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| 2. |
- Hellman, Stina, et al.
(författare)
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Equine enteroid-derived monolayers recapitulate key features of parasitic intestinal nematode infection
- 2024
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Ingår i: Veterinary research (Print). - : Springer Nature. - 0928-4249 .- 1297-9716. ; 55:1
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Tidskriftsartikel (refereegranskat)abstract
- Stem cell-derived organoid cultures have emerged as attractive experimental models for infection biology research regarding various types of gastro-intestinal pathogens and host species. However, the large size of infectious nematode larvae and the closed structure of 3-dimensional organoids often hinder studies of the natural route of infection. To enable easy administration to the apical surface of the epithelium, organoids from the equine small intestine, i.e. enteroids, were used in the present study to establish epithelial monolayer cultures. These monolayers were functionally tested by stimulation with IL-4 and IL-13, and/or exposure to infectious stage larvae of the equine nematodes Parascaris univalens, cyathostominae and/or Strongylus vulgaris. Effects were recorded using transcriptional analysis combined with histochemistry, immunofluorescence-, live-cell- and scanning electron microscopy. These analyses revealed heterogeneous monolayers containing both immature and differentiated cells including tuft cells and mucus-producing goblet cells. Stimulation with IL-4/IL-13 increased tuft- and goblet cell differentiation as demonstrated by the expression of DCLK1 and MUC2. In these cytokine-primed monolayers, the expression of MUC2 was further promoted by co-culture with P. univalens. Moreover, live-cell imaging revealed morphological alterations of the epithelial cells following exposure to larvae even in the absence of cytokine stimulation. Thus, the present work describes the design, characterization and usability of an experimental model representing the equine nematode-infected small intestinal epithelium. The presence of tuft cells and goblet cells whose mucus production is affected by Th2 cytokines and/or the presence of larvae opens up for mechanistic studies of the physical interactions between nematodes and the equine intestinal mucosa.
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| 3. |
- Ferrari, Desiree, et al.
(författare)
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Concentration of carprofen in the milk of lactating bitches after cesarean section and during inflammatory conditions
- 2022
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Ingår i: Theriogenology. - : Elsevier. - 0093-691X .- 1879-3231. ; 181, s. 59-68
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Tidskriftsartikel (refereegranskat)abstract
- Pain treatment of lactating bitches is a clinically relevant, but complicated issue. Published scientific studies regarding the excretion of drugs in canine milk are scarce. When considering the risk of side effects in their offspring, lactating bitches have traditionally received very restricted analgesic and anti-inflammatory therapy. Our aim was to quantify the concentrations of carprofen in milk from lactating bitches and relate those to potential risks for the puppies. A second aim was to evaluate the impact mastitis may have on the concentration of carprofen in milk. A population of 100 bitches was enrolled in the study, among which 88 were bitches treated with carprofen after cesarean section (Group CS), eight were bitches with painful inflammatory conditions (Group I) and four were bitches with mastitis (Group M). The patients enrolled in the study received carprofen 4 mg/kg sc at day 1 followed by 2 mg/kg po every 12 h for the following 2-5 days. Owners were instructed to collect milk once a day for five days. The concentration of carprofen in the milk was quantified with ultra-performance liquid chromatography-tandem mass spectrometry. The data obtained were statistically analyzed as repeated-measures data with a mixed-model approach. Data were used to calculate the theoretical maximum total daily intake of carprofen by the puppies in order to perform a computerized simulation of the plasma concentration of carprofen in the puppies. Follow-up telephone interviews to check the status of the enrolled bitches and their litters occurred at one week and three-six months after treatment with car-profen. The major finding of the study was that the concentration of carprofen in the milk was <700 ng/ mL from bitches undergoing CS or suffering painful conditions other than mastitis. In comparison, administration of 2 mg/kg of carprofen sc or po to adult dogs, results in mean maximal plasma con-centrations of 19480 +/- 5420 ng/mL (mean +/- SD). Moreover, data suggests that inflammation of the mammary gland results in a higher concentration of carprofen in milk (up to 1300 ng/mL). In the computerized simulation, the plasma concentrations of carprofen in puppies in group CS and in group I are one tenth of the concentration in adult dogs receiving carprofen at standard doses. Considering the low excretion into milk, carprofen provides an analgesic alternative to lactating bitches without mastitis.(c) 2022 Published by Elsevier Inc.
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| 4. |
- Weichert, Fabian, et al.
(författare)
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A multi-biomarker study on Atlantic salmon (Salmo salar L.) affected by the emerging Red Skin Disease in the Baltic Sea
- 2021
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Ingår i: Journal of Fish Diseases. - : Wiley. - 0140-7775 .- 1365-2761. ; 44:4, s. 429-440
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Tidskriftsartikel (refereegranskat)abstract
- For half a decade, the Atlantic salmon in the Baltic Sea has been facing severe health issues. Clinical signs like haemorrhage, erosions and ulcerative/necrotic skin conditions in returning adults have been reported from different Swedish rivers. These primary disease signs precede a secondary, terminal fungal infection. As initial investigations of the disease did not provide conclusive answers regarding the pathogenesis, this study was initiated to gain insight into a possible link between this so-called Red Skin Disease and anthropogenic influences. Therefore, returning salmon were caught in rivers along the Swedish coast and different tissues were sampled. The focus was put on the measurements of a battery of biomarkers as well as biochemical and haematological parameters, which were analysed using multivariate statistics. The main findings were a severe osmotic haemodilution, an immune response and an alteration of the carbohydrate metabolism in diseased fish. Furthermore, oxidative stress does not seem to be a likely factor in the pathogenesis. Concluding, certain changes in physiological parameters were shown to be indicative for the disease patterns, while others were ruled out as significant factors. Thus, this study contributes to the understanding of the Red Skin Disease and may act as a hypothesis generator for future studies.
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| 5. |
- Nystrom, S., et al.
(författare)
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The expression of nerve growth factor in healthy and inflamed equine chondrocytes analysed by capillary western immunoassay
- 2022
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Ingår i: Research in Veterinary Science. - : Elsevier BV. - 0034-5288 .- 1532-2661. ; 151, s. 156-163
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Tidskriftsartikel (refereegranskat)abstract
- Nerve Growth Factor (NGF) is a signalling molecule for pain and inflammation. NGF is increased in synovial fluid from osteoarthritic humans and animals, compared to healthy controls. Monoclonal antibody therapy directed against NGF has been approved to treat pain in osteoarthritic dogs but despite many years of trialling, therapy has not been approved for human use. One reason for this is that adverse reactions with rapidly progressing osteoarthritis has occurred in some individuals. More detailed knowledge of NGF expression in joints is needed. In this study, capillary-based Simple Western was used to analyse NGF in cultured equine chondrocytes. Chondrocytes were collected post mortem from three macroscopically healthy intercarpal joints and three intercarpal joints with mild osteoarthritic changes. The chondrocytes were expanded to passage one and seeded in chondrogenic medium to maintain the phenotype. On day four, cells were either stimulated with LPS or kept untreated in medium. All cells were harvested on day five. Wes analysis of lysates did not show mature NGF but two proforms, 40 and 45 kDa, were identified. Results were confirmed with western blot. The same proforms were expressed in chondrocytes from healthy and osteoarthritic joints. Acute inflammation induced by LPS stimulation did not change the forms of expressed NGF. Capillary Simple Western offers a sensitive and sample -sparing alternative to traditional western blot. However, confirmation of peaks is imperative in order to avoid misinterpretation of findings. In addition, in this case the method did not offer the possibility of quantification advertised by the manufacturers.
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| 6. |
- Vågsholm, Ivar
(författare)
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Challenges and opportunities in the implementation of new meat inspection systems in Europe
- 2021
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Ingår i: Trends in Food Science and Technology. - : Elsevier BV. - 0924-2244. ; 116, s. 460-467
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Forskningsöversikt (refereegranskat)abstract
- Background: The traditional meat inspection system is often found to be inefficient. Meat inspection in European countries is in a phase of modernisation to reflect improvements in livestock health and advances in understanding meat safety. The key point of progress is to replace the traditional meat inspection with risk-based meat inspection, including the elements of a meat safety assurance system (MSAS). Scope and approach: Modernisation of meat inspection was launched by the European Food Safety Authority's (EFSA) opinions and recommendations in the period 2011-2013 and consequent amendments to the EU meat inspection legislation in the period 2014-2019. For this study, the EU-funded RIBMINS COST Action conducted a comprehensive survey using an in-depth questionnaire to estimate the level of implementation of new risk-based meat inspection systems in Europe, stakeholders' confidence in the new systems and the main identified obstacles. Key findings and conclusions: The implementation of new meat inspection systems is still ongoing, as they have been fully implemented in just 61%, 42% and 38% of the surveyed countries in the pig, bovine, and poultry sectors, respectively. The main identified obstacles are existing trade agreements with 3rd countries, costs of implementation, inadequate food chain information and resistance from meat inspectors. Improvement of all components of the current meat inspection systems is a prerequisite for further modernisation.
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| 7. |
- Akula, Srinivas, et al.
(författare)
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Quantitative Transcriptome Analysis of Purified Equine Mast Cells Identifies a Dominant Mucosal Mast Cell Population with Possible Inflammatory Functions in Airways of Asthmatic Horses
- 2022
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Ingår i: International Journal of Molecular Sciences. - : MDPI. - 1661-6596 .- 1422-0067. ; 23:22
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Tidskriftsartikel (refereegranskat)abstract
- Asthma is a chronic inflammatory airway disease and a serious health problem in horses as well as in humans. In humans and mice, mast cells (MCs) are known to be directly involved in asthma pathology and subtypes of MCs accumulate in different lung and airway compartments. The role and phenotype of MCs in equine asthma has not been well documented, although an accumulation of MCs in bronchoalveolar lavage fluid (BALF) is frequently seen. To characterize the phenotype of airway MCs in equine asthma we here developed a protocol, based on MACS Tyto sorting, resulting in the isolation of 92.9% pure MCs from horse BALF. We then used quantitative transcriptome analyses to determine the gene expression profile of the purified MCs compared with total BALF cells. We found that the MCs exhibited a protease profile typical for the classical mucosal MC subtype, as demonstrated by the expression of tryptase (TPSB2) alone, with no expression of chymase (CMA1) or carboxypeptidase A3 (CPA3). Moreover, the expression of genes involved in antigen presentation and complement activation strongly implicates an inflammatory role for these MCs. This study provides a first insight into the phenotype of equine MCs in BALF and their potential role in the airways of asthmatic horses.
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| 8. |
- Dube, Faruk, et al.
(författare)
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Transcriptomics of ivermectin response in Caenorhabditis elegans : Integrating abamectin quantitative trait loci and comparison to the Ivermectin-exposed DA1316 strain
- 2023
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Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 18:5
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Tidskriftsartikel (refereegranskat)abstract
- Parasitic nematodes pose a significant threat to human and animal health, as well as cause economic losses in the agricultural sector. The use of anthelmintic drugs, such as Ivermectin (IVM), to control these parasites has led to widespread drug resistance. Identifying genetic markers of resistance in parasitic nematodes can be challenging, but the free-living nematode Caenorhabditis elegans provides a suitable model. In this study, we aimed to analyze the transcriptomes of adult C. elegans worms of the N2 strain exposed to the anthelmintic drug Ivermectin (IVM), and compare them to those of the resistant strain DA1316 and the recently identified Abamectin Quantitative Trait Loci (QTL) on chromosome V. We exposed pools of 300 adult N2 worms to IVM (10(-7) and 10(-8) M) for 4 hours at 20 degrees C, extracted total RNA and sequenced it on the Illumina NovaSeq6000 platform. Differentially expressed genes (DEGs) were determined using an in-house pipeline. The DEGs were compared to genes from a previous microarray study on IVM-resistant C. elegans and Abamectin-QTL. Our results revealed 615 DEGs (183 up-regulated and 432 down-regulated genes) from diverse gene families in the N2 C. elegans strain. Of these DEGs, 31 overlapped with genes from IVM-exposed adult worms of the DA1316 strain. We identified 19 genes, including the folate transporter (folt-2) and the transmembrane transporter (T22F3.11), which exhibited an opposite expression in N2 and the DA1316 strain and were deemed potential candidates. Additionally, we compiled a list of potential candidates for further research including T-type calcium channel (cca-1), potassium chloride cotransporter (kcc-2), as well as other genes such as glutamate-gated channel (glc-1) that mapped to the Abamectin-QTL.
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| 9. |
- Sharma, Garima, et al.
(författare)
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Antibiotic use, knowledge, and practices of milk vendors in India's informal dairy value chain
- 2022
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Ingår i: Frontiers in Sustainable Food Systems. - : Frontiers Media S.A.. - 2571-581X. ; 6
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Tidskriftsartikel (refereegranskat)abstract
- Background: Milk vendors play an important role in India's dairy value chain; however, their food safety practices are poorly understood. From the standpoint of milk safety, vendor behavior is significant because it has the potential to affect both consumer and producer behavior. This study investigates vendors' hygienic knowledge and practices toward milk safety, and antimicrobial resistance (AMR)Methods: A cross-sectional study was conducted in the states of Assam and Haryana (India). In selected villages, all the milk vendors identified at the time of the visit were interviewed. A questionnaire was used to assess the knowledge and practices on antibiotics, milk safety, and hygiene. The milk samples were tested for the presence of antibiotic-resistant bacteria using antibiotic susceptibility testing.Results: A total of 2444 milk vendors were interviewed during the survey. Out of these 156 (59.8%) of the vendors traded in raw milk while 40.2% traded in pasteurizes milk. Vendors were categorized depending on whom they supplied milk. Five categories were identified: (a) those who sold at grocery shops; (b) those who sold on the roadside; (c) those who sold from door to door; (d) those who sold to sweet makers/tea stalls; (e) those who sold from own home/other entity. The level of training among vendors on milk hygiene was non-existent and the knowledge related to antibiotics was low. Most of them (86.07%) agreed that boiled milk is always safer than raw milk but almost half (48.77%) of them admitted that sometimes they drink milk without boiling it. Most vendors believed that they could identify whether milk is safe or not for consumption just by its appearance and smell. Out of 124 milk samples collected from surveyed milk vendors and tested for the presence of antibiotic-resistant bacteria, 80 (64.52%) tested positive.Conclusion: This study highlights the low levels of knowledge regarding food safety among milk vendors. It shows the predominance of informal milk businesses in the surveyed states and the prevalence of AMR bacteria in milk traded by them. Training may be a beneficial strategy for addressing the issue.
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| 10. |
- Samad, Mohammed Abdus, et al.
(författare)
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Microbial Contamination and Antibiotic Resistance in Marketed Food in Bangladesh : Current Situation and Possible Improvements
- 2023
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Ingår i: Antibiotics. - : MDPI. - 0066-4774 .- 2079-6382. ; 12:3
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Tidskriftsartikel (refereegranskat)abstract
- Antimicrobial resistance (AMR) is a public health problem worldwide. Bangladesh, like its neighboring countries, faces many public health challenges, including access to safe food, inadequate food surveillance, as well as increasing AMR. This study investigated bacterial contamination and the AMR profile of pathogens in marketed food in Bangladesh and explored barriers to reducing AMR in the country. We collected 366 tomatoes, 359 chicken and 249 fish samples from 732 vendors in traditional markets in urban, peri-urban and rural areas in Bangladesh, as well as from 121 modern retails in Dhaka capital to analyse Vibrio cholerae and Escherichia coli in fish, Salmonella in chicken, and Salmonella and E. coli in tomatoes. Antibiotic susceptibility against 11 antibiotics was tested using a disc diffusion test and interpreted by an automated zone inhibition reader. In addition, a qualitative study using key informant interviews was conducted to explore antimicrobial use and AMR reduction potential in Bangladesh. We found E. coli in 14.21% of tomatoes and 26.91% of fish samples, while 7.38% of tomatoes and 17.27% of chicken were positive for Salmonella, and 44.98% of fish were positive for Vibrio cholerae. In total 231/319 (72.4%) of all pathogens isolated were multidrug-resistant (MDR) (resistant to three or more antibiotic groups). Qualitative interviews revealed an inadequate surveillance system for antibiotic use and AMR in Bangladesh, especially in the agriculture sector. To be able to fully understand the human health risks from bacterial hazards in the food and the AMR situation in Bangladesh, a nationwide study with a one health approach should be conducted, within all sectors, including AMR testing as well as assessment of the antimicrobial use and its drivers.
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