Intermediate Filament Proteins Expression and Carbohydrate Moieties in Trophoblast and Decidual Cells of Mature Cat Placenta

The aim of this study was to characterize cytoskeletal intermediate filament proteins and glycoconjugates of syncytiotrophoblast, cytotrophoblast and decidual cells of feline endotheliochorial placenta. Samples from 12 normal pregnant female cats, after 45 ± 5 days of gestation, were obtained removing the uterine horns by hysterectomy. Sections were processed for routine observation and for immunohistochemistry using anticytokeratin, antivimentin and antidesmin antibodies. In addition, lectin histochemistry was performed using a panel of several biotinylated lectins to characterize glycosides expression profile. Cytotrophoblast and syncytiotrophoblast showed immunoreactivity only with acidic and basic cytokeratins. Decidual cells were only positive to vimentin, consistent with their origin from endometrial fibroblasts. Trophoblast expressed a broad population of glycans, highly exposing terminal N‐acetyl glucosamine residues and non‐sialylated galactose and N‐acetyl galactosamine oligomers. Oligosaccharides bound by Phaseolus vulgaris erythroagglutinin were the only highly branched N‐linked residues evidenced in cats, and they were restricted to the syncytium. Unlike results reported on humans, mice and rats on lectin affinity of decidual cells, sialid acids and complex N‐linked oligosaccharides were not demonstrated in cats. Glycosylation of proteins determines many of their final properties, thus becoming essential for the embryo‐maternal dialogue during implantation and placentation. Changes in glycosylation pattern have been related to pathological pregnancies in other species. Hence, the knowledge about glycosylation profile of the normal cat placenta may lead to a better understanding of both normal and pathological reproductive events.     

Biomechanical Properties of Skin and Wounds in Ehlers-Danlos Syndrome

The biomechanical properties of wounded and nonwounded skin were studied in three dogs and three cats affected with type I Ehlers-Danlos syndrome. Three nonaffected dogs and one nonaffected cat served as controls. Samples of wounded skin and adjacent normal skin were harvested at days 75, 138, 141, 144, 147, and 150. Samples were subjected to uniaxial tensile strength testing. Tensile strength, energy absorbed, and site of failure were recorded. In the dogs with Ehlers-Danlos syndrome, there was an increase in tensile strength in samples containing a scar over adjacent intact skin. In nonaffected dogs, affected cats and the nonaffected cat, the nonwounded skin samples had greater tensile strength. The energy absorbed by the skin samples during testing was highly correlated with tensile strength.     

Haematological and biochemical reference values for the koala (Phascolarctos cinereus)

Haematological and biochemical reference values were established from 45 clinically healthy koalas. Statistical analysis revealed no significant differences for sex and season of sampling. Immature koalas had significantly higher alkaline phosphatase and inorganic phosphate values, and significantly lower total protein concentrations due to low globulins values. Enzyme reference values tended to be wide and could limit their usefulness in detecting disease. In the reference values for leukocytes, neutrophils and lymphocytes, the inclusion of low values which were not actually seen may interfere with the detection of reduced levels due to disease.     

Different Doses of Porcine Luteinizing Hormone in Precocious Puberty Induction in Gilts

The use of hormonal protocols in puberty induction and synchronization of oestrus has lead to an increase in the efficiency of replacement gilts. The aim of this study was to evaluate different doses of porcine LH in precocious puberty induction and oestrus synchronization in a homogeneous group of gilts. Sixty-seven homogeneous prepubertal gilts (Camborough 22) at 137 +/- 4 days of age and 87 +/- 7 kg were treated with three different hormonal protocols: T1--600 UI of equine chorionic gonadotrophin (eCG; Novormon) and after a 72-h period 5 mg of LH (Lutropin); T2--600 UI of eCG and a 72-h period 2.5 mg of LH; T3--600 UI of eCG and a 72-h period 1.25 mg of LH. The ovaries were examined at slaughter, on day 6 after the hormonal treatment. There were no statistical differences (p > 0.05) between the different LH doses in the percentage of the detected oestrus (T1 = 42.85%; T2 = 60.87%, T3 = 52.18%), oestrus duration (T1 = 41.44 +/- 16.30 h; T2 = 48.57 +/- 16.29 h, T3 = 39.33 +/- 11.42 h), number of corpora lutea (T1 = 9.61 +/- 5.43; T2 = 9.86 +/- 3.32, T3 = 8.13 +/- 5.52) and percentage of animals presenting ovarian cystic degeneration (T1 = 33.33%; T2 = 39.13%, T3 = 39.13%). The T2 (2.5 mg of LH) presented the lowest dispersion (p < 0.05) of the LH-ovulation interval (T1 = 37.17 +/- 4.07 h; T2 = 38.26 +/- 2.84 h; T3 = 36.25 +/- 5.69 h). The LH dose reduction to 2.50 and 1.25 mg presented equal results with the recommended dose of 5.0 mg, and could be used in the precocious induction of oestrus in gilts. The 2.5-mg LH dose showed the lowest dispersion of ovulation and it can be used in fixed-time artificial insemination programmes.     

Survival analysis of dogs with advanced primary lung carcinoma treated by metronomic cyclophosphamide,piroxicam and thalidomide

Unresectable or metastatic (advanced) primary pulmonary carcinoma (PPC) represents a therapeutic challenge where surgery may be contraindicated and the therapeutic role of maximum‐tolerated dose (MTD) chemotherapy remains uncertain. This study was undertaken to explore the impact of metronomic chemotherapy (MC) in dogs with advanced PPC. Previously untreated dogs with advanced (T3 or N1 or M1) PPC, with complete staging work‐up and follow‐up data, receiving MC (comprising low‐dose cyclophosphamide, piroxicam and thalidomide), surgery, MTD chemotherapy or no oncologic treatment were eligible for inclusion. For all patients, time to progression (TTP) and survival time (ST) were evaluated. Quality‐of‐life (QoL) was only evaluated in patients receiving MC. To assess QoL, owners of dogs receiving MC were asked to complete a questionnaire before and during treatment. Ninety‐one dogs were included: 25 received MC, 36 were treated with surgery, 11 with MTD chemotherapy and 19 received no treatment. QoL was improved in dogs receiving MC. Median TTP was significantly longer in patients receiving MC (172 days) than patients undergoing surgery (87 days), receiving MTD chemotherapy (22 days), or no oncologic treatment (20 days). Median ST was similarly longer in patients receiving MC (139 days) than those undergoing surgery (92 days), MTD chemotherapy (61 days) and no oncologic treatment (60 days). In dogs with advanced PPC, MC achieved a measurable clinical benefit without significant risk or toxicity. This makes MC a potential alternative to other recognized management approaches.     

Influence de la densité et de la période de concurrence de Solanum nigrum L. sur la tomate de semis direct, en relation avec le désherbage

Effects of density and period of competition by Solanum nigrum L. on direct seeded tomatoes in relation to weed control The effects of density and period of competition from Solanum nigrum L. were measured in direct seeded tomatoes given weed control treatments currently used in south-east France. S. nigrum emerging after a diquat treatment at the 2–3 leaf stage of the crop and thinned to low densities (<12.8 plants ha^?1) at the 5–6 leaf stage of the crop caused significant yield loss if left to compete with the crop until harvest. Yield reduction was smaller if the same weed densities were present only until the onset of flowering. The regression curves of yield on weed density differed as annual climatic variations affected sowing date and plant growth; a comparison between years was made using the relation ‘crop yield × weed biomass/crop biomass’. Significant interactions between weed density and period of competition were found with yield of both green and red fruit. For late sown crops with low densities of S. nigrum two weed control treatments at the 5–6 leaf stage and at the onset of flowering were sufficient to prevent yield loss.     

Evaluation of a Burdizzo Castrator for Neutering of Dogs

A Burdizzo castrator was evaluated for the neutering of dogs. Histological and morphological changes of spermatic cells and peripheral serum testosterone after challenge with a GnRH-analogue (gonadorelin) were assessed. There was a control group (G1), a surgically castrated group (G2) and a Burdizzo group (G3) divided in two, G3a receiving two crunches in each spermatic cord and G3b receiving one crunch in each spermatic cord. Sixteen days after application of the Burdizzo blood samples were taken from the dogs at 30 min interval during 2 h; after the second sample the dogs were treated with 1 mug/kg body weight of gonadorelin i.v. The same protocol of gonadorelin challenge was performed in G1 and G2 dogs. The G2 dogs were surgically castrated after the second blood sample, before the gonadorelin treatment, and the G1 dogs after the last blood sample. The excised gonads were examined histologically, and sperm smears were prepared from the caudae epididymidis. The testes and plexus pampiniformis of the G1 and G2 dogs had a normal histological appearance, and they had morphologically normal epididymal sperm cells. In all G3 dogs, there was an acute fibrosis with an inflammatory reaction in the plexus pampiniformis. The testes from the G3a dogs showed diffuse areas of infarction and degeneration of the parenchyma. Similar but less diffuse lesions were seen in group 3b dogs. The deferent ducts from all G3 dogs showed vasitis and/or sperm granulomas. Azoospermia or sperm malformations were observed in the epididymal smears from the G3 dogs. Testosterone concentration in the G1 dogs increased after gonadorelin application (p < 0.0001). The G2 dogs had basal testosterone levels after castration (p < 0.001) and did not respond to gonadorelin. Groups 3a and b showed a slight but non-significant increase in testosterone concentration after gonadorelin challenge, supposedly due to the reduction of testicular blood flow and loss of testicular interstitial tissue.     

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.     

Bovine thyroglobulin gene polymorphisms and their association with sexual precocity in Guzerat bulls

Puberty is a stage of sexual development determined by the interaction of environmental factors and genetic mechanisms. Among them, thyroid function plays a key role in sexual development and spermatogenic function and is under the control of several genes, including the well‐described thyroglobulin gene (TG). Previous reports have shown genetic association between thyroid function and selected single nucleotide polymorphisms (SNPs) in taurine cattle. Therefore, the identification of genetic mechanisms involved in the regulation of this trait can assist with the selection for early pubertal bulls, thus improving genetic progress in livestock breeding. The aim of this study was to validate the association between TG SNPs and age at puberty in zebuine bulls. Three SNPs (rs110406764, rs109662686, rs109057985) were genotyped in 159 Guzerat animals using SEQUENOM technology. Results showed a significant association (p < .05) between the studied SNPs and puberty age, in agreement with our previous reports in a taurine breed. Interestingly, allele frequencies were different from those already reported, being GAT the most favourable allele for age at puberty in Guzerat (94.4 days lower). Overall, our findings corroborate previous reports and reinforce the importance of genetic influence in the regulation of sexual development and puberty through a thyroid pathway in zebuine cattle.     

Effects of stocking density on reared Siberian sturgeon (Acipenser baerii) larval growth,muscle development and fatty acids composition in a recirculating aquaculture system

This study evaluated the effects of rearing density on muscle growth and development in Siberian sturgeon (Acipenser baerii) larvae. Three different stocking densities were tested: low (LD, 30 larvae/l), mid (MD, 80 larvae/l) and high (HD, 150 larvae/l) in a recirculating aquaculture system. Larvae were sampled at hatching (T0), schooling (T1) and complete yolk‐sac absorption (T2) stage and were weighed and processed for muscle tissue histometrical analyses and for qualitative morphological study analyses; fatty acid profile was also determined by Gas Chromatography—Flame Ionization Detector analysis. Low‐density larvae presented a higher weight than MD or HD at T2 (p < 0.05). Histometrical analysis revealed that total muscle area was similar at T1 and T2, but higher than T0, while it was lower at HD at schooling (p < 0.05). The fatty acid profile revealed no differences between densities while, during development, there was a selective consumption: sparing or increasing of essential fatty acids to the detriment of their precursors. Our study suggests that lower densities appear to be more suitable to rear Siberian sturgeon in this particular stage of development. Indeed, larvae reared at the lower density were heavier and longer while larvae reared at the higher density showed lower muscle proliferation rate. As a consequence, LD larvae may exert an increase of potential growth at a mid‐long term.