The energetic basis of specificity in the Eco RI endonuclease--DNA interaction

High sequence selectivity in DNA-protein interactions was analyzed by measuring discrimination by Eco RI endonuclease between the recognition site GAATTC and systematically altered DNA sites. Base analogue substitutions that preserve the sequence-dependent conformational motif of the GAATTC site permit deletion of single sites of protein-base contact at a cost of +1 to +2 kcal/mol. However, the introduction of any one incorrect natural base pair costs +6 to +13 kcal/mol in transition state interaction energy, the resultant of the following interdependent factors: deletion of one or two hydrogen bonds between the protein and a purine base; unfavourable steric apposition between a group on the protein and an incorrectly placed functional group on a base; disruption of a pyrimidine contact with the protein; loss of some crucial interactions between protein and DNA phosphates; and an increased energetic cost of attaining the required DNA conformation in the transition state complex. Eco RI endonuclease thus achieves stringent discrimination by both "direct readout" (protein-base contracts) and "indirect readout" (protein-phosphate contacts and DNA conformation) of the DNA sequence.     

Pleistocene extinction of genyornis newtoni: human impact on australian megafauna

More than 85 percent of Australian terrestrial genera with a body mass exceeding 44 kilograms became extinct in the Late Pleistocene. Although most were marsupials, the list includes the large, flightless mihirung Genyornis newtoni. More than 700 dates onGenyornis eggshells from three different climate regions document the continuous presence of Genyornis from more than 100,000 years ago until their sudden disappearance 50,000 years ago, about the same time that humans arrived in Australia. Simultaneous extinction of Genyornis at all sites during an interval of modest climate change implies that human impact, not climate, was responsible.     

辽宁省棉铃虫虫情划区的调查研究

研究表明,北方特早熟棉区的辽宁省各地棉铃虫的发生程度与气温（年均等温线８～９℃）、降水（年平均降水量５００～７００ｍｍ）及寄主植物的分布三个条件重合程度呈明显的正相关。据此,将本省棉铃虫的发生区域划分为：重发生区、轻发生区、可发生区、偶发生区、不发生区,并绘制出棉铃虫区域划分图。     

The relationship of training milestones with racing success in a population of Standardbred horses in New Zealand

AIM: To gather information on the repeatability of a faecal nematode egg count (FEC) reduction (FECR) test (FECRT), evaluating both different methods of calculating efficacy and variations within a method, in order to supply veterinarians and other advisors with sufficient information to apply some level of confidence around a diagnosis of anthelmintic resistance based on FECRT results.

METHODS: Two commercial sheep farms were selected on the basis of having previously recorded FECR <95% after treatment with ivermectin (Farm 1) or albendazole (Farm 2). On each farm at least 250 lambs, managed as a single mob, were individually ear-tagged and sampled for FEC. The resulting counts were used, 3—4 days later, to sort the lambs into 24 groups of 10. First, the animals were split into three groups of 80, having high, medium or low FEC. Second, within each of these groups the 80 animals were further divided into four replicate mobs of 20 (each with the same mean count). Third, each of these replicates was further split into two groups of 10: those that would be drenched and those that would remain as untreated controls. All animals were again faecal-sampled and those in the drenched groups were dosed, using a syringe, to their individual liveweight, with ivermectin (Farm 1) or albendazole (Farm 2). Ten days after treatment all animals were individually faecal- sampled again. FEC and larval cultures were undertaken for all 24 groups from both pre- and post-treatment samples. Ef- ficacy (FECR) of the undifferentiated FECRT was calculated using three different equations, and efficacy by genus was also calculated.

RESULTS: Calculated efficacies differed between equations, and the equation which did not utilise an untreated control yielded significantly lower efficacy estimates on both farms. Faecal cultures varied considerably in the proportions of parasite genera recovered. In general, this did not differ between FEC groups, except on Farm 1 where Haemonchus spp were more common and Cooperia spp less common in high-FEC samples. Estimated efficacies against individual genera varied considerably or very little, depending on the level of resistance. On both farms, differing proportions of tests against some genera passed or failed FECRTs based on a threshold pass mark of ≥95% FECR.

CONCLUSION: There was considerable variability in the outcomes of FECRTs and in larval culture results. Caution is warranted in interpreting the results of FECRTs when efficacy values fall into the 90—95% range. Further, the possibility of a test returning a false-negative result is raised, indicating that even an efficacy estimated ≥95% may not guarantee the absence of resistant parasites.     

Reproductive Performance of Gilts with Similar Age but with Different Growth Rate at the Onset of Puberty Stimulation

The aim of this study was to evaluate the reproductive performance of gilts that had a similar age but different weights at the onset of puberty stimulation by boar exposure at 144 days. Gilts were divided into two groups according to their lifetime growth rate from birth to approximately 144 days of age. Mean growth rates at this moment were 577 and 724 g/day for group 1 (G1; n = 58) and group 2 (G2; n = 58), respectively. After selection, gilts were weighed at approximately 155, 165 and 175 days of age, on the insemination day and at slaughter. Gilts were inseminated, on average, at 193 days of age and were slaughtered 32 days after insemination, when the number of corpora lutea and embryos were recorded. Higher growth rate gilts (G2) reached puberty earlier (155.3 vs 164.1 days; p < 0.01). More gilts of G2 group attained puberty by 190 days of age (p = 0.004) than G1 gilts (95%; 55/58 vs 76%; 44/58). The anoestrous rate, until 60 days after the onset of boar exposure was higher (p < 0.01) in G1 (19.0%; 11/58) than in G2 (3.4%; 2/58) group. However, there were no differences in the pregnancy rate (90.7 vs 94.5), ovulation rate (15.9 vs 16.5), total embryos (12.9 vs 11.7), viable embryos (12.0 vs 11.1) and embryo survival (73.7% vs 68.5%), between G1 gilts and G2 gilts, respectively (p > 0.05). High growth rate gilts attain puberty earlier and have a lower anoestrous rate than low growth rate gilts.     

Pyloric outflow obstruction secondary to sclerosing encapsulating peritonitis in a dog

A 6-year-old, male neutered mixed breed dog was presented emergently with a three-week history of hyporexia, vomiting, diarrhoea and weight loss. Upon examination, the patient was dull, had generalised muscle atrophy, moderate abdominal pain and a mild amount of peritoneal effusion. A fluid-filled, distended, corrugated small bowel with marked gastroparesis and moderate peritoneal effusion was noted on abdominal ultrasonography. Endoscopy revealed hyperaemic and friable mucosa and a subjectively narrowed pylorus. Emergency exploratory celiotomy was performed due to worsening patient condition and revealed thick, diffuse, fibrous adhesions of the abdominal cavity. Based on these findings, sclerosing encapsulating peritonitis (SEP) was suspected. A large mass of omentum adjacent to the greater curvature of the stomach had caused a pyloric outflow obstruction. Adhesiolysis was attempted but was unsuccessful due to the friability of the small intestines. The dog was humanely euthanased under anaesthesia. A diagnosis of SEP was confirmed via necropsy. No underlying cause was identified. This is the first known case of a pyloric outflow obstruction secondary to SEP in a dog. Although rare, this condition should be considered as a differential for dogs with signs of a pyloric outflow obstruction with concurrent ascites and abdominal pain, hyporexia, vomiting and diarrhoea.     

Fixed‐time post‐cervical artificial insemination in weaned sows following buserelin use combined with/without eCG

Fixed‐time post‐cervical artificial insemination (FTAI) drastically reduces labour requirements and increases the use of boars with higher genetic merit. This study evaluated the efficiency of eCG administration combined with/without the GnRH agonist buserelin for the induction and synchronization of ovulation in weaned sows submitted to FTAI. The sows were allocated into three groups. In the control group, the first artificial insemination was performed at the onset of oestrus and repeated every 24 hr. In the eCG+GnRH group, sows received 600 IU eCG at weaning and buserelin (10 μg) after 86–89 hr of eCG, and in the GnRH group, sows received only buserelin after 86–89 hr of weaning. The hormone‐treated sows received a single FTAI after 30–33 hr of buserelin application. All the sows were inseminated with homospermic doses (1.5 × 10⁹ sperm cells/50 ml). The interval between weaning and ovulation was shorter (p < .05) in the eCG+GnRH (133.3 hr) and GnRH (135.9 hr) groups than the control (141.5 hr) group. In the eCG+GnRH group, the sows ovulated earlier (p < .05) than those in the GnRH group (44.5 vs. 48.2 hr after buserelin administration). The reproductive performance of GnRH sows was not compromised when only sows exhibiting oestrus at the time of insemination were considered, but lower farrowing rate and smaller litter size were observed in eCG+GnRH sows. The reproductive performance of eCG+GnRH sows was primarily compromised because the insemination was performed outside the optimal time relative to ovulation; therefore, it is advisable to inseminate them before 116–122 hr after weaning.