Bacterial biofilms are structured communities of bacterial cells enclosed in a self-produced polymer matrix that is attached to a surface. Biofilms protect and allow bacteria to survive and thrive in hostile environments. Bacteria within biofilms can withstand host immune responses, and are much less susceptible to antibiotics and disinfectants when compared to their planktonic counterparts. The ability to form biofilms is now considered an attribute of many microorganisms. Diseases associated with biofilms require novel methods for their prevention, diagnosis and treatment; this is largely due to the properties of biofilms. Furthermore, the presence of biofilms on surfaces found at farms, slaughterhouses or food processing plants will have an impact on the efficacy of disinfection protocols. Surprisingly, biofilm formation by bacterial pathogens of veterinary or zoonotic importance has received relatively little attention. The objective of this brief Review article is to bring awareness about the importance of biofilms to animal health stakeholders.

Differences in the pathogenicity of atypical enteropathogenic Escherichia coli (EPEC) strains may be due, at least partially, to different expression patterns of some virulence genes. To investigate this hypothesis, the virulence gene expression patterns of 6 atypical EPEC strains isolated from healthy and diarrheic ruminants were compared using quantitative real-time reverse transcription polymerase chain reaction after growing the bacteria in culture medium alone or after binding it to HeLa epithelial cells. Some virulence genes in strains from diarrheic animals were upregulated relative to their expression in strains from healthy animals. When bacteria were cultured in the presence of HeLa cells, the ehxA and efa1/lifA genes, previously associated with the production of diarrhea, were expressed at higher levels in strains from diarrheic animals than in strains from healthy animals. Thus, the expression levels of some virulence genes may help determine which atypical EPEC strains cause diarrhea in ruminants.

During infection, nutrient deprivation can alter bacterial phenotype. This, in turn, may have implications for pathogenesis and prophylaxis. Actinobacillus pleuropneumoniae (biotype 1) and Haemophilus parasuis, respiratory tract pathogens of swine, are both V-factor-dependent. The concentrations of V factor in the extracellular fluids of pigs are unknown and may limit the growth of these bacteria in vivo. The aim of this study was to determine the concentrations of nicotinamide adenine dinucleotide (NAD) in select porcine body fluids and to compare the availability of NAD in vivo with the affinities of the organisms for this compound. Levels in plasma, tissue fluids (peritoneal, pleural, synovial, and cerebrospinal), and laryngeal, tracheal, and lung washings were determined with an enzymatic cycling assay. We concluded that, although the NAD supply in the respiratory tract is probably not growth-limiting, it may become limiting if the organisms are disseminated.

The innate immune system is a critical component directing the overall response of the immune system early in the inflammatory process. Evaluation of the innate immune system could offer a screening method for the selection of breeding stock from commercial chicken operations to improve flock health and prevent the loss of genes crucial to disease resistance. Three commercial broiler chicken lines (designated Lines A, B and C) were profiled for efficiency of their innate immunologic response. Oxidative burst and bactericidal functions of heterophils and monocytes, as well as heterophil degranulation, were analyzed. The birds were tested 1, 4, 8 and 15 days post-hatch. Individual lines differed in their ability to perform innate immunological responses during the first 15 days post-hatch. Although bactericidal capabilities were similar, oxidative burst responses by monocytes were low in comparison to that generated by heterophils. The fact that monocytes are not particularly adept at producing an oxidative burst at this age suggests that this is not a major avenue of innate defense by monocytes. Heterophil oxidative burst response was stronger in Line C than Line A during the first four days post-hatch. Line B showed no difference from Line C in heterophil oxidative burst response at 1 d, but produced a stronger response than Line C on 4 and 8 d post-hatch. Degranulation by heterophils showed significant differences in responses of Lines A and C depending on the day post-hatch, and stronger response in Line C vs Line B in the first four days post hatch. The first week post-hatch is an important time as chicks are particularly susceptible to infection as neonates. Mortality data of the commercial lines indicates that Line A is the most susceptible to demise, followed by Line C and then Line B. These results suggest that oxidative burst production efficiency is an important defensive function to monitor for immunoprofiling.

The effect that topical administration of cyclosporine would have on the number and type of microorganisms isolated from the corneal surface of dogs with keratoconjunctivitis sicca was studied. Schirmer tear tests were performed on and corneal swab specimens were collected from 61 eyes of 31 dogs with keratoconjunctivitis sicca prior to and after 3, 6, and 12 months of treatment with cyclosporine. In eyes that responded to cyclosporine treatment (Schirmer tear test value increased by greater than or equal to 5 mm/min, compared with pretreatment value), the percentage of eyes from which bacteria were isolated after 3, 6, and 12 months of treatment was significantly (P < 0.001) less than the percentage from which bacteria were isolated prior to treatment. However, among eyes that did not respond to treatment, we did not detect a significant change over time in prevalence of bacteria or type of bacteria isolated. The percentage of eyes from which fungi were isolated decreased during treatment; however, the small number of eyes in which fungal culture results were initially positive precluded demonstration of a significant change. For all eyes, we did not detect any significant differences over time in the frequency with which specific bacterial genera were isolated, with the exception of beta-hemolytic Streptococcus spp. Opportunistic corneal infections were not detected even though none of the dogs received antibiotics. An increase in production of tears, which contain anti-infection proteins, was believed to be the primary factor responsible for the decrease in the percentage of eyes from which microorganisms could be isolated.

Sixteen IgA-deficient German Shepherd Dogs with small intestinal bacterial overgrowth were randomized into 2 groups. One group was fed a chicken-based kibble diet; the other was fed the same diet, but with 1% fructo-oligosaccharides supplemented at the expense of cornstarch. After being exposed to the diets for 46 to 51 days, the group that ate the supplemented diet had significantly (P = 0.04) fewer aerobic/facultative anaerobic bacterial colony-forming units in fluid from the duodenum/proximal part of the jejunum, as well as in the duodenal mucosa. We could not detect significant differences in the species of bacteria found in the intestine of these 2 groups of dogs. We conclude that at least some dietary carbohydrates can affect small intestinal bacterial populations in dogs with small intestinal bacterial overgrowth.

Embryonating eggs were inoculated with filtered porcine ileal mucosa containing intracellular curved rods (ICR) and incubated for 4 to 6 days. Three of 12 pigs given the eggs per os developed microscopic lesions of proliferative enteritis (PE). Nonchallenge-exposed control pigs did not develop lesions of PE. Four of six positive control pigs given ileal mucosa from pigs with PE also developed microscopic lesions of PE. All of the PE lesions were found in pigs necropsied 10 to 29 days after challenge exposure. None of the swine in the study had clinical signs or gross lesions of PE. Campylobacter spp were isolated from pigs with and without exposure to the ileal mucosa from pigs with PE. There was no relationship between Campylobacter spp isolation and development of lesions. Deoxyribonucleic acids extracted from embryonating chicken eggs injected with the equivalent of 0.5 mg of mucosal lesions and incubated for 4 days hybridized to a DNA probe specific for the ICR whereas DNA extracted from 1.5 mg of mucosal homogenates of the same proliferative tissue did not hybridize with the same probe. Results of these experiments indicated that ICR injected into eggs remained infective for pigs and suggest replication of ICR in the first-passage eggs.

A protected catheter brush introduced by fiberoptic bronchoscopy was used to sample the tracheal and bronchial mucosa in 28 horses with small airway disease. Tracheal and bronchial brushings were examined for the presence of fungi, aerobic and anaerobic bacteria, and a cytoiogical evaluation was also done on fluid collected by the bronchoalveolar lavage (BAL) technique. Microorganisms (bacteria and fungi) were isolated more often in tracheal brushings (53.6%) than in bronchial brushings (10.7%). Anaerobic bacteria were not isolated. Results of this study indicate that fiberoptic bronchoscopy using a protected catheter brush is an easy and practical technique to obtain minimally contaminated samples for isolation of microorganisms from the lower respiratory tract of horses. However, no association was observed between isolation of high numbers of microorganisms from the bronchi and severity of small airway disease.

A double-blind randomized clinical trial was undertaken to determine the value of parenterally administered Streptococcus equi M-protein vaccine in foals during an epizootic of strangles. Weaned mixed-breed foals (n = 664) housed on 2 adjacent feed-lots (A and B) arrived over a 5-day period, 2 weeks before primary vaccination. Foals in lot B (n = 114) were randomly administered vaccine (n = 59) or saline solution (placebo; n = 55) on 3 occasions at biweekly intervals. Foals in lot A (n = 450) were given 1 dose of vaccine (n = 225) or placebo. The following clinical observations were scored blindly by a single observer for all foals in lot B and for 120 (randomly sampled) foals in lot A on a single day, 2 (lot B) and 6 (lot A) weeks after final vaccination: cervical lymphadenopathy, type of bilateral nasal discharge, and palpable swelling at injection site(s). Bacteriologic culture of nasal swab specimens or lymph node aspirates from selected foals with clinical disease yielded S equi. Cervical lymphadenopathy was observed in 17 of 59 (29%) vaccinates and 39 of 55 (71%) nonvaccinated controls in lot B and in 32 of 60 (53%) vaccinates and 29 of 60 (48%) controls in lot A. Contingency X2 analysis confirmed significantly lower cervical lymphadenopathy rate (X2 = 18.5; P < 0.001) and prevalence of mucopurulent nasal discharge (X2 = 11.4; P < 0.01) for vaccinates in lot B only. Swelling(s) at the vaccine injection site were palpated in 44% of lot B and 29% of lot A vaccinates vs < 2% of placebo controls. In the face of intense natural exposure, foals inoculated 3 times with M-protein vaccine were less than half as likely to have clinical signs of strangles as were nonvaccinated horses.

Chemical and cytologic effects and bactericidal activity of gentamicin in septic synovial fluid were evaluated in an experimental model of infectious arthritis in horses. Septic arthritis was induced by inoculation of approximately 7.5 x 10(6) colony-forming units of Escherichia coli into 1 antebrachiocarpal joint in each of 16 clinically normal adult horses. Clinical signs of septic arthritis were evident 24 hours after inoculation. Horses were allotted to 3 groups: group-1 horses (n = 5) each were given 150 mg of gentamicin (50 mg/ml; 3 ml) intra-articularly (IA); group-2 horses (n = 5) each were given 2.2 mg of gentamicin/kg of body weight, IV, every 6 hours; and group-3 horses (n = 6) each were given buffered gentamicin, consisting of 3 mEq of sodium bicarbonate (1 mEq/ml; 3 ml) and 150 mg of gentamicin (50 mg/ml; 3 ml), IA. Synovial fluid specimens were obtained at posttreatment hour (PTH) 0, 0.25, 1, 4, 8, 12, and 24 via an indwelling intra-articular catheter. Synovial fluid pH was evaluated at PTH 0, 0.25, and 24. Microbiologic culture and cytologic examination were performed on synovial fluid specimens obtained at PTH 0 and 24, and gentamicin concentration was measured in all synovial fluid specimens. At PTH 0, E coli was isolated from synovial fluid specimens obtained from all horses. Synovial fluid pH was lower (range, 7.08 to 7.16) and WBC count was higher (range, 88,000 to 227,200 cells/microliter) and predominantly neutrophilic (95 to 99%) at PTH 0 than before inoculation. Synovial fluid pH was lowered further (mean, pH 6.63) after IA administration of gentamicin in group-1 horses; mean pH remained unchanged (7.07) after buffered-gentamicin administration in group-3 horses. At PTH 0.25, mean peak synovial fluid gentamicin concentration in horses of groups 1 and 3 (4,745 and 6,190 microgram/ml, respectively) was 1,000 times greater than that in group-2 horses (5.1 microgram/ml) at the same time. Synovial fluid gentamicin concentration in group-1 and group-3 horses was always greater than that in group-2 horses and remained greater than a minimal inhibitory concentration of gentamicin (2 microgram/ml) against many common equine bacterial pathogens for at least 24 hours after injection. Further, the calculated apparent half-life and clearance of gentamicin in synovial fluid calculated after IA administration were similar in horses of groups 1 and 3. By PTH 24, E coli could not be isolated from synovial fluid specimens obtained from group-1 horses. However, moderate to heavy growth of E coli was isolated from synovial fluid specimens obtained at PTH 24 from horses in groups 2 and 3 (80 and 66%, respectively). In selected cases, IA administration of unbuffered gentamicin may be a useful supplement to drainage, lavage, and systemic antibacterial and anti-inflammatory treatment in horses with naturally acquired infectious arthritis.