Assessment of lipid peroxidation in dairy cows with subclinical and clinical mastitis

Mastitis is still one of the major causes of economic losses in dairy sector. The routine application of bacteriologic examination of milk samples is often insufficient and for this reason, alternative parameters are used to identify trends in the development of the udder health. Therefore, the objectives of this study were to determine the relationship of oxidative product levels, using malondialdehyde (MDA) as a marker on occurrence of mastitis and its causing pathogens. Dairy herd of 223 Slovak spotted cattle were tested for etiology and occurrence of mastitis based on assessment of clinical signs, abnormal udder secretions, Californian Mastitis Test (CMT) with subsequent collecting of milk samples for bacteriological examination. From 892 quarter milk samples were selected for MDA detection 51 subclinical (SM) and 26 clinical mastitis (CM) quarters with positive CMT score and positive bacteriological examination of Staphylococcus spp. and Streptococcus spp. as well 40 healthy quarters. Results showed that among the current pathogens of the mammary gland belong CNS, S. aureus, S. sanguinis, S. uberis and E. coli, which were the most frequently isolated from SM and CM. The highest MDA level was observed from clinical cases of mastitis however, increased MDA levels were detectable from subclinical cases. Bacterial isolates from subclinical quarter milk samples are different levels of MDA. In this study, we found that quarter milk samples infected with S. uberis were higher compared to other pathogens. In conclusion, differences in both severity of mastitis and mastitic pathogens were associated with differences of oxidative products in infected udders

Sheep mastitis caused by staphylococci and streptococci and their influence on oxidative status

Article Details: Received: 2020-10-14 | Accepted: 2020-11-27 | Available online: 2021-01-31https://doi.org/10.15414/afz.2021.24.mi-prap.53-57AbstractThe objectives of this study were to determine the relationship of oxidative product levels, using malondialdehyde (MDA) as a marker on occurrence of mastitis and its causing pathogens in two dairy flocks of ewes situated in east and north of Slovakia. The diagnosis of mastitis was performed on the basis of clinical examination of the udder, macroscopic evaluation of milk, with the evaluation of Californian mastitis test (CMT) and bacteriological analysis of individual raw milk samples. From total 537 and 444 halves ewe’s milk samples were 16.6% and 23.2% positive to CMT, respectively. The prevalence of mastitis caused by bacterial pathogenes in the monitored herds was 14.3% to 19.1%, respectively. In all monitored sheep flocks were confirmed predominantly subclinical forms (SM) of intramammary infection (IMI). The highest of etiological agents in all monitored herds had coagulase negative staphylococci and coagulase positive staphylococci especially Staphylococcus aureus. Except for staphylococci were Streptococcus uberis and Streptococcus spp. most frequently pathogens isolated from mastitic sheep. The highest MDA level was observed from clinical cases of mastitis however, increased MDA levels were detectable from subclinical cases. Bacterial isolates from mastitc halves milk samples are different in levels of MDA. In this study, we found that milk samples infected with S. aureus were higher compared to other pathogens. In conclusion, differences in both severities of IMI and mastitis pathogens were associated with differences of MDA in infected udders.Keywords: sheep, milking, mastitis, lipid peroxidation,S. aureus, coagulase negative staphylococciReferencesContreras, A. et al. (2007). Mastitis in small ruminants. Small Ruminant Research, 68(1-2), 145–153. https://doi.org/10.1016/j.smallrumres.2006.09.011Fthenakis, G. C. (1995). California mastitis test and White side test in diagnosis of subclinical mastitis of dairy ewes. Small Ruminant Research, 16(3), 271–276. https://doi.org/10.1016/0921-4488(95)00638-2Hariharan, H. et al. (2004). Bacteriology and somatic cell counts in milk samples from ewes on a Scottish farm. Canadian Journal of Veterinary Research, 68(3), 188–192.https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1142138/Malinowski, E. et al. (2008). Etiological agents of dairy cows’ mastitis in western part of Poland. Polish Journal of Veterinary Sciences, 9(3), 191-194. https://pubmed.ncbi.nlm.nih.gov/17020014/Mørk, T. et al. (2007). Clinical mastitis in ewes; bacteriology, epidemiology and clinical features. Acta Veterinaria Scandinavica, 49(1), 23. https://dx.doi.org/10.1186%2F1751-0147-49-23Pyörälä, S. and Taponen, S. (2009). Coagulase-negative staphylococci - Emerging mastitis pathogens. Veterinary Microbiology, 34(2), 3–8. https://doi.org/10.1016/j.vetmic.2008.09.015Sharma, N. (2011). Oxidative stress and antioxidant status during transition period in dairy cows. Asian-Australian Journal of Animal Science, 24(4), 479–484. https://www.ajas.info/upload/pdf/24-58.pdfSuriyasathaporn, W. (2006). Higher somatic cell counts resulted in higher malondialdehyde concentrations in raw cow´s milk. International Dairy Journal, 16(9), 1088–1091. https://doi.org/10.1016/j.idairyj.2005.11.004Turk, R. et al. (2017). The role of oxidative stress and inflammatory response in the pathogenesis of mastitis in dairy cows. Mljekarstvo, 67(2), 91–101. https://doi.org/10.15567/mljekarstvo.2017.020

New trends in the use of recycled manure solids in dairy housing

Article Details: Received: 2020-10-14 | Accepted: 2020-11-27 | Available online: 2021-01-31https://doi.org/10.15414/afz.2021.24.mi-prap.109-113The aim of the study was to compare improved bedding composition with conventional straw bedding under farm conditions, regarding its effects on the indicator microorganisms influencing hygiene levels. Dairy cows were housed in newly-built stables divided into two parts each with four sections and bedded cubicles arranged in three rows. In the first part, the bedded cubicle floors were layered with straw up to a height of 20 cm. In the second part, the cubicles were layered to a height of 20 cm with improved bedding composition consisting of recycled manure solids (RMS; 10 kg), ground limestone (200 kg), straw (40 kg) and water (110 l). After layering the bedding was treated with a concrete selector to provide strength and sufficient resistance. Samples for microbiological examination were taken from four sections according to the time interval of production and bedding. From three sections were taken bedding samples according to a new recipe in intervals of 1-3 months after its use. A control sample of bedding, consisting of straw, was taken from the last, fourth section. Comparing classical straw bedding with the improved recipe bedding, the total viable count (TVC) and coliforms bacteria (CB) in a first and second month after laying were found to be reduced. In addition to TVC and CB, decreased numbers of faecal coliforms (FC) and faecal streptococci (FS) were observed in the first, second and third months after layering. In addition to reducing the number of microorganisms, the improved bedding showed a stabilizing effect by keeping the litter thickness up to the bedding threshold (200 mm), which has a beneficial effect for increased purity of the body and udder.Keywords: dairy cows, housing, hygiene, improved bedding, microorganismsReferences Bradley, A.  et al. (2018). The impact of dairy cows’ bedding material and its microbial content on the quality and safety of milk–A cross sectional study of UK farms. International Journal of Food Microbiology, 23(269), 36–45. https://doi.org/10.1016/j.ijfoodmicro.2017.12.022Brouček, J. et al. (2015). Impact of gestation and lactation stage on the dairy cow response following removal to unfamiliar housing and milking system. Italian Journal of Animal Scince, 14(2), 233–237.https://doi.org/10.4081/ijas.2015.3410Fournel, S. et al. (2018). Production of recycled manure solids for bedding in Canadian dairy farms: I. Solid-liquid separation. Journal of Dairy Science, 102(2), 1832–1846. https://doi.org/10.3168/jds.2018-14966Heinonen-Tanski,  H. et al. (2006). Methods to reduce pathogen microorganisms in manure. Livestock Science, 102(3), 248–255. https://doi.org/10.1016/j.livsci.2006.03.024Ismaïl, R. et al. (2013). Methods for recovering microorganisms from solid surfaces used in the food industry: a review of the literature. International Journal of Environmental Research and Public Health, 10(11), 6169–6183.https://dx.doi.org/10.3390%2Fijerph10116169Maiorano, G. (2014). Livestock Production for a Sustainable Development. J. Microbiol. Biotechnol. Food Sci. 3, 34–38. Available online: https://www.jmbfs.org/wp-content/uploads/2014/01/10_jmbfs_maiorano_2014_fs.pdfLeach, K. A. et al. (2015). Recycling manure as cow bedding: Potential benefits and risks for UK dairy farms. Vet. J., 206, 123-130. https://doi.org/10.1016/j.tvjl.2015.08.013Popescu, S. et al. (2014). The effect of the housing system on the welfare quality of dairy cows. Italian Journal of Animal Scince, 13(1), 2940. https://doi.org/10.4081/ijas.2014.2940 Tančin, V. et al. 2013. Livestock farming in marginal areas (In Slovak), CVŽV, Nitra, Slovakia 2013. p. 70. ISBN 978-80-89418-26-8. Wolfe, T. et al. (2018). Effects of alternative deep bedding options on dairy cow preference, lying behavior, cleanliness, and teat end contamination. Journal of Dairy Science,101(1), 530-536. https://doi.org/10.3168/jds.2016-12358

Aging markers in equine red blood cells

Detection of hematopoietic activity in horses is a challenge due to the lack of cells carrying reticulocyte markers such as RNA remnants or CD71 in the circulation. In this study, we fractionated equine red cells according to their density and analyzed the cells forming low (L), medium (M), and high (H) density fractions for markers of aging such as membrane loss, oxidation, and alterations in the intracellular free Ca levels. Cells forming L and M fraction were highly heterogeneous in projected areas and shapes, and had higher propensity to swell in response to hypo-osmotic challenge than the cells from the H fraction. The densest cells were deprived of band 3 protein compared to the cells within L or M fraction. Furthermore, the equine red cells from the H fraction were hyper-oxidized compared to the cells within M and L fractions as follows from an increase in autofluorescence characteristic for oxidized damaged hemoglobin and from thiol oxidation as detected using monobromobimane. The lightest cells showed lower free thiol content compared to the red blood cells from the M fraction, but did not contain oxidized hemoglobin. Finally, the majority of red blood cells forming L, M, and H fraction prominently differed from each other in intracellular free Ca levels and its distribution within the cells. Based on the obtained findings, we suggest that intraerythrocytic Ca levels and its subcellular distribution, eosin-5-maleimide binding test for band 3 abundance, and autofluorescence of cells along with the changes in red blood cell indices, distribution width and creatine levels may become potential markers of regenerative erythropoiesis in horses. Validation of the power of these potential markers of red cell aging is pending

Low-Level Light Therapy Protects Red Blood Cells Against Oxidative Stress and Hemolysis During Extracorporeal Circulation

Aim: An activation of non-specific inflammatory response, coagulation disorder, and blood morphotic elements damage are the main side effects of the extracorporeal circulation (ECC). Red-to-near-infrared radiation (R/NIR) is thought to be capable of stabilizing red blood cell (RBC) membrane through increasing its resistance to destructive factors. We focused on the development of a method using low-level light therapy (LLLT) in the spectral range of R/NIR which could reduce blood trauma caused by the heart-lung machine during surgery.Methods: R/NIR emitter was adjusted in terms of geometry and optics to ECC circuit. The method of extracorporeal blood photobiomodulation was tested during in vivo experiments in an animal, porcine model (1 h of ECC plus 23 h of animal observation). A total of 24 sows weighing 90–100 kg were divided into two equal groups: control one and LLLT. Blood samples were taken during the experiment to determine changes in blood morphology [RBC and white blood cell (WBC) counts, hemoglobin (Hgb)], indicators of hemolysis [plasma-free hemoglobin (PFHgb), serum bilirubin concentration, serum lactate dehydrogenase (LDH) activity], and oxidative stress markers [thiobarbituric acid reactive substances (TBARS) concentration, total antioxidant capacity (TAC)].Results: In the control group, a rapid systemic decrease in WBC count during ECC was accompanied by a significant increase in RBC membrane lipids peroxidation, while in the LLLT group the number of WBC and TBARS concentration both remained relatively constant, indicating limitation of the inflammatory process. These results were consistent with the change in the hemolysis markers like PFHgb, LDH, and serum bilirubin concentration, which were significantly reduced in LLLT group. No differences in TAC, RBC count, and Hgb concentration were detected.Conclusion: We presented the applicability of the LLLT with R/NIR radiation to blood trauma reduction during ECC

Usefulness of Thulium-Doped Fiber Laser and Diode Laser in Zero Ischemia Kidney Surgery—Comparative Study in Pig Model

Background: The aim of this study was to evaluate the usefulness of a thulium-doped fiber laser and a diode laser in zero ischemia kidney surgery, by carrying out a comparative study in a pig model. Material and methods: Research was carried out on 12 pigs weighing 30 kg each. A thulium-doped fiber laser (TDFL) and a diode laser (DL) operating at wavelengths of 1940 and 1470 nm, respectively, were used. The cut sites were assessed both macroscopically and microscopically. The zone of thermal damage visible in the histopathological preparations was divided into superficial and total areas. Results: During partial nephrectomy, moderate to minimal bleeding was observed, which did not require additional hemostatic measures. All animals survived the procedure. On day 0, the total thermal damage depth was 837.8 µm for the TDFL and 1175.0 µm for the DL. On day 7, the depths were 1556.2 and 2301.7 µm, respectively. On day 14, the overall thermal damage depth for the DL was the greatest (6800 µm). The width of the superficial zone was significantly reduced on days 7 and 14 after TDFL application. Conclusion: Both lasers are suitable for partial wedge nephrectomy without ischemia in pigs. The TDFL produced similar or better hemostasis than the DL, with a smaller zone of thermal damage and, therefore, seems more suitable for application in human medicine

Applicability of FTIR-ATR method to measure carbonyls in blood plasma after physical and mental stress

Introduction. Oxidative stress is a state of imbalance between the production of reactive oxygen species and antioxidant defenses. It results in the oxidation of all cellular elements and, to a large extent, proteins, causing inter alia the formation of carbonyl groups in their structures. The study focused on assessment of changes in the plasma protein-bound carbonyls in police horses after combat training and after rest and the applicability of infrared spectroscopy with a Fourier transform, utilizing the attenuated total reflectance (FTIR-ATR) in detecting plasma protein oxidation. Methods. We evaluated the influence of both the different concentrations of hydrogen peroxide and combat training on protein carbonylation in horse blood plasma. The oxidation of plasma proteins was assessed using a spectrophotometric method based on the carbonyl groups derivatization with 2,4-dinitrophenylhydrazine (DNPH). The measured values were correlated with the carbonyl groups concentrations determined by means of the FTIR-ATR method. Results. The linear correlation between the DNPH and FTIR-ATR methods was shown. The concentration of plasma protein-bound carbonyls significantly deceased in police horses after one-day rest when compared to the values measured directly after the combat training (a drop by 23%, pThis work was financed under the program of the Minister of Science and Higher Education “Strategy of Excellence - University of Research” in 2018-2019 project number 0019/SDU/2018/18 in the amount of PLN 700 000 and partly by the statutory funds of Wrocław University of Science and Technology. This work was created in cooperation with the Ministry of the Interior of the Slovak Republic, Department of Police Kynology and Hippology. Tomasz Walski was supported by the European Union’s Horizon 2020 Research and Innovation Programme, under the Marie Skłodowska-Curie Grant Agreement No 713690, and by the Science Foundation Ireland (SFI) and the European Regional Development Fund (Grant Number 13/RC/2073)

Addendum: Żywicka, B., et al. Comparison of a 1940 nm Thulium-Doped Fiber Laser and a 1470 nm Diode Laser for Cutting Efficacy and Hemostasis in a Pig Model of Spleen Surgery. <i>Materials</i> 2020, <i>13</i>, 1167

The authors would like to add the following sentence to the “Funding” section of their article [...

Applicability of FTIR-ATR Method to Measure Carbonyls in Blood Plasma after Physical and Mental Stress

Introduction. Oxidative stress is a state of imbalance between the production of reactive oxygen species and antioxidant defenses. It results in the oxidation of all cellular elements and, to a large extent, proteins, causing inter alia the formation of carbonyl groups in their structures. The study focused on assessment of changes in the plasma protein-bound carbonyls in police horses after combat training and after rest and the applicability of infrared spectroscopy with a Fourier transform, utilizing the attenuated total reflectance (FTIR-ATR) in detecting plasma protein oxidation. Methods. We evaluated the influence of both the different concentrations of hydrogen peroxide and combat training on protein carbonylation in horse blood plasma. The oxidation of plasma proteins was assessed using a spectrophotometric method based on the carbonyl groups derivatization with 2,4-dinitrophenylhydrazine (DNPH). The measured values were correlated with the carbonyl groups concentrations determined by means of the FTIR-ATR method. Results. The linear correlation between the DNPH and FTIR-ATR methods was shown. The concentration of plasma protein-bound carbonyls significantly deceased in police horses after one-day rest when compared to the values measured directly after the combat training (a drop by 23%, p<0.05 and 29%, p<0.01 measured by DNPH and FTIR-ATR methods, respectively). These results were consistent with the proteins phosphorylation analysis. Conclusion. The FTIR-ATR method may be applied to measure the level of plasma proteins peroxidation