The effectiveness of peppermint and thyme essential oil mist in reducing bacterial contamination in broiler houses.

The antimicrobial properties of essential oils have been demonstrated by various in vitro studies, whereas their effect on poultry farm hygiene has not been thoroughly investigated, in particular with reference to aerial treatment. The present study aims to assess the antibacterial effects of natural essential oils in broiler houses. Two experimental rooms were fogged with aqueous solutions of peppermint and thyme oils. The control room was sprayed with pure water. The experiment was conducted on broilers aged 1 to 42 d. The rooms were fogged every 3 d. One day after fogging, the total counts of mesophilic aerobic bacteria, Enterobacteriaceae, and mannitol-positive staphylococci were determined. Samples were collected from the air, litter, walls, and drinkers. The results of the study demonstrate that essential oil mist may improve hygiene standards in broiler farms. During broiler growth, the mean total counts of mesophilic bacteria in the rooms treated with essential oils were lower (P < 0.01 or P < 0.05) in comparison with the control. Enterobacteriaceae and staphylococci counts were also higher in the control group. A single exception was noted in a litter sample where the mean count of Enterobacteriaceae in the room fogged with peppermint oil was higher than in the control. Both oils reduced bacterial counts, but thyme oil was more effective in reducing coliform bacteria, whereas peppermint oil had a higher inhibitory effect on the proliferation of staphylococci. These promising results encourage further research to determine the optimal doses and the effects of essential oils and their combinations on the living conditions and health status of broiler chickens.

Volatile gas concentrations in turkey houses estimated by Fourier Transform Infrared Spectroscopy (FTIR).

1. The aim of the present study was to estimate gas concentrations in commercial turkey houses by Fourier Transform Infrared Spectroscopy (FTIR). 2. The experiment was conducted in 5 buildings of a commercial turkey farm. The measurements of gases were carried out every 3 weeks of the growth cycle. 3. The results demonstrate that ammonia and carbon dioxide are the prevalent gases released during the entire production cycle in turkey houses. The mean concentrations of the above compounds ranged between 4-31 ppm and 220-2058 ppm, respectively. Thiols, nitriles, amines, aldehydes, hydrocarbons and other organic and inorganic compounds also occurred in turkey houses, but they were emitted periodically and their mean concentrations were significantly lower in comparison with CO2 and NH3. 4. Lower ventilation ratio and higher moisture of excreta in the first half of the growth period accelerated the release of some gases, whereas gradual faeces and urine accumulation contributed to an increase in the concentration of selected organic compounds. 5. A portable FTIR analyser is a useful device for measuring gas concentrations in commercial turkey farms, and it supports determinations of tolerable emission limits in turkey production.

Metal transport and homeostasis within the human body: toxicity associated with transport abnormalities.

In this work, latest reports about metal toxicity, transport and homeostasis have been thoroughly described and discussed. Although diseases associated with transport and homeostasis abnormalities are those of great interest, still a variety of the phenomena associated with these processes are under debate. In this paper, we try to summarize the newest theses on this topic, presenting contradictory points of view. We focus on toxic and essential metal pathways crossing and try to follow the exact metal binding molecules within the body and provide insight into the transport mechanism. Special attention is given to the mechanism of action of lately investigated metal transporters.

Toll-like receptor gene polymorphism and its relationship with somatic cell concentration and natural bacterial infections of the mammary gland in sheep.

Possible correlation between Toll-like receptor (TLR)-gene mutations and the susceptibility of the mammary gland to bacterial infections and also the associate breed-dependent aspects of somatic cell concentration (SCC), bacterial infection and TLR-gene mutations in sheep are described. In Polish Lowland Sheep (PLS), milk samples exceeding the level of 500/microL (i.e. 5 x 10(5) per mL) of SCC were recorded almost twice more frequently than in Polish Heath Sheep (PHS) (40 and 22.3%, respectively). The frequency of bacterial infections was also found in a similar ratio (20 and 12.7%, respectively). During detection of the TLR-gene mutation we recorded 2 alleles of TLR1, 6 alleles of TLR2 and 10 alleles of TLR4 genes in PHS sheep, while PLS sheep possessed 2, 4 and 6 alleles, respectively. Statistical analyses revealed a relationship between the specified TLR alleles, SCC and the frequency of incidence of bacterial inflammations of mammary gland. The data may serve as a benchmark for further study of TLR-gene mutation-dependent predisposition of mammary gland defensive cells to recognize the pathogen properly and initiate the immunological response, and may help in identifying one of the markers of natural resistance against sheep mastitis.

Serum antibodies to retinal antigens in lung cancer and sarcoidosis.

OBJECTIVE: Autoantibodies to various neuronal proteins frequently accompany lung cancer and their appearance may precede cancer symptoms. In this study we examined which retinal antigens (RAs) are recognized by sera of patients with lung cancer and whether the occurrence of serum antibodies to particular RAs is characteristic for cancer in comparison with a noncancer lung disease. METHODS: Sera of 72 patients with non-small-cell lung cancer (NSCLC), 29 with small-cell lung cancer (SCLC), 27 with sarcoidosis (S), and sera of 32 healthy donors were examined in immunoblotting using retinal extracts and purified RAs as antigens. RESULTS: 69.0% of SCLC, 45.8% of NSCLC, and 44.4% of S sera displayed anti-RAs reactivity. Significantly less (p < 0.05; chi(2) test) percent of healthy control sera reacted with RAs. Lung cancer sera recognized mainly 46-, 56-, and 36-kD and to a smaller extent also 96-, 72-, 43-, and 26-kD proteins. Most of them were recognized with about 2-fold lower frequencies by S and control sera. Only lung cancer sera contained very high-titer antibodies to 46- and 26-kD RAs, identified as alpha-enolase and recoverin, respectively. CONCLUSION: Antibodies to RAs occur more frequently and in higher titers in lung cancer (especially SCLC) than in sarcoidosis or control sera. Although antibodies to retinal alpha-enolase, recoverin and other RAs are present mainly or exclusively in lung cancer sera, none of them seems to be a specific marker of a particular disease.

Production of lytic enzymes by Trichoderma spp. and their effect on the growth of phytopathogenic fungi.

The production of beta-1,3-glucanases and chitinases by three strains of Trichoderma in submerged cultures was determined. The synthesis of enzymes was induced by cell wall biopolymers of phytopathogenic fungi (Botrytis cinerea, Fusarium culmorum and F. oxysporum). T. hamatum produced the highest beta-1,3-glucanase activity; the most effective inducer of enzyme synthesis was the biomass of F. oxysporum. All examined strains of Trichoderma inhibited phytopathogen growth in biotic tests. The diffusion tests showed that the lytic enzymes take part in growth inhibition of phytopathogenic fungi.

[Molecular mimicry of bacterial polysaccharides and their role in etiology of infectious and autoimmune diseases]. / Mimikra czasteczkowa bakteryjnych antygenów polisacharydowych i jej rola w etiologii chorób infekcyjnych i autoimmunologicznych.

Molecular mimicry is one of the most important pathogenic factor of microorganism and is defined as a structural similarity of microbial molecules to host tissue contributing to the pathogenicity. Mimicry can be observed at the molecular, serological and functional level. In the review the infectious diseases have been discussed where the mimicry phenomenon may occur, and also autoimmune disease where due to the molecular mimicry bacterial structures are potent to induce adverse immune reactions. The cross-reacting molecules mimicking the host structures comprise colominic acid, sialic acid containing capsular polysaccharides of Streptococcus group B, phosphocholine containing antigen, lipopolysaccharides of Campylobacter jejuni contributing in induction of Guillain-Barré syndrome or Lewis antigen containing lipopolysaccharides of Helicobacter pylori inducing gut carcinoma. Knowledge on the phenomenon of molecular mimicry is important when new conjugate vaccine has to be constructed, because great care should be paid not to induce autoantibodies with synthetic immunogen. Investigation of microbial factors reveal that many autoimmune diseases are of infection etiology.

[Fimbriae as a pathologic factor of bacteria and a carrier in conjugate vaccines]. / Fimbrie jako czynnik patogennosci bakterii i nosnik w szczepionkach koniugatowych.

Fimbriae play important role as pathogenic factors in many bacteria by their adhesive properties. Adhesin is located at the tip of fimbriae but also in other parts of fimbriae. Recent findings on structure of fimbriae genes and their expression for the biosynthesis and formulation of complete fimbriae have been described. Special attention was focused on the participation of fimbriae in the mechanism of pathogenesis and their specificity towards tissue receptors. Most recent studies have been performed on E. coli and Klebsiella and those data predominate in this work. Fimbriae can be used for the construction of vaccine as a proteinous carrier for haptenic carbohydrate epitopes. In conjugates fimbriae express distinct immunogenic, adjuvant and protective properties.

[Molecular mimicry of bacteria as a factor in bacterial pathogenicity]. / Mimikra czasteczkowa jako czynnik patogennosci bakterii.

Structural and functional homology between bacterial proteins and host antigens, called molecular mimicry, is considered as significant pathogenic factor involved in several autoimmune diseases. The most important examples of this phenomenon reviewed in this work, involve rheumatic fever, Graves' disease, ankylosing spondylitis, Reiter's syndrome and rheumatic arthritis caused by infections with Streptococcus, Yersinia, Klebsiella, Escherichia coli, respectively.

Structural heterogeneity of the sialic-acid-containing oligosaccharides from the lipopolysaccharide of Hafnia alvei strain 2 as detected by FABMS studies.

The structure of four oligosaccharide fractions from the Hafnia alvei strain 2 lipopolysaccharide (LPS) have been assigned by FABMS. This approach corroborates data previously established by NMR spectroscopy for the major oligosaccharides in these fractions [A. Gamian, E. Romanowska, U. Dabrowski, J. Dabrowski, Biochemistry 30 (1991) 5032-5038; E. Katzenellenbogen, A. Gamian, E. Romanowska, U. Dabrowski, J. Dabrowski, Biochem. Biophys. Res. Commun. 194 (1993) 1058-1064; N. Ravenscroft, A. Gamian, E. Romanowska, Eur. J. Biochem. 227 (1995) 889-896]. In addition, the MS/MS with B/E linked scan technique allowed the detection of an additional oligosaccharide with the structure: [formula: see text] lacking the branched O-6 linked glucopyranose residue at the 3-linked Gal unit, which indicates a structural heterogeneity for the major oligosaccharide fraction.

Calcium binding to recoverin: implications for secondary structure and membrane association.

Recoverin is an EF-hand calcium-binding protein reportedly involved in the transduction of light by vertebrate photoreceptor cells. It also is an autoantigen in a cancer-associated degenerative disease of the retina. Measurements by circular dichroism presented here demonstrate that the binding of calcium to recoverin causes large structural changes. increasing the alpha-helical content of the protein and decreasing its beta-turn, beta-sheet and 'other' structures. The maximum helical content (67%) was observed at 100 microM free calcium and, unlike calmodulin, decreased as the calcium concentration was modulated in either direction from this value. Fluorescence measurements indicated that recoverin may aggregate or undergo structural changes independent of calcium binding as the calcium concentration is increased above 100 microM. EGTA also appeared to affect the structure of recoverin independent of its chelation of calcium. While calcium-induced conformational changes have been proposed to alter the membrane binding of recoverin through association of its myristoylated amino terminus, in the experiments presented here the partitioning of recoverin between the cytoplasmic and membrane compartments of the rod photoreceptor outer segment was unaffected by the concentration of calcium, therefore it appears unlikely that a calcium-myristoyl switch acts alone to anchor recoverin directly to the membrane. These experiments were conducted with native recoverin which is heterogeneously acylated, but mass spectrometry confirmed that simple chromatographic methods could be devised to isolate the different forms of recoverin for further studies.

[Retinal calcium-binding proteins--their function and pathology]. / Bialka siatkówki wiazace wapn--ich funkcja i patologia.

Calcium is involved in most cellular processes through calcium-binding proteins, belonging to the EF-hand superfamily. Recently new members of this family were found in the retina of vertebrates. Functions of these proteins and their relations to some neurological diseases are reviewed. Other EF-hand proteins found in the retina are also described.

Effect of chlorfenvinphos, cypermethrin and their mixture on the intestinal transport of leucine and methionine.

We assessed the effect of 2-week oral treatment with chlorfenvinphos, cypermethrin and their mixture in a dose of 5% LD50 on the intestinal transport of L-leucine (Leu) and L-methionine (Met) in male Wistar rats. Both in vitro (jejunal slices) and in vivo (intestinal perfusion) methods were employed in the study, using Leu and Met labelled with 14C as markers of the investigated processes. Additionally, in the in vitro study, concentrations of two amino acids were measured in the whole blood, serum, liver and perfused segment of the intestine. In the in vitro study the kinetic constants describing the active and passive Leu and Met uptake were determined. The active uptake was found to be particularly affected by oral intoxication with chlorfenvinphos, cypermethrin and their mixture. This susceptibility was seen as major alterations in the parameters of active uptake, i.e. Jm and Ki constants in the pesticide-exposed groups as compared to controls. In the in vivo study we found a decreased rate of Leu and Met disappearance from the intestinal lumen, decreased blood concentrations of Leu and Met and decreased liver content of Leu in the pesticide-exposed groups as compared to controls. The findings allow the conclusion that oral administration of chlorfenvinphos, cypermethrin and their mixture in a dose of 5% LD50 impairs the intestinal transport of Leu and Met and alters their distribution in the rodent body.

Recoverin, a photoreceptor-specific calcium-binding protein, is expressed by the tumor of a patient with cancer-associated retinopathy.

Recoverin is a member of the EF-hand family of calcium-binding proteins involved in the transduction of light by vertebrate photoreceptors. Recoverin also was identified as an autoantigen in the degenerative disease of the retina known as cancer-associated retinopathy (CAR), a paraneoplastic syndrome whereby immunological events lead to the degeneration of photoreceptors in some individuals with cancer. In this study, we demonstrate that recoverin is expressed in the lung tumor of a CAR patient but not in similar tumors obtained from individuals without the associated retinopathy. Recoverin was identified intially by Western blot analysis of the CAR patient's biopsy tissue by using anti-recoverin antibodies generated against different regions of the recoverin molecule. In addition, cultured cells from the biopsy tissue expressed recoverin, as demonstrated by reverse transcription-PCR using RNA extracted from the cells. The immunodominant region of recoverin also was determined in this study by a solid-phase immunoassay employing overlapping heptapeptides encompassing the entire recoverin sequence. Two linear stretches of amino acids (residues 64-70, Lys-Ala-Tyr-Ala-Gln-His-Val; and 48-52, Gln-Phe-Gln-Ser-Ile) made up the major determinants. One of the same regions of the recoverin molecule (residues 64-70) also was uniquely immunopathogenic, causing photoreceptor degeneration upon immunization of Lewis rats with the corresponding peptide. These data demonstrate that the neural antigen recoverin more than likely is responsible for the immunological events associated with vision loss in some patients with cancer. These data also establish CAR as one of the few autoimmune-mediated diseases for which the specific self-antigen is known.

Lipopolysaccharide core region of Hafnia alvei: serological characterization.

Covalent glycoconjugates containing, as a ligand lipopolysaccharide core, oligosaccharides of Hafnia alvei standard strain ATCC 13337 and R mutant 1 M were used to produce anti-H. alvei core antibodies. The sera obtained were tested in rocket immunoelectrophoresis, immunoblotting and ELISA using H. alvei lipopolysaccharides of various strains. The experiments were carried out to study the antigenic relationships between lipopolysaccharide core regions in the H. alvei genus.

Recoverin: a potent uveitogen for the induction of photoreceptor degeneration in Lewis rats.

Recoverin is a calcium-binding protein identified as an autoantigen in a paraneoplastic degenerative disease of the human retina known as cancer-associated retinopathy (CAR). In this study we investigated whether recoverin could elicit an immune response leading to the degeneration of photoreceptor cells in a rodent retina, and whether an animal model of CAR could be developed. Injection of Lewis rats with recoverin caused degeneration of the photoreceptor cells. Several features of uveoretinitis were observed, including vitreous cells, perivasculitis, retinal lesions and complete loss of the photoreceptor cell layer. The first clinical signs of retinal inflammation were observed 10-14 days after immunization. The earliest histological changes in the retina also were observed 14 days after immunization. Infiltration of the photoreceptor cell layer and inner layers of the retina with lymphocytic and some polymorphonuclear cells was frequently observed. Photoreceptors were damaged and later fully degenerated. This sequence of events was associated with high antibody titers against recoverin in all animals tested. Cellular responses to recoverin assayed between days 7 and 28 after immunization showed strong in vitro proliferative activities to recoverin. In addition, all aspects of the degenerative events could be reproduced in naive animals by the adoptive transfer of stimulated lymphocytes obtained from animals previously immunized with recoverin. This study demonstrates the successful induction of photoreceptor degeneration using recoverin as an immunogen. We demonstrate that recoverin is both a potent antigen and uveitogen. These observations may be relevant to our understanding of CARs in humans.

Purification and primary structure of Capl, an S-100-related calcium-binding protein isolated from bovine retina.

Calcium protein placental homolog (Capl) is an S-100-related calcium-binding protein selectively expressed in cell lines that have been induced to grow or differentiate. In addition, the expression of Capl correlates with the induction of the metastatic phenotype in tumor cell lines and the transformation of normal cells by activated oncogenes or chemical carcinogens. Although not previously associated with the nervous system, in this study, Capl was purified from bovine neural retina by a combination of phenyl-Sepharose and organomercurial chromatography. The complete amino acid sequence of bovine Capl was established primarily by Edman degradation of peptides generated by cleavage of methionyl, lysyl, glutamyl, and aspartyl bonds. NH2-terminal methionyl and aspartyl peptides were analyzed by tandem mass spectrometry, which provided the sequence of the first 8 residues and identified the NH2-terminal blocking group as an acetyl moiety. The molecular mass of the intact protein determined by electrospray mass spectrometry (M(r) = 11,716.75 +/- 0.42) and the calculated molecular mass deduced from the amino acid composition (M(r) = 11,718) were in agreement, thus supporting the accuracy of the sequence assignment. Capl isolated from the retina was shown to be indistinguishable by mass and immunochemical properties from its counterpart in the bovine aorta, which previously was the only source of purified Capl. Northern analysis using cloned Capl cDNA revealed that Capl mRNA is present not only in the retina but the choroid as well. Further support for choroidal localization came from immunohistochemical experiments using specific anti-Capl antibodies. The physiological significance of Capl in ocular tissues and the aorta is discussed.

Effect of outer membrane proteins (OMP) of Shigella on interleukin 2 (IL-2) production by spleen cells of mice.

The kinetics of interleukin (IL-2) release from mouse spleen cells incubated with different doses of outer membrane proteins (OMP) from Shigella was investigated. OMP induced very low activity of IL-2 after 2 and 4 h, and only a slightly higher level of the cytokine was detected after 6 h. However, IL-2 activity increased markedly after 20 and 24 h of incubation, and doses of 5 and 10 micrograms of OMP were found to be the most effective. Spleen cells cultured with OMP for 48 h contained reduced concentration of IL-2.

Serological and structural features of Hafnia alvei lipopolysaccharides containing D-3-hydroxybutyric acid.

The serological heterogeneity of Hafnia alvei lipopolysaccharides from strains ATCC 13337, 1187, 1221, 114/60, 1211 and 1216, that contain D-3-hydroxybutyric acid, was analyzed by rocket immunoelectrophoresis, immunoblotting and passive hemagglutination. The significance of D-3-hydroxybutyric acid component for their cross-reactivity has been discussed. The results obtained allowed us to place four H. alvei strains (ATCC 13337, 1187, 1221 and 114/60) in one serotype (A) and to consider two other strains (1211 and 1216) as separate serotypes (B and C, respectively).