Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Background  

The establishment of mutant populations together with the strategies for targeted mutation detection has been applied successfully to a large number of organisms including many species in the plant kingdom. Considerable efforts have been invested into research on tomato as a model for berry-fruit plants. With the progress of the tomato sequencing project, reverse genetics becomes an obvious and achievable goal.     

Reproductive Performance and Reconception of Nellore Cows According to Their Pure‐ or Cross‐Bred Calves

The objective of the present research was to evaluate the effect of a calf's genetic group on the productive and reproductive efficiency of its Nellore dam. Fixed‐time artificial insemination was applied to 800 cows using semen extracted from Nellore, Simmental and Angus Red bulls. Four hundred eleven cows calved, producing 119 Nellore, 103 ½Simmental–½Nellore and 189 ½Nellore–½Angus Red calves. The second mating period, which paired Nellore cows with Simmental bulls, was initiated 10 days after parturitions began and lasted for 5 months. Based on the two successive parturitions, the cumulative parturition rate for calving periods of 3, 4 and 5 months was calculated. Although no significant difference was observed for birth weight among the genetic groups, cross‐bred calves weighed, on average, 10% more than did pure‐bred calves at the age of 205 days. Nellore dams experienced a gestation period that was 7 days longer than did the cross‐bred dams, and the former showed a higher parturition rate at 90 and 120 days of the calving season, but not at 150 days (calving rates of 80.6, 76.4 and 76.2% for mothers of Nellore, ½Nellore–½Angus Red and ½Nellore–½Simmental, respectively, p > 0.05). At 90 and 120 days, Nellore dams produced more kg of calf per mated dam. In conclusion, in a short breeding season, Nellore dams nursing pure‐bred Nellore calves were found to have a higher biological efficiency compared with Nellore dams nursing cross‐bred calves.     

Physiological and biochemical consequences of electroimobilisation in conscious sheep

An electroimmobilisation device has been developed to facilitate the automated shearing of sheep, but there is little information on its effects on the body. We have studied its effects on the cardiovascular system and on intermediary metabolism in sheep. Electroimmobilisation caused statistically significant increases in mean arterial pressure, heart rate, cardiac output, renal and hepatic and hindquarter glucose and lactate flux, organ and whole body oxygen flux, hindquarter blood flow and core temperature and decreases in arterial and posterior vena cava blood pH, renal and hepatic blood flow and PaCO2. Notably, no change occurred in PaO2. The metabolic changes demonstrated the capacity of sheep to respond to the increased muscular and cardiovascular work induced by electroimmobilisation. Pulmonary function was not compromised during electroimmobilisation as judged from blood gas changes, and the acid/base changes were rapidly reversed after electroimmobilisation. The recovery to control conditions for all perturbations generally took no longer than 30 min, consistent with a rapid and physiologically adequate reversal by the animal's homeostatic mechanisms.     

Imaging of intermittency in ripple-wave turbulence

The dynamics of a fluid surface filled with high-amplitude ripples were studied with a technique (diffusing light photography) that resolves the height at all locations instantaneously. Even when nonlinearities are strong enough to generate a (Kolmogorov) cascade from long wavelength (where energy is input) to shorter wavelength, the resulting turbulent state contains large coherent spatial structures. The appearance of these structures in a thermal equilibrium state (with the same average energy) would be highly improbable.     

A survey of Culicoides developmental sites on a farm in northern Spain,with a brief review of immature habitats of European species

Culicoides species (Diptera: Ceratopogonidae) belonging to the Obsoletus and Pulicaris groups are considered to be the main vectors of bluetongue virus (BTV) in non Mediterranean Europe. Selected terrestrial microhabitats (n = 17) on a farm in northern Spain were sampled repeatedly over a year-long period and characterized for use by Culicoides species for immature development. Concurrent use of CDC light traps showed the presence of 37 species and 66,575 specimens of adult Culicoides. A total of 28 species and 11,396 individuals emerged from laboratory-maintained soil samples. Culicoides obsoletus and Culicoides scoticus (pooled as Obsoletus complex) were particularly abundant (comprising 58.6% and 74.5% of the total collections in light traps and emergence traps respectively). Potential key vectors of animal viruses (such as BTV) were found in two main terrestrial types of microhabitats. In the case of C. obsoletus, different types of manure (old and composted manure, manure mixed with organic matter, and fresh manure) produced most of the specimens. In contrast, larvae of C. scoticus and Culicoides lupicaris were associated with soil substantially comprised of rotting leaf litter that included the parasitic plant Lathraea clandestina. Several species, Culicoides festivipennis, Culicoides punctatus and Culicoides brunnicans, were very common in mud at pond margins. Indeed, pond microhabitats and runoff below barn rooflines supported the greatest species richness. In the pond habitat, 49.4% of Culicoides specimens emerged from mud at the water edge, as opposed to 50 cm above (32.4%) and 1 meter above waterline (18%). Similar species richness, but statistically significant differences in abundance, were observed among the four pond microhabitats. Overall, the majority of the specimens were found in the upper layer (0–3 cm), except in manure, where they preferred deeper layers (>6 cm). Maximum peaks of abundance occurred in both light traps and soil samples in summer months, whereas increased captures in autumn were noticed only in light traps. Both trapping systems failed to collect adult Culicoides midges in the coldest months of December, January and February. The literature on immature habitats of species suspected in BTV transmission in Europe, the Pulicaris group and particularly the Obsoletus group, is briefly reviewed.     

Effect of Chemical or Electrical Activation of Bovine Oocytes on Blastocyst Development and Quality

Activation of in vitro‐matured (IVM) oocytes is essential for successful embryo production following nuclear transfer (NT) or intracytoplasmic sperm injection (ICSI). This study was designed to compare the rates of blastocyst production and embryo quality (as measured by numbers of viable cells) following parthenogenetic activation with electrical pulse or the use of two different calcium ionophores, A23187 (CA) or ionomycin (IO), with or without the addition of bovine serum albumin (BSA). IVM oocytes with a first polar body were randomly allocated to five treatment groups: CA (5 μm CA, 5 min; n = 88), CA + BSA (5 μm CA, 5 min; BSA, 5 min; n = 90), IO (5 μm IO, 5 min; n = 91), IO + BSA (5 μm IO, 5 min; BSA, 5 min; n = 86) and EL (two pulses of 1.5 kV/cm, 20 μs; n = 120). Blastocyst rates were higher (p < 0.05) for CA (54.4%), IO (51.4%) and EL (54.5%) than for IO + BSA (18.3%). Treatment CA + BSA (39.8%) did not differ from the others. There was no difference (p > 0.05) among treatments in total number of cells. However, the percentage of viable cells was reduced in CA (49.9%), CA + BSA (45.8%), IO (64.9%), IO + BSA (50.9%) compared with EL (82.7%). In summary, the addition of BSA to the IO treatment had an adverse effect on blastocyst production rates. Although there was no difference between electrical stimulation and chemical activation on blastocyst production rates, electrical activation resulted in blastocysts with a higher percentage of viable cells.     

Vitamin D3 supplementation of cull cows: effects on longissimus and semitendinosus muscle tenderness

Previous studies have shown that supplementation of vitamin D3 to cow diets for 4 to 10 d before slaughter lowers Warner-Bratzler shear force (WBSF) values and increases sensory tenderness scores in beef cuts. The present study was conducted to evaluate the effects of vitamin D3 supplementation on muscle calcium concentration, WBSF values, and sensory tenderness ratings of LM and semitendinosus (ST) muscles from cull, predominately Angus, cows (eight cows per treatment). Treatments included 0 (control), 5 million IU, or 7.5 million IU of vitamin D3 supplemented daily for 7 d preslaughter. Twenty-four hours after slaughter, 2.54-cm-thick LM and ST muscle steaks were cut; aged for either 0, 7, 14, or 21 d (ST steaks aged for 7 d only); and frozen at -20 degrees C for WBFS and sensory analysis. Mean values for LM calcium concentration tended to increase (P = 0.14) with vitamin D3 supplementation (154, 176, and 183 microig/g, fresh basis, for 0, 5, and 7.5 million IU/d, respectively). After 7 d of aging, LM steaks from cows fed 7.5 million IU had lower (P < 0.05) WBSF values than 7-d steaks from controls and cows fed 5.0 million IU/d aged 7 d; however, vitamin D3 supplementation had no (P > 0.05) effect on WBSF values of ST steaks aged 7 d. Vitamin D3 supplementation did not (P > 0.05) affect sensory tenderness ratings for either LM or ST steaks at any aging period. Aging, however, had a linear (P < 0.001) effect on tenderness, with an increase in tenderness as aging time increased from 0 to 21 d. Thus, results from the present study indicate that vitamin D3 supplementation, at these levels and duration before slaughter, provided little benefit to muscle tenderness of beef from cull cows.     

Color stability of semitendinosus,semimembranosus, and biceps femoris steaks packaged in a high-oxygen modified atmosphere

The objectives of this study were to evaluate visual and chemical attributes of beefsteaks from various USDA quality grades and muscles packaged in high-oxygen (80% O2/20% CO2) modified-atmosphere packaging (MAP). A total of nine carcasses were selected to represent Select (n = 3), low Choice (n = 3), and high Choice (n = 3) USDA quality grades. The semimembranosus (SM), semitendinosus (ST), and biceps femoris (BF) muscles were removed from each carcass and allotted to two packaging types (MAP or polyvinyl chloride over-wrap) and were displayed for up to 10 d, with evaluation on d 1, 3, 5, 7, and 10. Fifty-four steaks were evaluated on each day by a five-member trained panel for visual color (lean color and discoloration) and were also analyzed with a Minolta Chroma Meter CR-310 for L* and a* values (lightness and redness, respectively). Chemical properties measured included percentage of metmyoglobin formation and fat content. Visual color scores did not differ (P > 0.05) at d 1 and 3 with respect to all quality grades, but decreased after d 3, with a greater reduction (P < 0.05) in high Choice steaks for both lean color and discoloration. The low Choice steaks packaged in MAP displayedhigher (P < 0.05) lean color scores and less (P < 0.05) discoloration at d 7 and 10 than did Select and high Choice steaks. Redness (a*) values also decreased (P < 0.05) after d 3, whereas (lightness) L* values declined (P < 0.05) from d 1 to 5. The high Choice steaks had higher (P < 0.05) metmyoglobin content than low Choice and Select steaks, but packaging had no effect (P > 0.05) on metmyoglobin content. Muscle type did affect metmyoglobin content; however, the metmyoglobin content of the SM was greatest (P < 0.05), followed by the BF, with the ST having the lowest (P < 0.05) metmyoglobin formation. Results indicate that low Choice steaks react the best in MAP, and the ST maintained greater storage characteristics regardless of quality grade or packaging.     

Occurrence of Tomato spotted wilt virus (TSWV) in Jordan

Tomato spotted wilt virus was recorded for the first time in Jordan on tomato plants. Severe disease symptoms were observed in different tomato farms in the Jordan Valley. Using a specific primer pair a fragment of the capsid protein gene of the virus has been amplified by RT-PCR and IC-RT-PCR. The amplified PCR product was cloned and sequenced. Sequence analysis revealed that the Jordanian isolate of TSWV shared high nucleotide similarities with other isolates from different countries. The sequence of the capsid protein gene was deposited in GenBank under the accession number AY646682 . The response of different tomato breeding lines and hybrids, previously developed for resistance against Tomato yellow leaf curl virus (TYLCV) were tested for their reaction to TSWV infection. All tested lines and hybrids were susceptible to TSWV infection. This has been confirmed at the molecular level by using the SCAR 421 marker linked to the TSWV resistance gene Sw-5 .