An in vitro study of hematopoietic progenitor cells from peripheral blood of rabbits

The purpose of this study was to isolate and cultivate a subpopulation of pluripotent stem cells (PSCs) from the peripheral blood of rabbits, which are frequently used in veterinary research as an animal model. Pluripotent stem cells, as described in human beings, are fibroblast-like cells that exhibit a CD34 marker, specific from other hematopoietic stem cells. Commonly used human commercial media has been researched for culturing rabbit PSCs. These findings allow us to contemplate the direct application of this simple and standardized methodology in several areas of study, such as of the pharmacological effect of many drugs on hematopoietic cells, veterinary practice, and even the study of new strategies in cellular therapy for some human diseases.     

Renal handling of calcium and phosphorus in experimental renal hyperparathyroidism in dogs

Twenty-four hour urinary excretion, fractional excretion and the filtered load of calcium and phosphorus were monitored as hyperparathyroidism evolved in a model of progressive canine renal failure. Thirteen beagles of both sexes aged four and a half months were used. Nine of them were subjected to a renal damaging schedule (neomycine, 60 mg/kg/48 h, IM, 32 weeks) in order to induce chronic renal failure leading to secondary hyperparathyroidism (2HPT group). The remaining four were kept as the control group. The experiment was conducted over 32 weeks. Blood and 24 h urine were collected every four weeks. Calcium, phosphorus and creatinine were analyzed. Plasma parathormone and calcitonin were determined at weeks 0, 12, 24 and 32. The level of renal function in the 2HPT animals was reduced to 25% of that of the controls (endogenous creatinine clearance was 0.45 +/- 0.22 mL/min/kg as opposed to 1.81 +/- 0.54 mL/min/kg). Hyperparathyroidism was confirmed by a progressive increase in the levels of the parathyroid hormone. Calcitonin levels were not modified. A tendency to hypocalcaemia was observed, reaching statistically significant levels from the twenty-eighth week of the study, when hyperphosphataemia also became significant. Daily urinary excretion of calcium and phosphorus remained at values considered normal throughout the experiment with no alteration imputable to the impaired renal function. This is explained by the decrease in the filtered load of these elements (in both cases statistically significant from the 24th week on) being associated with an increase in their fractional excretion. Thus, calcium and phosphorus urinary excretion values could be maintained in a normal range up to the end of the experiment, showing that renal calcium handling in dogs with experimentally induced renal failure seems to differ from that observed in human patients.     

Humoral response (IgG) of goats experimentally infected with Fasciola hepatica against cysteine proteinases of adult fluke

The use of cysteine proteinases from Fasciola hepatica adult flukes for the serodiagnosis of caprine fasciolosis by means of an indirect ELISA test was studied. Two proteolytic fractions from adult fluke homogenates, with apparent molecular weights of 28 and 34 kDa (P28 and P34 respectively), were characterised as cysteine proteinases using azocasein assays and gelatin gel analysis. Both P28 and P34 fractions were electroluted and used as antigens in two different indirect ELISA tests. Serum IgG levels against P28 and P34 in goats given an experimental primary infection with 200 metacercariae or in goats given two experimental infections with 200 metacercariae were determined and compared with those observed in an uninfected control group. ELISA tests using both cysteine proteases showed a rapid and consistent detection of specific IgG in all experimentally infected goats. The IgG response to P28 was the first to be detected as early as 2-3 weeks post-infection and remained elevated throughout the experiment. The response to P34 was detected later (4-6 wpi) and disappeared in some animals at 18 wpi, while flukes were still present in the bile ducts. No significant differences were observed between the anti-P28 and anti-P34 IgG responses between animals receiving a primary or a challenge infection. The results of our study, although preliminary, are promising since the P28 ELISA described here may be a reliable method for the immunodiagnosis of F. hepatica infection in goats.     

SDS-PAGE and Western blot of urinary proteins in dogs with leishmaniasis

Canine leishmaniasis is an endemic disease in the Mediterranean area caused by the protozoan Leishmania infantum, which usually produces renal failure. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and Western blot using antibodies to IgG and IgA from dogs were carried out in the urine of 22 dogs with leishmaniasis diagnosed by ELISA and confirmed by PCR, and 20 healthy dogs. The results were compared to renal function laboratory tests and to those from a histopathological study of the kidneys from sick animals that died naturally or were euthanized. Five different bands with molecular weights ranging from 10 to 110 kDa were obtained from the electrophoresis of the urine of healthy dogs. 33.5% of total proteins corresponded to low molecular weight proteins and the other proteins had middle and high molecular weights. However, in the group with leishmaniasis, a maximum of 11 different bands with molecular weights ranging from 10 kDa to 150 kDa were displayed in the electrophoresis of the urine. The urine electrophoretic pattern in the sick dogs was classified as mixed (proteins with high and low molecular weights) because low molecular weight proteins made up 57.9% and the rest of the proteins had middle and high molecular weights. In Western blot, none of the healthy dogs showed excretion of IgG and/or IgA, whereas IgG and IgA were detected in the Western blot of urine of 68% and 55% respectively of dogs with leishmaniasis. The results obtained in the leishmaniasis group agreed with glomerular and tubular damage, which were confirmed by the histopathological findings.     

Chemical restraint of peccaries with tiletamine/zolazepam and xylazine or tiletamine/zolazepam and butorphanol

Objective To evaluate the use of a combination of tiletamine/zolazepam and xylazine (TZX) in collared and white‐lipped peccaries and to compare its efficacy as an anesthetic technique with that of tiletamine/zolazepam and butorphanol (TZB). Study design Prospective experimental trial. Animals Seven white‐lipped peccaries (Tayassu pecari) (four females and three males) and four collared peccaries (Tayasu tajacu) (two males and two females). Methods Animal immobilization was attempted with TZX and TZB (IM) on two different occasions. Heart and respiratory rates (HR, RR), rectal temperature (RT), sedation, muscle relaxation, posture, auditory response and analgesia were evaluated every 15 minutes during immobilization. Induction, anesthesia, standing and walking time were determined after drug administration. Results Doses for white‐lipped peccaries were 1.23 ± 0.26 mg kg^?1 (mean ± SD) of TZ and 1.23 ± 0.26 mg kg^?1 of X, and 1.46 ± 0.09 mg kg^?1 of TZ and 0.14 ± 0.008 mg kg^?1 of B; doses for collared peccaries were 1.51 ± 0.29 mg kg^?1 of TZ and 1.51 ± 0.29 mg kg^?1 of X and 1.68 ± 0.02 mg kg^?1 of TZ and 0.17 ± 0.002 mg kg^?1 of B. In white‐lipped peccaries, both drug combinations provided a smooth induction and good immobilization for more than an hour. Anesthesia and standing times were significantly longer in animals given TZB, whereas walking time was significantly longer in animals given TZX. A significant decrease in HR was observed with both treatments. Respiratory rate decreased significantly with TZX, but the rate remained higher than with TZB. Induction and recovery quality in white‐lipped peccaries was better with TZB than with TZX. Neither protocol provided adequate immobilization in collared peccaries. Conclusion and clinical relevance At the doses described, TZB is effective in providing a long period of immobilization, whereas TZX is adequate for short to medium immobilization in white‐lipped peccaries. Neither drug combination was effective in collared peccaries at the doses given.     

Fine structure of Henneguya pilosa sp. n. (Myxozoa: Myxosporea), parasite of Serrasalmus altuvei (Characidae), in Brazil

Henneguya pilosa sp. n., a new species of myxosporean from the gill filaments of the white piranha, Serrasalmus altuvei Ramirez, 1965 (Characidae), a freshwater teleost fish collected in the Zoological Garden of the city of Teresina (Piauí), Brazil, is described from light and transmission electron microscope observations. This myxosporean produced small plasmodia (up to 0.2 mm in diameter), each one containing all life-cycle stages of the parasite, including numerous spores. The spores, laterally compressed, averaged 54.2 (52.3-56.0) microm in total length and consisted of two unequal valves adhering together along the suture line and two caudal processes. The spore body measured 21.1 (20.0-23.1) microm in length, 5.9 (5.5-6.3) microm in width, and 2.2 (1.9-2.6) microm in thickness. The two equal ellipsoidal polar capsules of 7.4 (7.1-7.6) microm long and 1.2 (1.0-1.3) microm wide possessed a polar filament with 11-12 (rarely 13) turns. All surfaces of the spores were covered with a tightly adherent complex network of numerous densely ramified granulo-fibrillar masses, the longest measuring 1.5 microm long, observed around the caudal processes. The prevalence of infection was 30%. The taxonomic affinities of this parasite with other of the same genus in freshwater South American fish species are discussed.     

Component population study of Acanthocephalus tumescens (Acanthocephala) in fishes from Lake Moreno,Argentina

Seasonal samples of all fish species from Lake Moreno were taken in order to determine the presence of paratenia, to evaluate the status of the hosts and to characterise the transmission of Acanthocephalus tumescens (von Linstow, 1896) at the component population level. Prevalence, mean abundance, mean intensity, numbers of gravid females, relative abundance of the different fish species, relative output of eggs and relative flow rates for each host species were computed. Acanthocephalus tumescens showed low host specificity, successfully parasitizing six out of eight fish species present in the lake. No paratenic infection was registered. If prevalence, mean abundance, and number of gravid females are considered, host species can be placed in a continuum from the most to least suitable as follows: Galaxias platei Steindachner, Diplomystes viedmensis (Mac Donagh), Oncorhynchus mykiss (Walbaum), Salvelinus fontinalis (Mitchill), Percichthys trucha (Cuvier et Valenciennes) and Galaxias maculatus (Jenyns). However, when parasite flow rates and egg output were calculated, including relative abundance of each fish species, the continuum was rearranged as follows: P. trucha, O. mykiss, G. platei / G. maculatus, S. fontinalis and D. viedmensis. The first four species would be the main contributors to the population of A. tumescens in this lake, P. trucha being the major one. Different regulatory and non-regulatory mechanisms are suggested.     

Effect of rice growth stages and silicon on sheath blight development

ABSTRACT The objective of this study was to determine the effect of silicon (Si) and rice growth stages on tissue susceptibility to sheath blight (Rhizoctonia solani Kühn) under controlled conditions. Rice plants (cv. Rio Formoso) were grown in pots containing low-Si soil amended with Si at 0, 0.48, 0.96, 1.44, and 1.92 g pot(-1) and inoculated with R. solani at the following days after emergence: 45 (four-leaf stage), 65 (eight-leaf stage), 85 (tillering), 117 (booting), and 130 (panicle exsertion). For plants inoculated with R. solani at all growth stages, Si concentration in straw increased as rate of Si increased from 0 to 1.92 g pot(-1). Concentration of calcium in the straw did not differ among plant growth stages. Although incubation period was not affected by the amount of Si added to the soil, this variable was shorter at booting and panicle exsertion stages. As the rates of Si increased in the soil, the total number of sheath blight lesions on sheaths and total area under the relative lesion extension curve decreased at all plant growth stages. The severity of sheath blight was lower at booting and panicle exsertion stages as the rates of Si increased in the soil. In general, plants grown in Si-nonamended pots and inoculated with R. solani were more vulnerable to infection at all growth stages, but especially at 45 days after emergence. Plant dry weights for inoculated plants increased as the Si rates increased from 0 to 1.92 g pot(-1). The greatest dry weight increases occurred for plants inoculated at booting and panicle exsertion stages. Si fertilization is a promising method for controlling sheath blight in areas where soil is Si deficient and when cultivars that exhibit an acceptable level of resistance to sheath blight are not available for commercial use.     

Evaluation of the local analgesic effect of ketamine in the palmar digital nerve block at the base of the proximal sesamoid (abaxial sesamoid block) in horses

OBJECTIVE: To evaluate the local analgesic effect of ketamine in a palmar digital nerve block at the base of the proximal sesamoid (abaxial sesamoid block) in horses. ANIMALS: 36 mature healthy Andalusian horses. PROCEDURE: Horses were randomly assigned to 4 groups of 9 horses each and received an abaxial sesamoid block in a randomly chosen forelimb with 1 of the following: saline (0.9% NaCl) solution, 1% ketamine solution, 2% ketamine solution, or 3% ketamine solution. To determine analgesia, the radiant heat lamp-hoof withdrawal model was used as a noxious thermal stimulus. Before each nerve block, baseline hoof withdrawal reflex latency (HWRL, time between lamp illumination and withdrawal of the hoof) was determined; after the nerve block, local analgesic effects were determined by measuring HWRL at 2 and 5 minutes after injection and then every 5 minutes for a total period of 1 hour. RESULTS: Significant differences in HWRL were found between baseline values and values at 2 to 15 minutes following a nerve block with ketamine. Significant differences were found between HWRL values at every time point from 2 to 10 minutes following a nerve block with saline solution, compared with 1 or 2% ketamine solution. Similarly, significant differences were found between HWRL values at every time point from 2 to 15 minutes following a nerve block with saline solution, compared with 3% ketamine solution. CONCLUSIONS AND CLINICAL RELEVANCE: Abaxial sesamoid block with ketamine ensures adequate analgesia in horses with an onset of action of 2 minutes and a maximal duration of action of 15 minutes.