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Comparative expression analysis of inflammatory and immune-related genes in cattle during acute infection with foot-and-mouth disease virus in Egypt
2021
El Nahas, Abeer F. | Abd El Naby, Walaa S.H. | Khatab, Shymaa A. | Fergany, Al-Zahraa A. | Rashed, Rashed R.
Foot-and-mouth disease is a highly infectious viral disease affecting all cloven-footed domestic animals. The three foot-and-mouth disease virus (FMDV) serotypes A, O and SAT2 are at present the greatest threat to susceptible animals in Egypt. The aim of the present study was, for the host factors associated with different FMDV infections in cattle during the acute phase, to compare these factors’ influence on the expression of the IL-10, TLR-2, TNF-α, CXCL10, CD48, NFATC4 and IFNG inflammatory and immune-related genes. Vesicular fluid and epithelium samples were obtained from at least three infected cattle on the same affected farm during three different FMDV outbreaks and were used for serotyping of the virus and for expression analysis of host genes. A two-step RT-PCR was used for diagnosis of the virus with primers specific for each serotype. In quantitative PCR analysis, the expression patterns of TLR-2 and IFNG were prominent, while NFATC4 expression was absent in all FMDV-infected cattle. The highest expression of CD48 was associated with increased expression of other inflammatory and immune-related genes (IL-10, TLR-2, TNF-α and IFNG), which may be an indication of rapid virus clearance. The use of vesicular fluid and epithelium for investigation of viral and immune-related gene expression levels in acute FMDV infection is possible. Host-dependent variation in the expression of the studied genes was observed in different FMDV serotype outbreaks.
Mostrar más [+] Menos [-]Analysis of β-agonists in different biological matrices by liquid chromatography–tandem mass spectrometry
2021
Śniegocki, Tomasz | Sell, Bartosz | Posyniak, Andrzej
Wide use is made of β-agonists in therapy due to their smooth muscle–relaxant properties. They also have a side effect of increasing muscle mass. Besides improving oxygen utilisation as bronchodilators, β-agonists increase protein synthesis and promote fat burning. The growth- and performance-enhancing effects are often exploited in illegal use. The guiding objective of this study was to develop a procedure for the determination of β-agonists by a single method in different types of matrices. Five grams of homogenised samples were subjected to enzymatic hydrolysis with β-glucuronidase in ammonium acetate, pH 5.2. Purification was performed by solid phase extraction. Analytes were eluted with 10% acetic acid in methanol. The eluted β-agonists were analysed by high-performance liquid chromatography–tandem mass spectrometry. Validation results met the requirement of the confirmation criteria according to European Commission Decision 2002/657/EC in terms of apparent recoveries (93.2–112.0%), repeatability (3.1–7.1%) and intra-laboratory reproducibility (4.1–8.2%). The method can be successfully applied in the detection and determination of clenbuterol, salbutamol, mabuterol, mapenterol, terbutaline, brombuterol, zilpaterol, isoxsuprine and ractopamine in feed, drinking water, urine, muscle, lung and liver matrices.
Mostrar más [+] Menos [-]Characterisation of a new molecule based on two E2 sequences from bovine viral diarrhoea-mucosal disease virus fused to the human immunoglobulin Fc fragment
2021
González Pose, Alaín | Montesino Seguí, Raquel | Maura Pérez, Rafael | Hugues Salazar, Florence | Cabezas Ávila, Ignacio | Altamirano Gómez, Claudia | Sánchez Ramos, Oliberto | Roberto Toledo, Jorge
Proper conformational arrangement of the E2 molecules of bovine viral diarrhoea-mucosal disease virus (BVD-MDV) is crucial to obtain an effective recombinant vaccine candidate against the disease. In this study, we characterised a new molecule composed of two distinct sequences of the E2 glycoprotein of BVD-MDV and the Fc fragment of human immunoglobulin (BVDE2Fc). The chimaeric protein was expressed in mammalian cell lines of different species by adenoviral transduction and purified by immobilised metal-affinity chromatography. The N-glycans were profiled by HPLC, and the BVDE2Fc immunogenicity was assessed in male mice. The antigen-antibody reactions were evaluated by ELISA. The MDBK cell line was selected from among five for the final production of BVDE2Fc. After purification to over 90%, the N-glycan profile showed neutral and complex oligosaccharides. The mouse immunisation induced a strong humoral response, which produced antibodies able to attach to conformational epitopes on E2 molecules, while the Fc fragment barely contributed to the immune response. Additionally, BVDE2Fc attached to antibodies from bovine sera positive to distinct BVD-MDV subtypes, whereas the loss of BVDE2Fc structure during the deglycosylation process considerably diminished those interactions. These results demonstrate that the structure of E2 molecules arranged in tandem and attached to an Fc fragment could represent a viable design for future vaccine candidates against BVD-MD.
Mostrar más [+] Menos [-]Age and seasonal pattern of contagious agalactia in small ruminants in Ukraine
2021
Bohach, Dmytro Mykolaivych | Stegniy, Borys Tymofiyovych | Bohach, Mykola Volodymyrovych | Pavlov, Serhii Leonidovych | Bolotin, Vitaliy Igorovych
The aim of the study was to determine how the spread of contagious agalactia in sheep and goats in the Odesa region depended on the age of the animals and the season. From January 2016 to December 2018, 1,964 ewes and 1,484 nanny goats of different age groups were studied by ELISA for antibodies to Mycoplasma agalactiae. The highest incidence of contagious agalactia was registered in one-year-old animals and was 59.7‒83.0%, two-year-old ruminants showed 17.0‒40.3% prevalence, in livestock at the age of 3–4 years no serological evidence of the disease was registered and in ewes and nanny goats older than 5–6 years 1.5–3.6% were infected. The most susceptible were young animals at the age of one-month (11.6‒14.5%). The first peak of the disease was recorded in March‒April (21.0‒26.1%), in the lambing period, which coincided with the beginning of lactation and the suckling period, and the second peak occurred in June–July (28.9‒34.2%), the period of maximum lactation and of manual milking of sheep and goats. The results of serological investigations indicate the circulation of M. agalactiae in small ruminants in the south of Ukraine. To avoid greater dissemination of the pathogen, appropriate measures should be applied and strategies for its control need to be drawn up.
Mostrar más [+] Menos [-]The impact of selected risk factors on the occurrence of highly pathogenic avian influenza in commercial poultry flocks in Poland
2021
Gierak, Anna | Śmietanka, Krzysztof
Introduction of highly pathogenic avian influenza virus (HPAIV) into a country and its further spread may have a devastating impact on the poultry industry and lead to serious economic consequences. Various risk factors may increase the probability of HPAI outbreak occurrence but their relative influence is often difficult to determine. The study evaluates how the densities of selected poultry species and proximity to the areas inhabited by wild birds impacted HPAI outbreak occurrence during the recently reported epidemics in Poland. The analysis was developed using these risk factors in the locations of affected and randomly chosen unaffected commercial farms. Generalised linear and non-linear models, specifically logistic regression, classification tree and random forest, were used to indicate the most relevant risk factors, to quantify their association with HPAI outbreak occurrence, and to develop a map depicting spatial risk distribution. The most important risk factors comprised the densities of turkeys, geese and ducks. The abundance of these species of poultry in an area increased the probability of HPAI occurrence, and their farming intensity in several areas of central, western, eastern and northern Poland put these areas at the highest risk. The results may improve the targeting of active surveillance, strengthen biosecurity in the areas at risk and contribute to early detection of HPAI in outbreak reoccurrences.
Mostrar más [+] Menos [-]Microbiological safety of food of animal origin from organic farms
2021
Sosnowski, Maciej | Osek, Jacek
The organic food sector and consumer interest in organic products are growing continuously. The safety and quality of such products must be at least equal to those of conventional equivalents, but attaining the same standards requires overcoming a particular problem identified in organic food production systems: the occurrence of bacterial pathogens such as Salmonella, Campylobacter, Listeria monocytogenes, Staphylococcus aureus and pathogenic Escherichia coli. These food-borne microorganisms were detected in the production environments of such food. The prevalence of pathogenic bacteria in organic livestock and products may be higher, but may also be the same as or lower than in like material from conventional farms. Furthermore, the incidence of antimicrobial-resistant bacteria was more often detected in conventional than in organic production. The aim of this review was to present the recent information on the microbiological safety of food of animal origin produced from raw materials from organic farms.
Mostrar más [+] Menos [-]A pilot study to establish an ovalbumin-induced atopic dermatitis minipig model
2021
Kim, Yŏng-gyu | Lee, Ju Young | Hwang, Jeong Ho | Suh, Han Na
Because minipig skin is similar to human skin in anatomy and physiology, establishing an atopic dermatitis (AD) minipig model seems meaningful. We applied 1-fluoro-2,4-dinitrobenzene (DNFB) or ovalbumin onto the back skin of five Yucatan minipigs aged 8–10 months and 19 kg in median weight. Two minipigs with the same parameters served as controls. Both DNFB and ovalbumin mediated epithelial hyperplasia, spongiosis, and immune cell infiltration in the dermis, which is a typical histopathological feature of AD. Moreover, AD upregulated the Th1- and Th2-related cytokine expressions in DNFB- or in ovalbumin-treated skin. Notably, AD-induced minipigs exhibited greater cytokine serum concentrations. Histopathological finding and cytokine analysis revealed that DNFB or ovalbumin mediates AD. However, ovalbumin-treated minipig is a more reliable and precise AD model owing to the DNFB-induced severe skin damage. In summary, ovalbumin-treated skin shows similar AD as human in histopathological and molecular analysis.
Mostrar más [+] Menos [-]Evaluation of serum and urine neutrophil gelatinase-associated lipocalin and cystatin C as biomarkers of acute kidney injury in horses
2021
Siwińska, Natalia | Żak, Agnieszka | Pasławska, Urszula
Diagnosis of acute kidney injury (AKI) in horses is difficult at the subclinical stage, due to nonspecific clinical signs. The aim of this study was to evaluate the concentrations of selected serum and urinary biomarkers in healthy horses, horses at risk of AKI, and those with clinical AKI. Thirty healthy horses, 30 horses at risk of AKI and 11 horses with clinical AKI and azotaemia were included in the study. Serum and urinary neutrophil gelatinase-associated lipocalin (NGAL) and cystatin C were measured using commercially available enzyme immunoassay tests. The median and (in parentheses) first and third quartile concentrations of selected biomarkers in healthy horses, horses at risk of AKI and horses with AKI were respectively as follows: serum cystatin C – 0.25 (0.19–0.37), 0.23 (0.15–0.37) and 0.61 (0.37–1.13) mg/L; serum NGAL – 50.5 (38.8–58.8), 51.1 (40.4–66.9) and 98.1 (59.4–128.2) ng/mL; urinary NGAL – 20.7 (17.9–24.5), 32.3 (32.7–55.8) and 36.6 (26.8–89.9) ng/mL; and urinary cystatin C – 0.1 (0.07–0.13), 0.13 (0.1–0.2) and 0.34 (0.22–0.37) mg/L. There were significant differences in the concentration of all biomarkers between the healthy and AKI-affected horses. Horses with AKI all had biomarker concentrations higher than the healthy horses. None of the biomarkers made azotaemia recognisable in all affected horses. The obtained results indicate the need to create a serum and urinary biomarker panel to detect AKI.
Mostrar más [+] Menos [-]Comparison of chicken immune responses after inoculation with H5 avian influenza virus-like particles produced by insect cells or pupae
2021
Huang, Dean | Chao, Yu-Chan | Lv, Zhengbing | Jan, Jia-Tsrong | Yang, Yuzhi | Hsiao, Pei-Wen | Wu, Jiaying | Liao, Chiu-Hsun | Wu, Tzu-Hsien | Wang, Lih-Chiann
Novel clade 2.3.4.4 H5 highly pathogenic avian influenza virus (HPAIV) outbreaks have occurred since early 2015 in Taiwan and impacted the island economically, like they have many countries. This research investigates the immunogenicity of two HPAIV-like particles to assess their promise as vaccine candidates. The haemagglutinin (HA) gene derived from clade 2.3.4.4 H5 HPAIV and matrix protein 1 (M1) gene were cloned into the pFastBac Dual baculovirus vector. The resulting recombinant viruses were expressed in Spodoptera frugiperda moth (Sf)21 cells and silkworm pupae to generate Sf21 virus-like particles (VLP) and silkworm pupa VLP. Two-week-old specific pathogen–free chickens were immunised and their humoral and cellular immune responses were analysed. The silkworm pupa VLP had higher haemagglutination competence. Both VLP types elicited haemagglutination inhibition antibodies, anti-HA antibodies, splenic interferon gamma (IFN-γ) and interleukin 4 (IL-4) mRNA expression, and CD4⁺/CD8⁺ ratio elevation. However, chickens receiving silkworm pupa VLP exhibited a significantly higher anti-HA antibody titre in ELISA after vaccination. Although Sf21 VLP recipients expressed more IFN-γ and IL-4, the increase in IFN-γ did not significantly raise the CD4⁺/CD8⁺ ratio and the increase in IL-4 did not promote anti-HA antibodies. Both VLP systems possess desirable immunogenicity in vivo. However, in respect of immunogenic efficacy and the production cost, pupa VLP may be the superior vaccine candidate against clade 2.3.4.4 H5 HPAIV infection.
Mostrar más [+] Menos [-]Effects of topical treatment of foot rot in sheep using ozonated olive ointment
2021
Szponder, Tomasz | Zdziennicka, Joanna | Nowakiewicz, Aneta | Świeca, Michał | Sobczyńska-Rak, Aleksandra | Żylińska, Beata | Patkowski, Krzysztof | Junkuszew, Andrzej | Wessely-Szponder, Joanna
Foot rot in small ruminants is highly contagious, causes severe lameness, and impairs fertility and wool and meat production. It is usually treated with parenteral antibiotics, with attendant antibiotic resistance risk, and with bactericidal footbaths, potentially harmful to humans and the environment. An alternative treatment in sheep is proposed based on repeated topical ozonated ointment application. Its effectiveness and safety were evaluated by estimation of acute-phase response, biochemical indicators of organic damage, and antioxidant/oxidant balance (AOB). The study was conducted on ten sheep with Egerton scale 2–3 lesions. Ozone application was repeated every day for seven days. Blood was drawn first (T0) after foot cleaning and before ozonation, then (T1) seven days after the first ozone application, and finally (T2) four days after the last application. High clinical effectiveness was observed, with total recovery by 28 days from the start of treatment. A significant increase in antiradical activity was noted on the basis of a 2,2ʹ-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) assay from 1.16 ± 0.04 μmolTe/mL at T0 to 1.23 ± 0.03 μmolTe/mL at T1, with a slight decrease in oxidative stress. Calculated on the basis of antiradical capacity, AOB was higher at T1 (130 ± 19%) and decreased to 110 ± 16% at T2. Calculated on the basis of reducing power, it was 169 ± 22% at T1 and 131 ± 17% at T2. These results indicated that the AOB is efficient enough to prevent oxidative organ injury and the applied doses of ozone are safe for animals.
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