两株分离自鸡粪便费格森埃希菌的耐药性

【背景】费格森埃希菌(Escherichia fergusonii)是与大肠杆菌同属、近源的病原菌,目前耐药性鲜有报道。【目的】对在浙江省鸡粪便中分离到的2株费格森埃希菌EFCF053和EFCF056进行耐药性检测和分析。【方法】通过微量肉汤稀释法进行MIC测定,二代高通量测序获得全基因组序列,并通过ResFinder数据库预测获得性耐药基因。利用S1-PFGE和Southernblotting杂交进行质粒和耐药基因的确认。【结果】两种菌均对氨苄西林、庆大霉素、氟苯尼考、磺胺异噁唑、复方新诺明、四环素耐药,其中菌株EFCF056还对粘菌素、头孢噻呋、大观霉素、恩诺沙星、氧氟沙星耐药。预测到耐药基因β-内酰胺类blaTEM-1A、blaCTX-M-65、blaOXA-1、blaTEM-1B、blaCTX-M-55;氨基糖苷类aac(3)-IId、aph(3')-Ia、aph(3')-Ib、aph(6)-Id、rmtB、aac(6')-Ib-cr、aadA2;粘菌素mcr-1;喹诺酮类qnrS2、aac(6')-Ib-cr、oqxA、oqxB;磷霉素fosA3;大环内酯类mph(A);苯丙醇类catA1、floR、catB3;利福霉素ARR-3;磺胺类sul1、sul2、sul3、dfrA12、dfrA14;四环素类tet(A)。另外,含有mcr-1基因的质粒通过实验证实可发生接合转移。【结论】结果显示费格森埃希菌可能是重要耐药基因存储库,费格森埃希菌与大肠埃希菌要在抗药性流行病学中加以区分,深入研究其MIC频率分布、重要耐药基因mcr-1及ESBL等,保障临床检测的准确性。     

Bioinformatic discovery and initial characterisation of nine novel antimicrobial peptide genes in the chicken

Antimicrobial peptides (AMPs) are essential components of innate immunity in a range of species from Drosophila to humans and are generally thought to act by disrupting the membrane integrity of microbes. In order to discover novel AMPs in the chicken, we have implemented a bioinformatic approach that involves the clustering of more than 420,000 chicken expressed sequence tags (ESTs). Similarity searching of proteins—predicted to be encoded by these EST clusters—for homology to known AMPs has resulted in the in silico identification of full-length sequences for seven novel gallinacins (Gal-4 to Gal-10), a novel cathelicidin and a novel liver-expressed antimicrobial peptide 2 (LEAP-2) in the chicken. Differential gene expression of these novel genes has been demonstrated across a panel of chicken tissues. An evolutionary analysis of the gallinacin family has detected sites—primarily in the mature AMP—that are under positive selection in these molecules. The functional implications of these results are discussed.     

Morphological and functional adaptations of the female reproductive system in Veigaiidae (Acari: Gamasida) and implications regarding the systematic position of the family

Veigaiidae are predatory gamasid mites with a controversial systematic position. Some authors see a close relationship with Parasitina, others with Dermanyssina. Since the morphology of the genital system is of high relevance in terms of the systematics of Gamasida, we have carefully studied the ultrastructure of the female reproductive system. It consists of two parts, one which accepts spermatozoa (sperm access system), and the other which produces and delivers eggs. Since we have already published a detailed account of the sperm access system we add only some further information here on that system and focus instead on egg production and delivery. The unpaired gonad is divided into a germinal and a nutritive region, the latter of which is a compact structure as in Parasitina and does not show lateral arms as seen in most studied Dermanyssina. The oviduct (uterus, vaginal duct) is an unpaired structure that terminates in a vagina (genital atrium) covered by the genital plate. Two small vaginal glands open into the vagina, which is provided with a peculiar interlocking system. First observations on spermatozoa found in one female reveal similarities with those of Parasitina. Our results are discussed under morphological aspects and compared with other gamasid mites. In conclusion, the veigaiids show some peculiar features that are shared either with Parasitina or with Dermanyssina. These observations make it difficult to simply assume that the Veigaiidae belong either to the Dermanyssina or to the Parasitina. They appear instead to comprise an intermediate entity.     

The role of heat shock proteins in inflammatory injury induced by cold stress in chicken hearts

The aim of this study was to investigate the effects of cold stress on the expression levels of heat shock proteins (Hsps90, 70, 60, 40, and 27) and inflammatory factors (iNOS, COX-2, NF-κB, TNF-α, and PTGEs) and oxidative indexes in hearts of chickens. Two hundred forty 15-day-old male chickens were randomly divided into 12 groups and kept at the temperature of 12 ± 1 °C for acute and chronic cold stress. There were one control group and five treatment groups for acute cold stress, three control groups, and three treatment groups for chronic cold stress. After cold stress, malondialdehyde level increased in chicken heart; the activity of superoxide dismutase and glutathione peroxidase in the heart first increased and then decreased. The inflammatory factors mRNA levels were increased in cold stress groups relative to control groups. The histopathological analysis showed that heart tissues were seriously injured in the cold stress group. Additionally, the mRNA levels of Hsps (70, 60, 40, and 27) increased significantly (P < 0.05) in the cold stress groups relative to the corresponding control group. Meanwhile, the mRNA level and protein expression of Hsp90 decreased significantly (P < 0.05) in the stress group, and showed a gradually decreasing tendency. These results suggested that the levels of inflammatory factors and Hsps expression levels in heart tissues can be influenced by cold stress. Hsps commonly played an important role in the protection of the heart after cold stress.     

Immunolocalization of the hyaluronan receptor CD44 in the reproductive tract of the mare

Hyaluronan (HA), a glycosaminoglycan, is a major component of the pericellular matrix which envelopes mammalian cells. Binding of hyaluronan to one of its specific receptors, CD44, modulates transduction of intracellular signals which direct a variety of processes, including embryogenesis, wound healing, inflammation, and neoplasia. Since regulation of these processes is critical to equine reproductive success, localization of constitutive CD44 expression was evaluated by immunohistochemical methods in ovarian, oviductal, and uterine tissues from healthy mares. Ovarian stroma contained thecal cells with varying CD44 immunopositivity. Follicular and granulosa cells of some antral and atretic follicles were positive for CD44. In the oviduct, the luminal epithelium was variably positive for CD44, with overall decreasing intensity of immunostaining from the infundibulum to the isthmus. The CD44 molecule was expressed strongly by surface epithelial cells of the uterine endometrium, but was present only rarely among cells of uterine glands. In addition, CD44 was expressed by smooth muscle cells of vascular walls, oviduct, and uterus. Since CD44 is known to modulate cell movement and differentiation, and was present at multiple sites in the reproductive tract of normal mares, we inferred there may be an important role for the HA-CD44 signaling pathway in reproductive function and inflammation.     

Localization of the angiopoietin receptors Tie-1 and Tie-2 on the primary cilia in the female reproductive organs

Blood vessel homeostasis and endothelial cell survival depend on proper signalling through angiopoietin receptors such as the receptor tyrosine kinases Tie-1 and Tie-2. We have studied the presence and subcellular localization of these receptors in murine female reproductive organs using confocal microscopy analysis of antibody stained tissue sections of ovary and oviduct. We show that Tie-2 principally localizes to primary cilia of the surface epithelium of the ovary, bursa and extra-ovarian rete ducts as well as to plasma membranes of ovarian theca and endothelial cells. Primary cilia of follicular granulosa cells were negative. Further, Tie-1 and Tie-2 localized to motile cilia of the oviduct. Western blotting detection and immunolocalization of anti-Tie-2 in ovary and oviduct were abolished by administration of an anti-Tie-2 blocking peptide, confirming antibody specificity. In a series of immunohistochemical analysis on human ovarian tissues we also observed a unique localization of Tie-2 to the primary cilia of ovarian surface epithelium. These observations are the first to show ciliary localization of angiopoietin receptors. Our results support the hypothesis that cilia of the female reproductive organs play a novel and important sensory role in relaying physiochemical changes from the extracellular environment to epithelial cells of the oviduct, the ovary and extra-ovarian tissues.     

The effects of histones H1o,H5 and HMG proteins on cell division of cultured murine erythroleukemia cells

Summary In the present study the effect of histones H1^o and H5, and the nonhistone chromatin proteins HMG 1, 2, 14 and 17 (the high mobility group proteins), as well as the acidic peptide fragments of HMG 1 and 2 and polyglutamate, on cell division and differentation of cultured murine erythroleukemia (Friend) cells has been investigated. It was found that histones H1^o and H5, the acidic peptide fragments of HMG 1 and 2, HMG 14 and 17 and sodium polyglutamate stimulated cell division at a concentration of 10 g/ml. None of the H1^o, H5 or HMG protein preparations induced hemoglobin synthesis, as judged by benzidine staining.     

Cloning and expression analysis of androgen receptor gene in chicken embryogenesis

We cloned a full-length androgen receptor (AR) cDNA from chicken (Gallus gallus) gonads. The cDNA sequence has an open reading frame of 2109 bp encoding 703 amino acids. The chicken AR (cAR) shares high homology with ARs from other species in its amino acid sequences, in particular DNA binding domain (DBD) and ligand binding domain (LBD). RT-PCR analysis revealed that cAR mRNA is expressed in several embryonic tissues of both sexes, and relatively higher expression was observed in left ovary compared with testis. The immunoreactive signal of AR was co-localized within the ovarian cell nucleus, while such nuclear localization was not detected in those of testis. To get insight on the possible role of androgen-AR signaling during gonadal development, non-steroidal AR antagonist, flutamide, was administrated in ovo. The treatment induced the disorganization of sex cords in ovarian cortex at day 12 of incubation. The effect was restored by testosterone co-treatment, implying the possibility that AR mediated signaling may be involved in ovarian morphogenesis. Furthermore, co-treatment of flutamide with estradiol-17beta (E2) also restored the phenotype, suggesting androgen-AR signaling might activate aromatase expression that is necessary for estrogen synthesis. These findings suggest androgen-AR signaling might contribute to chicken embryonic ovarian development.     

Development of the calyx and lateral oviduct during oogenesis in Aedes aegypti

Summary The lateral oviduct and calyx of nulliparous Aedes aegypti on a sucrose diet are both flattened sacs, lacking a well defined lumen. Both are formed of an inner epithelial and an outer muscular layer, each one cell thick. The lateral oviduct is surrounded by a circular muscle sheath which is continuous with the ovarian sheath. Each ovariolar sheath is continuous with the outer layer of the calyx. The structure of both the lateral oviduct and the calyx is greatly modified after the initial blood meal. A distinct lumen develops; there is an extensive development of the outer muscular layers, and the inner epithelial layers become invaginated forming deep crypts lined with extensive microvilli. The follicular stem, which joins the primary follicle to the calyx in each ovariole, is not hollow and does not mark the opening into the calyx through which the mature egg can pass. The eggs gain access to the oviductal system after the calyx extends around the follicular epithelium of the primary follicle, when breaks appear in the calyx wall opposed to the follicular epithelium, until the mature eggs, eventually lie in a highly distended thin-walled sac of calyx from which they have direct and easy access to the lateral oviduct. After oviposition, this sac contracts to occupy once more a compact axial position in the ovary. Remnants of the follicular epithelium, containing many lysosomes are attached to the calyx at this time.     

线虫Distolabrellus veechi的生殖系统及配子发生

以线虫Distolabrellus veechi的卵巢和精巢为材料,用甲醇固定,DAPI荧光染料对细胞核进行染色,通过Normaski荧光显微镜对其生殖系统结构、性细胞染色体数目、体细胞染色体数目进行了观察.结果显示了该线虫雌虫和雄虫生殖细胞减数分裂Ⅰ前期的细胞核的变化特征;减数分裂Ⅰ前期丝球期雌、雄生殖细胞的二价体数目均为6;卵细胞染色体数目为6,精细胞染色体数目为5或6,其性别决定机制为XX/X0型;体细胞染色体数目为11(雄虫)或12(雌虫)[动物学报 54(3):500-509,2008].     

12 402例女性生殖道感染病原体分布以及耐药状况分析

目的了解宁波地区近年来女性生殖道感染病原体分布以及耐药状况,为临床用药提供实验依据。方法对2003年1月至2006年11月就诊于性病科、妇科、生殖中心、不孕不育科的拟诊生殖道感染患者,取宫颈分泌物进行病原体检测及药敏试验。结果12 402例中共检出生殖道感染患者4827例,感染率为38.92%,检出病原体6164株,1216人感染了2种或2种以上病原体,混合感染率为9.80%。结论感染病原体中,解脲脲原体占44.7%,且部分菌株耐药严重,临床医生治疗前应尽可能做药敏试验,做到合理用药,以提高妇女的生殖健康水平。     

Ovarian development and insulin-signaling pathways during reproductive differentiation in the queenless ponerine ant Diacamma sp.

In many social hymenopteran species, workers possess functional ovaries that are physiologically inactive in the presence of queens. We investigated the ovarian regulatory mechanism of workers and reproductives in a queenless ponerine ant, Diacamma sp., using histological and molecular techniques. In this ant, clear reproductive differentiation occurs via a highly sophisticated dominance behavioral interaction called “gemmae mutilation”. This clear and rapid bifurcation of reproductive physiology allows us to elucidate the detailed ovarian differentiation process. Histological characteristics of functional ovaries (fusomes and ring canals) were found in both workers and reproductives, suggesting that early oogenesis is not blocked in workers. Since insulin/insulin-like growth factor signaling (IIS) is known to control insect reproduction, orthologs of 2 positive IIS regulators, insulin receptor and serine-threonine kinase Akt (protein kinase B), were cloned in Diacamma (DiaInR, DiaAkt); their expression patterns during reproductive differentiation were examined by real-time quantitative polymerase chain reaction; DiaInR and DiaAkt were strongly expressed in the gasters of reproductives. Whole-mount in situ hybridization of ovaries indicated that DiaInR and DiaAkt were expressed in nurse cells, oocytes, and upper germarial regions of reproductives but not of workers. Our data suggest that the IIS pathway accounts for reproductive differentiation in late oogenesis.     

Ultra- and microstructure of the female reproductive system of Matsucoccus matsumurae

The ultra- and microstructure of the female reproductive system of Matsucoccus matsumurae was studied using light microscopy, scanning and transmission electron microscopy. The results revealed that the female reproductive system of M. matsumurae is composed of a pair of ovaries, a common oviduct, a pair of lateral oviducts, a spermatheca and two pairs of accessory glands. Each ovary is composed of approximately 50 telotrophic ovarioles that are devoid of terminal filaments. Each ovariole is subdivided into an apical tropharium, a vitellarium and a short pedicel connected to a lateral oviduct. The tropharium contains 8–10 trophocytes and two early previtellogenic oocytes termed arrested oocytes. The trophocytes degenerate after egg maturation, and the arrested oocytes are capable of further development. The vitellarium contains 3–6 oocytes of different developmental stages: previtellogenesis, vitellogenesis and choriogenesis. The surface of the vitellarium is rough and composed of a pattern of polygonal reticular formations with a center protuberance. The oocyte possesses numerous yolk spheres and lipid droplets, and is surrounded by a mono-layered follicular epithelium that becomes binucleate at the beginning of vitellogenesis. Accessory nuclei are observed in the peripheral ooplasm during vitellogenesis.     

Differential expression of p63 isoforms in female reproductive organs

p63 is the identity switch for uterine/vaginal epithelial cell fate, and disruption of p63 expression by diethylstilbestrol (DES) induces cervical/vaginal adenosis in mice. In this article, we report the expression patterns of p63 isoforms (TA, DeltaN, alpha, beta and gamma) in mice, focusing on the reproductive tract. We also present the reproductive tract phenotype of female p63-/- mice. Finally, to better evaluate the potential role of p63 in human development of DES-induced cervical/vaginal adenosis, we describe the ontogeny of p63 in human female fetuses. In adult mice, the DeltaN isoforms of p63 were expressed only in squamous/basal/myoepithelial cells of epithelial tissues, while TA isoforms of p63 were highly expressed in germ cells of the ovary and testis. In fetal mice, the DeltaN and alpha forms of p63 were expressed in the cloacal and urogenital sinus epithelia. In the female p63-/- mice, the sinus vagina developed, but p63-/- sinus vaginal epithelium failed to undergo squamous differentiation confirming an essential role of p63 in squamous epithelial differentiation. Although TAp63 was highly expressed in developing primordial germ cells/oocytes, p63-/- ovaries and oocytes developed normally. The ontogeny of p63 in female reproductive organs was essentially identical in mouse and human. In the human fetus at the susceptible stage for DES-induced cervical/vaginal adenosis, most cervical/vaginal epithelial cells were columnar and negative for p63. Therefore, inhibition of p63 expression by DES should change the cell fate of human Müllerian duct epithelial cells and cause cervical/vaginal adenosis as previously demonstrated in mouse.     

The effect of hCG treatment on Day 12 post-mating on ovarian function and reproductive performance of ewes and ewe lambs

The objectives of this study were to determine the effect of a single intramuscular injection of 200 IU hCG (Chorulon) on Day 12 post-mating on ovarian function and subsequent lambing performance in ewes and ewe lambs bred at synchronised oestrus during the breeding season and on the lambing performance of ewes induced to breed during late anoestrus. All animals were mated to rams at synchronised oestrus and on Day 12 post-mating given normal saline or 200 IU hCG.In Experiment 1, laparoscopic results showed that hCG treatment induced accessory corpora lutea in ewes (control = 0/7; hCG = 5/7) but not in ewe lambs (control = 0/7; hCG = 0/7).In Experiment 2, hCG treatment did not improve the lambing rate (control = 50; hCG = 57) or the litter size (control = 1.80; hCG = 1.96) in ewes (control = 100; hCG = 91). However, hCG treatment significantly (P > 0.05) improved the lambing rate (control = 29; hCG = 58; P < 0.05) in ewes conceiving at the first oestrus after treatment. hCG treatment (control = 42; hCG = 42) also failed to improve the lambing rate in ewe lambs (control = 48; hCG = 41).In Experiment 3, hCG treatment had no significant (P > 0.05) effect on the lambing rate (control = 72; hCG = 62) or the litter size (control = 1.59; hCG = 1.58) in ewes (control = 111; hCG = 115) induced to breed during anoestrus or on ewes returning to oestrus and conceiving after treatment (lambing rate: control = 86; hCG = 72; litter size: control = 1.44; hCG = 1.35). In conclusion, the data obtained in this study suggest that during the breeding season hCG may, by stimulating ovarian function, improve embryo survival in ewes conceiving at the first post-treatment oestrus. This effect, however was not observed in ewe lambs.     

Development of myofibrils in the gizzard of chicken embryos

Summary The intracellular distributions of major muscle proteins, myosin, actin, tropomyosin, -actinin, and desmin, in smooth muscle cells of chicken gizzard at various stages of embryogenesis were investigated by immunofluorescence-labeling of enzyme-dispersed cells cultured up to three hours. These muscle proteins, except some part of myosin, were organized into fibrous structures as soon as synthesis and accumulation of proteins started. As for myosin, a considerable amount of it was dispersed in soluble cytoplasm as well. On the other hand, Ca⁺⁺-dependent contractility was detected with detergent-extracted myoblasts and glycerinated tissue from embryos older than 7 days. Although the nascent myofibrils bear a resemblance to stress fibers, the former could be distinguished from the latter by their high stability in dispersed, spherical cells. The above findings, therefore, show that the synthesis of contractile proteins is followed by immediate assembly of them into functional myofibrils without undergoing any intermediate structure. Based on these findings, the mechanism of myofibril formation in developing smooth muscle cells is discussed.