Isolation, identification and antibiotic resistance of Campylobacter strains isolated from domestic and free-living pigeons.

1. The aim of this study was to evaluate the occurrence of Campylobacter spp. in domestic and free-living pigeons and to evaluate the antibiotic resistance profiles. 2. The material consisted of cloacal swabs obtained from 108 homing pigeons and fresh faeces from 72 wild birds from Lublin and its vicinity. The identification of strains isolated on differential/selective media for Campylobacter spp. was carried out by MALDI-TOF and PCR. The susceptibility to antibiotics was evaluated by minimum inhibitory concentration (MIC) in Mueller-Hinton broth. 3. A total of 35 strains of Campylobacter spp. were isolated; 27 were identified as Campylobacter jejuni and 8 as Campylobacter coli. Over half of the isolates were resistant to erythromycin and streptomycin, 40% of strains were resistant to tetracycline and ampicillin and 37% isolates were resistant to amoxicillin. Resistance to two or more antibiotics was observed in all strains tested. 4. The results indicate that both domestic and free-living pigeons are reservoirs for bacteria of the genus Campylobacter, which are characterised by varied and growing resistance to commonly used antibiotics.

Antibiotic susceptibility of Lactobacillus strains isolated from domestic geese.

The aim of this study was to determine the antibiotic susceptibility of 93 Lactobacillus strains isolated from domestic geese raised on Polish farms. The minimal inhibitory concentration (MIC) of 13 antimicrobial substances was determined by the broth microdilution method. All strains were sensitive to the cell wall inhibitors ampicillin and amoxicillin (MIC ≤ 8 µg/ml). Resistance to inhibitors of protein synthesis and to fluoroquinolone inhibitors of replication was found in 44.1% and 60.2% of isolates, respectively; 26.9% strains were resistant to neomycin (MIC ≥ 64 µg/ml), 23.6% to tetracycline (MIC ≥ 32 µg/ml), 15% to lincomycin (MIC ≥ 64 µg/ml), 18.3% to doxycycline (MIC ≥ 32 µg/ml), 9.7% to tylosin (MIC ≥ 32 µg/ml), 56% to flumequine (MIC ≥ 256 µg/ml) and 22.6% to enrofloxacin (MIC ≥ 64 µg/ml). Bimodal distribution of MICs indicative of acquired resistance and unimodal distribution of the high MIC values indicative of intrinsic resistance were correlated with Lactobacillus species. Eleven (11.8%) strains displayed multiple resistance for at least three classes of antibiotics. Data derived from this study can be used as a basis for reviewing current microbiological breakpoints for categorisation of susceptible and resistant strains of Lactobacillus genus and help to assess the hazards associated with the occurrence of drug resistance among natural intestinal microflora.

Detection of the bovine viral diarrhoea virus (BVDV) in young beef cattle in eastern and south-eastern regions of Poland.

In view of the scarcity of information concerning viral diarrhoea virus (BVDV) infections in beef cattle in Poland, the aim of this study was to evaluate the presence of the BVDV in young beef cattle from selected herds in eastern and south-eastern regions of Poland. The material consisted of 78 sera obtained from beef cattle from 15 farms, aged 6-12 months. The anti-BVDV antibody level in the sera was estimated with an ELISA kit, and detection of the BVDV was carried out by standard PCR and one step Real-Time RT-PCR. The ELISA results showed a high degree (80%) of positivity in 5 of the 78 samples. In 7 samples the degree of positivity was in the very low range: < 40%. Of the 78 cDNA samples, the presence of genetic material with a length of 288 bp was found by standard PCR in 3 sera. The genetic material of BVDV was also found in the sera of the same three calves by Real-Time HRM PCR. BVDV infection in young beef cattle in south-eastern Poland is not a significant problem. This was confirmed by the positive ELISA results for 6.4% of the animals and the positive PCR results for 3.9%. The percentage of positive beef herds was about 8.6%. However, due to the severe nature of the disease and rapid transmission of the virus, regular monitoring of BVDV should be carried out.

An electrophoretic characterization of iron-transporting proteins in Mannheimia haemolytica A1.

Iron-regulated outer membrane proteins (IROMPs) in Mannheimia haemolytica A1, which function as a receptor for complexes containing iron ions, are induced by iron deficiency in the growth environment of the bacteria. Densitometric analysis of SDS-PAGE separation showed expression of IROMPs of 71, 77, and 100 kDa in the case of bacteria grown in a medium with 2,2-dipyridyl. The electrophoregrams obtained in 2-DE separations confirmed the presence of protein fractions with these molecular weights and isoelectric points ranging from 5.4 to 6.4. The results of the study also confirmed the ability of M. haemolytica A1 proteins involved in iron uptake to induce a protective immune response. In Western blot with serum from convalescent calves naturally infected with M. haemolytica A1, distinct reactions were obtained for IROMPs of 71, 77, and 100 kDa.

Effect of conglutinin on phagocytic activity of bovine granulocytes.

In the present study we investigated the effect of bovine conglutinin on the phagocytic activity of leukocytes. We measured both the chemotactic activity of conglutinin and its effect on the internalization of zymosan particles and E. coli by granulocytes. We also assessed the binding of conglutinin to various microorganisms isolated from clinical cases in cattle. We showed that conglutinin binds strongly to the surface of yeast cells and to mannan-rich zymosan particles, while weak binding was observed in the case of the bacterial strains tested, including those whose O antigen is composed of mannan. Conglutinin (1-10 microg/ml) neither acts as a chemotactic factor for peripheral blood leukocytes nor affects ingestion of E. coli by granulocytes. However, as flow cytometry based assay showed, conglutinin (0.1-1 microg/ml) increased ingestion of zymosan expressed as mean fluorescence intensity (MFI) of positive cells.

Analysis of the culture-derived soluble Babesia canis canis antigens derived from the Polish strains of the parasites.

UNLABELLED: OBJECTIVE, MATERIAL AND METHODS: The aim of this study was to analyse the protein fractions of the soluble parasitic antigen (SPA) from in vitro cultures of the native Polish strains of Babesia canis canis and to determine their immunogenicity through Western blotting using the sera of dogs vaccinated with this antigen. RESULTS: Polyacrylamide gel electrophoresis revealed 21 protein fractions with molecular weights from 12 to 205 kDa. The most intense reaction in Western blotting was observed between the serum antibodies of the SPA-vaccinated dogs and the fraction with the molecular weight of 52 kDa. CONCLUSION: Detailed studies on the composition of SPA of Babesia canis canis and reactivity of its individual protein fractions can be a starting point for the development of subunit vaccines against babesiosis. Using a preparation with only some electrophoretic fractions of SPA in the production of vaccines would allow to avoid putting an unnecessary protein burden in the vaccine which could cause side effects.

Characterization of outer membrane proteins participating in iron transport in Pasteurella multocida serotype A3.

Iron-regulated outer membrane proteins (IROMPs) of P. multocida serotype A3, which function as receptors for complexes containing iron ions, are induced by iron deficiency in the bacterial growth environment. Analysis of an electrophoresis image of proteins isolated from bacteria grown on medium supplemented with 2,2'-dipyridyl revealed expression of 16 new proteins that were not noted in the case of the bacteria grown in standard conditions, with molecular weights from 30 to 160 kDa. Induction of IROMP expression occurred within 30 minutes after restricted iron conditions were established. In immunoblotting, distinct reactions were noted for proteins of molecular weight ranges of 25-49 kDa, 61-95 kDa, and 108-214 kDa. Proteins of the molecular weight of 68, 75 and 86 kDa were analysed using mass spectrometry and matched with the highest probability to proteins in the NCBI data base. Several dozen different proteins with similar amino acid sequences were matched to each sample.

Characterization of Hsp70 proteins in bovine leukocytes induced by the temperature 41 degrees C.

Heat shock proteins (Hsp) are the group of proteins observed in both prokaryotic and eukaryotic cell types. Hsp synthesis takes place in response to many environmental conditions, including ultraviolet radiation, heavy metal ions, hypoxia and toxic agents. Many authors have suggested that Hsp can be used in immunoprophylaxis, yet Hsp70 proteins expressed in bovine leukocytes have not been fully characterized. Hence the aim of this study was to evaluate the expression of Hsp70 proteins in bovine leukocytes exposed to temp. 41degrees C. The material for the study consisted of bovine white blood cells incubated at 41 degrees C for 2 hours. SDS-Page electrophoresis, Western blotting, and two-dimensional electrophoresis (2D) were performed to estimate the proteins obtained. The results of the study confirmed the influence of the temperature of 41 degrees C on induction of Hsp70 in bovine leukocytes. These proteins were mainly localized within molecular mass 70kDa. Some of the proteins with molecular mass from 20 to 50 kDa also showed positive reactions in Western blotting with anti-Hsp70 antibodies. Analysis of 2D electrophoresis showed a change in the localization of these proteins in the pH gradient. It can be postulated that analysis of Hsp70 expression in bovine leukocytes can be a very helpful marker for evaluating an organism's adaptation to environmental heat stress. The proteins obtained also showed immunological reactivity with rabbit antibodies in Western blotting reactions, indicating that they can be used as protective factors in the pathogenesis of many diseases.

Use of oromucosally administered interferon-alpha in the prevention and treatment of animal diseases.

Interferon-alpha (IFN-alpha) is well known as a clinically effective antiviral and antineoplastic therapeutic agent. It has also been shown to have immunoregulatory properties. IFN-alpha stimulates a cell-mediated innate immune response and then participates in the transition of the initial host innate response to an effective adaptive immune response. IFN-alpha is produced in small quantities in nasal secretions during viral infections, prompting many authors to suggest that low-dose oromucosal administration of IFN-alpha effectively mimics nature. Moreover, the injectable high-dose interferon therapy currently approved for various human disorders causes numerous side effects. By contrast, oromucosal administration of IFN-alpha is not associated with toxic effects. Another distinct advantage is ease of administration: the IFN can be dissolved in drinking water or administered by nebulization to the oral or nasal cavity. This review describes the current state of knowledge concerning orally administered IFN-alpha, of both human and animal origin, as a prophylactic or therapeutic agent in veterinary medicine. We present the effects of IFN-alpha in such animals as cattle, pigs, horses, cats, dogs and chickens, and attempt to explain its mechanism of action following oromucosal administration. It is hoped that this review of the medical literature on the use of IFN-alpha in animals will give practitioners a better understanding of the challenges and benefits of using this interesting cytokine in clinical practice.

Conglutinin, CL-43 and CL-46--three bovine collectins.

Conglutinin, collectin-43 (CL-43) and collectin-46 (CL-46) are serum proteins characteristic for Bovidae. They belong to collectins--family of oligomeric proteins composed of trimeric subunits containing collagen-like sequences joined to C-type lectin domains. The genes encoding conglutinin, CL-43 and CL-46 are located on the bovine chromosome 28, and phylogenetic analysis indicates their common origin--from the lung surfactant protein D gene. Northern blot or immunocytochemical analysis confirm biosynthesis of bovine collectins mainly in the liver (conglutinin, CL-43) and in the thymus (CL-46). The level of conglutinin in the serum of dairy cows depends on many factors such as breeding, the season of the year, the stage of the reproductive cycle and infection. The collectins are involved in the innate immune defense. They bind to microbial surface carbohydrates inducing aggregation and, thereby, impeding infectivity. On the other hand the destruction of pathogens occurs due to stimulation of effector cells. CL-43 as well as conglutinin, binds to the collectin receptor (C1qR) localized on many types of cells identified as a surface variant of calreticulin. Conglutinin and CL-43 show antiviral activities towards influenza A virus and rotaviruses. Conglutinin also displays protective activity against bacterial infections.

[Low frequency vibration and testosterone levels in blood serum and excretion of 17-ketosteroid in urine in exposed workers]. / Wibracja ogólna niskich czestotliwosci a stezenie testosteronu w surowicy krwi i wydalanie 17-ketosterydów z moczem u narazonych pracowników.

Serum testosterone concentrations and excretion of 17-Ketosteroid in urine were determined in 32 caterpillar vehicle drivers exposed to low frequency vibration of the whole body and to noise. Testosterone concentrations were assessed by the RIA method in fasting after 3 hrs of work. 17-Ketosteroid excretion was measured in the 8-hr collection of night urine and in urine collected during the day's work. No significant differences were found in exposed workers, either in mean testosterone concentration in fasting and after 3 hrs of work or in 17-Ketosteroid excretion, before and during the work, when compared with the control group.