Culture system and long-term storage of culture media in the in vitro production of bovine embryos

The optimum culture system for in vitro matured and fertilised oocytes still remains to be clarified. Culture media (CM) for mammalian embryos are routinely prepared fresh for use and preserved under refrigeration during one or two weeks. The purposes of this work were (1) to compare the efficiency of a synthetic oviduct fluid (SOF) with two different bovine serum albumin (BSA) concentrations (3 and 8 g/L) for the in vitro production of bovine blastocysts, (2) to test the effect of timing on adding fetal calf serum (FCS) to the SOF, and (3) to evaluate the effects on bovine embryo development of freezing and lyophilisation as procedures for preserving the SOF. Supplementation of SOF with 3 g/L BSA increased Day-7 blastocyst expansion rates (18.3 ± 1.6 vs. 14.4 ± 0.7; P < 0.05), although no differences in hatching rates were found. Addition of FCS to SOFaa (SOF with amino acids) medium supplemented with sodium citrate (SOFaaci) at 48 and at 72 h post-insemination (PI) allowed obtaining higher Day-6 embryo development rates than when FCS was added at 18 or 96 h PI (Day-6 morulae + blastocyst rate: 30.0 ± 1.1, 40.8 ± 1.1, 43.9 ± 2.3 and 39.3 ± 0.5 for FCS addition at 18, 48, 72 and 96 h, respectively). Hatching rates were significantly improved when serum was added at 72 h PI. Finally, both refrigeration and lyophilisation appeared as useful cryopreservation procedures for SOFaaci, although a significant loss of its ability to support embryo development, compared to the control fresh culture medium, was observed.     

Ultrasound-guided modified subcostal transversus abdominis plane block and influence of recumbency position on dye spread in equine cadavers

ObjectiveTo describe and evaluate an ultrasound-guided modified subcostal approach for the transversus abdominis plane (TAP) block in horse cadavers in lateral or dorsal recumbency.Study designProspective, experimental cadaveric study.AnimalsStudy of one preserved foal and eight fresh adult horse cadavers.MethodsThe lateral and ventral abdominal wall of a preserved cadaver was dissected to identify the muscles and nerves. A unilateral standard TAP block technique was performed (60 mL of methylene blue dye–bupivacaine) on a fresh cadaver in right lateral recumbency. A modified subcostal technique was performed on the opposite side using a linear ultrasound transducer and in-plane approach. Injection points (two 30 mL dye) were at the level of the TAP (between the rectus abdominis and transversus abdominis muscles and ventral to the cutaneous trunci muscle) perpendicular to: 1) the mid-point between the xiphoid cartilage and umbilical scar; and 2) at a point between the caudal and middle thirds of the abdomen measured from the first injection point to the umbilical scar. The modified subcostal approach was performed in seven additional cadavers in both hemiabdomens, with three cadavers in lateral and four cadavers in dorsal recumbency. Ultrasound guidance was used with all injections.ResultsThe standard approach stained the sixteenth to eighteenth thoracic nerves (T16–T18). The modified subcostal approach performed in lateral recumbency provided greater spread (T9–T17) than dorsal recumbency (T12–T18) (p = 0.016).Conclusions and clinical relevanceThe modified subcostal TAP approach resulted in extensive staining exceeding the standard approach. The nerves stained are consistent with production of ventral abdominal wall anesthesia in horses. Clinical studies are needed to verify these findings.     

Osteomalacia as a result of phosphorus deficiency in beef cattle grazing subtropical native pastures in Uruguay

We investigated 2 outbreaks of osteomalacia as a result of phosphorus (P) deficiency in herds of lactating beef cows grazing subtropical native pastures in Uruguay. Cows exhibited pica, difficulty to stand and walk, rib fractures, and body weight loss even with adequate forage availability. Osteopenia and severe osteomalacia were observed on gross and histologic examination. The concentrations of bicarbonate-extractable P in soil (4.0, 4.1 mg P/kg), total P in pasture (0.9, 1.1 g P/kg), inorganic P in serum (1.0, 0.71 mmol P/L), and P in bone (73 mg P/mL) were all low. Although injectable and mineral salt supplements provided additional P in both outbreaks, these supplementary amounts were insufficient to prevent P deficiency. The P ingested by the cows from the pasture and supplements would have provided 20–55% of their daily P requirements of ~21 g P/d. Osteomalacia occurred in cattle at the 2 ranches as a result of severe P deficiency in the soil and forage, and inadequate P supplementation. Following diagnosis, control of P deficiency in beef cattle requires estimation of the amount of pasture P ingested and provision of sufficient additional supplementary P to meet the animals’ requirements.     

Are complex carcinoma of the feline mammary gland and other invasive mammary carcinoma identical tumours? Comparison of clinicopathologic features,DNA ploidy and follow up

Feline mammary carcinomas are known for their unfavourable prognosis due to a strong tendency to local recurrence and metastasis. We studied 73 spontaneous primary mammary carcinomas and identified eight cases presenting a biphasic nature, with neoplastic epithelial and myoepithelial cells (complex carcinoma). These cases presented histopathologic features associated with a better prognosis; they were also associated with higher overall survival and disease-free survival rates compared to other common invasive mammary carcinomas of non-specified type. Complex carcinoma appears to be a low-grade malignancy.     

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.     

Disseminated pythiosis in three horses

Three cases of equine subcutaneous pythiosis with dissemination to the internal organs were investigated. The subcutaneous lesions were observed on the mammary gland, nostrils and limbs of the infected horses. Histopathological analysis of the infected tissues revealed a strong eosinophilic reaction, with macrophages, mast cells and giant cells. Sparsely septated hyphal filaments of 4-6 microm diameter were identified in the center of the eosinophilic areas. Specific fluorescent antibody against Pythium insidiosum confirmed the hyphae in the infected tissues in three examined horses. In one of the three cases, the DNA sequences amplified from the infected subcutaneous tissues and internal organs, revealed that P. insidiosum's 18S SSU rDNA amplicons shared 100% identity with those sequences deposit in GenBank. This is the first report confirming by immunochemical and genetic techniques that P. insidiosum can disseminated from superficial to deep structures.     

Larvicidal effects of several chemicals on Strongyloides infective larvae

The larvicidal effects of 11 anthelmintics, 7 pesticides and 4 disinfectants were evaluated with infective larvae of Strongyloides papillosus (SPL) and Strongyloides venezuelensis (SVZ). The lethal concentrations against SPL and SVZ were found to be similar. Three chemicals (dichlorvos, levamisole and trichlorfon) showed highest larvicidal effects. The 50% lethal concentration (LC(50)) values for the three compounds against SPL larvae were 0.08, 0.24, and 0.59 ppm, respectively.     

Retrospective study of 156 atrial septal defects in dogs and cats (2001-2005)

Atrial septal defect (ASD) is a common congenital heart disease (CHD) in humans, but considered relatively rare in veterinary medicine. However, modern echocardiographic and Doppler techniques currently offer a good view of the morphology of the interatrial septum, thus facilitating earlier detection of ASD in awake animals. In this context, we carried out a retrospective study of cases of recently diagnosed ASD in dogs and cats at the Cardiology Unit of Alfort (2001-2005) using echocardiography combined with colour Doppler mode. ASD was diagnosed in 156 animals and represented 37.7% of all canine and feline CHDs (n = 414). ASD was the most common CHD after mitral dysplasia in both species. Boxer and Domestic shorthair were the most common canine and feline breeds affected. Most defects (98.7%) were secundum-type ASD, without clinical signs in 73.7% of cases. The most common clinical signs included systolic murmur heard over the left heart base (20.2%), exercise intolerance (7.0%), syncope (5.3%), dyspnoea (2.6%) and cough (2.6%). Animals that presented a systolic heart murmur over the left base had a significantly larger ASD than others (P < 0.05). These data suggest that the incidence of ASD is higher than previously assumed. ASD should be suspected, for example, in instances of left basal systolic heart murmur, although its clinical and haemodynamic consequences are usually minor.     

Ocular lesions associated with Trypanosoma evansi in experimentally infected goats

Ocular lesions associated with Trypanosoma spp. infection have been described in man and many animal species. However, loss of vision has not been demonstrated in humans presenting Chagas disease or in animals affected by different trypanosome species. In order to assess the possible ocular disorders caused by Trypanosoma evansi infection, six goats were inoculated with 1 x 10(5) T. evansi and maintained for 12 months and four goats were used as control. The inoculated animals became positive at serological and parasitological tests at 1-month post-inoculation and showed a subclinical course of the disease. Unilateral superficial corneal ulceration and retinochoroiditis were observed in two inoculated animals. Data from ocular neurologic examination and electroretinography showed no significant differences between inoculated and non-inoculated goats. It could be concluded that Trypanosoma evansi can produce ocular lesion but without apparent loss of vision in goats.