Comparison of efficiency of in vitro cloned sheep embryo production by conventional somatic cell nuclear transfer and handmade cloning technique

Conventional somatic cell nuclear transfer (SCNT) technique of in vitro production of cloned embryos involves use of costly and complicated micromanipulators. Handmade cloning (HMC) technique has been applied as efficient and cost‐effective alternative in many livestock species. The aim of the present study was to compare the efficiency of in vitro production and in vitro development of cloned sheep embryos by the two techniques. Cloned embryos were produced by conventional SCNT using micromanipulator apparatus and by HMC technique. Enucleation efficiency and efficiency of fusion with somatic cell (nucleus donor) were compared. Cleavage percentage was observed on day 2 of in vitro culture (IVC), and morula and blastocyst percentages were calculated on day 7 of IVC. Higher enucleation efficiency (96.98 ± 1.01 vs. 93.62 ± 1.03; p > .05) as well as fusion efficiency was obtained with HMC technique than with conventional SCNT (96.26 ± 1.34 vs. 92.63 ± 0.70, p < .05); 181 cloned sheep embryos were produced in vitro by conventional SCNT and 92 by HMC. Cleavage percentage observed on day 2 of in vitro culture was higher in HMC than SCNT (66.92 ± 3.72 vs. 55.97 ± 2.5, respectively, p < .05). Morula percentage obtained was higher in SCNT than HMC (44.12 ± 2.93 vs. 30.43 ± 6.79, respectively, p < .05), whereas blastocyst percentage obtained by HMC was higher (12.46 ± 4.96) than SCNT (5.31 ± 2.25; p > .05). It was inferred that HMC technique provides a cost‐effective and efficient method of in vitro production of cloned sheep embryos with a comparatively simpler technique with a possibility of automation. Efficiency of cloned embryo production could be improved further by propagating and standardizing this technique.     

Maximum limits of organic and inorganic mercury in fish feed

The relatively high levels of mercury found in fish feeds might form a fish health and food safety risk. The present study aims to establish sublethal toxic threshold levels in fish and assess feed‐fillet transfer of dietary mercury. Atlantic salmon (Salmo salar L.) parr were fed for 4 months on fish meal‐based diets supplemented with mercuric chloride (0, 0.1, 1, 10 or 100 mg Hg kg^?1 dry weight (DW)) or methylmercuric chloride (0, 0.1, 0.5, 5 or 10 mg MeHg kg^?1 DW). At the end of the experiment, dietary inorganic mercury mainly accumulated in intestine (80% of body burden) and assimilation was low (6%). In contrast, methylmercury readily accumulated in internal organs and muscle (80% of body burden) and had a relatively high assimilation (23%). Highest accumulation of dietary inorganic mercury was observed in the gut and kidney. Fish fed 10 mg Hg kg^?1 had an early (after 2 months) significant increase in renal metallothionein (MT) level and intestinal cell proliferation, followed by intestinal pathological conditions after 4 months of exposure. At 100 mg Hg kg^?1, intestinal and renal function were reduced as seen from the significantly reduced protein and glycogen digestibility and increased plasma creatinine levels. For dietary methylmercury (MeHg), highest accumulation was found in blood and muscle. Intestinal cell proliferation and liver MT significantly increased at 5 mg MeHg kg^?1 after 2 months of exposure. At the end of the experiment, blood haematology was significantly affected in fish fed 5 mg MeHg kg^?1 and these fish exceeded the current food safety limit for mercury. Tissue MT induction and intestinal cell proliferation appeared to be useful and quantifiable early indicators of toxic mercury exposures. Based on the absence of induction of these early biological markers such as MT and cell proliferation, nonobserved effect levels (NOELs) could be set to 0.5 mg Hg kg^?1 for dietary methylmercury and 1 mg Hg kg^?1 for inorganic mercury. Lowest observed effect levels (LOELs) levels could be set to 5 mg kg^?1 for methylmercury and 10 mg Hg kg^?1 for inorganic mercury.     

Feeding Atlantic salmon diets with plant ingredients during the seawater phase – a full‐scale net production of marine protein with focus on biological performance,welfare, product quality and safety

By feeding Atlantic salmon diets with 64% of the fish oil (FO) replaced by vegetable oil, and with decreasing fishmeal (FM) inclusion levels from 213, 178 and 143 g kg⁻¹ (accumulated level during the seawater phase) in a full‐scale experiment producing 3.1 thousand tonnes fish, no significant negative effects on fish performance, health and product quality were observed. All dietary groups showed, however, moderate intestinal inflammation. Reduced growth and feed efficiency were seen with decreasing fishmeal inclusion levels. Two dietary groups demonstrated net marine protein production, while none of the groups showed net fish production (FIFO ≥1.65) due to the equal low FO inclusion. High plant oil level gave lower fillet level of persistent organic pollutants (POPs) compared with the levels surveyed on the Norwegian market. The study gave predictable incorporation rates of essential n‐3 long‐chain fatty acids in the fillet. Cooked salmon fillet from all dietary groups showed minor differences in sensory quality. Based on the present full‐scale production results, dietary FM inclusion down to 160 g kg⁻¹ (accumulated) during the seawater phase, concurrent to replacing ~70% of the FO with a suitable plant oil, is not regarded to represent any risk to fish performance, health or quality.     

Genetic screening of FecB,FecXG and FecXI mutations and their linkage with litter size in Barki and Rahmani sheep breeds

Characterization of fecundity genes offers the opportunity to improve production efficiency, and the consequent increase in litter size in livestock industry, through utilizing them in breeding programs. The main objective of this study was to detect the BMPR‐IB, BMP15 and GDF9 gene mutations and to investigate whether these mutations are associated with litter size in Egyptian sheep breeds. To achieve this goal, 73 adult ewes representing Barki (n = 33) and Rahmani (n = 40) breeds were used. Polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) screening approach was used to detect the presence of FecB, FecX^G and FecX^I mutations in the two selected breeds. Results of this study showed that the three different candidate gene mutations, namely FecB, FecX^G and FecX^I are not present among these selected populations of the Egyptian breeds. Further studies regarding other mutations and/or other genes, which may influence ovulation rate, should be carried out to determine the type and mode of inheritance of such genes in Egyptian sheep breeds.     

Lunar surface magnetic fields and their interaction with the solar wind: results from lunar prospector

The magnetometer and electron reflectometer experiment on the Lunar Prospector spacecraft has obtained maps of lunar crustal magnetic fields and observed the interaction between the solar wind and regions of strong crustal magnetic fields at high selenographic latitude (30 degreesS to 80 degreesS) and low ( approximately 100 kilometers) altitude. Electron reflection maps of the regions antipodal to the Imbrium and Serenitatis impact basins, extending to 80 degreesS latitude, show that crustal magnetic fields fill most of the antipodal zones of those basins. This finding provides further evidence for the hypothesis that basin-forming impacts result in magnetization of the lunar crust at their antipodes. The crustal magnetic fields of the Imbrium antipode region are strong enough to deflect the solar wind and form a miniature (100 to several hundred kilometers across) magnetosphere, magnetosheath, and bow shock system.     

化脓棒状杆菌的分离与坚定

北京动物园的2头斑羚和1头岩羊相继发病死亡。剖检发现肺部均有淤血和化脓灶。经取其肺组织作细菌培养,均分离到纯粹的杆菌。经细菌形态、培养特性和生化特性分析,鉴定为化脓棒状杆菌。     

Effects of herbivory on the reproductive effort of 4 prairie perennials

Background  

There is an urgent need to explore and utilize naturally occurring products for combating harmful agricultural and public health pests. Secondary metabolites in the leaves of the Tree of Heaven, Ailanthus altissima L. have been reported to be herbicidal and insecticidal. The mode of action, however, of the active compounds in A. altissima are not understood. In this paper, we report the chemical characteristics of the herbicidal and insecticidal components in this tree, and will discuss the effect of light on the bioactivity of the active components.     

Apoptosis‐Related Protein Expression During Pre‐ and Post‐Natal Testicular Development After Administration of Glucocorticoid in utero in the Sheep

Pre‐natal glucocorticoids are used in women at risk of preterm delivery to induce foetal lung maturation. However, glucocorticoids can produce negative outcomes for other tissues such as the reproductive system. We therefore tested the effects of pre‐natal betamethasone on testicular morphology and apoptotic protein immune expression during pre‐ and post‐natal development. Pregnant ewes (n = 42) bearing singleton male foetuses were randomly allocated to receive intramuscular injections of saline or betamethasone (0. 5 mg/kg) at 104, 111 and 118 days of gestation (DG). Testes were collected at 121 and 132 DG, and at 45 and 90 post‐natal days (PD) and subjected to morphometric analysis (volume densities of sex cords and interstitial tissues; sex cord diameter). Immunohistochemistry (% stained area) was used to assess active caspase‐3, Bax, Bcl‐2 and cell‐cycle proteins (PCNA). Compared with control values, betamethasone treatment decreased sex cord diameter at 121 DG, 45 and 90 PD, and sex cord volume at 90 PD. Active caspase‐3 was decreased by betamethasone at 121 DG and 90 PD, but Bax was increased in all betamethasone groups. Bcl‐2 and PCNA decreased in the betamethasone groups at 121 DG and 45 PD, but increased at 132 DG and 90 PD. We conclude that high levels of pre‐natally administered glucocorticoid reduce foetal testicular development, perhaps via changes in the balance between pro‐ and anti‐apoptotic proteins and cell‐cycle proteins. These outcomes could compromise the future spermatogenic potential of male offspring.     

Oestrogen and Progesterone Receptors and COX‐2 Expression in Endometrial Biopsy Samples During Maternal Recognition of Pregnancy in Llamas (Lama glama)

Endometrial expression of oestrogen receptor‐α (ERα), progesterone receptor (PR) and cyclooxigenase‐2 (COX‐2) was evaluated in non‐pregnant and pregnant llamas during the period when luteolysis/maternal recognition of pregnancy is expected to occur. Females (n = 28) were divided into two groups: non‐pregnant llamas were induced to ovulate with a Buserelin injection, and endometrial biopsies were obtained on day 8 (n = 5) or 12 (n = 5) post‐induction of ovulation. Animals of the pregnant group (n = 18) were mated with a fertile male. Pregnancy was confirmed by the visualization of the embryo collected by transcervical flushing in 5 of 9 animals on day 8 post‐mating and by progesterone profile on day 12 post‐mating in 4 of 9 animals, when endometrial biopsies were obtained. An immunohistochemical technique was used to evaluate receptors population and COX‐2 expression. Pregnant llamas showed a higher percentage of positive cells and stronger intensity for ERα than for non‐pregnant llamas in stroma on day 8 and in the luminal epithelium on day 12 post‐induction of ovulation, while a deep decrease in endometrial PR population was reported in pregnant llamas on that day in luminal and glandular epithelia and stroma. In the luminal epithelium, COX‐2 expression was lower in pregnant than in non‐pregnant animals. Briefly, the increase of ERα in pregnant llamas gives further support to the hypothesis that oestrogens are involved in the mechanism of maternal recognition of pregnancy. Endometrial PR decrease in pregnant llamas might be a necessary event to allow the expression of proteins involved in conceptus attachment, a mechanism widely accepted in other species. Moreover, embryo seems to attenuate maternal PGF_2α secretion during early pregnancy by decreasing the endometrial expression of COX‐2 in the luminal epithelium of pregnant llamas.     

Quantifying Transport Between the Tropical and Mid-Latitude Lower Stratosphere

Airborne in situ observations of molecules with a wide range of lifetimes (methane, nitrous oxide, reactive nitrogen, ozone, chlorinated halocarbons, and halon-1211), used in a tropical tracer model, show that mid-latitude air is entrained into the tropical lower stratosphere within about 13.5 months; transport is faster in the reverse direction. Because exchange with the tropics is slower than global photochemical models generally assume, ozone at mid-latitudes appears to be more sensitive to elevated levels of industrial chlorine than is currently predicted. Nevertheless, about 45 percent of air in the tropical ascent region at 21 kilometers is of mid-latitude origin, implying that emissions from supersonic aircraft could reach the middle stratosphere.