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.     

The clinical features and short-term treatment outcomes of scirrhous cord: A retrospective study of 32 cases

Scirrhous cord (SC) is an uncommon complication of castration, characterised by chronic infection of the spermatic cord remnant. It is reported that surgical excision of the infected tissue is the most effective means of treatment, but there are few published studies assessing the outcomes of horses treated for SC. The aims of this retrospective study were to describe the clinical features and short-term outcomes in horses treated for SC at two equine hospitals in the UK. The clinical records of horses diagnosed with SC over a 10-year period were reviewed. A diagnosis of SC was made if the gelding presented with typical clinical signs with confirmation at surgery. Thirty-two cases of SC were identified at the two equine hospitals. The mean age at presentation was 6 years (range 2–14 years, n = 22), and the median time from castration to presentation was 29.5 days (range 20–2500 days). Mean age at castration was 4.3 years (range 6 months to 10 years, n = 10). Clinical signs included scrotal swelling, discharging wounds, hindlimb lameness and pyrexia. Five horses demonstrated hyperfibrinogenaemia (n = 8). Microbial culture isolated various bacterial species. All 32 cases were treated with surgical excision of the infected tissue and discharged from the hospitals between 1 and 10 days post-operatively. A limitation of this study is that it was a retrospective study with no long-term follow-up available. It was concluded that the results of this study confirm that SC can present at variable time points following castration, even many years later, and that a variety of bacterial species may be involved. Surgical excision of infected tissue is a successful treatment with a good short-term prognosis for survival.     

A follow-up of Beagle dogs intradermally infected with Leishmania chagasi in the presence or absence of sand fly saliva

In this study, we compare the development of infection and/or disease in Beagle dogs intradermally infected with Leishmania chagasi, in the presence or absence of Lutzomyia longipalpis saliva, with those of intravenously infected animals.Spleen samples of all the animals inoculated with parasites had positive polymerase chain reaction tests for Leishmania DNA. Positive spleen cultures for Leishmania were detected earlier (P < or = 0.018) and were more frequent (five out of the five animals) in intravenously infected animals than in the intradermally infected animals, in presence (two out of the six animals) or absence (three out of the five animals) of salivary gland lysate of L. longipalpis. Significant increase in serum antibodies against Leishmania was observed only in the intravenously infected group (P = 0.004). In addition, dogs with infection confirmed by isolation of amastigotes or detection of parasite DNA were, nevertheless, negative for anti-Leishmania antibodies up to 5 months or more after infection. Only animals of the intravenously infected group developed progressive decreases in hematocrit (Pearson r = -0.8076, P = -0.0026) and hemoglobin (Pearson r = -0.8403, P = 0.0012) during the infection period. No significant difference in the course of infection was observed between groups of intradermally infected animals. The data presented herein confirms that the intradermal inoculation of dogs with Leishmania produces an asymptomatic form of infection. It also fails to show an advantage in using L. longipalpis saliva as an infection-enhancing agent in experimental canine leishmaniasis.     

A guinea pig model of bovine pneumonic pasteurellosis.

The induction of pneumonic pasteurellosis in guinea pigs (Cavia porcellus) was examined. Specific pathogen free male guinea pigs were anesthetized and a tracheostomy performed to introduce 10(5), 10(4) or 10(3) Pasteurella haemolytica-A1 into the left principal bronchus. The surgical site was closed with tissue adhesive and staples and the animals were monitored for signs of respiratory tract infection. Within 24 hours after inoculation they became depressed, anorectic, pyretic and dyspneic. Fibrinous pleuropneumonia with prominent areas of necrosis and hemorrhage was present. Pericardial effusion was a frequent finding. There was infiltration of the pleura and alveoli with degenerate heterophils and macrophages, a hyperplastic mesothelium and fibrin exudation on the pleura and within alveoli. Hemorrhage, congestion, consolidation, edema and fibrin exudation were prominent in the hilar region of the lungs. Bacterial colonies were evident in all airways. More bacteria were recovered from infected lungs than were inoculated (p less than 0.05) indicating P. haemolytica was actively multiplying in the lungs. Hematological and clinical chemistry data were consistent with fibrinous pneumonia, however, blood cultures were positive for P. haemolytica in 61% (11/18) of animals sampled. Examination of pneumonic pasteurellosis in guinea pigs may be useful in studying pathogenetic and pathological features applicable to bovine pneumonic pasteurellosis (shipping fever pneumonia).     

Diffusion of N-labelled N2O into soil columns: a promising method to examine the fate of N2O in subsoils

Nitrous oxide research has generally focused directly on measuring fluxes of N₂O from the soil surface. The fate of N₂O in the subsoil has often been placed in the ‘too hard’ basket. However, determining the production, fate and movement of N₂O in the subsoil is vital in fully understanding the sources of surface fluxes and in compiling accurate inventories for N₂O emissions. The aim of this study was to generate and introduce into soil columns ¹⁵N labelled N₂O, and to try and determine the consumption of the ¹⁵N₂O and production of ambient N₂O. Columns, 100 cm long by 15 cm diameter, were repacked with sieved soil (sampled from 0 to 5 cm depth) and instrumented with silicone rubber gas sampling ports. Nitrous oxide enriched with ¹⁵N was generated using a thermal decomposition process at 300 °C and then transferred to 2 l flasks. After equilibrating with SF₆ tracer gas the ¹⁵N₂O was introduced into the soil columns via passive diffusion. Gas samples from the soil profile and headspace flux were taken over a 12-day period. A watering event was simulated to perturb the ¹⁵N₂O gas composition in the soil profile. Using the measured ¹⁵N enriched fluxes and the rate of decline in ¹⁵N in the N₂O reservoir, from which the N₂O diffused into the soil, we calculated an N₂O sink (consumption plus absorption by water) equal to 0.48 ng N₂O g⁻¹ soil h⁻¹. The decrease in the ¹⁵N enrichment between successive soil depths indicated N₂O production in the soil profile and we calculated a net N₂O production rate of 0.88 ng N₂O g⁻¹ soil h⁻¹. This pilot study demonstrated the potential for simultaneously measuring both N₂O consumption and production rates, using the ¹⁵N enrichment of the N₂O measured. Further potential refinements of the methodology are discussed.     

Effects of contact dermatitis on hepatic gene expression in broilers

1.?Severe foot and hock dermatitis in broiler chickens can reduce growth rate and increase susceptibility to bacterial infection, affecting both profitability and welfare. However, little is known about the underlying physiological changes associated with foot and hock lesions.

2.?This study compared global hepatic gene expression in control birds and those with ammonia-induced foot and hock lesions using Agilent 44?K chicken oligonucleotide microarrays (8 birds per group).

3.?In total, 417 genes were differentially expressed of which 174 could be mapped onto the genome. Genes associated with energy metabolism, thyroid hormone activity and cellular control were affected, while there was also evidence of an up-regulation of genes linked to a pro-inflammatory response.

4.?It is conceivable that pain is the underlying cause for the observed changes in energy metabolism genes.

5.?Changes in hepatic gene expression provide new information on how a chicken's physiological mechanisms alter to cope with foot and hock lesions. The findings support other data indicating that birds with increased severity of lesions are likely to be in pain and that growth will be compromised. Reduction of the incidence of dermatitis by improved husbandry should therefore benefit both welfare and commercial performance.     

Host and environmental reservoirs of infection for bovine digital dermatitis treponemes

Bovine digital dermatitis (BDD) is a global infectious disease causing lameness of cattle and is responsible for substantial animal welfare issues and economic losses. The causative agents are considered to be spirochetal bacteria belonging to the genus Treponema, which have consistently been identified in BDD lesions worldwide. One potential means of controlling infection is the disruption of transmission; however, the infection reservoirs and transmission routes of BDD treponemes have yet to be elucidated. To address these issues, we surveyed for evidence of BDD treponeme presence in the dairy farm environment, in bovine tissues and in bovine gastrointestinal (GI) tract contents. A total of 368 samples were tested using PCR assays specific for each of three currently recognised, isolated phylotypes of BDD treponemes. All environmental samples, together with insects and GI tract content samples were negative for BDD treponeme DNA from the three phylotypes. However, we identified BDD treponemes in two non-pedal bovine regions: the oral cavity (14.3% of cattle tested) and the rectum (14.8% of cattle tested). Whilst only single phylotypes were detected in the oral cavity, two of the rectal tissues yielded DNA from more than one phylotype, with one sample yielding all three BDD treponeme phylotypes. Whilst it might be considered that direct skin to skin contact may be a major transmission route of BDD treponemes, further studies are required to characterise and determine the potential contribution of oral and rectal carriage to BDD transmission.