Prevalence of antimicrobial resistance in enteric Escherichia coli from domestic pets and assessment of associated risk markers using a generalized linear mixed model

Antimicrobial resistance (AMR) is a growing global public health problem, which is caused by the use of antimicrobials in both human and animal medical practice. The objectives of the present cross-sectional study were as follows: (1) to determine the prevalence of resistance in Escherichia coli isolated from the feces of pets from the Porto region of Portugal against 19 antimicrobial agents and (2) to assess the individual, clinical and environmental characteristics associated with each pet as risk markers for the AMR of the E. coli isolates.     

Case–control risk factor study of methicillin-resistant Staphylococcus pseudintermedius (MRSP) infection in dogs and cats in Germany

Methicillin-resistant Staphylococcus pseudintermedius (MRSP) has emerged as a highly drug-resistant small animal veterinary pathogen. Although often isolated from outpatients in veterinary clinics, there is concern that MRSP follows a veterinary-hospital-associated epidemiology. This study's objective was to identify risk factors for MRSP infections in dogs and cats in Germany. Clinical isolates of MRSP cases (n = 150) and methicillin-susceptible S. pseudintermedius (MSSP) controls (n = 133) and their corresponding host signalment and medical data covering the six months prior to staphylococcal isolation were analysed by multivariable logistic regression. The identity of all MRSP isolates was confirmed through demonstration of S. intermedius-group specific nuc and mecA. In the final model, cats (compared to dogs, OR 18.5, 95% CI 1.8–188.0, P = 0.01), animals that had been hospitalised (OR 104.4, 95% CI 21.3–511.6, P < 0.001), or visited veterinary clinics more frequently (>10 visits OR 7.3, 95% CI 1.0–52.6, P = 0.049) and those that had received topical ear medication (OR 5.1, 95% CI 1.8–14.9, P = 0.003) or glucocorticoids (OR 22.5, 95% CI 7.0–72.6, P < 0.001) were at higher risk of MRSP infection, whereas S. pseudintermedius isolates from ears were more likely to belong to the MSSP-group (OR 0.09, 95% CI 0.03–0.34, P < 0.001). These results indicate an association of MRSP infection with veterinary clinic/hospital settings and possibly with chronic skin disease. There was an unexpected lack of association between MRSP and antimicrobial therapy; this requires further investigation but may indicate that MRSP is well adapted to canine skin with little need for selective pressure.     

Establishment of a Conditional Transgenic System Using the 2A Peptide in the Female Mouse Germline

Transgenic mice are essential research tools in developmental biology studies. The 2A peptide allows multiple genes to be expressed simultaneously at comparable levels in somatic cells, but there are no reports of it being used successfully in germ cells. We constructed a Cre/loxP-based conditional vector containing the 2A peptide to significantly enhance the expression of a reporter and target gene from a constitutive promoter in oocytes. Mice with a transgene insertion containing the chicken β-actin promoter, floxed EGFP-polyA cassette, mCherry reporter, 2A peptide and target gene DNA methyltransferase 3A2 (Dnmt3a2) were crossed with TNAP- or Vasa-Cre mice to produce offspring, in which mCherry and DNMT3A2 proteins were highly expressed in oocytes upon Cre-mediated removal of EGFP-polyA. This novel transgenic mouse line based on the 2A expression system can serve as a useful tool for examining gene function during oogenesis.     

PRRSV特异性肽对CSF、PRRS免疫猪IFN-Y、IL-10分泌的影晌

应用PRRSV-GP5特异性肽刺激CSF、PRRS免疫猪外周血淋巴细胞,采用EI。IsA方法测定不同时间点淋巴细胞分泌IFN-7及IL-10的水平,分析PRRSv_GP5特异性肽刺激后,淋巴细胞培养上清中细胞因子的变化规律。结果显示,CSFV高抗PRRSV低抗体组于PRRSV免疫14d,肽刺激组中IFN-7水平显著高于对照组（P〈0．05）;其他组均差异不显著（P〉0．05）。PRRSV免疫14d,细胞培养24,72h后加PRRSV—GP5肽刺激,各组中试验组与对照组相比IL-10水平明显降低（P〈0．01）;PRRSV免疫28d,各组中肽刺激组与空白组相比,IL-10水平均无显著性差异（P〉0．05）。结果表明,PRRSV-GP5肽在PRRSV免疫早期可以促进IFN-7的分泌,并抑制IL-10的产生,但在免疫后期对淋巴细胞分泌的调节作用还需进-步研究。     

小肽营养研究进展

小肽(sp)是动物降解蛋白质为氨基酸过程中的中间产物,对动物的重要营养有重要的营养作用。本文就小肽的吸收机制和特点、影响小肽释放和吸收的因素、小肽的营养作用以及其在动物营养中的应用等作一综述。     

Natriuretic peptide inhibition of intestinal salt absorption in the Japanese eel: physiological significance

Electrophysiological studies in vitro demonstrated the significant inhibition by natriuretic peptides (NP) of short-circuit current across the eel intestine, an important osmoregulatory organ. Inhibitory potencies of several members of the NP family were assessed by voltage-clamp determination of net transepithelial salt absorption measured as the short-circuit current I_sc across the intestine of the freshwater-adapted (FW) and seawater-adapted (SW) Japanese eel (Anguilla japonica); the order of potency of synthetic eel peptides was: amidated atrial natriuretic peptide (ANP-NH₂) > ventricular natriuretic peptide (VNP) > atrial natriuretic peptide (ANP) >> C-type natriuretic peptide (CNP). Neither the order of potency nor the absolute potencies were effected by salinity adaptation. The observed potency sequence suggests that inhibition of intestinal absorption is mediated by A-type guanylyl cyclase-coupled NP receptors. The relatively low sensitivity of the intestinal response to NP compared with circulating NP concentrations suggests a role for intestinal regulation by NP which is independent of systemic delivery from cardiac sources. A novel model, incorporating the known immunohistochemical localization of NP-ergic cells and processes in the epithelial layer of the intestine and the dissipation of the Na⁺ electrochemical gradient along the alimentary tract, is developed in which local secretion of NP (in response to a bolus of food) inhibits salt absorption across the intestine regionally in favor of increased nutrient absorption.     

Biochemistry and physiology of a family of eel natriuretic peptides

Three natriuretic peptides with similar structures were isolated from eels and their amino acid sequences determined; atrial natriuretic peptide (ANP) from atria, ventricular natriuretic peptide (VNP) from ventricles, and C-type natriuretic peptide (CNP) from brains. All three hormones were circulating in eel blood, and their plasma levels invariably decreased when eels were transferred from fresh water (FW) to seawater (SW). Eel ANP and VNP inhibited drinking in FW and SW eels. Eel ANP inhibited water and Na⁺ absorption by the intestine of SW eels. The potency of these ANP effects was 2–3 orders greater than those of other hormones which are known to have similar effects. Eel ANP and VNP induced antidiuresis but not antinatriuresis in FW eels. Eel ANP increased plasma cortisol level in SW eels but not in FW eels. The antidiuretic effect and the stimulation of cortisol secretion in eels are opposite to the ANP effects reported in mammals. These data suggest that ANP plays a complex role in the eel osmoregulation.

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Résumé Trois peptides natriurétiques présentant des structures similaires ont été isolés chez l'anguille et leurs séquences en acides aminés a été déterminées; le peptide atrial natriurétique (ANP) dans l'atria, le peptide ventriculaire natriurétique (VNP) dans les ventricules et le peptide natriurétique de type C dans le cerveau. Ces trois hormones sont présents dans le sang d'anguille et leur niveau plasmatique decroit invariablement quand les anguilles sont transférées d'eau douce en eau de mer. L'ANP et le VNP d'anguille inhibent l'action de boire chez les anguilles d'eau douce et d'eau de mer. L'ANP d'anguille inhibe l'absorption d'eau et de Na⁺ par l'intestin d'anguille en eau de mer. Ces effets de l'ANP sont 2 à 3 fois plus grands que ceux observés avec d'autres hormones qui sont connues pour avoir des effets similaires. L'ANP et le VNP d'anguille induisent l'antidiurèse mais pas l'antinatriurèse chez les anguilles d'eau douce. L'ANP d'anguille augmente chez ce poisson les niveaux de cortisol plasmatique en eau de mer mais pas en eau douce. L'effet antidiurétique et la stimulation de la sécrétion de cortisol chez l'anguille sont contraire aux effets de l'ANP observés chez les mammifères. Ces résultats suggèrent que l'ANP joue un role complexe dans l'osmorégulation de l'anguille.

     

Structure and function of antimicrobial peptide penaeidin-5 from the black tiger shrimp Penaeus monodon

The gene for penaeidin-5, an antimicrobial peptide comprising 55 amino acids, was isolated from the hemocyte of black tiger shrimp (Penaeus monodon). RT-PCR expression tests revealed that penaeidin-5 was produced in hemocytes, gills, the intestine and muscle. Western blot analysis confirmed the panaeidin-5 was aboundantin hemocytes, the intestine and hemolymph. Immunohistochemistry revealedpenaeidin-5 in the cuticle and gills that are considered primary defense barriers. The deduced amino acid sequence of penaeidin-5 included a proline-rich N-terminal domain and a carboxyl-domain that contained six cysteine residues. Circular dichrosim analysis revealed an -helix in its secondary structure and the predicted 3D structure indicated two-disulfide bridges in the -helix. Based on the sequence of penaeidin-5 peptide cDNA, synthetic penaeidin-5 was prepared to carry out functional tests. The synthetic peptide had efficient bacteriostatic and bactericidal activity against Aerococcus viridans, and also inhibited the growth of two filamentous fungi, Fusarium pisi and Fusarium oxysporum. To measure penaeidin-5 in vivo, black tiger shrimp were challenged with Vibrio alginolyticus and A. viridans. At 3 h post-challenge, penaeidin-5 was induced and bacterial numbers decreased significantly by 12 h and 24 h.     

广西巴马小型猪GHRH成熟肽真核表达质粒对小鼠生长效应的影响

【目的】探讨广西巴马小型猪促生长激素释放激素（GHRH）成熟肽真核表达质粒对小鼠生长效应的影响,为进一步研究广西巴马小型猪GHRH成熟肽的促生长作用及GHRH基因疫苗的高效制备与安全使用提供参考依据。【方法】利用基因重组技术将广西巴马小型猪GHRH成熟肽序列与p EGFP-N1质粒连接构建重组真核表达质粒,然后按0.5μg/g的剂量肌肉注射经25%蔗糖溶液预处理的小鼠,观察记录小鼠体重变化情况,并分别提取重组质粒注射组小鼠的心脏、肝脏、肾脏及注射部位肌肉DNA,再以PCR检测是否存在基因质粒残留。【结果】以构建的p EGFP-GHRH重组质粒注射小鼠后,其外源GHRH成熟肽DNA可在肌肉中获得表达,但并未整合入小鼠基因组中。虽然重组质粒注射组小鼠的末重与空质粒注射组小鼠的差异不显著（P〉0.05）,但在注射后的10-25 d,重组质粒注射组小鼠体重均显著高于空质粒注射组（P〈0.05）。在相对日增重率方面,注射后的前15 d内,重组质粒注射组小鼠的相对日增重率均高于空质粒注射组。【结论】构建的广西巴马小型猪p EGFP-GHRH重组质粒能在一定时间内促进小鼠生长,即GHRH种属差异性小,在动物中有较高的保守性。