluated the treatment patterns and outcome of patients receiving ET for EOC in the Southeast medical region in Sweden.Method: Patients were identified through the population-based Southeast Quality Registry for gynaecological cancer. Inclusion criteria were: age 18 years, histologically verified EOC diagnosed 2000-2013, ET for 4 weeks. Coverage compared with the Swedish National Cancer Registry was 100%. Data extracted from medical records was collected by means of a study-specific Case Report Form. Last date of follow-up was February 1st, 2018. All statistics were descriptive.Results: Altogether 248 (18%) of 1414 patients were treated with ET. Most (49%) had received only one, and 34% two previous lines of chemotherapy. Time from last chemotherapy to ET was 4 months, range 0-55months. The reason for initiating ET was tumor progression (66%), chemotherapy related toxicity (29%) and maintenance (4%). Tamoxifen was prescribed in 94% of cases. Best response was partial (amp;lt; 5%) and stable disease (50%). No patient had a complete response. 194 (78%) patients received subsequent chemotherapy, of these 27% had 3-7 lines of chemotherapy. Duration of ET was a median 4 months (range 1-80 months). Median time from ET to subsequent chemotherapy was 5 months (range 0-79). The median overall survival was 45 months (range 9-173).Conclusion: In the Southeast region of Sweden, endocrine treatment for EOC was prescribed inconsistently and in various settings, usually initiated by a rising CA-125 level. Poorer documentation and irregular tumor response assessment were observed for endocrine treatment compared to chemotherapy.
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- Einarsson, Stig, et al.
(författare)
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Occurrence of bacteria and polymorphonuclear leukocytes in fetal compartments at parturition; relationships with foal and mare health in the peripartum period
- 2015
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Ingår i: Theriogenology. - : Elsevier BV. - 0093-691X .- 1879-3231. ; 84, s. 163-169
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Tidskriftsartikel (refereegranskat)abstract
- This study investigated the relationship of the health of the newborn foal and (1) number of polymorphonuclear leukocytes (PMNLs) in the amniotic fluid, (2) bacteria present in the amniotic fluid and the venous umbilical blood, and (3) bacteria present in the uterus of the newly foaled mare. A further aim was to investigate relationships between the bacteriologic findings in the amniotic fluid, umbilical blood, and uterus postpartum. Samples were taken from 50 Standardbred trotter foaling mares from a well-managed stud in Sweden. Parturition was spontaneous in all cases. Length of pregnancy, parturition and postpartum complications, health status of the foal, the time between foaling and the expulsion of the placenta, and the number of postfoaling mares becoming pregnant after insemination were recorded. Amniotic fluid was collected when the amniotic vesicle was clearly visible; it was analyzed for bacteriology and occurrence of PMNLs. Umbilical blood was analyzed for the presence of bacteria and the concentration of serum amyloid A. The uterus of themarewas swabbed for bacteriology 6 to 17 hours postpartum. A blood samplewas taken from the foal before administering plasma. The foals were divided into two groups: group 1 required up to 2 hours to rise after birth (2 hours; 31 foals) and group 2 requiredmore than two hours (>2 hours; 19 foals). The length of gestation varied between 332 and 356 days; there was no significant difference in gestation length between the two foal groups. Partus and postpartum complications occurred in a significantly higher proportion of mares giving birth to group 2 foals than group 1 foals (P ¼ 0.02), although uterine culture postpartum and the subsequent pregnancy rate per season were not different between the groups. Compromised health status was significantly higher among foals belonging to group 2 than group 1 (P ¼ 0.001). Most of the amniotic samples contained 5% or less PMNLs. Only three samples contained more than 30% PMNLs; group 2 foals had the highest percentage of PMNLs. Bacterial growth was found in both amniotic fluid (57%) and umbilical blood (35%) in mares irrespective of whether their foals were healthy or compromised. Coagulase-negative staphylococci were the most frequent bacteria. There were no differences in bacterial occurrence in amniotic fluid or in umbilical blood between the two foal groups
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| 9. |
- Ekman, Lisa, et al.
(författare)
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A shotgun metagenomic investigation of the microbiota of udder cleft dermatitis in comparison to healthy skin in dairy cows
- 2020
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Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 15
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Tidskriftsartikel (refereegranskat)abstract
- Udder cleft dermatitis (UCD) is a skin condition affecting the fore udder attachment of dairy cows. UCD may be defined as mild (eczematous skin changes) or severe (open wounds, large skin changes). Our aims were to compare the microbiota of mild and severe UCD lesions with the microbiota of healthy skin from the fore udder attachment of control cows, and to investigate whether mastitis-causing pathogens are present in UCD lesions. Samples were obtained from cows in six dairy herds. In total, 36 UCD samples categorized as mild (n = 17) or severe (n = 19) and 13 control samples were sequenced using a shotgun metagenomic approach and the reads were taxonomically classified based on their k-mer content. The Wilcoxon rank sum test was used to compare the abundance of different taxa between different sample types, as well as to compare the bacterial diversity between samples. A high proportion of bacteria was seen in all samples. Control samples had a higher proportion of archaeal reads, whereas most samples had low proportions of fungi, protozoa and viruses. The bacterial microbiota differed between controls and mild and severe UCD samples in both composition and diversity. Subgroups of UCD samples were visible, characterized by increased proportion of one or a few bacterial genera or species, e.g. Corynebacterium, Staphylococcus, Brevibacterium luteolum, Trueperella pyogenes and Fusobacterium necrophorum. Bifidobacterium spp. were more common in controls compared to UCD samples. The bacterial diversity was higher in controls compared to UCD samples. Bacteria commonly associated with mastitis were uncommon. In conclusion, a dysbiosis of the microbiota of mild and severe UCD samples was seen, characterized by decreased diversity and an increased proportion of certain bacteria. There was no evidence of a specific pathogen causing UCD or that UCD lesions are important reservoirs for mastitis-causing bacteria.
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| 10. |
- Eriksson, Helena, et al.
(författare)
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Erysipelothrix rhusiopathiae contamination in the poultry house environment during erysipelas outbreaks in organic laying hen flocks
- 2014
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Ingår i: Avian Pathology. - : Informa UK Limited. - 0307-9457 .- 1465-3338. ; 43, s. 231-237
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Tidskriftsartikel (refereegranskat)abstract
- This study investigated organic laying hen farms for the presence of Erysipelothrix rhusiopathiae in the house environment and from potential carriers (i.e. insects and mice) during ongoing erysipelas outbreaks, and compared the obtained isolates with those from laying hens. The samples were investigated by selective culture followed by species-specific polymerase chain reaction on cultures. E. rhusiopathiae was isolated from the spleen, jejunal contents, manure, dust and swabs from water nipples. Three more samples from the house environment tested positive by polymerase chain reaction compared with selective culture alone. Selected isolates were investigated by pulsed-field gel electrophoresis (PFGE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). One farm was represented by isolates from laying hens only, and one of these isolates differed in one PFGE band from the others. Different banding patterns were observed for isolates from laying hens and manure on one farm. On the remaining two farms, the isolates from the house environment and laying hens were identical but differed between farms. Outbreaks reoccurred in the next flock on two of the farms, and different PFGE types were isolated from consecutive flocks. Our results suggest an external source of infection, which would explain the previously reported increased risk of outbreaks in free-range flocks. Contaminated manure and dust may represent sources of transmission. For the isolates, MALDI-TOF MS and biochemical typing results were in agreement but, since the type strain of Erysipelothrix tonsillarum was typed as E. rhusiopathiae using MALDI-TOF MS, further studies into this method are needed.
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