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.

Histomorphologic/morphometric evaluation, leukocyte scintigraphy, and myeloperoxidase activity were used to determine whether neutrophils accumulate in the large colon of horses during low-flow ischemia and reperfusion. Twenty-four adult horses were assigned to 1 of 3 groups: group 1, sham-operated (n = 6); group 2, 6 hours of ischemia (n = 9); and group 3, 3 hours of ischemia and 3 hours of reperfusion (n = 9). Low-flow ischemia of the large colon was induced in horses of groups 2 and 3 by reducing colonic arterial blood flow to 20% of baseline. Radiolabeled (99mTc) autogenous neutrophils were injected at 175 minutes, which corresponded to 5 minutes prior to reperfusion in group-3 horses. Full-thickness biopsy specimens of the left ventral colon were collected at baseline and at 30-minute intervals for 6 hours; a portion of the biopsy specimen was placed in formalin for histologic examination, and the remainder was used to measure mucosal radioactivity and myeloperoxidase activity. There were no differences in baseline mucosal neutrophil index, mucosal neutrophil numbers, submucosal venular neutrophil numbers, mucosal radioactivity, or mucosal myeloperoxidase activity among groups, or over time in group-1 horses. Neutrophils accumulated in the colonic mucosa during ischemia and further increased at reperfusion, as indicated by neutrophil index (morphology) and mucosal neutrophil numbers (morphometry); mucosal neutrophil index was significantly (P < 0.05) greater in group-3 horses during reperfusion than at the corresponding periods of ischemia in group-2 horses. Neutrophil numbers were significantly (P < 0.05) increased in submucosal venules at 10 minutes of reperfusion in group-3 horses and were significantly (P < 0.05) greater in group-3 than in group-2 horses during the interval from 3 to 6 hours. Mucosal radioactivity significantly (P < 0.05) increased at reperfusion in group-3 horses; there was a trend (P = 0.076) toward greater mucosal radioactivity in group-3, compared with group-2 horses, throughout the 3- to 6-hour interval. There were no differences in mucosal myeloperoxidase activity among or within any of the 3 groups over time. Neutrophils accumulated in the large colon of horses during low-flow ischemia and reperfusion. Neutrophil infiltration was detected by histologic examination and leukocyte scintigraphy, but not by measurement of myeloperoxidase activity. The accumulation of neutrophils during ischemia and the further neutrophil infiltration during reperfusion indicate that neutrophils may contribute to reperfusion injury of the large colon.

A suction blister technique was used in 10 healthy dogs to remove the epidermis from the dermis in a standardized way. Collection chambers were attached to these skin windows and filled with autologous serum to attract exudative neutrophils. The chambers were emptied by fine-needle aspiration at 4-hour intervals and were refilled with serum for 24 hours after the Int aspiration. The collected cells were counted, differentiated, and stained, using the trypan blue dye-exclusion method to determine cell viability. Multiple skin biopsy specimens obtained during the procedure were examined histologically. The chamber fluid collected after 24 hours was cultured for bacteria. Increasing numbers of viable neutrophils were collected during the 24-hour period from the induced skin windows. In all but 1 dog, sufficient viable neutrophils could be collected to perform further functional tests in vitro. Our conclusion is that this technique might be useful to study chemotaxis in vivo and to perform functional tests on exudative neutrophils.

A protocol was developed for preparation of platelet concentrates (PC) to support thrombocytopenic dogs. Four clinically normal dogs with platelet counts that ranged from 200 to 330 X 10(9) platelets/L were used as donors. One unit (450 ml) of blood was collected by venipuncture into a double blood bag. Whole blood (WB) was centrifuged for 4 minutes at 1,000 X g (braking time = 2 minutes, 30 seconds) to prepare platelet-rich plasma (PRP). The PRP was expressed into the satellite bag and was centrifuged for 10 minutes at 2,000 X g (braking time = 2 minutes, 36 seconds). The platelet-poor plasma was expressed, leaving 40 to 70 ml of plasma and the pelleted platelets in the satellite bag. The resulting PC was left undisturbed for 60 minutes to promote disaggregation, and the platelets were then resuspended by gentle manual agitation. Forty-eight PC were prepared. Mean (+/- SD) platelet yield from WB to PRP was 78 +/- 13)% (range, 35 to 97%); yield from PRP to PC was 94 (+/- 6) % (range, 75 to 100%); and overall yield (PC from WB) was 74 (+/- 13) % (range, 36 to 91%). Mean PC platelet count was 8.0 (+/- 3.0) X 10(10) platelets/PC (range, 2.3 to 13.4 X 10(10) platelets/PC). The WBC content was 0.1 to 2.3 X 10(9) platelets/PC, representing 3 to 74% of WBC in the WB. Hematocrit was 0.1 to 26.2%. Results of bacterial and fungal culturing were negative.

Cardiorespiratory effects of abdominal insufflation were evaluated in 8 dogs during isoflurane anesthesia. Each dog was studied 3 times, in 1 of the following orders of insufflation pressures: 10-20-30, 20-30-10, 30-20-10, 10-30-20, 20-10-30, and 30-10-20 mm of Hg. Anesthesia was induced by use of a mask, dogs were intubated, and anesthesia was maintained by isoflurane in 100% oxygen. After instrumentation, baseline values were recorded (time 0), and the abdomen was insufflated with nitrous oxide. Data were recorded at 5, 10, 15, 20, 25, and 30 minutes after insufflation. The abdomen was then desufflated, with recording of data continuing at 35 and 40 minutes. Mean arterial pressure increased at 5 minutes during 20 mm of Hg insufflation pressure, and from 20 to 30 minutes during 30 mm of Hg pressure. Tidal volume decreased from 5 to 30 minutes during 10 and 20 mm of Hg pressures, and from 5 to 40 minutes during 30 mm of Hg pressure. Minute ventilation decreased at 10 and 20 minutes during 20 mm of Hg pressure. End-tidal CO2 concentration increased from 5 to 30 minutes during 20 and 30 mm of Hg pressure. The PaCO2 decreased at 40 minutes during 10 mm of Hg pressure, at 30 minutes during 20 mm of Hg pressure, and from 10 to 40 minutes during 30 mm of Hg pressure. Values for pH decreased from 10 to 30 minutes during 20 and 30 mm of Hg pressures. The PaO2 decreased from 20 to 40 minutes during 10 mm of Hg pressure, at 30 minutes during 20 mm of Hg pressure, and from 10 to 40 minutes during 30 mm of Hg pressure. Percentage decrease in tidal volume was greater at 5 and 15 minutes with 30 mm of Hg pressure. Differences in percentage increase in end tidal CO2 concentration were observed among the 3 pressures from 5 to 30 minutes. Although significant, these changes do not preclude use of laparoscopy if insufflation pressure > 20 mm of Hg is avoided.

The purpose of this study was to compare the thermodilution technique for estimation of cardiac output with the indocyanine green dye dilution technique at flows between 10 and 39 L/min in halothane-anesthetized horses. The estimation of area of dye dilution cardiac output curves was made by using the fore-'n-aft (FA) triangle method. This shorthand technique was compared with logarithmic exponential extrapolation and summation (extrapolated area), using 64 cardiac output curves. Then, 256 simultaneous thermodilution measurements were compared with dye dilution measurements calculated by use of the FA technique. Forty milliliters of iced 0.9% NaCl solution containing 15 mg of indocyanine green dye was used as the indicator. This was delivered in < 1 second to the right atrium, using a power injector. A thermistor positioned in the pulmonary artery detected the thermal indicator. Blood was withdrawn from the carotid artery through a densitometer cuvette to measure the dye concentration. The FA estimations of area were higher than those determined by use of extrapolated area. A multiplicative adjustment of 0.837 was estimated. On average, thermodilution estimates of cardiac output exceeded the adjusted FA determinations. Using a weighted linear regression, we determined the following calibration adjustment: thermal dilution cardiac output/1.048 = indocyanine green dye dilution cardiac output.

We developed a single-pedicle, axial pattern tubed skin flap that could be transferred to an in vitro perfusion apparatus. On the basis of results of prosections, angiography, contact radiography, and surviving-length studies, it was concluded that a single-pedicle, axial pattern skin flap measuring 4 cm X 12 cm incorporating the caudal superficial epigastric artery would survive to its entire length. Subsequently, a surgical (stage 1) procedure was developed for the routine preparation of single-pedicle, axial pattern tubed skin flaps. Healing after the stage-1 procedure was evaluated by visual inspection and fluorescein angiography. Stage-1 procedures were performed successfully 149 of 160 (93%) times. A second surgical (stage 2) procedure was developed for routine cannulation of the caudal superficial epigastric artery and harvest of the tubed skin flap. Stage-2 procedures were performed successfully 136 of 144 (94%) times.

The anatomy of each feature and structure of the laryngeal and adjacent regions, as perceived by palpation, is described for clinically normal standing horses. Visible skin contours produced by some of the superficial structures are also described. Concurrent dissection was performed on fresh cadavers to confirm initial findings. The procedure of systematic palpation in relation to clinical diagnosis and surgical procedure is discussed.

Three methods of determining glomerular filtration rate (GFR) were performed in adult ferrets, 9 months to 7 years old. Endogenous creatinine clearance was determined, using serum and urine creatinine values obtained during 24- and 48-hour collection periods from 27 ferrets housed in metabolic cages. Creatinine and radiolabeled inulin were administered to 12 female ferrets by constant IV infusion during isoflurane-induced anesthesia. Serial 20-minute urine collections, together with serum samples obtained at the midpoint of urine collection, provided measures for clearance calculations of these substances. Mean +/- SD endogenous creatinine clearance in ferrets for metabolic cage collections was 2.50 +/- 0.93 ml/min/ kg of body weight. There were no significant differences between the 24- and 48-hour clearance rates. Mean inulin clearance was 3.02 +/- 1.78, and mean exogenous creatinine clearance was 3.32 +/- 2.16 ml/ min/kg. Analysis of variance, using least-squared means adjustment, did not yield any significant differences between inulin and exogenous creatinine clearance rates. Exogenous creatinine clearance-to-inulin clearance ratio was 0.99 +/- 0.46, and there was significant correlation between the 2 methods (r = 0.82, P = 0.0001). Significant body temperature effects on inulin or exogenous creatinine clearance were not found. Infused inulin clearance, the generally preferred method for GFR calculation in mammalian species, was significantly (P = 0.0069) higher in younger (3.65 ml/min/kg) vs older ferrets (2.29 ml/min/kg). Results of this study indicate that inulin clearance is an adequate measure of GFR in ferrets as it is in other species. Compared with inulin clearance, exogenous creatinine clearance also provides a reliable estimate of GFR in ferrets.

Renal clearance procedures were performed on adult mixed-breed dogs with a wide range of renal function. Endogenous creatinine clearance was computed after analyzing plasma and urine for creatinine by use of 2 methods, PAP and kinetic Jaffe. For 20-minute clearance procedures, [14C]inulin clearance was measured simultaneously with endogenous creatinine clearance. For 111 twenty-minute clearance procedures performed on 24 dogs, [14C]inulin clearance was highly correlated with creatinine clearance for both methods of creatinine analysis (R2 = 0.979 for [14C]inulin-PAP; R2 = 0.943 for [14C]inulin-Jaffe). The absolute values for PAP and [14C]inulin clearance were nearly the same (PAP-to-[14C]inulin clearance ratio = 1.03 +/- 0.08), but those for Jaffe clearance were substantially less than those for [14C] inulin clearance Jaffe-to-[14C]inulin clearance ratio = 0.88 +/- 0.10). The Jaffe-to-[14C] inulin clearance ratio was inversely correlated with degree of renal function (R2 = 0.464), whereas the PAP-to-[14C]inulin clearance ratio was not correlated with degree of renal function (R2 = 0.060). Thus, Jaffe-determined creatinine clearance varied, in relation to [14C] inulin clearance, depending on degree of renal function. In 4 clinically normal dogs, 20-minute and 24-hour sample collections analyzed by use of the PAP method gave clearance values significantly greater, for both periods, than did Jaffe analyses. The PAP-determined creatinine clearance values were less than, but not significantly different from 20-minute exogenous creatinine clearance values determined 10 days after 24-hour collections. For 20-minute and 24-hour collections, the difference in clearance values between the PAP and Jaffe methods was attributable mostly to lower plasma creatinine values for the PAP method (mean +/- SEM, plasma PAP-to-Jaffe ratio = 0.798 +/- 0.053). However, urine creatinine values also were less by use of the PAP method (urine PAP-to-Jaffe ratio = 0.943 +/- 0.103). We conclude that PAP-determined creatinine clearance reliably measured glomerular filtration rate during 20-minute collections, and probably during 24-hour collections as well. By contrast, Jaffe-determined creatinine clearance underestimated glomerular filtration rate by a variable amount.