Enterovirus E infects bovine peripheral blood mononuclear cells. Implications for pathogenesis?

Introduction: Enterovirus E (EV-E) is a common viral pathogen endemic in cattle worldwide. Little is known, however, about its potential interactions with bovine immune cells. Material and Methods: The EV-E-permissiveness of bovine peripheral blood mononuclear cells (PBMCs) was evaluated. The infectious titres of extracellular virus were measured and the intracellular viral RNA levels were determined by reverse transcription quantitative PCR after cell inoculation. The effects of EV-E on cell viability and proliferative response were investigated with a methyl thiazolyl tetrazolium bromide reduction assay, the percentages of main lymphocyte subsets and oxidative burst activity of blood phagocytes were determined with flow cytometry, and pro-inflammatory cytokine secretion was measured with an ELISA. Results: Enterovirus E productively infected bovine PBMCs. The highest infectious dose of EV-E decreased cell viability and T-cell proliferation. All of the tested doses of virus inhibited the proliferation of high responding to lipopolysaccharide B cells and stimulated the secretion of interleukin 1ß, interleukin 6 and tumour necrosis factor α pro-inflammatory cytokines. Conclusion: Interactions of EV-E with bovine immune cells may indicate potential evasion mechanisms of the virus. There is also a risk that an infection with this virus can predispose the organism to secondary infections, especially bacterial ones.

No indication for SARS-CoV-2 transmission to pet ferrets, in five cities in Poland, 2021 - antibody testing among ferrets living with owners infected with SARS-CoV-2 or free of infection.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first identified in China by the end of 2019 and was responsible for a pandemic in the human population that resulted in millions of deaths worldwide. Since the beginning of the pandemic, the role of animals as spill-over or reservoir hosts was discussed. In addition to cats and dogs, ferrets are becoming increasingly popular as companion animals. Under experimental conditions, ferrets are susceptible to SARS-CoV-2 and it appears that they can also be infected through contact with a SARS-CoV-2 positive owner. However, there is still little information available regarding these natural infections. Here, we serologically tested samples collected from pet ferrets (n = 45) from Poland between June and September 2021. Of the ferrets that were included in the study, 29% (13/45) had contact with owners with confirmed SARS-CoV-2 infections. Nevertheless, SARS-CoV-2-specific antibodies could not be detected in any of the animals, independent of the infection status of the owner. The obtained results suggest that ferrets cannot be readily infected with SARS-CoV-2 under natural conditions, even after prolonged contact with infected humans. However, due to the rapid mutation rate of this virus, it is important to include ferrets in future monitoring studies.

The BCO2 Genotype and the Expression of BCO1, BCO2, LRAT, and TTPA Genes in the Adipose Tissue and Brain of Rabbits Fed a Diet with Marigold Flower Extract.

This study was undertaken to evaluate the effect of the BCO2 genotype and dietary supplementation with marigold flower extract on the expression of BCO1, BCO2, LRAT, and TTPA genes in the adipose tissue and brain of rabbits. The concentrations of lutein, zeaxanthin, ß-carotene, retinol, and α-tocopherol were determined in samples collected from rabbits. Sixty young male Termond White rabbits were allocated to three groups based on their genotype at codon 248 of the BCO2 gene (ins/ins, ins/del, and del/del). Each group comprised two subgroups; one subgroup was administered a standard diet, whereas the diet offered to the other subgroup was supplemented with 6 g/kg of marigold flower extract. The study demonstrated that the BCO2 genotype may influence the expression levels of the BCO2, LRAT, and TTPA genes in adipose tissue, and TTPA and BCO1 genes in the brain. Moreover, an increase in the amount of lutein in the diet of BCO2 del/del rabbits may increase the expression of BCO1, LRAT, and TTPA genes in adipose tissue, and the expression of the BCO2 gene in the brain. Another finding of the study is that the content of carotenoids and α-tocopherol increases in both the adipose tissue and brain of BCO2 del/del rabbits.

Staphylococcus aureus from Subclinical Cases of Mastitis in Dairy Cattle in Poland, What Are They Hiding? Antibiotic Resistance and Virulence Profile.

Bovine mastitis is a common disease worldwide, and staphylococci are one of the most important etiological factors of this disease. Staphylococcus aureus show adaptability to new conditions, by which monitoring their virulence and antibiotic resistance mechanisms is extremely important, as it can lead to the development of new therapies and prevention programs. In this study, we analyzed Staphylococcus aureus (n = 28) obtained from dairy cattle with subclinical mastitis in Poland. The sensitivity of the isolated strains to antibiotics were confirmed by the disc diffusion method. Additionally, minimum inhibitory concentration values were determined for vancomycin, cefoxitin and oxacillin. Genotyping was performed by two methods: PCR melting profile and MLVF-PCR (multiple-locus variable-number tandem-repeat fingerprinting). Furthermore, the presence of antibiotic resistance and virulence genes were checked using PCR reactions. The analyzed strains showed the greatest resistance to penicillin (57%), oxytetracycline (25%) and tetracycline (18%). Among the analyzed staphylococci, the presence of 9 of 15 selected virulence-related genes was confirmed, of which the icaD, clfB and sea genes were confirmed in all staphylococci. Biofilm was observed in the great majority of the analyzed bacteria (at least 70%). In the case of genotyping among the analyzed staphylococci (combined analysis of results from two methods), 14 patterns were distinguished, of which type 2 was the dominant one (n = 10). This study provides new data that highlights the importance of the dominance of biofilm over antibiotic resistance among the analyzed strains.

Does Aleutian Disease Occur among Domestic Ferrets in Poland? Results of Preliminary Studies Conducted in Two Regions of Poland.

Although ferrets are becoming increasingly popular as companion animals, their population in households is still far lower compared to cats or dogs. This results in a much smaller number of ferret specialists, and thus poorer diagnosis of various diseases, including the Aleutian disease. Aleutian disease is a slowly progressing viral disease which can cause different symptoms in these animals. The virus can also cause symptoms in different species of animals, but in the case of ferrets, there is relatively less information on about both the prevalence and symptoms of this disease. Therefore, the aim of this study was to determine the presence of antibodies and the virus itself in ferrets from two regions of Poland. Blood samples and rectal swabs were obtained from 61 domestic ferrets from Mazowieckie and Dolnoslaskie voivodships. The presence of antibodies was determined using serological methods and real-time PCR analysis was performed to determine presence of viral DNA. Serological analyses demonstrated that 49% (n = 30) of the ferrets had antibodies against Aleutian disease virus (ADV). No relationship was observed between the prevalence of antibodies and age, sex, habitual residence or origin of ferrets. The real-time PCR did not confirm DNA of the ADV in any of the blood and rectal swab samples. Obtained results suggest that ADV circulates in the analyzed population of ferrets, therefore further studies in this direction should be carried out.

The Effect of the BCO2 Genotype on the Expression of Genes Related to Carotenoid, Retinol, and α-Tocopherol Metabolism in Rabbits Fed a Diet with Aztec Marigold Flower Extract.

This study investigated the effect of the BCO2 genotype and the addition of Aztec marigold flower extract to rabbit diets on the expression of BCO1, BCO2, LRAT, and TTPA genes in the liver. The levels of lutein, zeaxanthin, ß-carotene, retinol, and α-tocopherol in the liver and blood serum of rabbits, as well as plasma biochemical parameters and serum antioxidant enzyme activities were also determined. Sixty male Termond White growing rabbits were divided into three groups based on their genotype at codon 248 of the BCO2 gene (ins/ins, ins/del and del/del); each group was divided into two subgroups: one subgroup received a standard diet, and the other subgroup was fed a diet supplemented with 6 g/kg of marigold flower extract. The obtained results indicate that the BCO2 genotype may affect the expression levels of BCO1 and BCO2 genes in rabbits. Moreover, the addition of marigold extract to the diet of BCO2 del/del rabbits may increase the expression level of the BCO2 gene. Finally, an increase in the amount of lutein in the diet of rabbits with the BCO2 del/del genotype contributes to its increased accumulation in the liver and blood of animals without compromising their health status or liver function.

Antiviral Effect of Bovine Lactoferrin against Enterovirus E.

Enterovirus E (EV-E), a representative of the Picornaviridae family, endemically affects cattle across the world, typically causing subclinical infections. However, under favorable conditions, severe or fatal disorders of the respiratory, digestive, and reproductive systems may develop. There is no specific treatment for enterovirus infections in humans or animals, and only symptomatic treatment is available. The aim of this study was to determine the in vitro antiviral effect of bovine lactoferrin (bLF) against enterovirus E using virucidal, cytopathic effect inhibition, and viral yield reduction assays in MDBK cells. The influence of lactoferrin on the intracellular viral RNA level was also determined. Surprisingly, lactoferrin did not have a protective effect on cells, although it inhibited the replication of the virus during the adsorption and post-adsorption stages (viral titres reduced by 1-1.1 log). Additionally, a decrease in the viral RNA level in cells (by up to 75%) was observed. More detailed studies are needed to determine the mechanism of bovine lactoferrin effect on enterovirus E. However, this highly biocompatible protein ensures some degree of protection against infection by bovine enterovirus, which is particularly important for young animals that receive this protein in their mother's milk.

High Seroprevalence against SARS-CoV-2 among Dogs and Cats, Poland, 2021/2022.

The coronavirus SARS-CoV-2 is responsible for a pandemic in the human population that has unfolded since the beginning of 2020 and has led to millions of deaths globally. Apart from humans, SARS-CoV-2 has been confirmed in various animal species, including felines, canines, mustelids, and primates. Of these species, dogs and cats are the most popular companion animals worldwide. Several seroprevalence studies have already been performed in these animal species; however, the results vary depending on the location and especially the time of sampling. Here, serum samples were collected from a total of 388 dogs and 243 cats from three veterinary clinics in two cities (Gdansk and Olsztyn) in Poland between October 2021 and February 2022, when the country was in the midst of the fourth wave of viral spread. All sera were tested for antibodies against SARS-CoV-2 by a multispecies ELISA based on the receptor-binding domain and by an indirect immunofluorescence assay (iIFA). Overall, 18.9% of the feline sera and 16.0% of the canine sera tested positive using ELISA and iIFA. This relatively high seroprevalence among randomly selected animals is most likely related to the high case numbers in the human population and indicates a continuous occurrence of transspecies virus transmissions from infected owners to their pets. Hence, dogs and cats should be included in monitoring studies and/or outbreak investigations for a better understanding of the epidemiology of this virus.

High cytotoxicity of betulin towards fish and murine fibroblasts: Is betulin safe for nonneoplastic cells?

BACKGROUND: Betulin, a natural pentacyclic triterpene with the lupane structure that is present in significant amounts in the outer bark of birch, is known for its broad array of biological and pharmacological properties. Betulin has attracted attention as a potential, natural-origin antimicrobial substance. The literature describes it as selectively toxic to neoplastic cells but safe for normal cells. The research aim was to evaluate the basal cytotoxicity of betulin towards fish (BF-2) and murine (NIH/3T3) fibroblasts. We used four colorimetric tests that provide a preliminary evaluation of possible mechanisms of the cytotoxicity of a compound to assess the degree of the toxicity of betulin after 24, 48 and 72 h of incubation with cells: the MTT assay (mitochondrial activity assessment), the NRU assay (lysosomal membrane integrity assessment), the LDH assay (cellular membrane integrity assessment) and the SRB assay (total cellular protein content determination). RESULTS: The results revealed an exceptionally high sensitivity of mitochondria to the effect of betulin, with the other endpoints being less sensitive. Although murine fibroblasts were more vulnerable to the toxic effect of betulin than fish fibroblasts, the betulin CC50 values for both cell lines were comparable with analogous IC50 values determined by other researchers in studies involving cancerous cells. CONCLUSIONS: The results indicate the need to verify the claim about the selective toxicity of betulin towards malignant cells and to conduct safety/toxicity tests before any potential therapeutic use of betulin in veterinary medicine.

The influence of maternally derived antibodies on protection against aMPV/A infection in TRT vaccinated turkeys.

Avian metapneumoviruses (aMPV) are a causative agent of turkey rhinotracheitis (TRT). Despite vaccination, cases of TRT outbreaks are frequently reported. Considering that there are aMPV-free areas, a part of turkey poults possess (MDA+) or do not possess (MDA-) maternally derived antibodies (MDA) in the first weeks of life, which is the time of TRT vaccination. Study was undertaken to establish the level of protection against homologue aMPV/A infection, in MDA+ and MDA- turkeys, vaccinated against TRT at the 0 or 14th d of life and infected 14 d later. With the use of ELISA test and qPCR techniques, we have established the level of immune system stimulation after the vaccination and how does it correlate with the level of protection against the aMPV infection. Vaccination of MDA+ turkeys (especially at 0 d of life) resulted in weaker IgA production in upper respiratory tract. In addition, we have demonstrated differences in both humoral and cell-mediated immunity stimulation after infection of vaccinated turkeys. Despite these differences, we have shown that all vaccinated birds were protected against the disease which was determined based on the clinical and histopathological scoring, as well as the level of aMPV/A replication and shedding. Nonvaccinated groups of turkeys displayed typical signs of TRT after infection which indicates that MDA alone are incapable of preventing the disease. Differences in TRT course were recorded between different age groups of nonvaccinated birds. Birds infected at the 28th d of life (especially MDA- birds) developed more severe signs, and the level of aMPV replication was higher than that in birds infected on the 14th d of life. Despite the minor role in alleviating TRT course, MDA seems not to interfere with the vaccination efficacy. It is hard to predict whether the observed immune system stimulation differences between MDA+ and MDA- birds after vaccination can influence the outcome of vaccination efficacy under the field conditions.

Nisin-A lantibiotic with immunomodulatory properties: A review.

Nisin, a member of class I bacteriocins known as lantibiotics, is produced by the lactic acid bacterium Lactococcus lactis and is characterized by a wide spectrum of antibacterial activity against gram-positive bacteria. This characteristic in conjunction with its low toxicity and safety of use in food has contributed to the worldwide success of nisin as a natural food preservative. This lantibiotic has attracted interest as a potential natural therapeutic agent for the control of bacterial infections. However, similar to other antimicrobial peptides of natural origin, the spectrum of biological activity of nisin surpasses its antibacterial properties, encompassing interesting and incompletely understood immunotropic characteristics. This paper is a systematic review of the current information about the potential immunomodulatory properties of nisin based on in vitro and in vivo studies in various experimental models. We also discuss the effect of potentially probiotic, nisin-producing L. lactis strains on the immune system of animals.

Can domestic pigeon be a potential carrier of zoonotic Salmonella?

Salmonellosis is one of the most important bacterial diseases in pigeons. This study aimed to estimate the prevalence of Salmonella spp. in domestic pigeons (Columba livia f. domestica) in Poland, its antimicrobial susceptibility (both phenotypic and genotypic), and its capability for biofilm formation. The presence of selected virulence genes, nucleotide homology of selected genes, and susceptibility to bacteriophages were investigated as well. From the 585 pigeons tested, 5.47% turned out positive. All isolated strains were recognized as Salmonella enterica ser. Typhimurium. The asymptomatic pigeons were carriers of 37.5% of the isolates. The dominant variants were as follows: 1,4,[5],12,:i:1,2 (53.13%) and 1,4,[5],12,:-:- (31.25%). Most of the strains analysed showed the ability to produce biofilm after 24 and 48 hr of incubation (59.38% and 53.13%, respectively). Over 90% of the strains were confirmed for lpfA, agafA, invA, sivH, and avrA virulence genes. Also, of the thirteen antimicrobial susceptibility genes, the following were confirmed: sul1, tet(A), blaTEM-1 , floR, strA, and strB. The most common were the strB (18%) and tet(A) (12%) genes that are responsible for coding resistance to aminoglycosides and tetracyclines, respectively. Most of the strains were phenotypically resistant to oxytetracycline (46.88%), neomycin (53.13%) and tylosin (100%). The susceptibility of the investigated Salmonella strains to the bacteriophages was between 33% and 100%. MLST, PCR MP and ERIC PCR analyses indicated a very high genetic similarity of the investigated strains (over 99%). Results of our study indicate that Salmonella enterica ser. Typhimurium is still an important agent in domestic pigeons and that its antimicrobial resistance increases. Alarming is also the confirmation of a single-phase variant 1,4,[5],12:i,-, which could have increased virulence and multi-drug resistance encoded on the plasmid. Most importantly, however, such strains have been isolated from humans with clinical symptoms of Salmonella infection.

The influence of dietary supplementation with the leucine metabolite ß-hydroxy-ß-methylbutyrate (HMB) on the chemotaxis, phagocytosis and respiratory burst of peripheral blood granulocytes and monocytes in calves.

BACKGROUND: A healthy immune system plays a particularly important role in newborns, including in calves that are far more susceptible to infections (viral, bacterial and other) than adult individuals. Therefore, the present study aimed to evaluate the influence of HMB on the chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit) and oxidative burst (BURSTTEST® kit) of monocytes and granulocytes in the peripheral blood of calves by flow cytometry. RESULTS: An analysis of granulocyte and monocyte chemotactic activity and phagocytic activity revealed significantly higher levels of phagocytic activity in calves administered HMB than in the control group, expressed in terms of the percentage of phagocytising cells and mean fluorescence intensity (MFI). HMB also had a positive effect on the oxidative metabolism of monocytes and granulocytes stimulated with PMA (4-phorbol-12-ß-myristate-13-acetate) and Escherichia coli bacteria, expressed as MFI values and the percentage of oxidative metabolism. CONCLUSION: HMB stimulates non-specific cell-mediated immunity, which is a very important consideration in newborn calves that are exposed to adverse environmental factors in the first weeks of their life. The supplementation of animal diets with HMB for both preventive and therapeutic purposes can also reduce the use of antibiotics in animal production.

The Effects of ß-Hydroxy-ß-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats.

The objective of this study was to determine the effect of ß-hydroxy-ß-methylbutyrate (HMB) on the chemotactic activity, phagocytic activity, and oxidative metabolism of peripheral blood granulocytes and monocytes in goats. Goat kids aged 30 ± 3 days were divided into two groups of 12 animals each: I-control, and II-experimental. Experimental group animals were fed a diet supplemented with HMB in the amount of 50 mg/Kg BW; whereas the diets of control goats were not supplemented. At the beginning of the experiment (day 0) and on experimental days 15, 30, and 60, blood was sampled from the jugular vein to determine and compare chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit), and oxidative metabolism (BURSTTEST® kit) of peripheral blood granulocytes and monocytes by flow cytometry. The analyses of the chemotactic and phagocytic activity of granulocytes and monocytes revealed statistically higher levels of phagocytic activity in the experimental group than in the control group, as expressed by the percentage of phagocytic cells and mean fluorescence intensity. HMB also enhanced the oxidative metabolism of both granulocytes and monocytes, expressed by the rate of oxidative metabolism and mean fluorescence intensity after stimulation with Escherichia coli bacteria and PMA (4-phorbol-12-ß-myristate-13-acetate).

Diurnal and circadian variations in intraocular pressure in goats exposed to different lighting conditions.

The effect of constant light and constant darkness on intraocular pressure (IOP) in goats has not been investigated. We hypothesized that IOP variations would differ between goats kept under a cycle of 12 hours of light and 12 hours of darkness (LD), constant darkness (DD), and constant light (LL). To test this hypothesis, goats were exposed to these conditions for five days (LD, 30 goats; DD, 10 goats; LL, 10 goats). IOP was measured by applanation tonometry at 9 a.m. (beginning of photophase in LD) and 9 p.m. (beginning of scotophase in LD) on the fourth and fifth days of exposure. We found that changes in mean IOP from 9 a.m. to 9 p.m. differed significantly between groups (χ2(2) = 23.04, p < .0001). Most goats in LD showed a regular pattern of higher IOP in the morning and lower IOP in the evening, whereas those in DD and LL did not follow this pattern. In LD conditions, mean IOP was 2.4 mm Hg lower at 9 p.m. than at 9 a.m. (95% confidence interval for the difference (CI): -2.8 to -1.9 mm Hg, p < .0001). In DD conditions, mean IOP did not differ between 9 p.m. and 9 a.m. (CI: -0.9 to 0.8 mm Hg, p = .90). In LL conditions, it was 0.6 mm Hg lower at 9 p.m. (CI: -1.5 to 0.2 mm Hg, p = .12). Our results indicate that IOP in goats kept in LD is higher in the morning than in the evening, and that IOP variations are reduced in goats kept in DD and LL. These results suggest that exposure to alternating periods of light and darkness is important for maintaining rhythmic variations in IOP in this species.

Antiviral effects of nisin, lysozyme, lactoferrin and their mixtures against bovine viral diarrhoea virus.

BACKGROUND: Bovine viral diarrhoea virus (BVDV), an enveloped, single-stranded, positive-sense RNA virus from the Flaviviridae family, is a globally distributed bovine pathogen. BVDV infection in cattle, despite having a wide range of clinical manifestations, is invariably responsible for significant economic losses. To counteract these losses, various schemes to control and eradicate BVDV have been implemented, although safe drugs effectively inhibiting the replication of the virus are still lacking. The purpose of this study was to characterize the antiviral effect of naturally occurring proteins and peptide, such as bovine lactoferrin, chicken egg lysozyme, and nisin from Lactococcus lactis, used both individually and in combination, against the cytopathic NADL strain of BVDV in vitro. After determining the cytotoxicity level of each protein or peptide to MDBK cells, its antiviral effects were evaluated using virucidal, cytopathic effect inhibition and viral yield reduction assays. In addition, the influence of the tested compounds on the intracellular viral RNA level was determined. RESULTS: The highest efficacy among the single treatments was achieved by bovine lactoferrin, which was effective both at the early stages of viral infection and during its entire course, although the effect weakened over time. Nisin and lysozyme were effective at later stages of infection, and the intensity of their effect did not diminish with time. Nisin+lactoferrin and lysozyme+lactoferrin combinations demonstrated stronger antiviral effects than did the single substances. The nisin+lactoferrin mixture present during the whole period of infection produced the strongest anti-BVDV effect in our entire research on both the extracellular viral titre (titre reduction up to 2.875 log ≈ 99.9%) and the intracellular viral RNA level (reduction up to 89%), and this effect intensified over the incubation time. CONCLUSIONS: The tested substances could be applied in bovine viral diarrhoea prevention and therapy, especially when used in combination.

In vitro immunomodulatory effect of nisin on porcine leucocytes.

Nisin, a lantibiotic bacteriocin, has been used for years as a natural food preservative. In addition to its antimicrobial activity, nisin also shows immunomodulatory properties, and the nisin-producing Lactococcus lactis strain has been successfully tested as a probiotic in weaned piglets. However, the impact of nisin on porcine immune cells has not yet been explored. The objective of the present study was to examine the in vitro immunomodulatory effect of nisin on porcine peripheral blood leucocytes. The whole heparinized blood samples or freshly isolated peripheral blood mononuclear cells (PBMCs) were incubated with different nisin concentrations (0, 1.56, 3.125, 6.25, 12.5, 25 or 50 µg/ml) for 1, 24, 48 or 72 hr. Escherichia coli bacteria were used to stimulate blood phagocytes, while concanavalin A and lipopolysaccharide from E. coli were used as mitogens. Control cells remained unstimulated. MTT colorimetric assay was used to evaluate PBMCs viability and mitogenic response. Phagocyte activity and T-cell proliferation were measured by flow cytometry. Flow cytometer was also used for immunophenotyping of T cells. Cytokine levels in the culture media were determined using commercial immunoassay (ELISA) kits. The highest concentration of nisin exhibited proliferative activity (p Ë‚ 0.05), stimulated interleukin-1 beta (IL-1ß) and interleukin-6 (IL-6) production (both at p Ë‚ 0.001), and increased the percentage of CD4+ CD8+ T cells (p Ë‚ 0.001) among unstimulated leucocytes. After cell stimulation, however, the highest nisin concentration showed antiproliferative activity (p Ë‚ 0.05), decreased phagocytic functions (p Ë‚ 0.05) and inhibited the synthesis of IL-6 (time- and concentration-dependent effect). As a typical bacterial product, nisin had a stronger impact on innate immune cells, and its effect on T cells was likely a consequence of the modulation of the activity of antigen-presenting cells. Nisin may be a good candidate as an immunomodulator in pig breeding.

Influence of trans-resveratrol on macrophage and lymphocyte activity in rainbow trout (Oncorhynchus mykiss) - in vitro study.

The objective of this study was to investigate the effect of trans-resveratrol, a potent antioxidant with anti-inflammatory and chemopreventive properties, naturally occurring in many fruits and plants on lymphocytes proliferation and also on macrophages metabolic and phagocytic activity. The aim of this study was to demonstrate the immunomodulatory effects of the compound on fish immunocompetent cells and determine the type of this interaction (immunosuppression or immunostimulation). Proliferative activity of lymphocytes was studied by MTT assay, and the respiratory burst was evaluated using the respiratory burst activity (RBA) test. Phagocytic killing was tested using the PKA test. The experiment have shown that trans-resveratrol suppressed blood B cells, while there was no significant influence on blood T lymphocytes. However, insignificant stimulatory effect occurred at the lowest concentration. In addition, the compound inhibited proliferation of T and B lymphocytes isolated from the organs. Importantly, trans-resveratrol caused stimulation of blood and organs macrophages phagocytic killing, and also increased the respiratory burst of macrophages isolated from organ. These results suggest a potential use of trans-resveratrol as an immunomodulator of innate immunity in fish. This is particularly important, as this kind of resistance plays leading role in protecting the body against infection. In comparison, adaptive immunity is slower and also much less precise.

Early Cytokine Response After Vaccination with Coxiella Burnetii Phase I in an Infected Herd of Dairy Cattle.

INTRODUCTION: Coxiella (C.) burnetii, the aetiological agent of Q fever, is able to modulate the macrophage/T-lymphocyte axis in an infected organism and impair synthesis of monokines and lymphokines. MATERIAL AND METHODS: The purpose of this research was to determine the levels of the cytokines that play a key role in the response to C. burnetii antigens (IL-1ß, IL-2, IL-6, IL-10, IFN-γ and TNF-α) in the serum of animals originating from an infected herd prior to vaccination (day 0) and at 1, 7, and 21 days afterwards. RESULTS: The vaccination of animals did not affect the production of IL-6, IL-1ß, or IL-2. The serum levels of these cytokines were too low to measure in most of the samples. The initial levels of TNFα, IFNγ, and IL-10 were higher in seropositive than in seronegative animals, although significant differences between seropositive shedders and seropositive nonshedders appeared only in the levels of IFNγ and IL-10. Additionally, the course of the post-vaccination response concerning these two cytokines was different among seronegative nonshedders, seropositive nonshedders, and seropositive shedders. CONCLUSION: It seems that analysis of the IFNγ and IL-10 concentrations in animal blood serum may have some practical value in an assessment of the health status of seropositive animals and post-vaccination response.