OIE report on meeting_ safety genetic engineered vaccine food animals_Paris 10

NB: Version adopted by the World Assembly of Delegates of the OIE in May 2010C H A P T E R2.4.13.I N F E C T I O U S B O V I N E R H I N O T R A C H E I T I S/I N F E C T I O U S P U S T U L A R V U L V O V A G I N I T I SSUMMARYDefinition of the disease: Infectious bovine rhinotracheitis/infectious pustular vulvovaginitis (IBR/IPV), caused by bovine herpesvirus 1 (BoHV-1), is a disease of domestic and wild cattle. The virus is distributed world-wide, but has been eradicated from Austria, Denmark, Finland, Sweden, Italy (Province of Bolzano), Switzerland, Norway and parts of Germany (the ‘Oberfranken’ and ‘Oberpfalz’ districts of Bavaria). Control programmes are running in several other countries, for example in Germany and Italy.Description of disease: The disease is characterised by clinical signs of the upper respiratory tract, such as a (muco)purulent nasal discharge, hyperaemia of the muzzle (red nose disease) and by conjunctivitis. Signs of general illness are fever, depression, inappetence, abortions and reduced milk yield. The virus can also infect the genital tract and cause pustular vulvovaginitis and balanoposthitis. Post-mortem examinations reveal rhinitis, laryngitis and tracheitis. Mortality is low, and most infections run a subclinical course. Secondary bacterial infections can lead to more severe respiratory disease, and BoHV-1 could play a role in multifactor diseases such as ‘shipping fever’.Identification of the agent: The virus can be isolated from nasal or genital swabs from animals with respiratory signs, vulvovaginitis or balanoposthitis, taken during the acute phase of the infection, and, in severe cases, from various organs collected at post-mortem. Following infection, BoHV-1 may persist in infected animals in a latent state in sensory neurons, e.g. in the trigeminal or sacral ganglia. The virus can be reactivated and this results in virus shedding (re-excretion) without exhibition of clinical disease. Therefore, antibody-positive animals have to be classified as infected with BoHV-1 (with two exceptions: serological responses induced by vaccination with an inactivated vaccine or by colostral antibodies).For virus isolation, various cell cultures of bovine origin are used, for example, secondary lung or kidney cells or the Madin–Darby bovine kidney cell line (MDBK). The virus produces a cytopathic effect in 2–4 days. It is identified by neutralisation or antigen detection methods using monospecific antisera or monoclonal antibodies. BoHV-1 isolates can be further subtyped by DNA restriction enzyme analysis (RFLP) into subtypes 1.1 and 1.2. BoHV-1.2 isolates can be further differentiated into 2a and 2b. Development of rhinotracheitis or vulvovaginitis/balanoposthitis depends more on the route of infection than on the subtype of the virus. The virus previously referred to as BoHV-1.3,a neuropathogenic agent, is now classified as BoHV-5.Viral DNA detection methods have been developed, and the polymerase chain reaction technique is increasingly used in routine diagnosis including real-time polymerase chain reaction (PCR).Serological tests: The virus neutralisation test and various enzyme-linked immunosorbent assays (ELISA; indirect or gB-blocking) are most widely used for antibody detection. With the ELISAs, antibodies can be detected in serum or plasma, and with lower sensitivity in milk or bulk milk samples.Requirements for vaccines and diagnostic biologicals: Inactivated and attenuated live vaccines are available. The vaccines protect cattle clinically in case of infection and markedly reduce the subsequent shedding of field virus. Although vaccination may not prevent field virus infection of individual animals, spreading of wild-type virus in infected herds is efficiently reduced. The vaccines must not induce disease, abortion, or any local or systemic reaction, and must be genetically stable.BoHV-1 glycoprotein E deleted mutant marker vaccines are now generally available (live orinactivated). The use of a gE-antibody-ELISA makes it possible to distinguish field virus infectedcattle from cattle vaccinated with such a marker vaccine (DIVA strategy).A. INTRODUCTIONInfectious bovine rhinotracheitis/infectious pustular vulvovaginitis (IBR/IPV), caused by bovine herpesvirus 1 (BoHV-1), is a disease of domestic and wild cattle. BoHV-1 is a member of the genus Varicellovirus in the subfamily Alphaherpesvirinae, which belongs to the Herpesviridae family, order Herpesvirales. The viral genome consists of double-stranded DNA that encodes for about 70 proteins, of which 33 structural and more than 15 nonstructural proteins have been identified. The viral glycoproteins, which are located in the envelope on the surface of the virion, play an important role in pathogenesis and immunity. BoHV-1 can be differentiated into subtypes 1.1, 1.2a and 1.2b (Metzler et al., 1985). The BoHV-1.2 subtypes may be less virulent than subtype 1.1 (Edwards et al., 1990). The former BoHV-1.3, which may act as a neuropathogenic agent in calves, has been re-classified as BoHV-5 (Magyar et al., 1993). BoHV-1 shares antigenic and genetic close relationships with other ruminant alphaherpesviruses: BoHV-5, caprine herpesvirus 1, cervid herpesvirus 1 (red deer), cervid herpesvirus 2 (reindeer), bubaline herpesvirus 1 and elk herpesvirus 1 (Thiry et al., 2006).After an incubation period of 2–4 days, serous nasal discharge, salivation, fever, inappetence, and depression become evident. Within a few days the nasal and ocular discharges change to mucopurulent. Where natural mating is practised, genital infection can lead to pustular vulvovaginitis or balanoposthitis. However, most infections run a very mild or subclinical course (Van Oirschot et al., 1993). Uncomplicated cases of respiratory or genital disease caused by BoHV-1 last about 5–10 days. Secondary bacterial or viral agents may contribute to a multifactor disease complex resulting in severe respiratory disease of young animals (‘shipping’ or ‘crowding fever’).After infection via the airborne route, BoHV-1 replicates to high titres in mucous membranes of the upper respiratory tract and in the tonsils. Subsequently, the virus disseminates to conjunctivae and reaches the trigeminal ganglia by neuronal axonal transport. After genital infection, BoHV-1 replicates in the mucous membranes of the vagina or prepuce, and becomes latent in the sacral ganglia. The viral DNA remains in the neurons of the ganglia, probably for the entire life of the host (status of latency). Stress, such as transport and parturition, but also the application of corticosteroids can induce reactivation of the latent infection. Consequently, the virus may switch between latent and lytic infection and may be shed intermittently into the environment and spread to contact animals.BoHV-1 infection elicits an antibody response and a cell-mediated immune response within 7–14 days. The immune response is presumed to persist life-long, although it may fall below the detection limit of some tests after a number of years. Maternal antibodies are transferred via colostrum to the young calf, which is consequently protected against BoHV-1-induced clinical disease (Mechor et al., 1987). Maternal antibodies have a biological half-life of about 3 weeks, but may be detected occasionally in animals up to 9 months old, and rarely in animals over this age.The virus is distributed world-wide, with the exception of the BoHV-1-free countries, paralleling the distribution of domestic cattle. Other Artiodactyla (e.g. goats, sheep, water buffaloes, camelids) may be infected with BoHV-1. After infection, nasal viral shedding is detected for 5–14 days, with peak titres of 108–1010 TCID50 (50% tissue culture infective doses) per ml of nasal secretion. The semen of an infected bull may contain BoHV-1, and the virus can thus be transmitted by natural mating and artificial insemination (Parsonson & Snowdon, 1975). Prevention and control of BoHV-1 infections are based on thorough farm management including hygienic measures, vaccination schedules and removal of infected animals. Ideally, a 4-week quarantine period is imposed for newly introduced cattle, if the cattle are not from certified BoHV-1-free farms. Only cattle that are BoHV-1-seronegative are then admitted to a free herd. Natural mating should be avoided and only semen from BoHV-1-negative bulls should be used.Vaccines usually prevent the development of clinical signs and markedly reduce the shedding of virus after infection, but do not completely prevent infection. Several eradication campaigns have been carried out or are currently running in different countries including test-and-removal programmes and/or vaccination campaigns (see Section C).BoHV-1 infection may be suspected on the basis of clinical, pathological and epidemiological findings. However, to make a definite diagnosis, laboratory examinations (serology and/or virus detection) are required. A complete diagnostic procedure in the laboratory is aimed at detecting the causative virus (or viral components) and the specific antibodies they induce. Nevertheless, because of latent infection induced by BoHV-1, detection of antibodies could be sufficient for the determination of the BoHV-1 status of individual animals.B. DIAGNOSTIC TECHNIQUES1. Identification of the agenta) Collection and processing of samplesNasal swabs are collected from several (from five to ten) affected cattle in the early phase of the infection.These cattle still have serous rather than mucopurulent nasal discharge. In cases of vulvovaginitis or balanoposthitis, swabs are taken from the genitals. The swabs should be vigorously rubbed against the mucosal surfaces. The prepuce can also be washed with saline; the washing fluid is then collected. The specimens are suspended in transport medium (cell culture medium containing antibiotics and 2–10% BoHV-1-free fetal bovine serum to protect the virus from inactivation), cooled at 4°C, and rapidly submitted to the laboratory.During necropsy, mucous membranes of the respiratory tract, and samples of the tonsil, lung and bronchial lymph nodes are collected for virus detection. In cases of abortion, the fetal liver, lung, spleen, kidney and placental cotyledons are examined. Samples should be kept on ice and sent to the laboratory as quickly as possible.After arrival at the laboratory, swabs are agitated at room temperature for 30 minutes in the transport medium to elute virus. Following removal of the swabs, the transport medium is clarified by centrifugation at 1500 g for 10 minutes. Tissues are homogenised to a 10–20% (w/v) suspension in cell culture medium before centrifugation at 1500 g for 10 minutes. The supernatants of these specimens are filtered through0.45 µm filters and used for virus isolation.The isolation of virus from semen needs some special adaptations, because the seminal fluid contains enzymes and other factors that are toxic to the cells and inhibit viral replication (see below).b) Virus isolationFor virus isolation, bovine cells of various origins can be used. Primary or secondary bovine kidney, lung or testis cells, cell strains derived from bovine fetal lung, turbinate or trachea, and established cell lines, such as the Madin–Darby bovine kidney cell line (MDBK), are suitable for BoHV-1 propagation. Cell cultures can be grown in glass or plastic tubes, plates or dishes. When 24-well plastic plates are used, a 100–200 µl volume of the supernatants described above is inoculated into these cell cultures. After a 1-hour adsorption period, the cultures are rinsed and maintenance medium is added. The serum used as a medium supplement in the maintenance medium should be free of antibodies against BoHV-1. The cell cultures are observed daily for CPE, which usually appears within 3 days after inoculation. It is characterised by grape-like clusters of rounded cells gathered around a hole in the monolayer; sometimes giant cells with several nuclei may be observed. Experience is needed to recognise this characteristic appearance. When, after7 days, no CPE has appeared, a blind passage must be made. The cell culture is freeze–thawed andclarified by centrifugation, and the supernatant is used for inoculation of fresh monolayers (Brunner et al., 1988; Edwards et al., 1983).To identify the recovered virus as BoHV-1, the supernatant of the culture should be neutralised with a monospecific BoHV-1 antiserum or neutralising monoclonal antibody (MAb). For this purpose, serial tenfold dilutions of the test supernatant are made, and to each dilution monospecific BoHV-1 antiserum or negative control serum is added. Following incubation at 37°C for 1 hour, the mixtures are inoculated into cell cultures; 3–5 days later, the neutralisation index is calculated. The neutralisation index is the virus titre (in log10) in the presence of negative control serum minus the virus titre in the presence of specific antiserum. If the neutralisation index is greater than 1.5, the isolate may be considered to be BoHV-1. To shorten the virus isolation procedure, two specimens may be inoculated into cell culture: one that has been pre-incubated with monospecific antiserum and another that has been preincubated with negative control serum.If the CPE is inhibited by the monospecific antiserum, the isolate can be considered to be BoHV-1, although definitive confirmation would require molecular characterisation to distinguish it from related ruminant alphaherpesviruses.An alternative method of virus identification is the direct verification of BoHV-1 antigen in cells around the CPE by an immunofluorescence or immunoperoxidase test (Kaashoek et al., 1994) with conjugated monospecific antiserum or MAb. Furthermore, the supernatant can be used as template for restriction endonuclease fragment length polymorphism (RFLP) (see Section B.1.e) and polymerase chain reaction (PCR) (see Section B.1.c) analyses.• Virus isolation from semen (a prescribed test for international trade)0.05 to 0.1 ml of raw semen should be tested with two passages in cell culture. Raw semen is generallycytotoxic and should be prediluted (e.g. 1/10) before being added to cell cultures. A similar problem maysometimes arise with extended semen. For extended semen, an approximation should be made to ensure that the equivalent of a minimum of 0.1 ml raw semen is examined (e.g. a minimum of 0.5 ml extended serum). Multiple diluted samples may need to be tested with this procedure to reach a volume equivalent to0.1 ml raw semen (e.g. 5 × 1 ml of a 1/10 diluted sample of extended semen). A suitable test procedure isgiven below. See also Brunner et al. (1988).procedure• Testi) Dilute 200 µl fresh semen in 2 ml fetal bovine serum (free from antibodies against BoHV-1) withantibiotics.ii) Mix vigorously and leave for 30 minutes at room temperature.iii) Inoculate 1 ml of the semen/serum mixture into a monolayer of susceptible cells (see virus isolation above) in a six-well tissue culture plate.iv) Incubate the plates for 1 hour at 37°C.v) Remove the mixture, wash the monolayer twice with 5 ml maintenance medium, and add 5 ml maintenance medium to each well.vi) Include BoHV-1 negative and positive controls in the test. Special caution must be taken to avoid accidental contamination of test wells by the positive control, for example always handling the control last, and using separate plates.vii) Observe plates under a microscope daily for the appearance of a CPE. If a CPE appears, confirmatory tests for BoHV-1 are made by specific neutralisation or immunolabelling methods (see above).viii) If there is no CPE after 7 days, the cultures are frozen and thawed, clarified by centrifugation, and the supernatant is used to inoculate fresh monolayers.ix) The sample is considered to be negative, if there is no evidence of a CPE after 7 days’ incubation of the passaged cultures.c) Nucleic acid detectionDuring the past decade, various methods for the detection of BoHV-1 DNA in clinical samples have been described, including DNA–DNA hybridisation and the PCR. The PCR is also increasingly used in routine diagnostic submissions (Moore et al., 2000). Compared with virus isolation, the PCR has the primary advantages of being more sensitive and more rapid: it can be performed in 1–2 days. It is also possible to detect episomal DNA of non-replicating virus in sensory ganglia (Van Engelenburg et al., 1993), such as the trigeminal ganglion, in the latent phase of infection. The disadvantage is that PCR analyses are prone to contamination and therefore precautions have to be taken to prevent false-positive results. Risk of contamination is markedly reduced by new PCR techniques, such as real-time or quantitative PCR (see below) (Abril et al., 2004; Lovato et al., 2003).So far PCR has been used mainly to detect BoHV-1 DNA in artificially (Kramps et al., 1993) or naturally (Van Engelenburg et al., 1993) infected semen samples. It is important to thoroughly optimise the PCR conditions, including the preparation of the samples, the concentration of Mg2+, primers and polymerase, and the cycle programmes. The target region for amplification must be present in all BoHV-1 strains, and its nucleotide sequence must be conserved. The TK, gB, gC, gD and gE genes have been used as targets for PCR amplification. In addition, PCRs based on detection of gE sequences can be used to differentiate between wild-type virus and gE-deleted vaccine strains (Fuchs et al., 1999; Schynts et al., 1999). Discrimination between infection with virulent IBR strains and infection with live attenuated strains is not possible with the PCR technique, and RFLP is used for this purpose. Specific PCRs have been developed that are able to discriminate between BoHV-1, BoHV-5 and other related alphaherpesviruses (Ashbaugh et al., 1997; Ros et al., 1999).Experimentally, PCR was found to be more sensitive than virus isolation: in egg yolk-extended semen samples obtained from experimentally infected bulls, PCR detected five times as many positives as did virus isolation (Van Engelenburg et al., 1995). The detection limit of validated PCR assays amounts to only a few genome copies per PCR reaction. Nevertheless, false-negative results cannot be excluded. To identify possible false-negative results, it is recommended to spike an internal control template into the reaction tube of the semen sample to be amplified by the same primers. Such a control template may be constructed by inserting, for example, a 100 base-pair fragment into the target region. This control template also makes it possible to semi-quantify the amount of DNA that is detected (Ros et al., 1999; Van Engelenburg et al., 1993). When using an internal control, extensive testing is necessary to ensure that PCR amplification of the added internal control does not compete with the diagnostic PCR and thus lower the analytical sensitivity (see also Chapter 1.1.5 Validation and quality control of PCR methods used for the diagnosis of infectious diseases). DNA extraction and quality of the DNA preparations can also be controlled by amplification of cellular sequences (housekeeping genes) or by addition of ‘artificial’ DNA sequences prior to extractionprocedures (e.g. green fluorescent protein, non-BoHV-related viruses) as internal controls. To enhance the sensitivity and specificity of the BoHV-1 PCR, real-time PCR systems are the methods of choice.• Real-time polymerase chain reaction (a prescribed test for international trade)The following real-time PCR test method has been developed to detect BoHV-1 in extended bovine semen destined for trade. The method has been validated according to Chapter 1.1.5, and includes a comprehensive international inter-laboratory comparison involving six collaborating laboratories with specialist status in IBR testing (Wang et al., 2008).A number of studies have shown that PCR assays are more sensitive than virus isolation (Smits et al., 2000;Van Engelenburg et al., 1995; Vilcek et al., 1994; Wang et al., 2008; Wiedmann et al., 1993). Real-time PCR has been used for the detection of BoHV-1 and BoHV-5 in experimentally infected cattle and mice (Abril et al., 2004; Lovato et al., 2003) and a series of conventional PCR assays have been used for the detection of BoHV-1 DNA in artificially or naturally infected bovine semen samples (Deka et al., 2005; Grom et al., 2006;Masri et al., 1996; Van Engelenburg et al., 1993; Weiblen et al., 1992; Wiedmann et al., 1993; Xia et al., 1995). Conventional detection of amplified PCR products relies on gel electrophoresis analysis (Rola et al., 2003). Sequence-specific primers have been selected to amplify different parts of conserved glycoprotein genes of the BoHV-1 genome, including glycoprotein B (gB) gene (Grom et al., 2006; Santurde et al., 1996), gC gene (Smits et al., 2000; Van Engelenburg et al., 1995), gD gene (Smits et al., 2000; Wiedmann et al., 1993), gE gene (Grom et al., 2006), and the thymidine kinase (tk) gene (Moore et al., 2000; Yason et al., 1995).Real-time PCR differs from standard PCR in that the amplified PCR products are detected directly during the amplification cycle using a hybridisation probe, which enhances assay specificity. Real-time PCR assays have several advantages over the conventional PCR methods. Real-time PCR assays are able to provide sensitivity close or equal to nested PCR methods with a much lower risk of contamination. The amplification and detection of target is conducted simultaneously, and tubes have not to be opened for product analysis on agarose gels. There is no post-amplification PCR product handling, which significantly reduces the risk of contamination, and it is possible to perform quantitative analysis.The real-time PCR described here uses a pair of sequence-specific primers for amplification of target DNA and a 5’-nuclease oligoprobe (TaqMan) for detection of amplified products. The oligoprobe is a single, sequence-specific oligonucleotide, labelled with two different fluorophores, the reporter/donor, 5-carboxyfluorescein (FAM) at the 5’ end, and the acceptor/quencher 6-carboxytetramethylrhodamine (TAMRA) at the 3’ end. This real-time PCR assay is designed to detect viral DNA of all BoHV-1 strains, including subtype 1 and 2, from extended bovine semen. The assay selectively amplifies a 97 basepair sequence of the glycoprotein B (gB) gene. Details of the primers and probes are given in the protocol outlined below.• Sample preparation, equipment and reagentsi) The samples used for the test are, typically, extended bovine semen stored in liquid nitrogen. Thesemen samples can be transported to the laboratory in liquid nitrogen, or shipped at 4°C, and stored inliquid nitrogen or at –70°C (for long-term storage) or 4°C (for short-term storage). Storing semen at 4°Cfor a short period (up to 7 days) does not affect PCR test result.ii) Three straws from each batch of semen should be processed. Duplicate PCR amplifications should be carried out for each DNA preparation (six amplifications in total) to ensure the detection of DNA in samples containing low levels of virus.iii) A real-time PCR detection system, and the associated data analysis software, is required to perform the assay. A number of real-time PCR detection systems are available from various manufacturers. Inthe procedure described below, a RotorGene 3000, Corbett Research Ltd, Australia, was used. Otherreal-time PCR detection systems can also be applied. Other equipment required for the test includes amicro-centrifuge, a heating block, a boiling water bath, a micro-vortex, magnetic stirrer and micropipettes. Real-time PCR assays are able to detect very small amounts of target nucleic acid molecules therefore appropriate measures are required to avoid contamination1. Furthermore, a minimum of one negative sample should be processed in parallel to estimate the risk of low level contamination.iv) The real-time PCR assay described here involves two separate procedures. Firstly, BoHV-1 DNA is extracted from semen using Chelex-100 chelating resin, along with proteinase K and DL-Dithiothreitol(DTT). The second procedure is the PCR analysis of the extracted DNA template in a real-time PCR 1 Sources of contamination may include product carry-over from positive samples or, more commonly, from cross-contamination by PCR products from earlier experiments. Samples and reagents should be handled in separate areas, with separate equipment for reagent and sample preparation and amplification/detection.reaction mixture: Platinum Quantitative PCR SuperMix-UDG, Invitrogen Technologies (note that thereare a number of other commercial real-time PCR amplification kits available from various sources andthe particular kits selected need to be compatible with the real-time PCR platform selected). Therequired primers and probes can be synthesised by various commercial companies. In this protocol, allthe primers and probes used were supplied by Sigma-Genosys.• Extraction of DNAi) In a screw top 1.5 ml tube, add:Chelex 100 sodium (Sigma) (10% w/v in distilled deionised water) 100 µl.Proteinase K (10 mg/ml, Sigma) 11.5 µlDL-Dithiothreitol (1 M, Sigma) 7.5 µlNuclease-free water 90 µlSemen sample 10 µlMix gently by pipetting2.ii) The samples are incubated at 56°C for 30 minutes and then vortexed at high speed for 10 seconds.iii) Subsequently, the tubes are incubated in a boiling water bath for 8 minutes and then vortexed at high speed for 10 seconds.iv) The tubes are centrifuged at 10,000 g for 3 minutes.v) Thesupernatant3 is transferred into a new microtube and can be used directly for PCR, or stored at –20°C.• Preparation of reagentsThe real-time PCR reaction mixture (Platinum Quantitative PCR SuperMix-UDG, or other reaction mixture) is normally provided as a 2 × concentration ready for use. The manufacturer’s instructions should be followed for application and storage.Working stock solutions for primers and probe are made with nuclease-free water at the concentration of4.5 µM and 3 µM, respectively. The stock solutions are stored at –20°C and the probe solution should bekept in the dark. Single-use aliquots can be prepared to limit freeze-thawing of primers and probes and extend their shelf life.• Real-time PCR test procedurei) Primers and probe sequencesSelection of the primers and probe are outlined in Abril et al. (2004) and described below.Primer gB-F: 5’-TGT-GGA-CCT-AAA-CCT-CAC-GGT-3’ (position 57499–57519 GenBank®, accessionAJ004801)Primer gB-R: 5’-GTA-GTC-GAG-CAG-ACC-CGT-GTC-3’ (position 57595–57575 GenBank®, accessionAJ004801)TaqMan Probe: 5’-FAM-AGG-ACC-GCG-AGT-TCT-TGC-CGC-TAMRA-3’ (position 57525–57545 GenBank®, accession AJ004801)reaction mixturesofii) PreparationThe PCR reaction mixtures are prepared in a separate laboratory room. For each PCR test, appropriatecontrols should be included. As a minimum, a no template control (NTC, reagents only), appropriatenegative controls, i.e. 1 per 10 test samples, and two positive controls (moderate and weak positive)should be included. Each test sample and control is tested in duplicate. The PCR amplifications arecarried out in a volume of 25 µl.a) PCR reagent mixtures are added in a clean room (no viral cultures, DNA extracts or post-amplification products should be handled here)2 × Platinum Quantitative PCR SuperMix-UDG 12.5 µlROX reference dye (optional) 0.5 µlForward primer (gB-F, 4.5 µM) 1 µlReverse primer (gB-R, 4.5 µM) 1 µlProbe (3 µM) 1 µlNuclease free water 4 µl2 It is important that the Chelex 100 solution is homogeneous while pipetting, as Chelex 100 sodium is not soluble. This canbe achieved by putting the vessel containing Chelex-100 solution on a magnetic stirrer while pipetting.3 Some DNA samples can become cloudy and a thin white membrane may form occasionally after freezing and thawing.This appears to have no influence on the PCR performance. No heating or re-centrifuging of the samples is necessary.。

Installation and Maintenance Manual Bar Type Ionizer Series IZS40/41/421 Safety InstructionsThis manual contains essential information for the protection of users and others from possible injury and/or equipment damage.∙ Read this manual before using the product, to ensure correct handling, and read the manuals of related apparatus before use. ∙ Keep this manual in a safe place for future reference.∙ These instructions indicate the level of potential hazard by label of “Caution”, “Warning” or “Danger”, followed by important safety information which must be carefully followed.∙ To ensure safety of personnel and equipment the safety instructions in this manual and the product catalogue must be observed, along withWarning∙ The compatibility of pneumatic equipment is the responsibility of thepersonwho designsthepneumaticsystem ordecides its specifications.Since the products specified here canbe used in various operating conditions, their compatibility with the specific pneumatic system must be based on specifications or after analysis and/or tests to meet specific requirements.∙ Only trained personnel should operate pneumatically operated machinery and equipment.Compressed air can be dangerous if an operator is unfamiliar with it. Assembly, handling or repair of pneumatic systems should be performed by trained and experienced personnel.∙ Do not service machinery/equipment or attempt toremove components until safety is confirmed.1) Inspection and maintenance of machinery/equipment should only be performed after confirmation of safe locked-out control positions.2) When equipment is to be removed, confirm the safety process as mentioned above. Switch off air and electrical supplies and exhaust all residual compressed air in the system.3) Before machinery/equipment is re-started, ensure all safety measures to prevent sudden movement of cylinders etc. (Supply air into the system gradually to create back pressure, i.e. incorporate a soft-start valve).∙ Do not use this product outside of the specifications. Contact SMC if it is to be used in any of the following conditions:1) Conditions and environments beyond the given specifications, or if the product is to be used outdoors.2) Installations in conjunction with atomic energy, railway, air navigation, vehicles, medical equipment, food and beverage, recreation equipment, emergency stop circuits, press applications, or safety equipment.3) An application which has the possibility of having negative effects on people, property, or animals, requiring special safety analysis.1.1 SelectionWarning∙ These products are intended for use in general factory automation equipment.Consult SMC before hand when using this product for other intentions. ∙ Use within the specified voltage and temperature range. Operation with a voltage other than that specified can cause malfunction, damage to the product, electric shock or fire. ∙ Use clean compressed air as fluid.Never use flammable or explosive gas as fluid. This may lead to fire or explosion. If fluid other than compressed air is used, consult SMC.1 Safety Instructions (Continued)∙ The product is not designed to be explosion proof.Never use in an atmosphere of potential dust explosion, flammable gas or explosive gas. It may cause fire.Caution∙ Clean specification is not available with this product.This product has not been cleaned. When using this product in a clean room environment, flush and confirm the product’s purification level before use. A minute amount of particles are generated due to wearing of the electrodes while the ionizer is operating.1.2 InstallationWarning∙ Secure enough space for maintenance, inspection and wiring.When routing cables and tubings, secure sufficient maintenance space for the installation and removal of connector and One-touch fitting. Consider the minimum bending radius of the cables and tubings and avoid bending them at an acute angle so that unreasonable stress is not applied to the mounting parts of the connectors and One-touch fittings. Position the connectors and One-touch fittings as close as possible.Routing of the wiring and cables in unreasonable positions may cause malfunction, broken cables, and fire.[Minimum bending radius] Power supply cable: 38mmTransition wiring cable: 38mm Sensor cable: 25mmNote:This is the minimum bend radius at 20o C. If the installation is at a lower temperature, the radium will be higher. When the cables are bent at a lower temperature than 20 o C, it may cause unreasonable force to be applied to the connectors.Refer to the tubing operation manual for minimum bending radius of tubing.∙ Mount to a flat surface.Mounting on an uneven surface will apply excessive force to the housing and bracket, which may lead to damage or failure. Do not drop the product or subject it to a strong impact. This may cause an injury or accident.∙ Install the product so that the entire bar does not have an excessive deflection.For a bar length of 820mm or more, support the bar at both ends and in the middle by using brackets (IZS40-BM). If the bar is held only at the both ends, self-weight of the bar causes deflection, resulting in damage to the bar.∙ Avoid using in a place where noise (electromagnetic wave and surge) is generated.It may cause malfunction, deterioration or damage to internal components. Take measures to prevent noise at its source and avoid power and signal lines from coming into close contact. ∙ Use a correct tightening torque.If the screws are tightened in excessive of the specified torque range, it may damage the mounting screws, mounting brackets, etc. If the tightening torque is insufficient, the mounting screws and brackets may become loose.∙ Do not directly touch the electrodes with your finger or tools. Do not directly touch the electrode with your finger. If the electrode sticks to your finger, or electrical shock makes an instantaneous rapid body motion to escape from the shock, your body may touch the equipment around you, causing injury. If electrode or cartridge isdamaged by tools, etc., it may interfere with the specified function and performance, and may also cause operation failure and accident.1 Safety Instructions (Continued)∙ Do not adhere tape or sticker onto the product body.If the tape or sticker contains conductive adhesive or reflective paint, it is possible that due to the dielectric effect, charge could build up causing an electro-static discharge or electrical leakage.∙ Be sure to remove power supply and air supply to the product before starting the product installation.Caution∙ Install the IZS4* series ionizer maintaining distance from a wall, etc. as shown in the Fig. below.When there is a wall or an object within the area shown in the Fig. below, generated ions may not reach the workpiece effectively, resulting in deterioration of efficiency.∙ Make sure to confirm the effect of de-ionization after installation. The effect of the ionizer varies depending on the surrounding installation and operating conditions. Confirm the effect of static electricity elimination after installation.∙ When installing IZS41 or IZS42 in proximity with an ionizer which operates in DC mode, they should be positioned at least 2 meters away from each other. When IZS41 or IZS42 isused close to an ionizerwhich operates in DCmode, separate theionizers at least 2 meters.Ion balance may not be adjusted by the internal sensor due to the ions which are discharged from the DC mode ionizer.1.3 Wiring and PipingWarning∙ Ensure that the power supply capacity is large enough, and thatvoltage is within specification before wiring.∙ To maintain product performance, a DC power supply shall be connected per UL listed Class 2 certified by National Electric Code (NEC) or evaluated as a limited power source provided by UL60950. ∙ To maintain the product performance, ground the product with an earth ground cable with a resistance of 100 ohm or less according to this manual.∙ Remove the power supply before wiring (including the connector plug in/out).∙ Use a cable with sensor for connection of the ionizer, feedback sensor or auto balance sensor (high accuracy type), and do NOT disassemble or retrofit.∙ Ensure the safety of wiring and surrounding conditions before supplying power.∙ Do not connect or disconnect the connectors (including power source) while the power is being supplied. The ionizer may malfunction.∙ Malfunctions induced by noise may occur if the wire is installed in the same route as that of power or high-voltage cable. Wire the ionizer independently.∙ Confirm that there is no error in wiring before operation. Incorrect wiring will lead to a malfunction or breakage of the product.∙ Flush the piping before connecting. Verify that all dust, moisture, oil, etc. are eliminated from the piping before connecting.1.4 Operating and Storage EnvironmentWarning∙ Operate the product in the specified fluid temperature range and ambient temperature range.Fluid temperature and ambient temperature ranges are; 0 to 40 o C for Ionizer, 0 to 50 o C for feedback sensor and auto balance sensor (high accuracy type), 0 to 40 o C for AC adapter, and 0 to 45 o C for remote controller. Avoid sudden temperature change even within specified temperature range, as it may cause condensation.1 Safety Instructions (Continued)∙ Do not use this product in an enclosed space.This product utilizes the corona discharge phenomenon. Although the amount is very small, Ozone and NOx are generated. Do not use in an enclosed space.∙ Environments to avoidNever use or store under the following conditions, as these cause product failure.a. Areas where ambient temperature exceeds the operating temperature range.b. Areas Where ambient humidity exceeds the operating humidity range.c. Areas where abrupt temperature changes may cause condensation.d. Areas where corrosive gas, flammable gas or other volatile flammable substances are stored.e. Areas where the product may be exposed to conductive powder such as iron powder or dust, oil mist, salt, organic solvent, machining chips, particles or cutting oil (including water and any liquids), etc.f. Paths of direct air flow, such as air conditioners.g. Enclosed or poorly ventilated areas.h. Locations that are exposed to direct sunlight or heat radiation.i. Areas where strong electromagnetic noise is generated, such as strong electrical and magnetic fields or supply voltage spikes.j. Areas where the product is exposed to static electricity discharge. k. Locations where strong high frequency is generated. l. Locations that are subject to potential lightning strikes.m. Areas where the product may be exposed to direct impact or vibration. n. Areas where the product may be subjected to forces or weight that could cause physical deformation.∙ Do not use air containing mist and/or dust.Air containing mist and/or dust may cause performance deterioration, and reduce the maintenance cycle.Install a dryer (IDF series), air filter (AF/AFF series), or mist separator (AFM/AM series) to obtain clean compressed air (air quality of Class 2.6.3 or higher according to ISO 8573-1: 2001 is recommended for operation). ∙ Ionizer, feedback sensor, auto balance sensor (high accuracy type), remote controller, and AC adapter are not resistant to lightening surge.1.5 Maintenance and InspectionWarning∙ Perform maintenance regularly to keep the electrodes clean.Perform regular maintenance of the product to prevent undetected failures. The maintenance must be carried out by an operator who has sufficient knowledge and experience. If the product is used for an extended period of time with dust is present on the electrodes, the product’s abili ty to eliminate static electricity will be reduced.If the electrodes become worn and the product’s ability to eliminate static electricity is not restored after cleaning, replace the cartridge.∙ Make sure to remove power and air supply from the product before cleaning the electrodes or replacing the cartridges.If the electrodes are touched while the product is energized, this may cause an electric shock or accident.If an attempt to replace the cartridges is performed before removing air supply, the cartridges may eject unexpectedly due to presence of the supply air. Remove air supply before replacing the cartridges. If cartridges are not securely mounted to the bar, they may eject or release when air is supplied to the product. Securely mount or remove the cartridges referencing the instructions shown below.∙ Perform contamination detection of the electrode without workpiece. (IZS41 and IZS42)While electrode detects contamination, ionizer discharges positive ions and negative ions for contamination detection. ∙ Do not disassemble or modify the product.This may lead to accidents such as electric shock, failure, fire or etc. If the product is disassembled and/or modified, the functions andperformance in the specifications may not be achieved and the product will not be guaranteed.∙ Do not operate the product with wet hands. This may cause an electric shock or accident.Unit: mm 150 150 200 200 High voltage is applied to the electrodes. Never touch thegenerating High voltage cautionM4 screwM4 screwEnd bracket Intermediate bracket Part A2.1 Mounting and installation of the bracket1) End bracketMount an end bracket to both ends of the ionizer body using the M4 screws supplied as accessories.Tightening torque: 1.3 to 1.5Nm2) Intermediate bracket (for bar lengths of 820mm or more)Match the groove of the ionizer body and protrusion of the intermediate bracket, and slide the bracket from the end of the ionizer body. Intermediate brackets should be mounted at the same intervals.3) Installation of the ionizer (when using brackets)Tap (M5) screws at the bracket mounting positions for installation of the Ionizer and fix the ionizer body and brackets with M5 screws.IZS40 and IZS41 are constructed such that the brackets at the bracket mounting positions on both ends of the bar are shared with F.G. Use caution to avoid short-circuit with the +24V power supply when installing and supplying power.4) Mounting angle adjustmentAdjust the angle of the ionizer body for effective de-ionizing and fix the ionizer with the rotating set screw (M4) at each bracket.End bracket tightening torque: 1.3 to 1.5 NmIntermediate bracket tightening torque: 0.73 to 0.75Nm2.2 InstallationWarning∙Do not install the product unless the safety instructions have been readand understood.2.3 PipingCaution∙Before piping make sure to clean up chips, cutting oil, dust etc.∙When installing piping or fittings, ensure sealant material does not enterinside the port. When using seal tape, leave 1.5 to 2 threads exposed onthe end of the pipe/fitting.∙Tighten fittings to the specified tightening torque.2.4 EnvironmentWarning∙Do not use in an environment where corrosive gases, chemicals, saltwater or steam are present.∙Do not use in an explosive atmosphere.∙Do not expose to direct sunlight. Use a suitable protective cover.∙Do not install in a location subject to vibration or impact. Check theproduct specifications.∙Do not mount in a location exposed to radiant heat.3 WiringWire cables according to the circuitry and wiring chart.3.1 Grounding of F.G. cableMake sure to ground the F.G. cable (green) with a resistance of100 ohms or less.The F.G. cable is used as a reference electric potential for de-ionization. If the ground terminal F.G. is not grounded, the ionizer willnot be able to achieve the optimal ion balance.3.2 Grounding during operation in DC modeApplicable models:IZS40 and IZS41When an ionizer is used in DC mode, make sure to ground the F.G.cable (green) and GND cable (blue) of the input power supply with aresistance of 100 ohms or less. Without grounding the GNDterminal, the ionizers and/or power supply may be damaged.3.3 Circuit (Wiring of IZS40)e-con is adopted for the connector of IZS40. Connector with cableor without cable maybe selected when placing an order for the powersupply cable.When only an e-con is required, place an order for it as a part.Cable is not supplied.)Wiring1) Cut the cable as shown in theFigure to the right.applicable wire size.Applicable wire3.4 Circuit (Wiring of IZS41 and IZS42)2) Insert the cable which was cut intothe back of the connector.3) Confirm that the cable is insertedinto the back of the connector andpress part A with your finger tohold tentatively.4) Use a tool such as pliers to firmlytighten the center of Part A.5) The connector cannot be reusedonce crimped. If cable insertionfails, use a new connector.DescriptionRB5B1A5A1When an ionizer is used in DC mode, make sure to ground the F.G. cable (green) and GND cable (blue) of the input power supply with aresistance of 100 ohms or less. Without grounding the GND terminal, the ionizers and/or power supply in connection may be damaged.If cables are prepared by the user, the cable colors shown in the diagram may change according to the cable colors by the user.Signal name Signal direction1) NPN typeRefer to the operation manual for this product.5 Settings5.1 Descriptions and Functions of the Panel (IZS40)When an ionizer (IZS41) is used in DC mode, make sure to ground the F.G. cable (green) and GND cable (blue) of the input power supply with a resistance of 100 ohms or less. Without grounding the GND terminal, the ionizers and/or power supply in connection may be damaged. When an ionizer (IZS41) is used in DC mode, make sure to ground the F.G. cable (green) and GND cable (blue) of the input power supply with a resistance of 100 ohms or less. Without grounding the GND terminal, the ionizers and/or power supply in connection may be damaged.2 1 53 45.2 Descriptions and Functions of the Panel (IZS41, IZS42)Refer to the operation manual for this product.11 10 9 8 7 6 5 4 3 2 1 14 13 12"N", and order a part (Part No.: ZS-28-C) separately.option number. (A cable is attached to the AC adapter)- Input/ Output function cannot be used when the AC adapter isbeing used.- To use AC adapter, specify "N", and select AC adapter with theRefer to the operation manual for this product. 7.1 IZS407.2 IZS41 / IZS428.1 General MaintenanceCaution∙ Not following proper maintenance procedures could cause the product to malfunction and lead to equipment damage.∙ If handled improperly, compressed air can be dangerous. Maintenance of pneumatic systems should be performed only by qualified personnel.∙ Before performing maintenance, turn off the power supply and be sure to cut off the supply pressure. Confirm that the air is released to atmosphere. ∙ After installation and maintenance, apply operating pressure and power to the equipment and perform appropriate functional and leakage tests to make sure the equipment is installed correctly. ∙ Do not make any modification to the product.∙ Do not disassemble the product, unless required by installation or maintenance instructions.8.2 Detection and cleaning of contamination on the electorodeCaution∙ If the ionizer is used for a long time, contamination such as dust can stick to the electrodes, reducing the static electricity elimination performance. For this reason, IZS41 and IZS42 have a contamination detecting function. ∙ Dirt detection is performed when a contamination detection signal from an electrode is input. When the electrode requires cleaning due to deterioration of de-ionizing ability, the maintenance signal turns ON and maintenance LED also turns ON to notify the timing of cleaning. When the maintenance LED turns ON, make sure to clean the electrode. (Ionizer keeps operating even after the maintenance signal and maintenance LED turn ON.)∙ Dirt detection of electrodes should be performed without a workpiece, as it is performed with ions discharged from the ionizer at a regular cycle and this may electrify the workpiece.∙ Clean the electrodes with the electrode cleaning kit [IZS30-M2] or a cotton swab soaked in alcohol.∙ In cases where the electrode contamination detecting function is not used or when the IZS40 does not have a contamination detecting function,, as contamination on the electrodes may vary depending on the installation environment and supply pressure, etc., confirm the product performance and set a maintenance cycle for a periodic cleaning.∙ Make sure to turn OFF the power and air supply before cleaning the electrodes. If the electrodes are touched while the product is energized, it may cause an electric shock or accident. Do not touch the end of the electrodes. As electrodes have a sharp end, touching them directly with your fingers may cause injury.∙ If the maintenance signal is output upon completion of cleaning the electrode, it may not have been cleaned sufficiently or it may have been worn or damaged. If wear or damage of the electrode is detected, replace the electrode cartridge with a new one. (If the electrode is worn out or damaged, the static electricity elimination performance will decrease.)∙ Refer to the Fig. below for mounting, removal and cleaning of an electrode cartridge.Warning∙ Do not exceed any of the specifications laid out in section 4 of this document or the specific product catalogue.10 Disposal informationThe remote controller IZS41-RC used for this ionizer is a product sold separately. For this reason it is classified as Waste Electrical or Electronic Equipment according to the WEEE Directive 2012/19 / EU and should not be disposed of as municipal waste, in order to reduce the impact on human health and the environment. Remove and dispose of old batteries and the remaining electrical or electronic equipment according to local environmental regulations.11 ContactsAUSTRIA (43) 2262 62280-0 LATVIA (371) 781 77 00 BELGIUM (32) 3 355 1464 LITHUANIA(370) 5 264 8126 BULGARIA (359) 2 974 4492 NETHERLANDS (31) 20 531 8888 CZECH REP. (420) 541 424 611 NORWAY (47) 67 12 90 20 DENMARK (45) 7025 2900 POLAND (48) 22 211 9600 ESTONIA (372) 651 0370 PORTUGAL (351) 21 471 1880 FINLAND (358) 207 513513 ROMANIA (40) 21 320 5111 FRANCE (33) 1 6476 1000 SLOVAKIA (421) 2 444 56725 GERMANY (49) 6103 4020 SLOVENIA (386) 73 885 412 GREECE (30) 210 271 7265 SPAIN (34) 945 184 100 HUNGARY (36) 23 511 390 SWEDEN(46) 8 603 1200 IRELAND (353) 1 403 9000 SWITZERLAND (41) 52 396 3131 ITALY(39) 02 92711UNITED KINGDOM(44) 1908 563888URL : http// (Global) http// (Europe) Specifications are subject to change without prior notice from the manufacturer. © 2010 SMC Corporation All Rights Reserved.counter-clockwise direction. Removal of electrode cartridge 2) Pull to remove. IZS30-M2 Cleaning of electrode 1) Insert the cartridge into the barso that the longer side of the cartridge is mounted at a right angle to the bar.Mounting of electrode cartridge2) Rotate the cartridge degrees in the clockwise and ma tch the ma rkings on the the ca rtridge to fix.。

全面护理安全事故报告指南英文版Comprehensive Care Safety Incident Report GuideIn the fast-paced healthcare environment, it is crucial to have a systematic approach to handling safety incidents. This guide aims to provide a comprehensive overview of the steps to take when encountering a safety incident in a care setting.Reporting Procedure:1. Immediate Response: Ensure the safety of the individual involved and any others in the vicinity.2. Document Details: Record the date, time, location, and nature of the incident.3. Notify Supervisor: Inform the appropriate authority about the incident.4. Complete Incident Report Form: Fill out a detailed report outlining the sequence of events.Investigation Process:1. Gather Information: Collect statements from witnesses and those directly involved.2. Review Policies: Refer to organizational protocols and procedures related to the incident.3. Identify Root Cause: Determine the underlying factors that contributed to the incident.4. Develop Corrective Action Plan: Implement measures to prevent similar incidents in the future.Communication Strategy:1. Internal Communication: Notify relevant staff members about the incident and any necessary follow-up actions.2. External Communication: Inform regulatory bodies and other stakeholders as required.3. Maintain Confidentiality: Respect the privacy of individuals involved in the incident.Follow-Up Actions:1. Monitor Progress: Track the implementation of corrective measures and evaluate their effectiveness.2. Provide Support: Offer assistance to those affected by the incident, including counseling if necessary.3. Review and Learn: Conduct a post-incident analysis to identify areas for improvement and prevent recurrence.Conclusion:By following this comprehensive guide, healthcare providers can ensure a proactive and effective response to safety incidents in care settings. Remember, the goal is to prioritize the well-being of individuals and maintain a culture of safety and accountability.。

跨境动物疫病的预防和控制王华;吴晓东;彭冬;王淑娟;王志亮【摘要】跨境动物疫病对全球养殖业和食品安全造成严重威胁,危及到国际贸易.主要跨境动物疫病的暴发会造成毁灭性的经济损失.本文概述了TADs的危害和防控机制,阐述了优先防控一些主要的跨境动物疫病,为减少对动物和人类健康以及经济发展的危害,提出了预防控制和管理的措施.【期刊名称】《中国动物检疫》【年(卷),期】2014(031)006【总页数】4页(P36-38,42)【关键词】跨境动物疫病;预防;控制;经济损失【作者】王华;吴晓东;彭冬;王淑娟;王志亮【作者单位】中国动物卫生与流行病学中心,山东青岛266032;中国动物卫生与流行病学中心,山东青岛266032;中国动物卫生与流行病学中心,山东青岛266032;中国动物卫生与流行病学中心,山东青岛266032;中国动物卫生与流行病学中心,山东青岛266032【正文语种】中文【中图分类】S851.34跨境动物疫病（Transboundary Animal Disease，TADs）是对大多数国家的经济、贸易或者食品安全产生重大影响的一类疾病统称，这些疾病很容易传播到其他国家并且流行起来。

预防和控制需要多个国家之间建立联合防控的合作机制。

TADs主要是原先OIE列出的A类动物疫病，例如其中一些病毒性疾病禽流感、猪瘟、非洲猪瘟、小反刍兽疫、口蹄疫、裂谷热和新城疫[1-6]。

这些疾病共同的特点是能引起严重的经济损失；通过动物和动物产品运输进行远距离的传播；采取有效的免疫和限制流动措施对疫病能够进行有效控制；采用一些有效的诊断方法能够检测出这些疾病。

总之，尽管此类疾病属于烈性传染病，但是通过有效的方法能预防和控制它们的传播[6]。

从历史的角度观察，TADs引起战争、饥荒、政权动荡和人类的贫困。

目前，TADs出现非常大规模的暴发情况虽然罕见，但是仍然需要高度重视，需要投入大量的资金、人力、物力和财力等资源来控制和消除。

凡事必有报告英文翻译Everything Must Be ReportedIn order to ensure transparency and accountability, it is essential that every event, action, or occurrence be meticulously documented and reported. Proper reporting allows for effective communication, analysis, and decision-making, benefiting individuals, organizations, and society as a whole.First and foremost, reporting provides a clear and accurate account of events. Whether it is a daily activity log, a financial report, or a project update, documenting the details allows for a comprehensive understanding of what has transpired. This information can then be shared with relevant parties, ensuring that everyone is on the same page and can make informed decisions based on accurate data.Reporting also serves as a valuable tool for analysis. By systematically recording and categorizing information, patterns, trends, and anomalies can be identified. Such insights enable individuals and organizations to gain a deeper understanding of their operations, identify areas for improvement, and make data-driven decisions. For example, analyzing sales data can help a company identify its most profitable products and create targeted marketing strategies.Furthermore, reporting plays a crucial role in accountability and compliance. By documenting actions and outcomes, individuals and organizations can be held responsible for their conduct and performance. This is especially important in areas such asgovernance, finance, and law enforcement. Through thorough reporting, misconduct, fraud, or non-compliance can be detected and addressed, ensuring ethical practices and adherence to regulations.Additionally, reporting facilitates effective communication. By sharing accurate and comprehensive reports with stakeholders, important information can be disseminated in a standardized and organized manner. This allows for efficient communication across various levels and departments, promoting collaboration and ensuring that everyone is well-informed.Reporting also contributes to transparency. Whether it is a public announcement, a financial statement, or an audit report, making information available to the public fosters trust and credibility. This is particularly crucial for government entities, non-profit organizations, and corporations, as transparency is an essential characteristic of responsible and accountable governance.In conclusion, reporting is a vital aspect of any endeavor, be it personal or professional. The act of documenting and sharing information not only ensures transparency and accountability but also enables analysis, communication, and compliance. Embracing the practice of reporting will undoubtedly lead to better decision-making, improved efficiency, and increased trust among individuals, organizations, and society as a whole.。

A. 经常困扰食品公司的一个问题是,如何控制对食物造成破坏的老鼠和昆虫。

B. 经常困扰食品公司的一个问题是,如何控制老鼠和昆虫，同时又不会对食物造成破坏。

C. 经常困扰食品公司的一个问题是,如何控制老鼠和昆虫，虽然它们不会对食物造成破坏。

aspects of promoti ng good health.; Chronic sick ness.; The family.; From a n ewbor nbaby all the way up to the end of one's life.; Information.};;二、完型填空（每空10分）。

操作提示：通过题目中的下拉选项框选择恰当的词语补全填空。

Have you heard about Gran dpare nts Week? This importa nt com mun ity event bega n{as; with; in} a one day celebration in 2012 to commemorate National Grandparents Day. In 2014, the ACT Government expa nded the program to a whole week to fully {capturing; capture; captures} the true worth of grandparents in all their variousforms.Gran dpare nts Week is an annual eve nt to honor and recog nize the valuablecontribution grandparents make, and the valuable role they play, in their families and the wider ACT com mun ity.This year's festival in cludes an excit ing range of eve nts, programs andactivities for young and old {plus; add; including} those in between! Sing some oldtime tunes or shake a move on the dance floor. Join a fun game of bowls or table tennis. ; ;Play your hand at cards. Atte nd a Senior Club Open Day. ; ;Partake in delicious morning teas. ; ;{Whoever; However; Whatever} your vin tage there'ssometh ing for every one duri ng Gran dpare nts Week!Does your com mun ity group have a great event or activity {out mind; in mind;of mind} for seniors? Would you like to include it as part of the Grandparents Week 2016 program?;;二、阅读理解阅读下面的文章，根据文章容，完成相应的选择题。

E2F Development Safety Update ReportU.S. Department of Health and Human ServicesFood and Drug AdministrationCenter for Drug Evaluation and Research (CDER)Center for Biologics Evaluation and Research (CBER)August 2011ICHE2F Development Safety Update ReportAdditional copies are available from:Office of CommunicationsDivision of Drug Information, WO51, Room 220Center for Drug Evaluation and ResearchFood and Drug Administration110903 New Hampshire Ave.Silver Spring, MD 20993Phone: 301-796-3400; Fax: 301-847-8714druginfo@/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/default.htmand/orOffice of Communication, Outreach andDevelopment, HFM-40Center for Biologics Evaluation and ResearchFood and Drug Administration1401 Rockville Pike, Rockville, MD 20852-1448(Tel) 800-835-4709 or 301-827-1800/BiologicsBloodVaccines/GuidanceComplianceRegulatoryInformation/Guidances/default.htm U.S. Department of Health and Human ServicesFood and Drug AdministrationCenter for Drug Evaluation and Research (CDER)Center for Biologics Evaluation and Research (CBER)August 2011ICHContains Nonbinding RecommendationsTABLE OF CONTENTSI.INTRODUCTION (1) (1)A.Background (1.1) (2)B.Objectives (1.2) (2)C.Scope of the DSUR (1.3) (3)D.Relation of the DSUR to the Periodic Safety Update Report (1.4) (3)E.Recipients of the DSUR (1.5) (4)II.GENERAL PRINCIPLES (2). (4)A.Single DSUR for an Active Substance (2.1) (4)B.Periodicity and DSUR Data Lock Point* (2.2) (4)C.Duration of DSUR Submissions (2.3) (5)D.Responsibilities for Preparing and Submitting a DSUR (2.4) (5)1.Sponsor’s Responsibilities (2.4.1) (5)2.Responsibilities of Multiple Parties (2.4.2) (5)E.DSURs for Combination Therapies (2.5) (6)F.Reference Safety Information (2.6) (7)G.Format and Presentation of DSUR (2.7) (7)1. Format (2.7.1) (7)2. Presentation (2.7.2) (7)III.GUIDANCE ON CONTENTS OF A DSUR (3) (8)1.Introduction (3.1) (9)2.Worldwide Marketing Approval Status (3.2) (9)3.Actions Taken in the Reporting Period for Safety Reasons (3.3). (10)4.Changes to Reference Safety Information (3.4) (11)5.Inventory of Clinical Trials Ongoing and Completed During the Reporting Period (3.5) (11)6.Estimated Cumulative Exposure (3.6) (12)6.1.Cumulative Subject Exposure in the Development Program (3.6.1) (12)6.2.Patient Exposure From Marketing Experience (3.6.2) (13)7.Data in Line Listings and Summary Tabulations (3.7) (13)7.1.Reference Information (3.7.1) (14)7.2.Line Listings of Serious Adverse Reactions During the Reporting Period (3.7.2) (14)7.3.Cumulative Summary Tabulations of Serious Adverse Events (3.7.3) (15)8.Significant Findings from Clinical Trials During the Reporting Period (3.8) (15)Contains Nonbinding Recommendationspleted Clinical Trials (3.8.1). (15)8.2.Ongoing Clinical Trials (3.8.2) (16)8.3.Long-Term Follow-up (3.8.3) (16)8.4.Other Therapeutic Use of Investigational Drug (3.8.4) (16)8.5.New Safety Data Related to Combination Therapies (3.8.5) (16)9.Safety Findings From Noninterventional Studies (3.9) (16)10.Other Clinical Trial/Study Safety Information (3.10) (17)11.Safety Findings From Marketing Experience (3.11) (17)12.Nonclinical Data (3.12) (17)13.Literature (3.13) (17)14.Other DSURs (3.14) (17)ck of Efficacy (3.15) (18)16.Region-Specific Information (3.16) (18)te-Breaking Information (3.17) (19)18.Overall Safety Assessment (3.18) (19)18.1.Evaluation of the Risks (3.18.1) (19)18.2.Benefit-Risk Considerations (3.18.2) (20)19.Summary of Important Risks (3.19) (21)20.Conclusions (3.20) (21)IV.APPENDICES TO THIS GUIDANCE. (23)APPENDIX A — Glossary (23)APPENDIX B — Examples of Tables and Table Headings for Clinical Trial Listings (26)APPENDIX C — Examples of the Summary of Important Risks (29)Guidance for Industry1E2F Development Safety Update ReportI.INTRODUCTION (1)2The development safety update report (DSUR) proposed in this guidance is intended to be a common standard for periodic reporting on drugs under development (including marketed drugs that are under further study) among the ICH regions. U.S. and European Union (EU) regulators consider that the DSUR, submitted annually, would meet national and regional requirements currently met by the U.S. investigational new drug application (IND) annual report and the EU annual safety report, respectively, and can therefore take the place of these existing reports.3 This guidance defines the recommended content and format of a DSUR and provides an outline of points to be considered in its preparation and submission.本指南中提出的开发安全性更新报告（DSUR）旨在成为ICH区域内正在开发的药物（包括正在进一步研究的上市药物）定期报告的通用标准。

0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status1.0 Q U A L I T Y M A N A G E M E N T1.1The quality system is documented,controlled, and maintained to clearly describe current practice.Documented procedures required.Records required.Quality manual and all QSprocedures show revision control (sign-offs & dates), history of changes, quality organization's responsibilities1.0 Q U A L I T Y M A N A G E M E N T1.2Quality reports, trend charts and data analysis identify areas of opportunity and are used bymanagement on a routine basis.Records required.Product quality yield data, top problems and corresponding improvement actions, status of preventive/corrective actions taken, internal audit results1.0 Q U A L I T Y M A N A G E M E N T1.3Quality performance targets are clearly defined, included in the business plan and monitored for improvements.Strategic and tactical objectives,goals, action plans, etc.1.0 Q U A L I T Y M A N A G E M E N T1.4Executive management participates in periodic quality system reviews that address quality related feedback from customers and internal quality metrics. Records required.Analysis of field failures,inspection yields, resource needs, internal audit results,corrective action status, etc.2.0 C O N T I N U O U S I M P R O V E M E N T2.1Preventive actions are taken based on the analysis of significant business trends, design reviews,customer satisfaction surveys or other meaningful inputs.Documented procedures required.Records required.Management review meetings,goal setting, performance measurement, internal audits,action plans, customer surveys2.0 C O N T I N U O U S I M P R O V E M E N T2.2A formal approach is used to actively pursue cost containment and other continual improvement activities throughout the organization. Documented procedures required. Records required.Employee involvement /recognition program, Lean, Six Sigma, kaizen, SPC, 5-S, cost reduction program, preventive actions0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)StatusS0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence AUDIT FINDINGS & OBSERVATIONSN o t A p p l i c a b l eN o t V e r i f i e dSupplier Self-Audit SCORE SupplierCA-PA Req'd?(Y / N)On-Site Audit SCORE After CAPA Verif. SCORECompletionDate (mm/dd/yy)Status0-Jan-00RequirementsTypical Objective Evidence 1.0 Q U A L I T Y M A N A G E M E N T1.1The quality system is documented,controlled, and maintained to clearly describe current practice.Documented procedures required.Records required.Quality manual and all QSprocedures show revision control (sign-offs & dates), history of changes, quality organization's responsibilities1.0 Q U A L I T Y M A N A G E M E N T1.2Quality reports, trend charts and data analysis identify areas of opportunity and are used bymanagement on a routine basis.Records required.Product quality yield data, top problems and corresponding improvement actions, status of preventive/corrective actions taken, internal audit results1.0 Q U A L I T Y M A N A G E M E N T1.3Quality performance targets are clearly defined, included in the business plan and monitored for improvements.Strategic and tactical objectives,goals, action plans, etc.1.0 Q U A L I T Y M A N A G E M E N T1.4Executive management participates in periodic quality system reviews that address quality related feedback from customers and internal quality metrics. Records required.Analysis of field failures,inspection yields, resource needs, internal audit results,corrective action status, etc.2.0 C O N T I N U O U S I M P R O V E M E N T2.1Preventive actions are taken based on the analysis of significant business trends, design reviews,customer satisfaction surveys or other meaningful inputs.Documented procedures required.Records required.Management review meetings,goal setting, performance measurement, internal audits,action plans, customer surveys2.0 C O N T I N U O U S I M P R O V E M E N T2.2A formal approach is used to actively pursue cost containment and other continual improvement activities throughout the organization. Documented procedures required. Records required.Employee involvement /recognition program, Lean, Six Sigma, kaizen, SPC, 5-S, cost reduction program, preventive actions0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence S0-Jan-00Requirements Typical Objective Evidence0-Jan-00Requirements Typical Objective Evidence。