Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.     

Larval traits of the Caribbean amphidromous goby Sicydium punctatum (Gobioidei: Sicydiinae) in Guadeloupe

Amphidromous fish are the biggest contributor to the diversity of fish communities in river systems of Caribbean islands. Among them, Sicydium punctatum Perugia, 1896, which is endemic to the West Indies, represents the vast majority of fish in these rivers. The otolith microstructure and the biometry of S. punctatum postlarvae from Guadeloupe were investigated to explore the dispersal modalities of this species through an appreciation of the growth pattern, the pelagic larval duration (PLD) and the size‐at‐recruitment. The study was made on one cohort of 83 recruited postlarvae, fished at the Capesterre River's mouth on 2 November 2011. The mean (±SD) size‐at‐recruitment of the postlarvae was 24.6 ± 1.3 mm (range of 20.5–28.1 mm, n = 83). We found a mean (±SD) PLD of 72.2 ± 10.5 days (range of 54–101.5 days, n = 67). The growth rate estimated from the otolith increments showed a globally decreasing pattern during the marine larval phase. Growth rates at the beginning of the larval stage were significantly different between hatching periods, suggesting a relation between the hatching period and the growth rate in S. punctatum. This study adds on to the general understanding of the life cycle of S. punctatum in Guadeloupe that will help implement strategies to manage amphidromous fish populations in the Caribbean region.     

Seed dormancy and germination in the summer annual Galeopsis speciosa

This study examined germination and dormancy in Galeopsis speciosa (Lamiaceae), a common summer annual weed in cold‐temperate areas. Seeds collected in southern Sweden were subjected to several experiments. The seeds were dormant at maturity. Seeds sown outdoors after collection produced a small number of seedlings that emerged early in the spring. After long cold stratification or stratification outdoors over two winters, the maximum germination was 40–50%; germination occurring over a wide range of temperatures. Warm stratification preceding cold stratification had no effect on germination, but repeated warm and cold periods seemed to promote germination. Gibberellic acid (GA) stimulated germination, but full germination was only achieved after more than 2 months of incubation at the most suitable temperature regime tested. Excised embryos grew and developed into normal seedlings. With these results, the species does not fit into the currently used system for seed dormancy classifications. The response to GA and the growth of excised embryos indicate non‐deep or intermediate physiological dormancy, but dormancy alleviation by stratification was not in line with the guiding principles for these classifications. Galeopsis speciosa has a strong dormancy that is sufficiently alleviated during the winter to allow germination of only part of a seed batch each year; hence a stepwise germination pattern occurs over a period of several years.     

Multi#ex—CAPS技术及其在番茄遗传鉴定中的应用

 ：应用Multiplex—PCR和CAPS(Cleaved amplified polymorphic sequence)原理研究建立了Mutiplex—CAPS技术。该技术包括：(1)在PCR扩增反应体系加入2对、3对或多对核苷酸引物，经30～40个扩增循环后得到每对引物的特异扩增片段；(2)应用同一种限制性内切酶消化这些不同引物经扩增后获得的特异片段；(3)琼脂糖凝胶电泳溴化乙锭染色检测酶切片段长度的多态性。利用该技术对番茄(Lycopersicon es—cdentum)TA517及其近等基因系的分析表明，Multiplex—CAPS能同时检测2个、3个或多个CAPS标记的多态性，其效果等同于每个CAPS标记的多态性的叠加，重复性好，分析效率高，降低成本数倍。     

Unilateral scrotal pyocele in ram caused by Staphylococcus capitis

A massive unilateral scrotal pyocele caused by Staphylococcus capitis in a 6‐year‐old ram is reported. Ultrasound examination of the right hemiscrotum showed an irregular hyperechoic mass in an anechoic fluid. A dense exudate was collected from the scrotum for microbiological analysis. Grossly, there was an extensive greenish purulent exudate and a completely atrophied right testis. Coagulase‐negative S. capitis was isolated in pure culture. To the authors' knowledge, this is the first report of genital infection by S. capitis in rams. This microorganism should be included in the differential diagnosis of ovine genital infections.     

Association between cattle herd Mycobacterium avium subsp. paratuberculosis (MAP) infection and infection of a hare population

Paratuberculosis has long been considered a disease of domestic and wild ruminants only. The known host range of Mycobacterium avium subsp. paratuberculosis (MAP) was recently extended to include non-ruminant wildlife species believed to be exposed to spillover of MAP from infected domestic cattle herds. The aim of the present study was to assess the association between cattle herd MAP infection pressure level and the infection level of a hare population in two dairy farms of southern Chile. Fifty hares from a herd A and 42 hares from herd B were captured and sampled for MAP culture. The results showed a statistically significant association between the cattle herds’ infection prevalence and the hare infection prevalence.