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.     

Non-bioequivalence of various trademarks of enrofloxacin and Baytril in cows

Including Baytril, in various parts of the world many commercial preparations of enrofloxacin for parenteral administration are being employed for the treatment of bacterial diseases in cows. To optimize clinical responses and to minimize development of bacterial resistance to this agent, the copied pharmaceutical preparations must comply with some key pharmacokinetic features when bioequivalence studies are performed. To assess whether or not there was bioequivalence among nine commercial preparations of enrofloxacin and the original one, a controlled pharmacokinetic study was carried out. These was done utilizing the microbiological agar-diffusion test as quantitative/qualitative analytical method. A non-compartmental model defined kinetic variables. Results for Baytril revealed that maximal serum concentration (Csmax) was only matched by one preparation while area under the curve (AUC) of the serum concentration/activity of enrofloxacin and metabolites in time was not matched by any preparation. Time to Csmax (Tmax), elimination half-life, and shape of the time-serum concentrations of enrofloxacin profiles obtained for the nine generic preparations differ significantly somehow from the corresponding data obtained for the reference enrofloxacin. The need for studies to demonstrate bioequivalence becomes mandatory if similar preparations of enrofloxacin become commercially available. Enrofloxacin should be used selectively and cautiously to limit development of bacterial resistance. Non-bioequivalence of relevant pharmacokinetic values, such as Csmax and bioavailability (AUC) would facilitate development of bacterial resistance and limit the useful life span of this antibacterial agent.     

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.     

Pharmacokinetics of an injectable long‐acting parenteral formulation of doxycycline hyclate in pigs

Based on its ideal PK/PD ratios, doxycycline hyclate (DOX‐h), a time‐dependant antibacterial, is ideally expected to achieve sustained plasma drug concentrations at or slightly above the MIC level for as long as possible between dosing intervals. Pursuing this end, a poloxamer‐based matrix was used to produce a 10% long‐acting injectable preparation (DOX‐h‐LA) and its serum concentrations vs. time profile investigated after its injection to pigs in the pericaudal s.c. by parallel design. Results were compared with the forced oral bolus dose and i.v. pharmacokinetics of DOX‐h. For this study, 12 recently weaned pigs per group were included in this trial, and a dose of 20 mg/kg was injected in all cases. DOX‐h‐LA showed the greatest values for bioavailability (115.38%); maximum serum concentration (Cmax) value was 1.5 ± 0.2 with a time to reach Cmax of 3.41 ± 0.04 h and an elimination rate constant of 70.93 ± 0.87 h. Considering minimum effective serum concentration of 0.5 μg/mL, a dose interval of at least 5 days can be achieved for DOX‐h‐LA, whereas p.o. and i.v. dosing of DOX‐h may only last 11 and 15 h, respectively. Pigs were slaughtered on day 30 after this trial, and no visible remnants of the preparation were detected neither fibrosis was observed after a thorough macroscopic and histopathological analysis.     

Viability of Extended Goat Semen Stored at Refrigerated Condition

Due to the small volume of goat semen ejaculate, just a few doses of goat semen were produced when the sperm concentration is 100 × 10^/mL. The study was aimed to determine the viability of extended goat semen at refrigerated condition at 5 ℃ using varying sperm concentrations and evaluated if sperm concentration lower than 100 × 10^6/mL would affect the motility, viability and sperm morphology at refrigerated condition. Using an artificial vagina, ejaculated goat semen was collected from goat semen donor aged 1.5 year. Physical evaluation of the collected semen showed an average volume of 0.54 mL, mean pH of 6.8 and a milky white color with thick consistency indicative of high concentration. Fresh goat semen had an initial average of 76% with an average initial sperm concentration of 128 × 10^/mL. The semen was divided into four treatments： sperm concentration of 100× 10^/mL, 75 × 10^/mL, 50 × 10^/mL and 25 × 10^/mL, and were stored at 5 ℃ for a period of 10 d. The semen evaluation was performed for each of the four treatments every other day. Results showed that the sperm concentration of spermatozoa affected the duration of storage based on the sperm motility percentage, viability and morphology of spermatozoa. The extended goat semen with sperm concentration of 25 and 50 million sperm/mL is optimal for storage within 6 d that gave satisfactory percentage motile, viable and morphologically normal spermatozoa.     

Susceptibility and effectiveness of vesicular-arbuscular mycorrhizae in wheat cultivars under different growing conditions

Summary Wheat cultivars assumed to be non-susceptible to vesicular-arbuscular (VA) mycorrhizae became colonized, and this effect persisted under different growth conditions. Colonization of all cultivars was similar regardless of the amount of inoculum and the time interval of inoculation. Different plant growth temperatures and the support given by the culture media, inoculation with different endophytes, and inoculation with sterilized and unsterilized spores affected VA colonization levels, although the level of colonization reached in cv. Champlein was similar to that reached in cv. 7-Cerros under each condition. VA mycorrhizal colonization was also affected by different plant growth conditions. After VA reinoculation, the plant dry weight of Castan and 7-Cerros increased, but not Negrillo and Champlein cultivars. VA mycorrhizae increased the shoot dry weight of 7-Cerros only, but not of Champlein, when grown at 35/24°C, and had no effect on the dry weight of either cultivar grown at 18/12°C and 42/24°C. Inoculation with Glomus mosseae increased the dry weight of the cultivars more than inoculation with G. fasciculatum or G. agregatum. The effect on the plant dry weight was greater in plants grown in soil than in sand/vermiculite pots. Inoculation with sterilized and unsterilized spores of G. mosseae, either in soil pots or in sand/vermiculite tubes, did not increase the plant dry weight. Our results indicate that there was no close relationship between the level of root colonization and the effect on plant growth. The effects of accompanying microorganisms in the VA inoculum on VA mycorrhizal symbiosis are discussed.     

Endolithic blue-green algae in the dry valleys: primary producers in the antarctic desert ecosystem

Endolithic unicellular blue-green algae occur under the surface of orthoquartzite rocks in the dry valleys of southern Victoria Land, Antarctica. This report of primary producers in the Antarctic desert ecosystem suggests that, in future efforts to detect life in extraterrestrial (for example, martian) environments, scientists should consider the possible existence of endolithic life forms.     

Effect of a new thermal treatment in combination with saprobic fungal incubation on the phytotoxicity level of alperujo

Byproducts generated from food industries, such as olive oil mills, have been studied to decrease harmful pollution and their environmental consequences. In this work, a new thermal pretreatment and saprobic fungal incubation to detoxify alperujo (two-phase olive mill waste) have been evaluated in view of its use as fertilizer in agriculture. The sequential use of both methods simplifies the thermal conditions and incubation times of the fungal treatment. Optimization of the thermal treatment from 150 to 170 °C for 45 and 15 min, respectively, reduced the incubation time with Coriolpsis rigida from 20 to 10 weeks needed to reduce phytotoxic effects on tomato plants. Therefore, the combination of thermal and biological treatments will allow the development of the potential benefits of alperujo to improve nutrients in agricultural soil.