Flow cytometry follow-up analysis of peripheral blood leukocyte subpopulations in calves experimentally infected with field isolates of Mycoplasma bovis

Changes in peripheral blood leukocyte subpopulations were investigated in calves challenged intratracheally with three different Mycoplasma bovis isolates in Groups E1, E2, and E3. The controls received a placebo. Blood samples were collected before challenge and then at days 1 to 7, 14, 21 and 28. White blood cells (WBC), polymorphonuclear leukocytes (PMNLs), lymphocytes (LYMs), monocytes, eosinophils and basophils (mid-size cells, MID), as well as CD2+, CD4+, CD8+, WC4+ lymphocyte subsets with CD4:CD8 ratio were also analysed. A transient increase of WBC and PMNLs in all challenged calves was observed on day 1. Increased LYM counts were observed in E1 throughout the study, whereas in E2 the LYM counts were higher only between days 14 and 28, and consistently lower in E3. The MID count had broadly comparable values for all groups. Stimulation of the CD2+ response was observed in E2 and E3 in contrast with E1 which had a lower CD2+ throughout. The CD4+ response was dominant in E1 and E2, whereas in E3 a parallel CD4+ and CD8+ stimulation was observed. The B-cell response (WC4+) and an increased CD4:CD8 ratio was most apparent in E1. The main host responses to M. bovis infections are a stimulation of CD4+ cells and an enhancement of the WC4+ response

Preliminary study on Se-enriched Lentinula edodes mycelium as a proposal of new feed additive in selenium deficiency

The presence of selenium in European soil is low and this causes its deficiency in livestock and, in consequence, in humans. This study aimed to obtain Lentinula (L.) edodes mycelium with the maximum content of selenium. This species was used for experiment based on its documented medicinal properties. Calves were fed with selenium-enriched L. edodes mycelium, and serum selenium concentration, average daily weight gains and selected immune parameters were estimated. The selenium-enriched mushroom was found to be safe based on cytotoxicity tests (MTT and LDH tests) and for this reason it was used for further experiments. The mean quantity of selenium in the serum of calves fed with selenium-enriched L. edodes mycelium was significantly higher than that of control calves. Additionally, the calves fed with selenium-enriched L. edodes mycelium had higher body weight gains than those of control calves. White blood cell counts and subpopulations of lymphocytes in the experimental and control calves were within the reference range. The administration of L. edodes enriched with selenium had a beneficial effect on state of health of the calves

The Effect of Pegbovigrastim Injection on Phagocytic and Oxidative Burst Activities of Peripheral Blood Granulocytes and Monocytes in Calves Challenged with Mycoplasma bovis

Mycoplasma bovis (M. bovis) is an important pathogen affecting cattle, causing various diseases including pneumonia which mainly occurring in calves. Control of M. bovis infections is difficult due to the lack of commercial vaccines in most parts of the world and increasing trends of antimicrobial resistance in field isolates of the pathogen; therefore, it seems reasonable to look for new solutions for the prevention of the infection. Pegbovigrastim is a pegylated form of naturally occurring circulating cytokine in cattle that affects bovine leukocytes and some cell functions. Most studies on pegbovigrastim have focused on reducing the occurrence of mastitis and other diseases occurring during the periparturient period in cows, while this study attempts to use pegbovigrastim in the prevention of respiratory diseases in calves, which are largely caused by M. bovis. Based on previous observations on the immunostimulatory properties of pegbovigrastim in cattle, for the first time, the effect of its injection on the number and phagocytic and oxidative burst activities of peripheral blood granulocytes and monocytes in calves experimentally infected with M. bovis was investigated. Pegbovigrastim administration in the calves significantly stimulated an increase in peripheral blood granulocyte and monocyte counts and phagocytic activity of the cells, especially granulocytes, which was also generally expressed in the course of M. bovis infection. In response to pegbovigrastim administration, a general increase in the oxygen burst activity of the cells was observed. This effect was also shown despite ongoing infection with M. bovis which, taken together, may indicate a beneficial effect of pegbovigrastim injection on the immunity of the affected animals

Prevalence of pathogens from Mollicutes class in cattle affected by respiratory diseases and molecular characteristics of Mycoplasma bovis field strains

Introduction: Mycoplasma bovis is one of the main pathogens involved in cattle pneumonia. Other mycoplasmas have also been directly implicated in respiratory diseases in cattle. The prevalence of different Mycoplasma spp. in cattle affected by respiratory diseases and molecular characteristics of M. bovis field strains were evaluated. Material and Methods: In total, 713 nasal swabs from 73 cattle herds were tested. The uvrC gene fragment was amplified by PCR and PCR products were sequenced. PCR/DGGE and RAPD were performed. Results: It was found that 39 (5.5%) samples were positive for M. bovis in the PCR and six field strains had point nucleotide mutations. Additionally, the phylogenetic analysis of 20 M. bovis field strains tested with RAPD showed two distinct groups of M. bovis strains sharing only 3.8% similarity. PCR/DGGE analysis demonstrated the presence of bacteria belonging to the Mollicutes class in 79.1% of DNA isolates. The isolates were identified as: Mycoplasma bovirhinis, M. dispar, M. bovis, M. canis, M. arginini, M. canadense, M. bovoculi, M. alkalescens, and Ureaplasma diversum. Conclusion: Different Mycoplasma spp. strains play a crucial role in inducing respiratory diseases in cattle

Effect of supplementation with the combination of se-enriched Lentinula edodes mycelium, exogenous enzymes, acidifiers, sodium butyrate and silicon dioxide nanoparticle feed additives on selected parameters in calves

During the initial months of calves’ lives, the young animals are exposed to bacterial and viral infections, and during this period, crucial physiological changes take place in their organisms. Offering calves feed additives that will have a beneficial influence on their organisms and improve their growth while reducing the morbidity rate is the optimal task of feeding. This is the first study to investigate the effect of experimental supplementation for calves with the combination of two feed additives—one containing Lentinula edodes enriched with selenium (Se), and the second containing pancreatic-like enzymes, fat-coated organic acids, sodium butyrate, and silicon dioxide nanoparticles—on the serum Se concentration, selected immune parameters, and the average daily gains in the calves. During the study, the serum Se concentration was examined by means of inductively coupled plasma mass spectrometry, and the immunoglobulin and cytokine concentrations with ELISA assays. The white blood cell (WBC) count with leukocyte differentiation was examined with the use of a hematological analyzer, and the percentages of subpopulations of T lymphocytes and monocytes, phagocytic activity, and oxidative burst of monocytes and granulocytes with the use of a flow cytometer. The average daily gains of the calves were also evaluated. In summary, the supplementation of the experimental calves with the combination of two feed additives resulted in significantly higher serum Se concentrations, and the immune systems of the calves were not suppressed while the examined feed additives were being delivered. Although not statistically significant, some positive effects on the calves were seen: a tendency towards the improvement of some of the immune parameters evaluated, and a tendency for higher average daily gains in the calves