Promoter anchored amplified polymorphism based on random amplified polymorphic DNA (PAAP-RAPD) in cotton

Non-coding sequences account for a majority of the higher plant genome, some of which have important effects in gene regulation and plant development. In an effort to develop molecular marker systems to search for polymorphisms associated with high fiber yield and quality in cotton, we have developed a methodology that could specifically target the regulatory regions of the cotton genome. In this study we designed 10-nucleotide degenerate promoter primers based on conserved core promoter sequences and tested their applicability in PCR amplifications in combination with 10-mer random amplified polymorphic DNA (RAPD) primers. The amplified markers are called promoter anchored amplified polymorphism based on RAPD (PAAP-RAPD). Forty cotton genotypes with diverse genetic and geographical backgrounds were used to test the PAAP-RAPD system using polyacrylamide gel electrophoresis. Based on PAAP-RAPD markers amplified from 12 primer combinations, the 40 genotypes were classified into five distinctive groups: two Upland cotton (Gossypium hirsutum) groups from China, another two Upland cotton groups from the USA, and one group from American Pima cotton (G. barbadense). The groupings are in general consistent with their genetic and geographical origins. Thirty-six PAAP-RAPD and RAPD fragments were cloned and four of them were further subjected to sequence analysis. Signal scanning using software PLACE confirmed that they contained an array of cis-regulatory sequences in addition to the core promoter sequences. The results demonstrate the potential application of PAAP-RAPD as a new marker system specifically targeting regulatory regions of the plant genome.     

An investigation of the effects of intratesticular alfaxalone and lidocaine during castration in piglets

Objective

To determine whether intratesticular injection of an alfaxalone–lidocaine combination can induce anesthesia and provide a rapid recovery in piglets undergoing surgical castration.

Adiponectin links adipose tissue function and monocyte inflammatory responses during bovine metabolic stress

The periparturient period of dairy cows is characterized by intense lipid mobilization from adipose tissue leading to increased plasma concentrations of nonesterified fatty acids (NEFA). High NEFA are a predisposing factor for inflammatory based diseases. A major component of these diseases is uncontrolled macrophage/monocyte inflammatory responses. Changes in the endocrine activity of adipose tissue during the periparturient period could impact macrophage function by modifying the secretion of adipokines including adiponectin. Currently, the effects of adiponectin on monocyte activation in dairy cattle are unknown. In humans and rodents, this adipokine regulates monocyte phenotype and alterations in its plasma levels are linked with the development of inflammatory diseases. The objectives of this study were to establish associations between plasma adiponectin expression dynamics and different markers of lipid mobilization during the periparturient period of dairy cows and to characterize the effects of adiponectin on the inflammatory response of bovine monocytes. Plasma adiponectin, NEFA, BHB, albumin, and subcutaneous and retroperitoneal fat depots depth were measured during the periparturient period of dairy cows. In vitro, bovine monocytes were cultured with adiponectin to assess changes in pro-inflammatory responses following LPS stimulation. Results from this study demonstrate that alterations in plasma adiponectin levels in periparturient cattle are inversely correlated with the concentrations of plasma NEFA, an important marker of lipid mobilization. Furthermore, adiponectin exposure significantly decreased monocyte expression of TNFα after LPS stimulation thus markedly reducing their inflammatory response. Reduced plasma adiponectin during the periparturient period could predispose dairy cows to the development of uncontrolled monocyte inflammatory responses.     

Recreational sandboxes for children and dogs can be a source of epidemic ribotypes of Clostridium difficile

Different studies have suggested that the sand of public playgrounds could have a role in the transmission of infections, particularly in children. Furthermore, free access of pets and other animals to the playgrounds might increase such a risk. We studied the presence of Clostridium difficile in 20 pairs of sandboxes for children and dogs located in different playgrounds within the Madrid region (Spain). Clostridium difficile isolation was performed by enrichment and selective culture procedures. The genetic (ribotype and amplified fragment length polymorphism [AFLP]) diversity and antibiotic susceptibility of isolates was also studied. Overall, 52.5% (21/40) of samples were positive for the presence of C. difficile. Eight of the 20 available isolates belonged to the toxigenic ribotypes 014 (n = 5) and 106 (n = 2), both regarded as epidemic, and CD047 (n = 1). The other 12 isolates were non‐toxigenic, and belonged to ribotypes 009 (n = 5), 039 (n = 4), and 067, 151 and CD048 (one isolate each). Nevertheless, all isolates (even those of a same ribotype) were classified into different AFLP genotypes indicating non‐relatedness. In conclusion, our results revealed the presence of epidemic ribotypes of C. difficile in children's and dog's sandboxes located nearby, which constitutes a major health risk.     

A Syndrome Resembling Idiopathic Noncirrhotic Portal Hypertension in 4 Young Doberman Pinschers

We describe 4 young male Doberman Pinschers (3 littermates and 1 unrelated dog) with a syndrome resembling idiopathic or noncirrhotic portal hypertension of humans. Each dog was evaluated for a hepatopathy resulting in portal hypertension, development of portosystemic collateral vessels, and hepatic encephalopathy. These dogs differ from previous reports of young dogs with hepatic insufficiency associated with portal hypertension and acquired portal systemic shunting by their lack of intrahepatic arteriovenous fistulae, portal vein atresia, or intrahepatic fibrosis. Clinicopathologic features included erythrocyte microcytosis, normal to mildly increased liver enzyme activities, increased concentrations of serum bile acids, reduced plasma indocyanine green clearance, and normal total bilirubin concentration. Abdominal ultrasonography disclosed a small liver and portosystemic collateral vessels. Radiographic imaging studies confirmed hepatofugal portal circulation and discounted hepatic arteriovenous fistulae. Histopathologic features in liver tissue from each dog were similar and consistent in all sections examined. Common findings included increased cross-sectional views of hepatic arterioles; hepatic lobular atrophy; scanty increase in connective tissue around some large portal triads; and absence of inflammation, disturbed lobular architecture, bile duct proliferation, or intrahepatic cholestasis.     

Nutritional Contribution of Biofloc within the Diet of Growout and Broodstock of Litopenaeus vannamei,Determined by Stable Isotopes and Fatty Acids

The relative contributions of feed sources were determined through the isotopic signal (δ¹³C and δ¹⁵N) and fatty acid profile of feed items, shrimp muscle, and eggs of Litopenaeus vannamei reared in a biofloc system. In the growout phase, the isotope analysis showed the biofloc particle size class ≥250 μm contributed 55–100%; size ≥50 < 250 μm contributed 0–22%; and artificial feed contributed 0–45%. Principal component analysis applied to fatty acid profiles showed that biofloc ≥250 μm and artificial feed were the most important items in shrimp growout. For the egg production, isotope analysis suggested that the most important feed sources according to their relative contributions were polychaetes (0–100%), followed by artemia biomass (0–86%) and semi‐moist feed (0–66%), with lower contributions from squid, mussel, and the muscle of L. vannamei broodstock that had been cultured in biofloc. In terms of fatty acids, the most important items were artemia, polychaetes, and semi‐moist feeds. This work clarified the importance of feed sources for shrimp during culture in biofloc systems and during reproduction. Analysis of stable isotopes and fatty acids can be successfully used to trace the assimilation of nutrients during the nutrition of shrimp.     

木材加工用计算机软件的设计体会

介绍设计木材加工用计算机软件所需的技术准备、软件的设计思想与应用环境，以及编程语言选择对软件应用性的影响。     

Postnatal piglet husbandry practices and well-being: the effects of alternative techniques delivered separately

The aim of this study was to evaluate stress responses evoked by 2 alternative methods for performing the following processing procedures: 1) teeth resection-clipping vs. grinding; 2) tail docking-cold vs. hot clipping; 3) identification-ear notch vs. tag; 4) iron administration-injection vs. oral; 5) castration-cords cut vs. torn. Eight to 10 litters of 8-, 2-, and 3-d-old piglets were assigned to each procedure. Within each litter, 2 piglets were assigned to 1 of 4 possible procedures: the 2 alternative methods, a sham procedure, and a sham procedure plus blood sampling. Blood was sampled before processing and at 45 min, 4 h, 48 h, 1 wk, and 2 wk postprocedure and assayed for cortisol and beta-endorphin. Procedures were videotaped and analyzed to evaluate the time taken to perform the procedure and the number of squeals, grunts, and escape attempts. Vocalizations were analyzed to determine mean and peak frequencies and duration. Piglets were weighed before the procedure and at 24 h, 48 h, 1 wk, and 2 wk afterward. Lesions were scored on a scale of 0 to 5 on pigs in the identification, tail docking, and castration treatments at 24 h, 1 wk, and 2 wk postprocedure. For teeth resection, grinding took longer than clipping and resulted in greater cortisol concentration overall, poorer growth rates, and longer vocalizations compared with pigs in the control treatment (P<0.05). For tail docking, hot clipping took longer, and hot-clipped piglets grew slower than cold-clipped piglets (P<0.05). Hot clipping also resulted in longer and higher frequency squealing compared with pigs in the control treatment (P<0.01). For identification, ear notching took longer than tagging, and ear-notched piglets had worse wound scores than tagged piglets (P<0.05). Cortisol concentrations at 4 h also tended to be greater for ear-notched piglets (P<0.10). Ear notching evoked calls with higher peak frequencies than the control treatments. For iron administration, oral delivery took numerically longer than injecting, but there were no significant differences between injecting and oral delivery for any of the measures. For castration, tearing took longer than cutting the cords (P<0.05), but beta-endorphin concentrations at 45 min postprocedure were greater for cut piglets. When measures of behavior, physiology, and productivity were used, the responses to teeth resection, tail docking, and identification were shown to be altered by the procedural method, whereas responses to iron administration and castration did not differ. The time taken to carry out the procedure would appear to be an important factor in the strength of the stress response.