转基因牛多重PCR检测方法的建立

为建立一种快速检测转基因牛的多重PCR方法,针对牛线粒体16 S rRNA基因(BOS mtD-NA16 S rRNA,BOS)、转基因动物常用标记基因新霉素磷酸转移酶基因(NPT Ⅱ)、人乳铁蛋白编码基因(human laetoferrin gene,hLF)、人-α-乳清蛋白编码基因(human α-lactalb...     

Biofilm production by pathogens associated with canine otitis externa,and the antibiofilm activity of ionophores and antimicrobial adjuvants

Otitis externa (OE) is a frequently reported disorder in dogs associated with secondary infections by Staphylococcus, Pseudomonas and yeast pathogens. The presence of biofilms may play an important role in the resistance of otic pathogens to antimicrobial agents. Biofilm production of twenty Staphylococcus pseudintermedius and twenty Pseudomonas aeruginosa canine otic isolates was determined quantitatively using a microtiter plate assay, and each isolate was classified as a strong, moderate, weak or nonbiofilm producer. Minimum biofilm eradication concentration (MBEC) of two ionophores (narasin and monensin) and three adjuvants (N‐acetylcysteine (NAC), Tris‐EDTA and disodium EDTA) were investigated spectrophotometrically (OD_570nm) and quantitatively (CFU/ml) against selected Staphylococcus and Pseudomonas biofilm cultures. Concurrently, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of planktonic cultures were assessed. 16/20 of the S. pseudintermedius clinical isolates were weak biofilm producers. 19/20 P. aeruginosa clinical isolates produced biofilms and were distributed almost equally as weak, moderate and strong biofilm producers. While significant antibiofilm activity was observed, no MBEC was achieved with narasin or monensin. The MBEC for NAC ranged from 5,000–10,000 µg/ml and from 20,000–80,000 µg/ml against S. pseudintermedius and P. aeruginosa, respectively. Tris‐EDTA eradicated P. aeruginosa biofilms at concentrations ranging from 6,000/1,900 to 12,000/3,800 µg/ml. The MBEC was up to 16‐fold and eightfold higher than the MIC/MBC of NAC and Tris‐EDTA, respectively. Disodium EDTA reduced biofilm growth of both strains at concentrations of 470 µg/ml and higher. It can be concluded that biofilm production is common in pathogens associated with canine OE. NAC and Tris‐EDTA are effective antibiofilm agents in vitro that could be considered for the treatment of biofilm‐associated OE in dogs.     

Economic evaluation of artificial insemination of sex‐sorted semen on a Brown Swiss dairy farm—A case study

Artificial insemination using sex‐sorted semen is employed to efficiently increase the number of female dairy calves born. Previous studies have determined that using sex‐sorted semen is beneficial to improve the management, but the mechanism by which it increases cattle numbers through objective indices of breeding remains unclear. This study focused on a Brown Swiss cattle herd in which frozen female sex‐sorted semen was systematically employed to increase the number of cattle. We analyzed the correlation between the increase in the number of cattle and the screening accuracy of sex‐sorted semen, measuring indices such as pregnancy rate and birth rate of female calves. Study revealed that: (1) production cost for female calves is influenced by the pregnancy rate, rate of female calves, and using sex‐sorted semen is less expensive than using nonsorted semen; (2) improvements in screening accuracy nearly doubled the number of cows and tripled the number of heifers in 5 years; and (3) use of sex‐sorted semen improved milk quality. The pregnancy rate was lower when sex‐sorted semen was used, but the birth rate of heifers was improved. Results suggest that artificial insemination using sex‐sorted semen is beneficial because it economically produces offspring to increase the herd.     

Which source and level of dietary sodium is appropriate for broiler chickens reared in a high‐altitude area?

This study investigated effect of increasing level of dietary sodium using sodium bicarbonate or sodium chloride on growth performance, mortality, characteristics of carcass, organs and tibia, calcium and phosphorus of serum in broilers reared in a high‐altitude area (1,700 m above sea level). A total of 588 Ross 308 male broiler chicks were used in seven treatments, six replicates per treatment of 14 birds per each from 1 to 38 d of age. Seven dietary treatments consisted of a basal diet (with 0.16% sodium and 0.23% chloride), top‐dressed for six diets to give three supplementary levels of sodium (0.07%, 0.14% and 0.21%) from sodium bicarbonate (respectively by 0.26%, 0.52% and 0.78%) or sodium chloride (respectively by 0.18%, 0.36% and 0.54%), resulting in seven diets with total sodium and chloride levels of 0.16% and 0.23%, 0.23% and 0.23%, 0.30% and 0.23%, 0.37% and 0.23%, 0.23% and 0.33%, 0.30% and 0.44%, 0.37% and 0.55% respectively. Increasing sodium level improved feed conversion ratio (FCR) linearly and quadratically. However, when FCR was calculated without adjusting for feed intake of mortalities, the enhanced sodium level did not improve this parameter. Increasing sodium level via sodium chloride enhanced ascites mortality, total mortality, relative weight of heart and right ventricle linearly. Increasing sodium level reduced serum calcium and enhanced serum phosphorus linearly; however, there was a linear tendency to increase tibia ash when sodium level was enhanced by sodium bicarbonate (p = 0.08) or sodium chloride (p = 0.07). Increasing sodium level via sodium bicarbonate tended (p = 0.08) to reduce tibia strength linearly. In conclusion, a diet with 0.16% sodium and 0.23% chloride is enough for broiler chicken reared in a high‐altitude area, and increasing dietary sodium level via sodium chloride has detrimental effect on survivability of broiler in such condition.     

Impressions and purchasing intentions of Japanese consumers regarding pork produced by ‘Ecofeed,’ a trademark of food‐waste or food co‐product animal feed certified by the Japanese government

Impressions and purchasing intentions of Japanese consumers regarding pork produced by ‘Ecofeed’, a trademark of food‐waste or co‐product animal feeds certified by the Japanese government, were investigated by a questionnaire on the Internet. ‘Ecofeed’ did not elicit specific impressions as compared to domestic, imported, Kurobuta (in Japan), and specific pathogen‐free (SPF) pork. Purchasing intent for ‘Ecofeed’ pork was the second lowest of the five pork products. Knowledge and purchasing experience regarding ‘Ecofeed’ pork was the lowest of the five pork products. Respondents were classified into four categories according to their impressions of ‘Ecofeed’ pork. The largest category of respondents did not have any specific impression of ‘Ecofeed’ pork and had little knowledge of pork farming. A category that had a positive impression for ‘Ecofeed’ pork had high knowledge of the pork farming system. In order to establish ‘Ecofeed’ pork in Japan, our results suggest that information disclosure and education about ‘Ecofeed’, its certification system, environmental benefits and the current self‐efficiency ratio of animal feed, are needed.     

应用PCR技术检测鹅细小病毒

根据鹅细小病毒GPVH1株结构蛋白VP1与VP3编码基因非重叠核苷酸序列设计了一对引物GPR1 /GPR2 ,用这对引物对感染器官内的GPR和接种鹅胚增殖的GPV进行PCR扩增 ,其结果引物GRP1 /GPR2能特异性地扩增GPVVP1 VP3区段核苷酸序列 ,扩增出与设计核苷酸片段大小相符的 0 6kb的序列 ,而对照的鹅副粘病毒cDNA及鸭瘟病毒 (DPR)DNA对照组均出现阴性结果。     

Detection of genomic DNA of the crayfish plague fungus Aphanomyces astaci (Oomycete) in clinical samples by PCR

A diagnostic procedure, based on a polymerase chain reaction method (PCR) was developed to detect infection of crayfish with the Oomycete Aphanomyces astaci. A set of oligonucleotide primers was designed to specifically amplify A. astaci DNA in the ITS region surrounding the 5.8S rDNA gene. The PCR amplifies a 115 bp amplicon. The specificity of the primers was demonstrated by testing on 27 A. astaci strains and against 20 non-A. astaci Oomycetes and 5 fungal species. Most of the non-A. astaci Oomycete or fungal species included in the study are either known parasites of freshwater crayfish cuticle or can be found in their natural environment. Specificity was also tested against crayfish tissue and some known parasites and bacteria infecting crayfish.

A protocol for the extraction of A. astaci DNA from infected crayfish tissue was developed. The optimised method allows the detection of two genome equivalents of purified A. astaci genomic DNA.

The method was tested on noble crayfish (Astacus astacus), artificially infected with A. astaci. Detection of A. astaci was possible at the very first time of sampling, which was 2 days after the beginning of spore exposure.