Plasma and Tissue Amikacin Concentrations Following Regional Limb Perfusion of Chickens (Gallus gallus domesticus).

Intravenous regional limb perfusion (IVRLP) has been used in the treatment of pododermatitis and distal limb infections, which are significant causes of morbidity in avian species. This intravenous drug administration technique is designed to achieve high drug tissue concentrations while minimizing systemic toxic effects. Amikacin is commonly used for IVRLP in veterinary medicine, but dosing guidelines have not been established for its use in birds. The current study aimed to determine the tissue concentration of amikacin after a single IVRLP administration in healthy, euhydrated leghorn hen chickens (Gallus gallus domesticus). Chickens received a single IVRLP dose of 10 mg/kg amikacin and were euthanatized posttreatment at 1 hour (n = 6), 12 hours (n = 6), and 24 hours (n = 6) to assess tissue and synovial fluid concentrations of amikacin in the injected leg. Mean tissue concentrations were highest 1 hour post-IVRLP (synovial fluid = 153.0 µg/mL, metatarsal pad tissue = 26.05 µg/mL) before declining at the 12- and 24-hour time points. This indicates that administration of amikacin via IVRLP can reach minimum inhibitory concentrations of common bacterial isolates in tissues after a single treatment with 10 mg/kg amikacin. Regional limb perfusion every 24 hours is recommended, although the minimum days of treatment may be case dependent and vary based on response to therapy.

Abdominal perfusion in canine patients with pyometra and sepsis evaluated by Doppler and contrast-enhanced ultrasound.

BACKGROUND: Sepsis is a condition characterized by organic dysfunction, leading to hemodynamic instability and high morbidity and mortality rates in humans and animals. Early identification of perfusion changes and appropriate management of sepsis are crucial for improving patient prognosis. Currently, the Systemic Inflammatory Response Syndrome (SIRS) and Sequential Organ Failure Assessment (SOFA) scores are widely studied for sepsis identification and evaluation of organ dysfunction. However, these scores do not assess gastrointestinal involvement, which is common in this condition. Contrast-enhanced ultrasound (CEUS) and Doppler have been considered promising diagnostic techniques for detecting changes in vascularization and microcirculation in a non-invasive and safe manner, particularly in the gastrointestinal system. This study aimed to evaluate duodenal perfusion using CEUS, as well as abdominal aortic and cranial mesenteric artery blood flow using Doppler ultrasound, and systolic arterial pressure (SAP) in 17 bitches with pyometra and in 10 healthy animals. RESULTS: The variables were compared between the pyometra and control groups, as well between patients with and without sepsis determined by the SOFA or SIRS scores. Pyometra was found to cause a reduction in abdominal aortic blood flow volume, aortic peak systolic velocity, and resistivity index as evaluated by Doppler ultrasound. Patients with sepsis according to the SOFA criteria only presented lower SAP. In contrast, sepsis animals identified by the SIRS score exhibited lower SAP, aortic peak systolic velocity, aortic blood flow volume, and aortic resistivity index and additionally, higher peak intensity of contrast in the duodenal wall. CONCLUSIONS: Pyometra causes a reduction in abdominal aortic blood flow, which is more pronounced in animals with sepsis identified by the SIRS criteria. These animals also exhibited a decrease in systolic blood pressure and an increase in duodenal perfusion, as evident by CEUS. However, these changes were not observed in patients with sepsis identified by the SOFA criteria. The alterations in intestinal perfusion observed in animals with sepsis indicate the presence of inflammation or dysfunction. In this regard, CEUS proves to be a valuable technique for detecting subtle changes in tissue hemodynamics that may not be apparent in conventional exams.

Effects of a 10% dimethyl sulfoxide solution on radiocarpal joint amikacin pharmacokinetics during intravenous regional limb perfusion in standing sedated horses.

OBJECTIVE: To determine the effect of a 10% dimethyl sulfoxide (DMSO) solution on the peak concentration (CMAX ) of amikacin in the radiocarpal joint (RCJ) during intravenous regional limb perfusion (IVRLP) compared with 0.9% NaCl. STUDY DESIGN: Randomized crossover study. ANIMALS: Seven healthy adult horses. METHODS: The horses underwent IVRLP with 2 g of amikacin sulfate diluted to 60 mL using a 10% DMSO or 0.9% NaCl solution. Synovial fluid was collected from the RCJ at 5, 10, 15, 20, 25, and 30 minutes after IVRLP. The wide rubber tourniquet placed on the antebrachium was removed after the 30 min sample. Amikacin concentrations were quantified by a fluorescence polarization immunoassay. The mean CMAX and time to peak concentration (TMAX ) of amikacin within the RCJ were determined. A one-sided paired t-test was used to determine the differences between treatments. The significance level was p < .05. RESULTS: The mean ± SD CMAX in the DMSO group was 1361.8 ± 593 µg/mL and in the 0.9% NaCl group it was 860 ± 481.6 µg/mL (p = .058). Mean TMAX using the 10% DMSO solution was 23 and 18 min using the 0.9% NaCl perfusate (p = .161). No adverse effects were associated with use of the 10% DMSO solution. CONCLUSION: Although there were higher mean peak synovial concentrations using the 10% DMSO solution no difference in synovial amikacin CMAX between perfusate type was detected (p = .058). CLINICAL SIGNIFICANCE: Use of a 10% DMSO solution in conjunction with amikacin during IVRLP is a feasible technique and does not negatively affect the synovial amikacin levels achieved. Further research is warranted to determine other effects of using DMSO during IVRLP.

Contrast-enhanced ultrasonography for evaluation of the blood perfusion of sciatic nerves in healthy dogs.

Blood supply to the peripheral nerves is essential for fulfilling their structural and functional requirements. This prospective, experimental, exploratory study aimed to assess the feasibility of contrast-enhanced ultrasonography (CEUS) for evaluating blood perfusion of the sciatic nerve in normal dogs. Contrast-enhanced ultrasonography examinations were performed on the bilateral sciatic nerves after bolus injection of Sonazoid™ (0.015 mL/kg) in 12 healthy Beagles for 150 s. Then, qualitative assessment of the wash-in timing, degree and enhancement patterns, and quantitative measurement of the peak intensity and time to peak intensity were performed from the sciatic nerve. The results were compared to those obtained from the adductor muscle around the nerve and caudal gluteal artery. After contrast agent injection, the sciatic nerve was enhanced at approximately 13-14 s, immediately after wash-in of the caudal gluteal artery. The peak intensity of the sciatic nerve was significantly lower than that of the caudal gluteal artery and higher than that of the adductor muscle. The time to peak intensity was significantly slower than that of the caudal gluteal artery; but was not significantly different from that of the adductor muscle. There were no significant differences in the peak intensity and time to peak intensity between the left and right sciatic nerves. These results demonstrate the feasibility of CEUS to assess blood perfusion of the sciatic nerve in healthy dogs qualitatively and quantitatively. This result from healthy dogs could serve as a reference for further studies that evaluate the sciatic nerve under pathological conditions.

Computed tomographic evaluation of pancreatic perfusion in 10 dogs with acute pancreatitis.

Severe canine acute pancreatitis can be fatal; imaging features that can predict the clinical course of disease are useful for clinicians. On computed tomography (CT), both pancreatic heterogeneous contrast enhancement and portal vein thrombosis have been correlated with poorer outcome. Perfusion CT is used in human medicine to evaluate pancreatic microcirculation to predict the future development of severe sequela to pancreatitis; this technology has yet to be explored in dogs with acute pancreatitis. The objective of this prospective, case-control study is to evaluate pancreatic perfusion using contrast-enhanced CT in dogs with acute pancreatitis and compare it with previously established values obtained in healthy dogs. Ten client-owned dogs preliminarily diagnosed with acute pancreatitis received a full abdominal ultrasound, specific canine pancreatic lipase (Spec cPL), and perfusion CT. Computer software calculated pancreatic perfusion, peak enhancement index, time to peak enhancement, and blood volume for 3-mm and reformatted 6-mm slices. The data was analyzed using Shapiro-Wilk test, linear mixed model, and Spearman's rho. Values for 3-mm slices were similar to 6-mm slices (all P < 0.05). Dogs with acute pancreatitis had a faster time to peak enhancement than healthy dogs (P = 0.04-0.06). Dogs with acute pancreatitis and homogeneous pancreatic enhancement had higher perfusion, faster time to peak enhancement, and greater blood volume compared to healthy dogs and dogs with acute pancreatitis and heterogeneous pancreatic enhancement (all P = / < 0.05). Pancreatic perfusion decreased with increased pancreatitis severity. No correlation was identified between Spec cPL and pancreatic perfusion (all P > 0.05). These findings preliminarily support perfusion CT in dogs with acute pancreatitis.

Treatment of Septic Tenosynovitis and Osteoarthritis in an American White Pekin (Anas platyrhynchos domesticus) with Repeated Intravenous Regional Limb Perfusion and Antibiotic-Impregnated Calcium Sulfate Beads.

A 2-year-old female American white pekin (Anas platyrhynchos domesticus) was assessed for a 2-month history of chronic lameness and swelling involving the left leg. Radiographic images of the left leg showed soft tissue swelling of the digits and tarsometatarsus with osteolysis of the tarsometatarsal-phalangeal joint. A complete blood count revealed marked leukocytosis and hyperproteinemia. A Streptococcus species was isolated from a bacterial culture of fluid obtained from the left tarsometatarsal-phalangeal joint. Biweekly intravenous regional limb perfusions of the left leg with ampicillin-sulbactam and amikacin were performed on the patient. Despite initial improvement in left leg lameness and swelling, follow-up radiographic images showed progressive osteolysis of the tarsometatarsal-phalangeal joint and associated digits. Surgical placement of antibiotic-impregnated calcium sulfate beads into the left tarsometatarsal-phalangeal joint was next performed with concurrent, repeated intravenous regional limb perfusion using the same antibiotic. Following the placement of antibiotic-impregnated beads and continued intravenous regional limb perfusion, the duck had decreased lameness and swelling of the left leg. Repeated antibiotic treatment through intravenous regional limb perfusion and concurrent placement of antibiotic-impregnated calcium sulfate beads is a practical treatment option for complicated distal limb infections in avian species. This therapeutic protocol has great potential in treating aggressive distal leg infections in many avian species because regional limb perfusion alone may not penetrate the joint adequately to achieve complete resolution of infection.

Ovarian, uterine, and luteal vascular perfusions during follicular and luteal phases in the adult cyclic female rabbits with special orientation to their histological detection of hormone receptor.

Understanding the does reproductive hemodynamic changes during the estrous cycle is crucial for improving reproductive competence and fertility potential in this species. The objective of this study is to investigate the hemodynamic variations in ovarian (OA) and uterine (UA) arteries, histological and morphometric changes in ovarian and uterine tissues throughout the follicular (FP) and luteal (LP) phases in rabbits and determine estrogen (ER), progesterone (PR) receptors, and vascular endothelial growth factor (VEGF) distributions using immunohistochemistry.Fourteen adults pluriparous New Zealand rabbits were divided into rabbits at the FP (Day - 1; n = 7) and those at the LP (Day 9; n = 7). Animals were subjected to Doppler, hormonal (estrogen [E2], progesterone [P4], insulin-like growth factor [ILGF], and VEGF), histological, and immunohistochemical analyses. In LP, OA Doppler indices were significantly increased, whereas peak systolic velocity (PSV) was decreased compared with that in FP. UA Doppler indices were significantly decreased in the LP, whereas PSV was increased (P < 0.05). E2 levels were increased in the FP, whereas P4 levels were increased in the LP. The morphometric analysis of uterine tissues during the LP revealed an increase in the mean uterine endometrium length, endometrial connective tissue area percentage (%), endometrial glands number, myometrial area (%) and thickness. Furthermore, ovarian follicles and corpus luteum (CL) displayed strong positive immunoreactivity for ER, PR, and VEGF-A during both phases. The ovarian sections displayed a substantial (P < 0.05) increase in the area % of VEGF-A in the ovarian follicles during FP while in the CL during LP. Conversely, area percentage of VEGF-A immunoreactivity in the uterine luminal and glandular epithelia during the FP and LP revealed no differences. However, the number of VEGF-A-stained blood capillaries revealed an increase during LP than FP. In conclusion, this study demonstrated for the first time the changes in both ovarian and uterine arteries during two different phases of the rabbit cycle in relation to the histo-morphometric analysis and distribution of ER, PR, and VEGF-A, which regulate uterine functions that play a role in reproduction.

The effects of aging and gestational month on uteroplacental vascular perfusion, and umbilical artery hemodynamics in pregnant jennies.

This study aimed to correlate the pulsed wave spectral indices of the middle uterine artery at both sides with placental development in jenny within mid-late pregnancies, and establish umbilical Doppler values for different ages and different gestational months. Twenty Equus Asinus pregnant jennies 260-450 kg (average, 320 ± 10 kg) were examined from 5 to 9 months of pregnancy with different ages (4-14 years). Monthly B-mode ultrasound examination was performed on both the combined thickness of the uterus and placenta (CTUP; mm) and umbilical artery cross-sectional diameter, and Doppler mode examination was performed on both the middle uterine (MUA at right [R] and left [L] sides) and umbilical arteries to measure both Doppler indices that expressed by resistance (RI) and pulsatility indices (PI), and blood flow rate. CTUP was elevated within pregnancy time at different ages (P < 0.05). L. PI was significantly declined throughout different ages (P < 0.05), but this declining trend was not observed in L. RI. The L. blood flow rate (R; bpm) was elevated among different ages and different months (P < 0.05). Both RI and PI were significantly decreased from 5 to 9 month of gestation period in jennies (P < 0.05).. The umbilical arteries cross-sectional diameter (Umb A; mm), was elevated among different ages and different months, while both Doppler indices were declined. A positive correlation was found (between both Doppler indices of both umbilical and uterine arteries P < 0.001). There was elevated vascular perfusion in uterine and umbilical arteries associated with reduced both Doppler indices along the course of pregnancy at different ages.

Perfusion with 10% neutral-buffered formalin is equivalent to 4% paraformaldehyde for histopathology and immunohistochemistry in a mouse model of experimental autoimmune encephalomyelitis.

Intravascular (IV) perfusion of tissue fixative is commonly used in the field of neuroscience as the central nervous system tissues are exquisitely sensitive to handling and fixation artifacts which can affect downstream microscopic analysis. Both 10% neutral-buffered formalin (NBF) and 4% paraformaldehyde (PFA) are used, although IV perfusion with PFA is most commonly referenced. The study objective was to compare the severity of handling and fixation artifacts, semiquantitative scores of inflammatory and neurodegenerative changes, and quantitative immunohistochemistry following terminal IV perfusion of mice with either 10% NBF or 4% PFA in a model of experimental autoimmune encephalitis (EAE). The study included 24 mice; 12 were control animals not immunized and an additional 12 were immunized with PLP139-151 subcutaneously, harvested at day 20, and fixed in the same fashion. Equal numbers (4 per group) were perfused with 10% NBF or 4% PFA, and 4 were immersion-fixed in 10% NBF. NBF-perfused mice had less severe dark neuron artifact than PFA-perfused mice (P < .001). Immersion-fixed animals had significantly higher scores for oligodendrocyte halos, dark neuron artifact, and perivascular clefts than perfusion-fixed animals. Histopathology scores in EAE mice for inflammation, demyelination, and necrosis did not differ among fixation methods. Also, no significant differences in quantitative immunohistochemistry for CD3 and Iba-1 were observed in immunized animals regardless of the method of fixation. These findings indicate that IV perfusion of mice with 10% NBF and 4% PFA are similar and adequate fixation techniques in this model.

Blood perfusion and diameter of bovine corpus luteum as predictors of luteal function in early pregnancy.

A feasible and non-invasive luteal function assessment which enables timely intervention to improve progesterone (P4) support at early pregnancy is not well-established. The aim of this study was to determine the correlation among morphological and functional assessment methods of corpus luteum (CL) on Day 5 (D5) following timed-artificial insemination effect on luteal blood perfusion (LBP), CL diameter and serum P4 concentration. Beef heifers (n = 89) were synchronized and inseminated (D0). On D5, CL was scanned by colour-Doppler ultrasonography using an LBP score (0 = absent; 3 = high blood perfusion); CL diameter was obtained and blood was collected. Diameter of CL had a positive linear effect on P4 concentration (p < .001); and larger CL diameter increased the probability to become pregnant (p < .05; odds ratio =1.21). Heifers with a CL larger than 14.95 mm had a higher pregnancy rate than heifers with a 14.95 mm or smaller CL (p < .05). Animals with LBP 2 and 3 had larger CL when compared to scores 0 and 1 (p < .001). Scores 1, 2 and 3 can accurately estimate heifers with higher P4 (accuracy =0.79, 0.72 and 0.61 respectively). In conclusion, LBP on Day 5 post-TAI allows to estimate heifers with higher P4 and larger CL size, and a larger CL diameter was positively associated with pregnancy rate.

Exogenous L-arginine administration improves uterine vascular perfusion, uteroplacental thickness, steroid concentrations and nitric oxide levels in pregnant buffaloes under subtropical conditions.

Heat stress (HS) during pregnancy adversely affects uterine vascular perfusion and foetal development. L-arginine (L-Arg), a nitric oxide (NO) precursor, has been proven to enhance an organ's vascular perfusion. Therefore, this study aimed to examine the effect of L-Arg administration on the pregnant buffaloes' uterine haemodynamics and uteroplacental thickness under environmental HS conditions. For this purpose, pluriparous (n = 12) HS-pregnant buffaloes (mid-gestation, 180-190 days; temperature humidity index >85) were haphazardly assigned into two groups, either administered a single intravenous (IV) bolus of 5 mg/kg BW of L-Arg Hcl (n = 6; ARG) or received normal saline (0.9%; IV; 25 ml) that served as a control group (n = 6; CON). Combined uteroplacental thickness (CUPT, mm), middle uterine arteries' (MUA) diameter (mm) and uterine Doppler haemodynamics (ipsilateral and contralateral to the pregnant horn) were examined utilizing B-mode and colour Doppler ultrasonography, respectively. The serum levels of progesterone (P4), estradiol 17ß (E2), nitric oxide (NO) and total antioxidant capacity (TAC) were assayed using commercial kits. The experimental time points were - 1, 0, 2, 4, 24, 48, 72, 96 and 120 h post-L-Arg administration. The ARG group showed higher (p < .05) CUPT and MUA diameter values starting at 24 h post-treatment and onward than the control buffalo cows. In addition, there were improvements (p < .05) in the blood flow parameters in the ipsilateral MUA after L-Arg treatment evidenced by lower values of pulsatility and resistive indices starting at 48 and 4 h, respectively and onward, and higher values of peak systolic velocity, coloured areas towards the pregnant uterine horn (both 24-72 h). Increases in the NO levels were found during the period between 4 and 120 h in the ARG group compared to the CON group. Moreover, significant increases in the E2 and P4 means were noted in the ARG group, especially at 24 h onward, respectively compared to the CON group. Concerning the TAC status, neither the administration nor the hours affected serum TAC levels in the HS pregnant buffaloes (CON and ARG). In conclusion, L-Arg administration improved uteroplacental thickness and enhanced uterine haemodynamics, NO levels, and steroids production in mid-pregnant buffalo cows under environmental heat stress conditions which could improve foetal growth and development.

Concentration of amikacin sulphate in synovial fluid when given in combination with dexamethasone phosphate in intravenous regional limb perfusion in standing horses.

Eight horses underwent IVRLP at two occasions through a 23-gauge 2 cm long butterfly catheter. Regional anaesthesia of the ulnar, median and medial cutaneous antebrachial nerves was performed prior, and an 8 cm rubber tourniquet was placed on the proximal radius for 30 minutes following the infusion. The first infusion consisted of 2 g of amikacin sulphate and 10 mg of dexamethasone phosphate diluted with 0.9% NaCl to a total volume of 100 ml. The second perfusion was performed after a 2-week washout period, the same protocol was used but without dexamethasone phosphate. Synovial fluid samples were collected from the metacarpophalangeal joint at T = 0, 0.5, 2, 12, 24 and 36 h post-infusion. Synovial fluid amikacin sulphate concentrations were determined by use of liquid chromatography/tandem mass-spectrometry. All horses (n = 8) remained healthy throughout the study, and no adverse effects associated with the study were encountered. No statistically significant differences were found in synovial fluid amikacin sulphate concentrations between the treatment and the control group at any of the time points. In conclusion, dexamethasone phosphate can be used in IVRLP concomitantly with amikacin sulphate in cases of distal limb inflammation and pain without decreasing the synovial fluid concentration of amikacin sulphate.

Effect of perfusate volume on amikacin concentrations after saphenous intravenous regional limb perfusion in standing, sedated horses.

OBJECTIVE: To determine the influence of perfusate volume on synovial fluid amikacin concentrations in the joints of the hind limb after standing saphenous intravenous regional limb perfusion (IVRLP). STUDY DESIGN: Randomized crossover design. ANIMALS: Six adult horses. METHODS: Saphenous IVRLP was performed in 6 standing horses with 1 g of amikacin diluted with 0.9% NaCl to volumes of 10 ml, 60 ml, and 120 ml. Samples of synovial fluid from the tarsocrural, metatarsophalangeal, and hind limb distal interphalangeal joints were collected at 15 and 30 min after perfusate administration. Concentrations of 40 µg/ml and 160 µg/ml were considered therapeutic for susceptible and resistant pathogens, respectively. RESULTS: No difference in synovial fluid amikacin concentrations was detected between volumes in any joint (P = .4). All synovial fluid amikacin concentrations were higher at 30 min compared to 15 min (P = .003). All median synovial fluid amikacin concentrations at 30 min were > 40 µg/ml using the 60 ml and 120 ml volumes. Synovial fluid amikacin concentrations >40 µg/ml were only reached in the hind limb distal interphalangeal joint when the 10 ml volume was used. All median synovial fluid amikacin concentrations observed were < 160 µg/ml. CONCLUSIONS: Target concentrations for pathogens that were considered susceptible were consistently reached with perfusate volumes of 60 ml and 120 ml. However, median synovial fluid amikacin concentrations did not reach target levels for resistant pathogens. CLINICAL SIGNIFICANCE: Perfusate volumes of 60 ml or 120 ml are recommended to treat infections due to susceptible pathogens in the joints of the distal hind limb. These results justify investigation of saphenous IVRLP with different perfusate volumes using higher doses of amikacin.

Clinicopathologic Findings in Chickens (Gallus gallus domesticus) Administered Amikacin Through Intravenous Regional Limb Perfusion.

Regional limb perfusion (RLP) has been used to treat cases of distal limb infections in avian species. Potentially nephrotoxic drugs, such as amikacin, may increase the risk of nephrotoxicity with RLP because of the presence of the renal portal system and direct venous blood flow from the pelvic limbs to the kidneys. In a randomized, blinded, placebo-controlled study, the safety of repeated amikacin administration (20 mg/kg q24h for 3 doses) via RLP was evaluated in healthy female chickens (Gallus gallus domesticus; n = 8 treatment, n = 8 saline control group). Plasma uric acid concentrations were not significantly elevated in treated birds compared with the control group at any time point following RLP. One week following the final RLP, birds were necropsied and the kidneys evaluated grossly and histologically. There was no significant difference in renal pathology scores between treated and control birds or between kidneys ipsilateral to the perfused limb and contralateral kidneys. This study concludes that RLP of amikacin at high doses produced no discernable renal pathology in healthy euhydrated chickens.

Evaluation of totally implantable catheters in healthy horses.

BACKGROUND: For horses requiring prolonged daily cephalic intravenous regional limb perfusion (IVRLP), the use of a totally implantable catheter (TIC) could be indicated to reduce complications associated with frequent venipuncture or external catheterization. This study aims to evaluate the implantation technique of the TIC in the cephalic vein of horses for IVRLP, describe the complications associated with the device's placement and use, and assess its viability up to 60 days after implantation. Totally implantable catheters, cut to 15 cm (n = 5) and 46 cm (n = 5) in length, were implanted into one cephalic vein in ten adult horses (n = 10). Twenty-four hours following placement, IVRLP with contrast was performed via the TIC and evaluated with radiography. Physical examinations, lameness evaluation, hematologic assessment, and the catheter patency tests were performed at scheduled intervals for the duration of catheterization (7-60 days). RESULTS: Catheters were implanted without difficulty and allowed for IVRLP 24 h post implantation. Complications resulted in removal of the catheters, with four maintained for 7 days, three in place for 15 days, and three catheters maintained for 60 days. Complications included lameness, limb swelling, catheter kinking, and venous thrombosis. CONCLUSIONS: The implantation technique of the TIC in the cephalic vein of horses is feasible and requires minimal technical effort. Although TIC allows venous access without the need for repeated venipuncture, its long-term use presents complications. For horses requiring prolonged daily cephalic IVRLP, the use of a TIC could be indicated. However, the high incidence of venous thrombosis may limit clinical application.

Early inflammatory events of mastitis-a pilot study with the isolated perfused bovine udder.

BACKGROUND: Bovine mastitis is an important health and cost factor in the milk industry. To elucidate whether isolated perfused bovine udders can be used to study early inflammatory events of mastitis, 1 mg of lipopolysaccharide (LPS) was instilled into quarters of 10 isolated perfused bovine udders. Three hours and 6 h after LPS instillation, tissue samples were taken from the gland cistern and base of the udder, subsequently stored in RNAlater and processed for the determination of inflammation-dependent gene regulation by real-time RT-qPCR. Gene expression analysis was performed using delta-delta Ct method. To translate mRNA results to protein, IL-1ß and IL-6 were determined in tissue homogenate by ELISA. RESULTS: The instillation of 1 mg LPS lead to an increased expression of pro-inflammatory cytokines and chemokines like TNF-α, CCL20, CXCL8 as well as of IL-1 ß, IL-6 and IL-10, lingual antimicrobial peptide (LAP) and S100A9. However, the degree of elevation differed slightly between gland cistern and udder base and markedly between 3 and 6 h after instillation, with a distinct increase in mediator expression after 6 h. IL-1ß protein increased in a time-dependent manner, whereas IL-6 was unchanged within 6 h of LPS instillation. CONCLUSION: Compared to in vivo studies with instillation of LPS into udders of living cows, a similar inflammation-dependent gene regulation profile can be mimicked in the isolated perfused bovine udder, indicating a supplementation of animal experiments.

Pharmacokinetics of ceftiofur in the metacarpophalangeal joint after standing intravenous regional limb perfusion in horses.

This study investigated the pharmacokinetics of ceftiofur after intravenous regional limb perfusion (IVRLP). Six horses were involved in 3 IVRLP sessions. For each session, operators with varying clinical experience placed the tourniquet. A wide-rubber tourniquet was applied in the antebrachium as 2 g of ceftiofur in a total volume of 100 mL was injected into the cephalic vein. Plasma and metacarpophalangeal synovial fluid samples were obtained to evaluate perfusate leakage and synovial fluid concentrations of ceftiofur over 24 h. Overall, mean plasma concentrations were not significantly different before and after tourniquet removal. Mean synovial fluid ceftiofur concentrations were significantly higher 5 min and 8 h after tourniquet removal versus 24 h, after which values above the minimum inhibitory concentration (MIC) (1 µg/mL) were not detected. Concentrations above the MIC were detected in 72% and 50% of the horses at 5 min and 8 h, respectively. Overall, higher synovial fluid concentrations were obtained for the operator with the most recent clinical experience performing IVRLP.

Pharmacocinétique du ceftiofur dans l'articulation métacarpophalangienne après perfusion intraveineuse loco-régionale chez le cheval debout. Cette étude a examiné la pharmacocinétique du ceftiofur après perfusion intraveineuse loco-régionale des membres (PLR). Six chevaux ont participé à trois séances PLR. Pour chaque séance, un opérateur d'expérience clinique différente a placé le garrot. Un garrot large en caoutchouc a été appliqué dans l'avant-bras tandis que 2 g de ceftiofur dans un volume total de 100 mL ont été injectés dans la veine céphalique. Des échantillons de plasma et de liquide synovial métacarpophalangien ont été prélevés pour évaluer les fuites de perfusion et les concentrations de liquide synovial de ceftiofur au fil du temps. Dans l'ensemble, les concentrations plasmatiques moyennes n'étaient pas significativement différentes avant et après le retrait du garrot. Les concentrations moyennes de ceftiofur dans le liquide synovial étaient significativement plus élevées 5 min et 8 h après le retrait du garrot par rapport à 24 h, où les valeurs supérieures à la CMI (1 µg/mL) n'étaient pas détectées. Des concentrations supérieures à la CMI ont été détectées chez 72 % et 50 % des chevaux à 5 min et 8 h, respectivement. Des concentrations plus élevées de liquide synovial ont été obtenues pour l'opérateur avec une expérience clinique plus récente sur PLR.(Traduit par les auteurs).

Orthopedic Infections-Clinical Applications of Intravenous Regional Limb Perfusion in the Field.

For the equine veterinarian, orthopedic emergencies are a common occurrence in clinical practice, with traumatic wounds of the distal limb and penetrating injuries of the hoof being some of the most common medical conditions to affect horses. Intravenous regional limb perfusion is a technique widely used for the treatment of orthopedic infections in horses. The objectives of this review are to discuss some of the clinical applications for this treatment modality in the field and to review the technique for the practitioner.

MRI perfusion analysis using freeware, standard imaging software.

BACKGROUND: Perfusion-weighted imaging is only scarcely used in veterinary medicine. The exact reasons are unclear. One reason might be the typically high costs of the software packages for image analysis. In addition, a great variability concerning available programs makes it hard to compare results between different studies. Moreover, these algorithms are tuned for their usage in human medicine and often difficult to adapt to veterinary studies. In order to address these issues, our aim is to deliver a free open source package for calculating quantitative perfusion parameters. We develop an "R package" calculating mean transit time, cerebral blood flow and cerebral blood volume from data obtained with freely imaging software (OsiriX Light®). We hope that the free availability, in combination with the fact that the underlying algorithm is open and adaptable, makes it easier for scientists in veterinary medicine to use, compare and adapt perfusion-weighted imaging analysis. In order to demonstrate the usage of our software package, we reviewed previously acquired perfusion-weighted images from a group of eight purpose-breed healthy beagle dogs and twelve client-owned dogs with idiopathic epilepsy. In order to obtain the data needed for our algorithm, the following steps were performed: First, regions of interest (ROI) were drawn around different, previously reported, brain regions and the middle cerebral artery. Second, a ROI enhancement curve was generated for each ROI using a freely available PlugIn. Third, the signal intensity curves were exported as a comma-separated-value file. These files constitute the input to our software package, which then calculates the PWI parameters. RESULTS: We used our software package to re-assess perfusion weighted images from two previous studies. The clinical results were similar, showing a significant increase in the mean transit time and a significant decrease in cerebral blood flow for diseased dogs. CONCLUSION: We provide an "R package" for computing the main perfusion parameters from measurements taken with standard imaging software and describe in detail how to obtain these measurements. We hope that our contribution enables users in veterinary medicine to easily obtain perfusion parameters using standard Open Source software in a standard, adaptable and comparable way.

Subtourniquet pressures generated by application of wide-rubber tourniquets in standing, sedated horses.

OBJECTIVE: To determine the influence of location and movement on subtourniquet pressure (STP) generated by application of a wide-rubber tourniquet (WRT) on equine limbs. STUDY DESIGN: Randomized experimental cross-over design. ANIMALS: Six standing, sedated horses. METHODS: Horses were sedated with detomidine hydrochloride (0.2 µg/kg IV), and 4 investigators applied WRTs to the antebrachium (AB), gaskin (GK), and midmetacarpus (MC) of each horse in a predetermined, randomized order. Subtourniquet pressure was consequently measured at 10-minute intervals (T0, T10, T20, T30) for 30 minutes. Indirect systolic blood pressure (SBP) was measured presedation, postsedation, and throughout the tourniquet application period. Target STP was established as SBP + 100 mm Hg. Limb movements at each location were classified as none, low, moderate, or high, on the basis of counts and magnitude. RESULTS: Mean STP did not change with time (P = .93) and exceeded SBP by 163 mm Hg (95% CI 122-205), 185 mm Hg (95% CI 156-214), and 402 mm Hg (95% CI 351-454) at the AB, GK, and MC, respectively. Mean STP at each location exceeded the target STP in 59 of 70 (81%) of the trials. Limb movements affected STP generated by tourniquets at the AB (P = .04) and MC (P < .0001) but not at the GK (P = .67). CONCLUSION: Wide-rubber tourniquets applied at the AB, GK, and MC generated STP >100 mm Hg above SBP for 30 minutes in standing, sedated horses. CLINICAL SIGNIFICANCE: Wide-rubber tourniquets as applied in this study can achieve and maintain the current recommended STP (SBP + 100 mm Hg) for equine IV regional limb perfusion. Number and magnitude of limb movement can decrease STP over time, potentially reducing the efficacy of a WRT.