Study of the nervous tissue development in rainbow trout (Oncorhynchus mykiss) embryos treated with oxytetracycline

The purpose of this study was to discover whether the use of different doses of oxytetracycline causes any alteration in the development of nervous tissue in rainbow trout embryos. Five thousands eggs of females rainbow trout were divided into five groups. One group acted as control and the other four were administered with one of four doses of oxytetracycline, 0.025, 0.050, 0.100, or 0.201 microM, at the moment of fertilization. The eggs were incubated under pisciculture conditions to just before being ready to spring off. From the 10th day, 10-egg samples were taken regularly and fixed. Five were processed for histological techniques and stained with haematoxylin and eosin, cresyl fast violet and silver, the other five were homogenized for antibiotic detection. Histological alterations appeared in 37-day-old embryos, with an abnormal migration of the neuroblasts to the marginal layer of the neural cord, and alterations in the development of the lens and eye layers. Some embryos showed abnormal curvature of the spinal cord but these changes were not statistically significant.     

Successful transcatheter retrieval of embolized Amplatz Canine Duct Occluders in two dogs

Transcatheter occlusion of patent ductus arteriosus (PDA) using an Amplatz® Canine Duct Occluder (ACDO) is routinely performed in dogs. Pulmonary embolization of the device is a rarely reported complication of this procedure. We report here the first two cases of successful transcatheter retrieval of an embolized ACDO. An 8-month-old, 21-kg, German shepherd (case 1) was referred for pulmonary edema secondary to a large PDA with left-to-right shunting. After medical stabilization, an emergency procedure for PDA closure was recommended. An 8-mm ACDO was deployed under general anesthesia. The device was released after confirming its stability with a gentle tug test but migrated into the pulmonary trunk. A 10-mm ACDO was subsequently successfully deployed and released. Vascular access was then obtained from the right jugular vein so that a vascular snare could be used to capture the ACDO waist and pull it back toward the right ventricle and then the right atrium. It was then removed through the jugular vein. The dog recovered uneventfully and was discharged after confirmation of complete ductal occlusion. The same complication occurred in a second case (case 2, asymptomatic 6-month-old, 7.9-kg, cocker spaniel), and a similar procedure was used to successfully retrieve the embolized device. Both dogs are still doing well 9 and 21 months, respectively, after the procedure. These cases illustrate that transcatheter retrieval of ACDO devices embolized in the pulmonary artery using vascular snares is technically feasible in the dog.     

Effects of IGF‐1 on In Vitro Culture of Bovine Preantral Follicles are Dose‐Dependent

This study aimed at assessing the effect of different concentrations of the growth factor similar to insulin 1 (IGF‐1) in the development, survival and ultrastructure of the bovine preantral follicles cultured in situ. Fragments of bovine ovarian cortical tissue were cultured during 1 and 7 days in 1 ml of α‐MEM⁺, supplemented with different concentrations of human recombinant IGF‐1 (0, 30, 70 and 100 ng/ml), in an incubator at 37°C and 5% of CO₂ in 24‐well plates with total replacement of the medium every 2 days. Non‐cultured ovarian fragments (control) and ovarian fragments cultured during 1 and 7 days were processed for classic histology, mechanical isolation and electron transmission microscopy (ETM). Parameters such as normality, viability, activation, development, diameter and ultrastructure were evaluated. All statistical analyses were carried out using sas Version 9.2. The results showed that the percentage of follicles morphologically normal in the IGF‐1 30 ng/ml treatment was similar to the fresh control (p > 0.05) both on the day 1 and on the day 7 of in vitro culture. In the viability analysis, the cultured treatments maintained the percentage of viable follicles during the entire culture period (p > 0.05). After 7 days of culture, the IGF‐1 30 ng/ml treatment showed higher percentages of developing follicles (48.33%) than those of the fresh control (22.22%) and the cultured treatments (p < 0.05). Also, after 7 days of culture, IGF‐1 30 ng/ml presented a higher follicular diameter when compared to the control and other concentrations of IGF‐1 tested. Ultrastructurally, the non‐cultured control and IGF‐1 30 ng/ml, after 7 days of culture, showed conserved oocytes, nuclei and organelles. Hence, it is concluded that IGF‐1 30 ng/ml was the most efficient concentration for the development of bovine preantral follicles cultured in vitro.     

Status of Macrophomina phaseolina on Strawberry in California and Preliminary Characterization of the Pathogen

ABSTRACT

Macrophomina crown and root rot has become a significant soil-borne disease issue in California. For many locations in the state, the disease is associated with fields that are no longer pre-plant, flat field fumigated with methyl bromide + chloropicrin. Inoculation experiments indicated that some differences in strawberry cultivar susceptibility to Macrophomina phaseolina were seen a short time after the inoculation, but as disease progressed such differences did not persist. Preliminary characterization studies of Macrophomina phaseolina isolates from strawberry indicated that such isolates may have a host preference for strawberry. Macrophomina phaseolina isolates from watermelon, thyme, and apple failed to cause disease in strawberry. Five cover crop species, which can be rotated with strawberry, did not develop disease when inoculated with strawberry isolates. In preliminary analysis using simple sequence repeat markers, isolates obtained from strawberry formed a separate group compared to isolates recovered from other known Macrophomina phaseolina hosts.     

Progress in the development of Fasciola hepatica vaccine using recombinant fatty acid binding protein with the adjuvant adaptation system ADAD

Fatty acid binding proteins (FABP) have been designed as a potential vaccine against fasciolosis. In this work, the immunoprophylaxis of the recombinant Fh15 FABP from F. hepatica (Fh15) in adjuvant/immunomodulator ADAD system was evaluated using mice and sheep challenged with F. hepatica. The ADAD system combines the Fh15 antigen with an immunomodulator (hydroalcoholic extract of Polypodium leucotomos; PAL) and/or an adjuvant (saponins of Quillaja saponaria; Qs) in a water/oil emulsion (30/70) with a non-mineral oil (Montanide). All the infected control mice died by 41-48 days post-infection. The mice vaccinated with ADAD only with PAL+Fh15 present a survival rate of 40-50% and those vaccinated with ADAD containing PAL+Qs+Fh15 had a survival rate of 50-62.5%. IgG1 antibodies were lower in surviving mice in comparison with non-surviving mice. The sheep vaccinated with ADAD PAL+Qs+Fh15 showed lower fluke recovery (43%), less hepatic lesions and higher post-infection daily weight gain than F. hepatica infected control animals. Thus, the ADAD system using recombinant fatty acid binding proteins from F. hepatica could be a good option to develop vaccines against F. hepatica.     

Effect of sperm pretreatment with glutathione and membrane destabilizing agents lysolecithin and Triton X‐100, on the efficiency of bovine intracytoplasmic sperm injection

Intracytoplasmic sperm injection (ICSI) is an assisted reproduction tool with several applications. Its effectiveness in bovines is lower than that in other species, mainly because of difficulties in the decondensation of the sperm nucleus after injection, and the presence of the acrosome and the plasma membrane which remain intact in this procedure. In this study, we assessed the effect of lysolecithin (LL) and Triton X‐100 (TX), in combination with glutathione (GSH) as sperm pretreatments prior to ICSI. The GSH‐LL and GSH‐TX groups showed 0% of spermatozoa with intact membrane (SYBR 14+/PI), in comparison with the control (63.3%) and GSH (65.7%) groups. The proportions of spermatozoa with damaged acrosome membrane in the GSH‐LL, GSH‐TX, GSH and control groups were 46%, 35.9%, 10.5% and 7.5%, respectively. Sperm chromatin decondensation analysis showed that the groups incubated for 3 hr with GSH presented greater decondensation (p < .05). Although fertilization was improved in all treatment groups evaluated, no differences were observed in the cleavage rate 72 hr after activation in the GSH (73.7%), GSH‐LL (80.2%) and GSH‐TX (77.8%) groups compared to the control (66.3%), neither in the blastocyst rate on day 8 (24.0%, 26.2%, 27.1% and 28.4% for the control, GSH, GSH‐LL and GSH‐TX groups, respectively). No differences were also observed in the total number of cells in all groups. In conclusion, although these sperm treatments promoted nuclear decondensation and induced plasma membrane disruption, these effects were not sufficient to improve bovine embryonic development after ICSI.     

Phenotypic Resistance to Disinfectants and Antibiotics in Methicillin‐Resistant Staphylococcus aureus Strains Isolated from Pigs

The aim of this research was to study the phenotypic resistances to disinfectants and antibiotics in strains of methicillin‐resistant Staphylococcus aureus (MRSA) obtained from Canary black pigs. Analyses were performed on 54 strains of MRSA, isolated in Canary black pigs from the province of Tenerife (Spain); all of them carried the mecA gene. The strains were isolated by means of nasal swab samples of healthy pigs, collected under veterinarian supervision. Bactericidal activity of antiseptics and disinfectants was tested by means of the dilution–neutralization method. Susceptibility to the disinfectants glutaraldehyde, peracetic acid and silver nitrate was assessed, as well as to the antiseptics chlorhexidine, benzalkonium chloride and povidone iodine. Susceptibility to a wide array of antibiotics representing the main groups was determined by means of the disc diffusion method. All the strains demonstrated susceptibility to the disinfectants tested at the recommended concentration, and even to dilutions equal to or lesser than 1/16. The most effective antiseptic and disinfectant were, respectively, chlorhexidine and silver nitrate. With regard to the antibiotics, the strains proved to be multiresistant. All presented phenotypic resistance to the β‐lactam antibiotics ampicillin, penicillin and cefoxitin, as well as to numerous aminoglycosides, tetracycline and trimethoprim–sulfamethoxazole. It was also observed that 61.1% of the strains were carriers of plasmids. Our results underline that in the strains such as MRSA, which show multiple resistances to antibiotics, the antiseptics and disinfectants show great efficacy. Moreover, as other authors also suggest, for the treatment and prevention of infections caused by MRSA, the use of β‐lactam and aminoglycoside antibiotics may be less effective.     

Vergleichende histologische Studie des weiblichen Geschlechtsapparates beim Lama (Lama guanicoe glama). II. Eileiter, Uterus, Cervix, Vagina

The histological characteristics of the oviduct, uterus, cervix and vagina of the llama are described and compared with those of domestic mammals. There are differences between llama and domestic mammals in the type of placentation and the epithelium of the vagina, which seems to be a transitional epithelium.     

Effects of iron and organic matter on the porosity and structural stability of soil aggregates

Knowledge of the soil components controlling aggregate formation and stability is fundamental to the conservation of soil structure. In this work, the effects of Fe and organic matter (OM) on the porosity and structural stability of aggregates <4 mm of two cropped soils from Galicia (NW Spain) were determined. Porosity was estimated directly, by mercury intrusion porosimetry, and indirectly, from moisture characteristic and shrinkage curves. The three porosities obtained were similar and indicated that Soil 1, with the highest Fe and OM contents, had lower total porosity and a wider pore-size distribution than Soil 2. As regards structural stability, Hénin and Monnier's test, simulated rainfall and dispersion experiments, and determination of textural tensile strength all indicated Soil 1 to be the more stable soil. Oxidation of OM and selective extraction of Fe, which were most concentrated in the clay and silt fractions, indicated both these components to be important soil aggregants. It is suggested that the higher content of Soil 1 in Fe and OM is responsible for its greater stability.