Pig Leukocytes Research Articles

Evaluating the impact of influenza infection on pig lungs by single-cell RNA sequencing (scRNA-seq)

Abstract Despite being the most widely consumed meat worldwide, the pig industry faces economic challenges from respiratory viruses including influenza. Pigs are also used as a reliable translational model for human respiratory disorders due to their similar respiratory tracts. However, limited pig-specific reagents have hindered understanding the pig’s immune system. Hence, here we used single cell RNA sequencing (scRNAseq) for unbiased transcriptional profiling of pig immune responses during influenza infection and oseltamivir antiviral therapy. Fourteen six-week-old mixed-breed pigs were intratracheally infected with pandemic 2009 H1N1 influenza virus. Seven of these pigs were treated for 5 days with oseltamivir. A control group was mock-infected. Lung samples were taken 5 days post-infection for scRNAseq, histopathology, and virus titers. We identified 24 pig leukocytes populations which were integrated with previously published human and mouse lung cells. Influenza virus infection/oseltamivir treatment altered the cellular composition, transcriptional networks, and cell-cell interactions in the lung. γδ T cells that are unique to pigs and abundant in the lung responded strongly to infection. Oseltamivir reduced antiviral and inflammatory responses. Our data provide a resource to better understand the pulmonary immune responses of pigs and advances using swine as a model for studying human pulmonary immune responses.

Profile of Leukocytes in Pig (Sus scrofa domestica) Before and After Heart Infarct Surgery

Pigs are the most appropriate species in cardiovascular research, one of which is myocardial infarction. This study aimed to obtain data on the number of leukocytes in domestic pigs (Sus scrofa domestica) before and after heart infarct surgery. A total of six four-month-old pigs at the Teaching Animal Hospital, IPB University, had their blood samples taken through the jugular vein, then put into a vacuum tube containing an anticoagulant (K3EDTA) for further testing using a hematology analyzer. Data were analyzed descriptively. The results of the study from 6 samples analyzed that the average number of leukocytes before and after heart infarct surgery were 16.33 ± 3.62 x 103/μL and 16.97 ± 4.59 x 103/μL. The increase in leukocytes in the blood cannot be used as an independent predictor to diagnose cardiac infarction. This is because many events can trigger an increase in the number of leukocytes in the blood, such as inflammation during heart infarct surgery.

APPLICATION OF VERMIGHUMUS IN FEEDING PIGS

The article analyzes a new method of fattening young pigs by introducing a biologically active additive into the diet - vermighumus, which is a product of pig manure processing by California worms. In addition to the standard diet, the youngsters of the research groups consumed vermighumus in a dose of 130, 180, 230 g per head per day. The young of the control group consumed standard compound feed. It was established that the application of vermighumus into the diet of young pigs improved their fattening indicators. In particular, the youngsters of the second and third experimental groups exceeded the control analogues in terms of live weight at the end of fattening by 3.73 and 5.53%, respectively. Youngsters of the second and third experimental groups also had higher growth energy by 4.80 and 7.40%, respectively, and better feed payment (by 0.43 and 071 kg). In terms of half-carcass length, young animals of the experimental group 3 did not significantly exceed the counterparts of the control group. In the 2nd and 3rd experimental groups, the fat thickness at the level of the 6th-7th thoracic vertebra was smaller (by 5.85-7.50%). They prevailed over the control analogues in the area of the "muscle eye" (by 2.83-5.45%). The weight of the rear third half of the carcass and the content of meat in the carcass was the highest in young animals of the 3 experimental group (by 7.85% and 4.91%, respectively), and the fat content in it was lower than in the control group (by 14.52%). The smallest fat thickness at the level of the 6-7 thoracic vertebra, as well as the content of fat in the carcass, was in young animals of the third experimental group. Among the indicators of the physical and chemical properties of the muscle tissue of young pigs of the experimental groups, no statistically significant differences were found. However, there is a certain trend towards a decrease in such indicators as the level of acidity, tenderness and losses during cooking, and an increase in the moisture-holding capacity and intensity of coloring in the young of the control group. The tasting evaluation revealed minor differences among the experimental animals. The relative number of lymphocytes (% of the total number of leukocytes) in pigs treated with vermighumus was higher (by 4.44-6.66% Р<0.05) than in counterparts fed with conventional compound feed, which indicates an improvement in resistance animalsEnrichment of the ration of young pigs with vermihumus as a biologically active additive contributed to the improvement of fattening and meat qualities and ensures obtaining additional basic products in the amount of UAH 320.38-354.34 per head.

Study of Sporo-Lex probiotic effectiveness on pigs

Food sorbents are veterinary drugs and feed additives designed to bind toxic substances in the gastrointestinal tract and remove them from the body of animals. The goal of the work is to conduct a study of the effectiveness of the veterinary drug Sporo-Lex for use on pigs. Materials and methods. Probiotic Sporo-Lex is a mixture of probiotic cultures of Bacillus licheniformis VK-25 and Bacillus subtilis MK-3 on a natural standardized sorbent (montmorillonite rock of the Volodymyretsk city deposit). The research was carried out in the conditions of a private pig farm on 74-day-old landrace pigs, while 76 animals were involved in the experiment (36 heads - experimental and 36 heads - control), groups were formed according to the principle of analogs. The number of erythrocytes, hemoglobin content, bactericidal activity of blood serum (BABS), level of phagocytic activity of leukocytes, phagocytic index, as well as the concentration of Escherichia coli bacteria in feces were determined. Results of research and discussion. As a result of the study of the dynamics of changes in the number of erythrocytes in the blood of experimental and control subjects, it was established that on the 13th day, the number of erythrocytes in the blood of the animals treated with Sporo-Lex significantly increased on the 25th day of the study. The results of the quantitative determination of the level of hemoglobin in the blood of the experimental and control groups of pigs indicate a significant increase in the level of hemoglobin in the experimental group, on the 13th day, in accordance with the control. At the same time, the probability level was 99.6% for experimental group 1 and 99.9% for group 2. A significant increase in the BABS of pigs was recorded from the 13th day of the experiment. At the same time, the level of probability was 98%. According to the obtained results of the study of the influence of the probiotic Sporo-lex on the level of phagocytic activity of blood leukocytes of pigs, it was established that its significant increase was observed on the 4th day of the experiment, in the experimental group it was (42±1.75)%, while in the control group – (34±1.75) %.Probiotic Sporo-Lex when applied to pigs, according to the obtained data, in the experimental group, a significant increase in the phagocytic blood index was observed on the 4th day, and amounted to (2.8±0.1)%, while in the control this indicator was (2.5±0.12) %. Regarding the influence of the probiotic Sporo-Lex on the concentration of bacteria of the Escherichia coli group in feces, it was established that in the animals of the experimental group, on the 25th day, a significant decrease in the concentration of coli group bacteria was observed to (3.2±0.7)×105, while in the animals of the control group group their concentration was (9.8±0.3)×105. According to the research results, it was established that the probiotic Sporo-Lex in the animals of the experimental group increases the average statistical weight by 4.94 kg on the 84th day and by 4.71 kg on the 112th day in comparison with the control. Conclusions. It has been experimentally proven that the drug Sporo-lex reliably increases the level of BABS, an increase in the phagocytic activity of leukocytes is observed and induces an increase in the phagocytic index of leukocytes compared to the control. It is advisable to use the drug as a biological feed additive with a directed adaptogenic and immunocorrective effect on the fattening of young farm animals. Key words: Sporo-lex, probiotic, sorbent, efficiency, animals, piglets, bacteria of the group of Escherichia coli.

Skin in the Game: An Assay to Monitor Leukocyte Infiltration in Dermal Lesions of a Guinea Pig Model for Tick-Borne Rickettsiosis.

Intact, the skin typically serves as an effective barrier to the external world; however, once pathogens have breached this barrier via a wound, such as a tick bite, the surrounding tissues must recruit immune cells from the blood to neutralize the pathogen. With innate and adaptive immune systems being similar between the guinea pig and human systems, the ability of guinea pigs to show clinical signs of many infectious diseases, and the large size of guinea pigs relative to a murine model, the guinea pig is a valuable model for studying tick-borne and other pathogens that invade the skin. Here, we report a novel assay for assessing guinea pig leukocyte infiltration in the skin. Briefly, we developed an optimized six-color/eight-parameter polychromatic flow cytometric panel that combines enzymatic and mechanical dissociation of skin tissue with fluorescent antibody staining to allow for the immunophenotyping of guinea pig leukocytes that have migrated into the skin, resulting in inflammation. We designed this assay using a guinea pig model for tick-borne rickettsiosis to further investigate host–pathogen interactions in the skin, with preliminary data demonstrating immunophenotyping at skin lesions from infected ticks. We anticipate that future applications will include hypothesis testing to define the primary immune cell infiltrates responding to exposure to virulent, avirulent tick-borne rickettsiae, and tick-borne rickettsiae of unknown virulence. Other relevant applications include skin lesions resulting from other vector-borne pathogens, Staphylococcus aureus infection, and Buruli ulcer caused by Mycobacterium ulcerans.

Characterization of circulating porcine immune cells using single-cell RNA-sequencing and comparison to human datasets

Abstract Pigs are part of global food security and serve as a valuable human biomedical model. Immunoreagent unavailability limits resolution of pig leukocyte populations by protein markers; thus, single-cell RNA-sequencing (scRNA) was used to characterize pig peripheral blood mononuclear cell (PBMC) populations and infer function based on transcriptional profiles and comparison to human datasets. Across seven PBMC samples partitioned and sequenced, 28,810 cells distributed across 36 clusters were classified into 13 general cell types including plasmacytoid dendritic cells (DC), conventional DCs, monocytes, more than ten B cell clusters including plasmablasts, conventional CD4 and CD8 αβ T cells, NK cells, and four clusters of γδ T cells. Signature gene sets for a publicly available human bulk RNA-seq dataset were assessed for relative enrichment in genes expressed in pig cells and substantial overlap in gene expression between specific pig and human PBMC populations was detected. Integration of pig scRNA with a public human scRNA dataset provided further validation for similarity between human and pig. Of four pig γδ T cell clusters, two had high prediction and mapping scores with human γδ T cells. The other two pig γδ T cell clusters had gene expression profiles predictive of human γδ T cells, but also human CD4 T central memory (Tcm) and innate lymphoid cells, suggesting innate and adaptive function of some pig γδ T cells. Pigs have a population of circulating double-positive (DP) CD4+CD8αα+ αβ T cells, and putative DP clusters had high prediction scores to human CD4 Tcm cells, supporting their classification in literature as memory cells. Overall, the data provided the first single-cell atlas for porcine PBMCs and comparison to human datasets.

Bacteria-induced expression of the pig-derived protegrin-1 transgene specifically in the respiratory tract of mice enhances resistance to airway bacterial infection

About 70% of all antibiotics produced in the world are used in the farm animal industry. The massive usage of antibiotics during farm animal production has caused rapid development of antibiotic resistance in bacteria, which poses a serious risk to human and livestock health when treating bacterial infections. Protegrin-1 (PG-1) is a potent antimicrobial peptide (AMP). It was initially identified in pig leukocytes with a broad-spectrum antibacterial and antiviral activity, and a low rate of inducing bacterial resistance. To develop a genetic approach for reducing the use of antibiotics in farm animal production, we produced transgenic mice carrying a bovine tracheal AMP gene promoter-controlled PG-1 transgene. The PG-1 transgene was specifically expressed in the respiratory tract of transgenic mice upon induction by bacterial infection. These PG-1 transgenic mice exhibited enhanced resistance to nasal bacterial infection as the transgenic mice showed a higher survival rate (79.17% VS. 34.78%), lower bacterial load and milder histological severity than their wild-type control littermates. The improved resistance to bacterial infection in the PG-1 transgenic mice could be resulted from the direct bacteria-killing activities of PG-1, and the immunomodulatory effects of PG-1 via stimulating interleukin 1 beta secretion. The present study provides a promising genetic strategy to prevent airway bacterial infections in farm animals by bacteria-inducible tissue-specific expression of PG-1 transgene. This approach may also be helpful for decreasing the possibility of inducing bacterial resistance during farm animal production.

Research of toxicological safety of production and operation of products from sulfur gypsum composite

By spent researches is established, that the products from a composite material on a basis gypsum and sulfur do not include in the structure potentially dangerous for organism of the person and environment of connections of mercury, thallium, selenium, arsenic; are chemically stable and do not allocate in inducing air, water and acid of environment unstable of inorganic connections. By manufacture of products on air environment of a working zone can be allocated hydrogen sulphide and sulphureous anhydride, that requires the appropriate organization of labour of the workers with maintenance of a premise by compulsory ventilation and measures of fire-prevention safety.
 Toxicological and hygienic studies of powdered sulfur gypsum material in case of cutaneous admission (50% vaseline emulsion) allowed to establish the following: after the first application of the paste, no visible changes were observed on the skin of animals. After ten applications in 2/3 of the experimental animals, pale pink erythema was observed throughout the paste application area. With further applications, the intensity of skin hyperemia did not increase. The resulting changes can be classified as a moderately pronounced skin irritating effect of powdered sulfur gypsum material.
 Analysis of the information obtained during the formulation of basophil degranulation reactions and specific agglomeration of blood leukocytes of guinea pigs sensitized with powdered sulfur gypsum material indicates the absence of statistically significant changes in the parameters of the studied reactions in animals of the experimental and control groups. This gives grounds for concluding that sulfur gypsum materials do not possess allergenic activity.
 Based on the conclusion of the Chief Sanitary Doctor of Ukraine, a composite material based on gypsum and sulfur is recommended for the manufacture of tiles that can be used in the construction of buildings of “B” and “V” groups (industrial and public buildings).

Photoperiodic Effects on Diurnal Rhythms in Cell Numbers of Peripheral Leukocytes in Domestic Pigs.

The photoperiod is known to modulate immune cell number and function and is regarded essential for seasonal disease susceptibility. In addition, diurnal variations in the immune system are regarded important for immune competence. Whereas few studies investigated the influence of season, none investigated the specific effect of the photoperiod on these diurnal immune rhythms until now. Therefore, the present study compared diurnal rhythms in cell numbers of peripheral leukocyte types in domestic pigs held either under long day conditions (LD) or short day conditions (SD). Cosinor analyses of cell numbers of various peripheral leukocyte subtypes investigated over periods of 50 h revealed distinct photoperiodic differences in diurnal immune rhythms. Relative amplitudes of cell numbers of total leukocytes, NK cells, T cells, and monocytes in blood were higher under SD than LD. In addition, cell counts of total leukocytes, NK cells, T cells including various T cell subtypes, and eosinophils peaked earlier relative to the time of lights-on under SD than LD. In contrast, diurnal rhythms of neutrophil counts did not show photoperiodic differences. Mesor values did not differ in any leukocyte type. Generalized linear mixed model analyses revealed associations of leukocyte counts with plasma cortisol concentration and activity behavior in most investigated cell types. Moreover, the present study demonstrated photoperiodic effects on diurnal rhythms in plasma cortisol concentrations and activity behavior, which is in agreement with human and primate studies. The results of the present study imply stronger rhythmicity in leukocyte counts in general under SD. Common intrinsic mechanisms seem to regulate photoperiodic effects on diurnal rhythms in leukocyte counts, except for neutrophils, in domestic pigs. Our results reveal considerable insights into the regulation of immune rhythms in diurnally active species.

Allergic sensitization increases the amount of extracellular ATP hydrolyzed by guinea pig leukocytes.

Increased levels of ATP have been found in the bronchoalveolar lavage of patients with asthma, and subjects with this disease, but not healthy subjects, develop bronchospasm after nebulization with ATP. Because the main mechanism for controlling the noxious effects of extracellular ATP is its enzymatic hydrolysis, we hypothesized that allergic sensitization is accompanied by a decreased functioning of such hydrolysis. In the present study, peripheral blood leukocytes from sensitized and non-sensitized guinea pigs were used for determining the extracellular metabolism (as assessed by inorganic phosphate production) of ATP, ADP, AMP, or adenosine, and for detecting possible changes in the expression (qPCR and Western blot) of major ectonucleotidases (NTPDase1, NTPDase3, and NPP1) and purinoceptors (P2X1, P2X7, P2Y4, and P2Y6). Contrary to our hypothesis, we found that leukocytes from allergic animals produced higher amounts of inorganic phosphate after stimulation with ATP and ADP, as compared with leukocytes from non-sensitized animals. Although at first glance, this result suggested that sensitization caused higher efficiency of ectonucleotidases, their mRNA and protein expressions were unaffected. On the other hand, after sensitization, we found a significant increase in the protein expression of P2X7 and P2Y4, two purinoceptors known to be responsible for ATP release after activation. We concluded that allergic sensitization increased the amount of ATP hydrolyzed by ectonucleotidases, the latter probably not due to the enhanced efficiency of its enzymatic breakdown, but rather due to an increased release of endogenous ATP or other nucleotides, partly mediated by enhanced expression or P2X7 and P2Y4 receptors.

Influence of polyphenols from olive mill wastewater on the gastrointestinal tract, alveolar macrophages and blood leukocytes of pigs

In the last years, great importance has been given to the beneficial effects of polyphenols. Among the most relevant health promoting effects, there is the capacity to reduce the amount of free radicals and stimulate the immune response. In this study, polyphenols extracted from olive mill wastewater (OMWW), were fed to adult ‘Casertana’ pigs during the finishing period. No significant differences in the length of the jejunum-ileum villi and the depth of the colon crypts were detected between control and polyphenols fed pigs. Instead, intra-epithelial and lamina propria leukocytes were more abundant in pigs fed polyphenols (p < .05). Cyclooxygenase-2 immunoreactivity in the gastrointestinal tract, employed as marker of inflammation, was more intense in the control group. Superoxide anion production in primary cell cultures of both blood leukocytes and alveolar macrophages was lower in pigs fed polyphenols (p < .05). Taken together these data indicate that, according to our in vitro studies, OMWW polyphenols seem to be potent antioxidants, while the interpretation of the in vivo experiments is more problematic and further studies are necessary on the interactions between bioactive feed compounds and intestinal status. Such studies can contribute to a better understanding of both positive and negative interactions in vivo and to the identification of new functional feeds.HIGHLIGHTSThe effects of polyphenols extracted from olive mill wastewater (OMWW) have been studied in pigs.Gut morphology, inflammation and immune response were investigated.OMWW polyphenols act as potent antioxidants.

Influence of the hormonal background on the morphological replacement piglets

The puberty period in pigs depends on the functional activity of the endocrine system, which has an effect on the blood through the connective tissue. It has been established that steroid and thyroid hormones regulate the processes of metabolism and proliferation, differentiation and apoptosis of cells. The aim of the research was to establish the peculiarities of formation of the morphological composition of blood under the influence of steroid and thyroid hormones in the process of puberty in different directions of productivity in the pigs. Experiments were performed on two groups of clinically healthy pigs of Pietrain breed and Large White breed. To assess the hormonal and morphological blood was taken from pigs from the anterior hollow vein in the 4-, 5-.6-, 7-month-old age (when their live weight reaches 100 kg). The predominance of hemoglobin content and the amount of erythrocytes in the blood of piglets of Pietrain breed over a Large White breed on the 120–150th day of development (P &lt; 0.05) was observed, against the background of the general decrease in leukocytes in pigs of both breeds at the age of 5 months with the next gradual increase. The age-related changes in the leukocyte formula were directly related to the direction of animal productivity. In the Large White breed, from the beginning of the experiment, there was an increase in the number of rodenuclear neutrophils to 210-day-olds, followed by a sharp decreasing on 36.4%. In Pietrain breed, the level of the rodenuclear neutrophils decreased on 28.6% (P &lt; 0.01) at the age of 5 months, followed by a sharp increasing over the coming months. From the 5th to the 7th months of the development of animals, the concentration of estradiol decreased in the first genotype 2 times (P &lt; 0.05); in the second genotype – 1.9 times (P &lt; 0.01). The content of thyroxine and triiodothyronine increased in the blood serum in Pietrain breed pigs (P &lt; 0.01) and Large White breed (P &lt; 0.001) in the period from 120th to 180th day of development. It was found that the amount of progesterone in Large White breed was higher relatively to Pietrain breed in all investigated periods. The content of estradiol in pigs of both breeds from the 120th and the 150th day of development tended to decrease – by 2.8 times (P &lt; 0.001) in Large White breed, 1.4 times (P &lt; 0.05) in Pietrain breed, and testosterone increases, respectively, 2.1 (P &lt; 0.05) and 1.9 (P &lt; 0.05) times. The significant influence of progesterone on the number of lymphocytes and common leukocytes in Pietrain breed pigs was established. The level of estradiol significantly influenced the number of granulocytes in Pietrain breed pigs at the age of 5 months (r = 0.92 ... 0.99), and in animals of Large White breed the maximum correlation level was observed at the achievement of the 150th (r = 0.61) and 180 days of development (r = 0.94).

Short- and Long-Term Effects of Weaning Age on Pig Innate Immune Status

The study was conducted to evaluate short- and long-term effects of pig wean age on innate immunity and cortisol. Seventy-two white crossbred pigs from 12 litters were randomly assigned to a weaning age of 14 or 28 d-of-age. Pigs were weaned at assigned treatment age and kept as littermates until 20 wk-of-age. Blood samples were taken prior to weaning (d 0) and days 1, 7, and 14 post-weaning (short-term), and at 8, 12, 16, and 20 wk-of-age (long-term). Prior to weaning (d 0), total WBC and lymphocyte numbers were greater for 28-d weaned pigs than 14-d and 28-d pigs had greater lymphocyte numbers at d 1, 7, and 14 post-weaning. At d 0, cytotoxicity and phagocytosis were greater for 14-d than 28-d weaned pigs. Regardless of age, at d 1 and 7 post-weaning all pigs had greater WBC counts, neutrophils, and phagocytosis, but reduced lymphocytes and NK cytotoxicity compared with d 0. Cortisol was decreased at d 7 and increased at d 14 post-weaning in 28-d weaned pigs. These pigs also had greater cortisol at d 0, 1 and 14 post-weaning than 14-d weaned pigs. Effects of weaning on leukocyte profile and N:L ratio were longer-lasting in 14-d weaned pigs than 28-d with effects still apparent at d 14 post-weaning for lymphocytes, neutrophils, N:L ratio, NK, phagocytosis, and IgG. These data imply that weaning age differentially affected pig leukocyte populations and innate immunity in response to weaning stress in both short- and long-term. More specifically, pigs weaned at 14 d-of-age had a more profound and longer-lasting stress response to weaning and 14-d weaned pigs had a more profound innate response, especially NK cytotoxicity while 28-d weaned pigs had more profound antibody response (IgG) in the long-term and these responses were still evident at 20 wk-of-age.

Release of pig leukocytes and reduced human NK cell recruitment during ex vivo perfusion of HLA-E/human CD46 double-transgenic pig limbs with human blood.

In pig-to-human xenotransplantation, interactions between human natural killer (NK) cells and porcine endothelial cells (pEC) are characterized by recruitment and cytotoxicity. Protection from xenogeneic NK cytotoxicity can be achieved in vitro by the expression of the non-classical human leukocyte antigen-E (HLA-E) on pEC. Thus, the aim of this study was to analyze NK cell responses to vascularized xenografts using an ex vivo perfusion system of pig limbs with human blood. Six pig forelimbs per group, respectively, stemming from either wild-type (wt) or HLA-E/hCD46 double-transgenic (tg) animals, were perfused ex vivo with heparinized human blood for 12hours. Blood samples were collected at defined time intervals, cell numbers counted, and peripheral blood mononuclear cells analyzed for phenotype by flow cytometry. Muscle biopsies were analyzed for NK cell infiltration. In vitro NK cytotoxicity assays were performed using pEC derived from wt and tg animals as target cells. Ex vivo, a strong reduction in circulating human CD45 leukocytes was observed after 60minutes of xenoperfusion in both wt and tg limb groups. NK cell numbers dropped significantly. Within the first 10minutes, the decrease in NK cells was more significant in the wt limb perfusions as compared to tg limbs. Immunohistology of biopsies taken after 12hours showed less NK cell tissue infiltration in the tg limbs. In vitro, NK cytotoxicity against hCD46 single tg pEC and wt pEC was similar, while lysis of double tg HLA-E/hCD46 pEC was significantly reduced. Finally, circulating cells of pig origin were observed during the ex vivo xenoperfusions. These cells expressed phenotypes mainly of monocytes, B and T lymphocytes, NK cells, as well as some activated endothelial cells. Ex vivo perfusion of pig forelimbs using whole human blood represents a powerful tool to study humoral and early cell-mediated rejection mechanisms of vascularized pig-to-human xenotransplantation, although there are several limitations of the model. Here, we show that (i) transgenic expression of HLA-E/hCD46 in pig limbs provides partial protection from human NK cell-mediated xeno responses and (ii) the emergence of a pig cell population during xenoperfusions with implications for the immunogenicity of xenografts.

Assessment of the Degree of Allergenic Effects of Various Drugs of Francisella tularensis

The evaluation of sensitization conducted on guinea pig leukocytes immunized with the cell wall preparations of F. tularensis. In this research was shown that the cell wall preparations F tularensis subsp mediaasiatica A-61 and F tularensis subsp tularensis В-399 A-Cole do not caused allergization in contrast with cell wall prepararions of F. tularensis 15 (extracted by Research Institute of Epidemiology and Hygiene) and F. tularensis subsp. holarctica 306.

Fractionation of guinea pig leukocyte by flow cytometry using a novel MIL4/SSC parameter

Fractionation of guinea pig leukocyte by flow cytometry using a novel MIL4/SSC parameter

Effects of stress factors on the performance of non-specific resistance and per-formance of piglets

Feeding «feed mixture STO HA» piglets had a positive impact on their resistance and performance in the early postnatal period and the first week after weaning. Research has established that the content of total protein in pigs 4 day age 2nd and 3rd groups did not significantly different from the control group, while their peers from the 4th group born from sows that received «feed mixture STO HA» specified an indicator was higher by 2.8%. The level of total protein in piglet 2 nd, 3 rd, 4 th groups of 16-day age was higher by 5.8, 6.2, 8.8% (P &lt; 0.05), respectively, in the control group, the 40th day age – significantly (P &lt; 0.05) higher than same indicator 9.9, 8.6, 9.6% respectively. The difference in levels of immunoglobulin A in pigs receiving «feed mixture STO HA» (2nd and 4th groups) in 16 aged and performance of piglets 1st (control) group was 54.5 – 81.8% and was statistically significant (P &lt; 0.05 – P &lt; 0.01). The 28-day age and concentration of Ig A serum pigs 2nd and 4th groups significantly exceeded by 72.2 and 77.7% rate in the control group, the 40th day age at 60.6 and 54,5%, respectively (P &lt; 0.05). The bactericidal activity of blood serum piglets 40 days old all research groups significantly (P &lt; 0.05 – P &lt; 0.01) greater than 15.9, 11.3, 14.8% performance of their peers in the control group. Lysozyme activity of blood serum pigs 2nd, 3rd, 4th groups and 28 days old dominated his peers likely group 1 15.3, 11.7, 15.6% in the 40-day age – to 19.0; 16.7; 19.8% respectively. Phagocytic activity of leukocytes in pigs 4 days old 4th group born from sows that received «feed mixture STO HA» was higher by 5.7, 9.5, 10.4% compared to piglets 1 – 3rd groups. Phagocytic activity of leukocytes pigs 2nd, 3rd, 4th groups 16 days old and has been above 18.3 (P &lt; 0.05); 11.5; 16.8% (P &lt; 0.05), respectively group 1, the 28-day age – 17.4 (P &lt; 0.05); 12.9 (P &lt; 0.05); 15.2%. Phagocytic activity of leukocytes in piglets 40th age of daily study groups was significantly (P &lt; 0.05) higher respectively pigs in the control group. The difference between the performance of piglets second and the first group was 20.5%, the first and third – 16.0%, fourth and first – 18.6%. The intensity of phagocytosis (FI) in pigs 16-day age 2 nd, 3 rd, 4 th group was significantly higher at 16.5 (P &lt;0.05), 12.9, 22.4% (P &lt; 0.01), the 40th day age – to 23.6, 20.1, 21.3%, respectively (P &lt; 0.05, P &lt; 0.01) compared with the control group. The average daily gain of the piglets 4th group of from 31 th to 40 th day of life superior piglets of group 1 to 58.3%, and body weight in 40-day age – 15.5% (P &lt; 0.01).

Antimicrobial Peptides Share a Common Interaction Driven by Membrane Line Tension Reduction

Antimicrobial peptides (AMPs) are a class of host-defense molecules that neutralize a broad range of pathogens. Their membrane-permeabilizing behavior has been commonly attributed to the formation of pores; however, with the continuing discovery of AMPs, many are uncharacterized and their exact mechanism remains unknown. Using atomic force microscopy, we previously characterized the disruption of model membranes by protegrin-1 (PG-1), a cationic AMP from pig leukocytes. When incubated with zwitterionic membranes of dimyristoylphosphocholine, PG-1 first induced edge instability at low concentrations, then porous defects at intermediate concentrations, and finally worm-like micelle structures at high concentrations. These rich structural changes suggested that pore formation constitutes only an intermediate state along the route of PG-1’s membrane disruption process. The formation of these structures could be best understood by using a mesophase framework of a binary mixture of lipids and peptides, where PG-1 acts as a line-active agent in lowering interfacial bilayer tensions. We have proposed that rather than being static pore formers, AMPs share a common ability to lower interfacial tensions that promote membrane transformations. In a study of 13 different AMPs, we found that peptide line-active behavior was not driven by the overall charge, and instead was correlated with their adoption of imperfect secondary structures. These peptide structures commonly positioned charged residues near the membrane interface to promote deformation favorable for their incorporation into the membrane. Uniquely, the data showed that barrel-stave-forming peptides such as alamethicin are not line-active, and that the seemingly disparate models of toroidal pores and carpet activity are actually related. We speculate that this interplay between peptide structure and the distribution of polar residues in relation to the membrane governs AMP line activity in general and represents a novel, to our knowledge, avenue for the rational design of new drugs.

Increase in chemokines CXCL10 and CCL2 in blood from pigs infected with high compared to low virulence African swine fever virus isolates

Modulation of the expression of chemokines and chemokine receptors in whole blood was compared following infection of pigs with high and low virulence isolates of African swine fever virus. Levels of mRNAs for CCL2, CCL3L1, CCL4, CXCL10, CCR1 and CCR5 were significantly increased in at least one time point following infection in two experiments and CCL5, CCR9 and CXCR4 mRNA were significantly increased in one of the experiments. The results showed that greatest fold increases in mRNAs for CXCL10 and CCL2 were observed following infection of pigs. CXCL10 mRNA was increased by up to 15 fold in infected compared to uninfected pigs. CXCL10 protein was also detected in serum from pigs infected with the high virulence Benin 97/1 isolate. Levels of CCL2 mRNA were increased in pigs infected with high virulence Benin 97/1 isolate compared to low virulence OURT88/3 isolate and this correlated with an increase of greater than 30 fold in levels of CCL2 protein detected in serum from pigs infected with this isolate. An increase in overall chemotaxis active compounds in defibrinated plasma samples from Benin 97/1 infected pigs was observed at 3 days post-infection (dpi) and a decrease by 7 dpi as measured by chemotaxis assay using normal pig leucocytes in vitro. Increased levels of CXCL10 may either contribute to the activation of lymphocyte priming toward the Th1 phenotype or induction of T lymphocyte apoptosis. Increased levels of CCL2, a chemoattractant for macrophages, may result in increased recruitment of monocytes from bone marrow thus increasing the pool of cells susceptible to infection.

Immunological characterization of peripheral blood leukocytes using vaccine for mycoplasmal pneumonia of swine (MPS) in swine line selected for resistance toMPS

This study was conducted to evaluate immunological changes in peripheral blood leukocytes in pigs that were genetically selected for their improved resistance to mycoplasmal pneumonia of swine (MPS), using MPS vaccine as an antigen. Twelve castrated MPS-selected Landrace pigs were compared with the same number of pigs from a nonselected line by using a time-course analysis at the hematological level. After the second sensitization with MPS vaccine, the percentages of B cells, CD4(+) T cells, and natural killer (NK) cells in total leukocytes were lower in the selected line than in the nonselected line, whereas the percentage of granulocytes in total leukocytes increased in the MPS-selected line. We also assessed the proliferative ability of peripheral blood mononuclear cells (PBMCs) stimulated with Mycoplasma hyopneumoniae, lipopolysaccharide or concanavalin A, and found that although the proliferative ability of the PBMC was not different between the two lines at a steady state, the nonselected line showed a significantly higher proliferative ability after sensitization with MPS vaccine than the selected line regardless of antigens used. These results thus indicate that the selection of pigs on the basis of MPS resistance changes their immunophenotype, and would give us beneficial information for the prevention of MPS infection.