Determination of the MIC, based on the activities of antibiotics against planktonic bacteria, is the standard assay for antibiotic susceptibility testing. Adherent bacterial populations (biofilms) present with an innate lack of antibiotic susceptibility not seen in the same bacteria grown as planktonic populations. The Calgary Biofilm Device (CBD) is described as a new technology for the rapid and reproducible assay of biofilm susceptibilities to antibiotics. The CBD produces 96 equivalent biofilms for the assay of antibiotic susceptibilities by the standard 96-well technology. Biofilm formation was followed by quantitative microbiology and scanning electron microscopy. Susceptibility to a standard group of antibiotics was determined for National Committee for Clinical Laboratory Standards (NCCLS) reference strains: Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, andStaphylococcus aureus ATCC 29213. Growth curves demonstrated that biofilms of a predetermined size could be formed on the CBD at specific time points and, furthermore, that no significant difference (P > 0.1) was seen between biofilms formed on each of the 96 pegs. The antibiotic susceptibilities for planktonic populations obtained by the NCCLS method or from the CBD were similar. Minimal biofilm eradication concentrations, derived by using the CBD, demonstrated that for biofilms of the same organisms, 100 to 1,000 times the concentration of a certain antibiotic were often required for the antibiotic to be effective, while other antibiotics were found to be effective at the MICs. The CBD offers a new technology for the rational selection of antibiotics effective against microbial biofilms and for the screening of new effective antibiotic compounds.

To define the pattern of change at the molecular and cellular levels during the healing of excisional skin wounds in the skeletally immature pig, mRNA levels for relevant molecules were assessed by semiquantitative RT-PCR using porcine specific primer sets and RNA isolated from normal skin and samples at various time post-wounding. Analysis of cellular change was assessed by DNA quantification and histology of tissue sections. The results demonstrated that the changes in the pattern of RNA and DNA content of the scar tissue were consistent with the observed increasing cellularity. The mRNA levels for collagen I, III, HSP47, IL-1, TGF-beta, MMP-1, -2 and -9, TIMP-1, -2, and-4, PAI-1, versican were significantly elevated during healing; levels for biglycan and fibromodulin were not significantly altered; and the mRNA levels for TIMP-3 were depressed. These findings suggest that skin wound healing is a series of complex matrix-cell interactions that involve cellular migration and inflammation, followed by proliferation of fibroblasts with new collagen synthesis, and lastly tissue remodeling of the scar.

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Giardiasis and cryptosporidiosis are gastrointestinal diseases caused by protozoan parasites that may infect domestic animals, wildlife and human beings. The ability of cysts and oocysts of these parasites to persist in the environment was determined because agricultural fecal waste has the potential to contaminate municipal water supplies. The degradation rate and viability of Giardia cysts and Cryptosporidium oocysts in water, cattle (Bos taurus) feces, and soil was evaluated at temperatures of −4, 4, and 25°C for up to 12 wk. Cysts and oocysts were enumerated after staining samples with a specific fluorescent monoclonal antibody and the viability was determined using propidium iodide dye exclusion and mouse infectivity assays. Giardia cysts were noninfective in water, feces, and soil following 1 wk of freezing to −4°C and within 2 wk at 25°C. At 4°C Giardia cysts were infective for 11 wk in water, 7 wk in soil, and 1 wk in cattle feces. Cryptosporidium oocysts were more environmentally resistant. At −4 and 4°C, the oocysts could survive in water and soil for >12 wk but degradation was accelerated at 25°C. Cryptosporidium oocysts also were degraded more rapidly in feces and in soil containing natural microorganisms. Contaminated cattle feces should be distributed on fields during warmer weather and after 12 wk of storage to reduce potential waterborne transmission following heavy runoffs.

The bacterial biofilm theory which describes bacterial populations in natural and pathogenic ecological systems in terms of a free-floating or 'planktonic' population of bacteria interacting with a more important matrix enclosed 'sessile' population of bacteria associated with or adherent to a surface, may help explain some of the problems linked to our understanding the nature of urinary tract infections. This paper reviews the role of bacterial biofilm formation in catheter-associated infection, prostatitis and struvite (infected stone) calculogenesis stressing, the importance of bacterial biofilms in the pathogenesis, persistence and hence the treatment of urinary tract infection.

BackgroundCastration is one of the most common procedures performed on beef and dairy cattle. The objective of the study was to determine the efficacy of meloxicam oral suspension in reducing pain and inflammation in calves following band or surgical castration.MethodsTwo identical trials with the exception of the method of castration (Band Castration Study 1 and Surgical Castration Study 2) were conducted. Sixty (60) healthy Holstein calves 4 to 5 months of age (138–202 Kg) were used. Animals received either Meloxicam Oral Suspension at a dose of 1 mg/kg BW (n = 15 Study 1 and 15 Study 2) or Saline (n = 15 Study 1 and 15 Study 2) 2 h before castration. Physiological (Heart Rate, Plasma Cortisol and Plasma Substance P) and Behavioral (Visual Analog Scale (VAS), Accelerometers and tail Pedometers) evaluations were conducted before (day -1) and after Castration (Day 0, 1, 2, 3). Inflammation was evaluated daily by providing an individual animal score (Study1) or with a measurement of scrotal thickness (Study 2).ResultsHeart rates were significantly greater in control animals following band and surgical castration. Plasma cortisol and substance P were significantly reduced in animals receiving Meloxicam Oral Suspension. Control animals had significantly greater VAS scores. Accelerometers showed that meloxicam treated animals had a significantly greater motion index and number of steps as well as less % time lying and number of lying bouts. The scrotal inflammation (based on scrotal swelling) was significantly decreased in the meloxicam treated animals compared to the control animals on day 1, day 2 and 3.ConclusionMeloxicam Oral Suspension was able to significantly reduce the display of painful behaviors and physiological responses to pain in band castrated and surgical castrated calves for up to 72 h following a single oral treatment of 1 mg/kg body weight. Meloxicam Oral Suspension was able to significantly reduce scrotal inflammation in band castrated and surgical castrated calves.

Xylella fastidiosa colonizes the xylem of various host plants, causing economically important diseases such as Pierce's disease in grapevine and citrus variegated chlorosis (CVC) in sweet oranges. The aggregative nature of this bacterium has been extensively documented in the plant xylem and the insect's foregut. Structured communities of microbial aggregates enclosed in a self-produced polymeric matrix and attached to a surface are defined as biofilms. In this study, we characterized biofilm formation by X. fastidiosa through the use of a novel in vitro assay for studying biofilm growth in a potential mimic system of what might occur in planta. We used wood, a xylem rich material, as a surface for bacterial attachment and biofilm formation, under shear force. We demonstrated that X. fastidiosa strains isolated from various hosts formed biofilm on wood in this in vitro assay. Different biofilm morphology was detected, which seems to vary according to the strain tested and microenvironmental conditions analyzed. We observed that strains from different hosts could be grouped according to three parameters: biofilm morphology, the ability to form clumps in liquid culture, and the ability to attach to glass surfaces. We hypothesize that biofilm formation is likely a major virulence factor in diseases related to X. fastidiosa, bringing a new perspective for disease treatment.

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