Biofilm bacteria: formation and comparative susceptibility to antibiotics

The Calgary Biofilm Device (CBD) was used to form bacterial biofilms of selected veterinary gram-negative and gram-positive pathogenic bacteria from cattle, sheep, pigs, chicken, and turkeys. The minimum inhibitory concentration (MIC) and minimum biofilm eradication concentration (MBEC) of ampicillin, ceftiofur, cloxacillin, oxytetracycline, penicillin G, streptomycin, tetracycline, enrofloxacin, erythromycin, gentamicin, tilmicosin, and trimethoprim-sulfadoxine for gram-positive and -negative bacteria were determined. Bacterial biofilms were readily formed on the CBD under selected conditions. The biofilms consisted of microcolonies encased in extracellular polysaccharide material. Biofilms composed of Arcanobacterium (Actinomyces) pyogenes, Staphylococcus aureus, Staphylococcus hyicus, Streptococcus agalactiae, Corynebacterium renale, or Corynebacterium pseudotuberculosis were not killed by the antibiotics tested but as planktonic bacteria they were sensitive at low concentrations. Biofilm and planktonic Streptococcus dysgalactiae and Streptococcus suis were sensitive to penicillin, ceftiofur, cloxacillin, ampicillin, and oxytetracycline. Planktonic Escherichia coli were sensitive to enrofloxacin, gentamicin, oxytetracycline and trimethoprim/ sulfadoxine. Enrofloxacin and gentamicin were the most effective antibiotics against E. coli growing as a biofilm. Salmonella spp. and Pseudomonas aeruginosa isolates growing as planktonic populations were sensitive to enrofloxacin, gentamicin, ampicillin, oxytetracycline, and trimethoprim/sulfadoxine, but as a biofilm, these bacteria were only sensitive to enrofloxacin. Planktonic and biofilm Pasteurella multocida and Mannheimia haemolytica had similar antibiotic sensitivity profiles and were sensitive to most of the antibiotics tested. The CBD provides a valuable new technology that can be used to select antibiotics that are able to kill bacteria growing as biofilms.     

Experimental studies on cold survival of enchytraeid cocoons

The effect of different culture methods, cold acclimation and desiccation on the supercooling point (SCP), the melting point (MP) of fluids and the quantity of water freezing (osmotically active water) was investigated in individual cocoons of Enchytraeus crypticus and an undescribed Enchytraeus species using differential scanning calorimetry (DSC) techniques. Both species can be easily cultured in the laboratory in agar where the development and hatching of the worms can be observed. Culture methods (agar with nettle leaves or oats as food and wet filter paper without food supply) had a significant effect on fresh weight and SCP of E. crypticus cocoons. The water content (as a proportion of fresh weight) was slightly lower in the cocoons from the wet filter paper cultures. Acclimation at —3 °C did not affect the supercooling ability of E. crypticus cocoons, whereas the SCP of E. sp.1 cocoons was lowered from a mean of —8.7 to —12.4 °C. The supercooling ability of cocoons of E. crypticus was only slightly increased by desiccation and cold acclimation, but the quantity of frozen water was significantly reduced with acclimation and desiccation (2 and 3 min) at room temperature. The MP values of the cocoon fluids reflected these changes in both species. No cocoons of E. crypticus and E. sp.1 survived desiccation and freezing in our experiments and no worms of Enchytraeus albidus, E. crypticus and Henlea perpusilla survived freezing to —10 or —20 °C.     

Effects of Christmas-tree plantations on phytodiversity: implications for conservation

New Forests - Perennial crops such as Christmas trees have increasingly been cultivated throughout Europe. However, knowledge on the influence of Christmas-tree plantations (CTP) on biodiversity is...     

Abnormal ocular pigment deposition and glaucoma in the dog

The combined occurrence of ocular pigment deposition and glaucoma has been described in Cairn Terriers. Recently, this condition was also observed in two other breeds: the Boxer (two cases) and the Labrador Retriever (one case). Six dogs were referred to the Ophthalmology section of the Department of Clinical Sciences of Companion Animals and to a private referral clinic because of glaucoma or blindness in one or both eyes. In five cases ophthalmic examination showed pigment depositions in the sclera around the entire circumference of the perilimbal zone. Eight enucleated eyes (four eyes of two Cairn Terriers, three eyes of two Boxers and one eye of a Labrador Retriever) were examined microscopically. All eyes showed the same findings: an extensive infiltration of large melanin-containing cells with an eccentric nucleus, located in the iris, ciliary body, retina, choroids and sclera. Transmission electron microscopy of two of the examined eyes revealed that the morphology of most of these cells was consistent with melanophages. While reports in the veterinary literature concerning this condition are limited the cells concerned have been described to be melanocytes. Further research is needed to conclusively identify the cell type. As described in the present report, the histologic and transmission electron microscopic findings suggest a different etiology of the ocular pigment deposition and glaucoma compared with the pigment dispersal syndrome in humans.     

Identification of bluetongue virus serotype 2 (Corsican strain) by reverse-transcriptase PCR reaction analysis of segment 2 of the genome

In October 2000, bluetongue virus was detected on the French island of Corsica. The disease was also reported in Sardinia, Calabria, Sicily and on the Spanish islands of Majorca and Minorca. This paper describes the use of molecular techniques for a rapid identification and serotype determination of serotype 2 of the virus. The nucleotide sequences of segments 2 and 7 of the genome of the Corsican strain were determined and its phylogenetic relationships are described.     

Giardiasis and cryptosporidiosis in ruminants.

Although they differ considerably with respect to their biology, both Giardia duodenalis and Cryptosporidium parvum are common in ruminants, whereas Cryptosporidium andersoni is not. G. duodenalis infections are acquired during the first few months of life, tend to be chronic, and may be a production-limiting disease of ruminants. C. parvum infections remain an important cause of diarrhea in neonatal ruminants. Abomasal cryptosporidiosis, caused by C. andersoni, is an emerging disease of cattle that may affect both beef and dairy herds. This article reviews the life cycles, production impacts, treatments, controls, and zoonotic potentials of these important ruminant parasites.     

Einfluß von Schwefeldioxid auf Fichten (Picea abies [L.] Karst.) in verschiedenen Bodensubstraten

Zusammenfassung Drei Jahre alte Fichten wurden nach einer sechsmonatigen Anzuchtsphase in verschiedenen Bodensubstraten sieben Wochen lang mit 140 μg SO₂/m³ begast. Dabei kam ein Begasungsprinzip zur Anwendung, das auch bei Dauerbelastung konstante Gaskonzentrationen, unabh?ngig von sich ?ndernden Depositionsraten gew?hrleistet und Gaswechselmessungen in der geschlossenen Anzuchts- und Begasungskammer erlaubt. Es zeigte sich, da? sowohl n?hrstoffarme B?den als auch eine SO₂-Belastung im Kronenbereich zu Beeintr?chtigungen des Mineralstoffhaushaltes der Pflanzen führen. Insbesondere in den Wurzeln kam es nach einer siebenw?chigen Begasungsdauer zu einer starken Verminderung der Calcium- und Magnesiumgehalte. Auch die Photosyntheseleistung der Pflanzen und der Chlorophyllgehalt der Nadeln wurde durch die kombinierte Belastung mit sauren, n?hrstoffarmen Substraten und Schwefeldioxid st?rker beeintr?chtigt als durch den jeweiligen Einflu? der einzelnen Faktoren.

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Effects of sulfur dioxide on spruces (Picea abies [L.] Karst.) growing in different soil substrata Exploratory investigations on photosynthesis and mineral nutrient supplies following long-term application of low SO₂ concentrations

Summary Three-year-old spruce plants were grown in different soils for six months and then treated with 140 μg SO₂/m³ for seven weeks. The fumigation system used provides constant gas concentrations during a long-term fumigation period. Information on the combined growth and fumigation chamber is presented. It can be shown that not only nutrient deficiency in the soil but also dry deposition of SO₂ affects the mineral supply of plants. In particular the contents of calcium and magnesium in the roots were drastically reduced after the 7-week exposure period. The combination of acid soil and sulfur dioxide had a greater effect on both net photosynthetic rates of the plants, and chlorophyll contents of the needles as compared with the effect of only one of these stresses.

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Herrn Prof. Dr.W. Liese zum 60. Geburtstag gewidmet.     

Content of polyphenolic compounds in the Nigerian stimulants Cola nitida ssp. alba, Cola nitida ssp. rubra A. Chev, and Cola acuminata Schott & Endl and their antioxidant capacity

Varieties of kola nuts (Cola nitida alba, Cola nitida rubra A. Chev, and Cola acuminata Schott & Endl), a group of popular Nigerian and West African stimulants, were analyzed for their content of secondary plant metabolites. The three varieties of the kola nuts contained appreciable levels of (+)-catechin (27-37 g/kg), caffeine (18-24 g/kg), (-)-epicatechin (20-21 g/kg), procyanidin B 1 [epicatechin-(4beta-->8)-catechin] (15-19 g/kg), and procyanidin B2 [epicatechin-(4beta-->8)-epicatechin] (7-10 g/kg). Antioxidant capacity of the extracts and purified metabolites was assessed by two HPLC-based and two colorimetric in vitro assays. Extracts of all varieties exhibited antioxidant capacity with IC 50 values in the range 1.70-2.83 and 2.74-4.08 mg/mL in the hypoxanthine/xanthine oxidase and 2-deoxyguanosine HPLC-based assays, respectively. Utilization of HPLC-based assays designed to reflect in situ generation of free radicals (e.g., HO(*)), as opposed to general assays (DPPH, FRAP) in common use which do not, indicate that, of the major secondary plant metabolites present in kola nut extracts, caffeine is potentially the more effective cancer chemopreventive metabolite in terms of its antioxidant capacity.