Infection of a canine macrophage cell line with leishmania infantum: determination of nitric oxide production and anti-leishmanial activity

We have previously shown that resistance to Leishmania infantum in dogs is associated with a Th1 type of immune response. In this study, we use a canine macrophage cell line (030-D) that can readily be infected with this protozoan parasite. Our aim is to further characterize the effector mechanisms involved in killing of Leishmania parasite in dogs. We observed that activation of 030-D cells by incubation with a supernatant derived from a Leishmania-specific T cell line containing IFN-gamma, TNF-alpha and interleukin-2 (IL-2) resulted in enhanced nitric oxide (NO) production by these cells. In addition, we observed enhanced anti-leishmanial activity of infected 030-cells after activation. Both, NO production and anti-leishmanial activity were abrogated by addition of L-N(G)-nitroargininemethyl ester (L-NAME), an analogue of L-arginine. Thus, NO play an important role in the anti-leishmanial activity of these canine macrophages. We propose the infection of the 030-D cell line as a good in vitro model to further investigate parasite-host cell interactions in dogs, a natural host of Leishmania parasites.     

Efficacy of non-acaricidal containing otic preparations in the treatment of otoacariasis in dogs and cats

Eighty-nine cats and 38 dogs naturally infested with the ear mite Otodectes cynotis were randomly allocated into two treatment groups. One group was treated with a product containing miconazole nitrate, polymyxin B sulphate and prednisolone acetate, the other with a combination of diethanolamine fusidate, framycetin sulphate, nystatin and prednisolone. The treatment (five drops in each ear) was applied twice daily for 14 days, and its efficacy was evaluated on days 7, 14 and 21 on the basis of an otoscopic examination of the external ear canal, a microscopical examination of scrapings for the presence of ear mites and clinical signs of pruritus, pain, erythema and/or exudate. Both treatments were highly effective, and there were no significant differences between the two products, either in efficacy or in the clinical improvements observed. Apart from an allergic reaction in one cat treated with the second product, no adverse effects were observed.     

Effects of a hydrolyzed collagen dressing on the healing of open wounds in dogs

OBJECTIVE: To determine the effects of a hydrolyzed bovine collagen dressing (HBCD) on healing of open wounds in healthy dogs. ANIMALS: 9 female Beagles. PROCEDURES: 2 full-thickness skin wounds were made bilaterally on the trunk of each dog. Wounds on 1 side were treated with powdered HBCD covered with a semiocclusive nonadherent bandage. Wounds on the other side (control wounds) were covered with a semiocclusive nonadherent bandage only. Wound healing was subjectively assessed, and percentage increase in tissue perfusion was assessed by use of laser Doppler perfusion imaging (LDPI). Planimetry was performed to determine the percentages of contraction, epithelialization, and total wound healing. Biopsy specimens were examined microscopically to evaluate histologic changes. RESULTS: The HBCD did not induce a strong inflammatory reaction, as reflected by results of LDPI and histologic examination. Moreover, HBCD appeared hydrophilic and provided an environment to keep wounds clean and enhance early epithelialization. After treatment for 7 days, treated wounds had a significantly greater percentage of epithelialization than control wounds (12.13 vs. 7.03%). CONCLUSION AND CLINICAL RELEVANCE: The hydrophilic property of HBCD may cleanse contaminated wounds with the body's homeostatic fluids and enhance early wound epithelialization.     

Immunoblot analysis of calpastatin degradation: evidence for cleavage by calpain in postmortem muscle.

A negative correlation exists between calpastatin activity and meat tenderness. Therefore, it is important to determine the mechanism of calpastatin inactivation in postmortem skeletal muscle. Western immunoblot analysis was performed to determine the protease(s) responsible for degradation of muscle calpastatin during postmortem storage. To accomplish this, purified calpastatin was digested with different proteases in vitro, and their pattern of calpastatin degradation was compared with that of calpastatin degradation in postmortem muscle. Polyclonal antibodies raised in mice against recombinant bovine skeletal muscle calpastatin were used to monitor calpastatin degradation. Lamb longissimus was stored at 4 degrees C and sampled at 0, 6, 12, 24, 72, 168, and 336 h postmortem. Postmortem storage produced a discrete pattern of calpastatin degradation products that included immunoreactive bands at approximately 100, 80, 65, 54, 32, and 29 kDa. Undegraded calpastatin (130 kDa) was barely detectable after 72 h of postmortem storage at 4 degrees C, and no immunoreactive calpastatin was observed by 336 h postmortem. For in vitro proteolysis, lamb longissimus calpastatin (0 h postmortem) was purified using Affi-Gel Blue chromatography. Calpastatin was digested with m-calpain, mu-calpain, cathepsin B, proteasome, trypsin, or chymotrypsin. Each of these enzymes degraded calpastatin. Immunoreactive fragments resulting from digestion of calpastatin with m- and mu-calpain were similar to each other and closely resembled those observed during postmortem aging of lamb longissimus at 4 degrees C. Digestion of calpastatin with mu-calpain reduced calpastatin activity. Degradation of calpastatin by other proteases resulted in unique patterns of immunoreactive fragments, distinct from that observed in longissimus. Thus, m- and(or) mu-calpain seem to be responsible for calpastatin degradation during postmortem storage of meat.     

Sevoflurane anesthesia in psittacines.

Duration of anesthesia onset (time to intubation) and recovery (time to extubation, sternal and standing) and quality of recovery were compared for sevoflurane and isoflurane in 10 adult psittacines. Both agents were initially administered at an equal volume percentage (2%) rather than at equal minimum alveolar concentrations (MACs), therefore the initial concentration was above the isoflurane MAC for dogs and birds (1.3%) but below the sevoflurane MAC for dogs (2.3%). The time to intubation was significantly longer with sevoflurane because of initially delivering the sevoflurane below suspected MAC for birds. Although recovery times (time to extubation, sternal, and standing) were not significantly different, birds recovering from sevoflurane were less ataxic. Sevoflurane is a suitable inhalant agent for use in these psittacines and merits further study.     

Disease-induced alterations in plasma drug-binding proteins and their influence on drug binding percentages in dogs

Disease-induced variations of plasma albumin (ALB) and alpha 1-acid glycoprotein (AAG) levels were investigated in dogs. Lower ALB (sometimes > 50% reduction) and higher AAG (sometimes > 10-fold increase) levels were observed in dogs with various diseases. Drug binding was determined at therapeutic concentrations using normal, low-ALB and high-AAG dog plasma. The binding percentages of the ALB-binding drugs decreased in low-ALB plasma, resulting in a large increase in unbound drug, particularly for naproxen (a 13-fold increase). The binding percentages of all AAG-binding drugs investigated in this study increased in high-AAG plasma, resulting in a large decrease in unbound drug, particularly for quinidine (99% decrease). The fluctuation in the unbound fraction of drugs could affect their efficacy or could cause side-effects. Veterinary clinicians should monitor the ALB and AAG levels in the plasma of patients and correct dosage regimens according to these levels, where field conditions permit this, in order to ensure the proper usage of drugs with high affinity for ALB or AAG.     

Analysis of the right ventricular function in the exercising horse: use of the Fourier Transform

The objective of this study was to develop and test a technique to allow dynamic cardiac function to be studied during exercise in the horse. Blood pressure waveforms in the exercising horse are difficult to interpret because of the large influence of stride and respiration. A method has been devised to study dynamic right ventricular variables during high-speed exercise in the horse. A Fast Fourier Transform was performed on the digitised pressure waveforms and the frequency components associated with stride and respiration were removed. An inverse Fourier Transform was then performed to generate a time-domain pressure signal. Several dynamic right ventricular variables were calculated using the derived signal. Various parameters associated with removing frequencies from the frequency-domain pressure signal were changed to determine their influence on the variables. Most of the variables were not sensitive to these parameters. When compared during separate exercise bouts, some variables differed among runs, while others were not significantly different. Using the signal separation technique described here, right ventricular function of an exercising horse can be critically analysed.     

Comparison Between a γ-IFN Assay and Intradermal Tuberculin Test for the Diagnosis of Bovine Tuberculosis in Field Trials in Brazil

Bovine tuberculosis is a major problem in Brazil. The intradermal tuberculin test is the standard test for detection of bovine tuberculosis in Brazil but can lack both sensitivity and specificity. The purpose of this study was to compare a bovine γ-IFN assay with the tuberculin test under field conditions in Brazil. A total of 1632 animals from 13 dairy farms were tested using the single cervical tuberculin test (SCTT). Among those animals, about 15% of each herd, 220 in total, represented a high-risk group and were selected to be tested using the γ-IFN test. Of the 1632 animals tested, 207 presented significant reactions representing 12.7% of the cattle studied. In the selected group the number of animals positive by the γ-IFN assay was 126/220 (57.3%) and the total number of reactive cows on SCTT was 106/220 (48.2%). The real number of infected cattle, following standards, was 120/220 (54.5%). From these results the relative sensitivity rate of γ-IFN test was 100% including the false-positive results and 88.3% for the SCTT — a significant (P < 0.01) difference in favour of the γ-IFN test of 11.7%. The γ-IFN assay also identified some positive animals 60–120 days earlier than the SCTT. In conclusion, we believe that the γ-IFN assay can be used alone or in combination with the SCTT, as a valuable tool for the control of bovine tuberculosis in the Brazilian national herd.