These six subspecies are distinguishable by certain biochemical characteristics drugs for erectile dysfunction philippines buy levitra professional australia, and some of them correspond to what Kauffmann caUed sub-genera lipo 6 impotence cheap levitra professional 20 mg on-line. Salmonellae of subspecies I cause transmissible infection both in man and many domesticated and wild animal species erectile dysfunction 18-25 buy generic levitra professional 20 mg on line. The course of infection impotence natural cures order levitra professional 20mg free shipping, the clinical signs what causes erectile dysfunction cure buy levitra professional 20mg on-line, the post-mortem findings and epidemiological patterns vary according to the serovar and animal species involved (2) erectile dysfunction endovascular treatment cheap levitra professional 20 mg with amex. Septicaemia causes juvenile erectile dysfunction purchase levitra professional 20mg without prescription, genital infections with sporadic or endemic abortions and affections of joints are often other signs of a salmonella infection gas station erectile dysfunction pills cheap levitra professional 20 mg. The isolation and subsequent identification of the pathogen involved will enable epidemiological markers including phage type and antibiotic sensitivity to be determined. The isolation and identification of salmonellae depend not only on the quality of the samples but also on the culture medium and growth characteristics of the serovars, particularly those adapted to a host species (1,3,4,5). Identification of the agent the frequency of sampling and the type of samples obtained will depend largely on the clinical signs, epidemiological data, laboratory facilities available and the objectives. Samples are collected preferably during the acute phase of the disease or as soon as possible after death. Precautions should be taken to avoid contamination between samples before they are received at the laboratory. For small animal species, it may be preferable to transport 1-2 sick or recently dead animals to the laboratory if it is feasible to do so. Isolation media for salmonellae Salmonellae are isolated by using four different types of medium, as follows: a) Pre-enrichment media these are liquid and non-selective. They assist in revitalising bacteria in Salmonellosis (B31,47) 411 samples that have been frozen, heated, or have become dried. They are mainly employed in food and feed-stuff bacteriology, but also in environmental sampling of animal housing. The formulation of the medium, temperature and duration of incubation, and the volume of the sample used to seed the medium, all may serve to improve the isolation rate. These variables should be taken into account to obtain optimal adaptation to specific situations, particularly for the detection of serovars adapted to a host species, although these are not characteristic of all host-adapted species. Examples of selective enrichment media are sodium tetrathionate as in Muller-Kauffman broth, selenite F, brilliant green, and Rappaport-Vasiliadis broths. They inhibit growth of other bacteria and give information on some of the principal differential biochemical characteristics of salmonellae. Salmonellae form characteristic colonies on such media and are usually distinguishable from the colonies of other bacteria that may not have been inhibited. It should be noted that lactose fermentation by salmonellae may occasionally be seen. Examples of these isolation or separation media include brilliant green, deoxycholate-citrate, SalmonellaShigella and bismuth sulphite agars. In differential and selective media such as Salmonella-Shigella or Hektoen, colonies observed after 36 or 48 hours may remain white, without black centres indicating H2S production. They are used for obtaining pure cultures prior to further characterisation tests. The use of a magnifying glass and sub-culturing on non-selective ordinary agar media to check the specificity of isolated colonies for further examination is recommended, as well as a slide agglutination of suspected colonies with polyvalent salmonella antiserum. If necessary, additional culture can be performed, for example on blood agar and MacConkey agar. This is appropriate for clinical samples such as blood after incubation in non-selective broth. The interval between sampling and testing, the nature of any treatment carried out prior to sampling, the presumed numbers of viable salmonellae in relation to contaminants, and a knowledge of the characteristics of strains previously isolated in an area are all factors which require a variety of techniques. Samples that may be contaminated with other bacteria, such as faeces, mucus or genital lochia, are treated separately and are seeded in both enrichment and isolation media. The samples of normally sterile fluids or tissues are seeded directly into isolation and non-selective media. The homogeneity of a bacterial population, its morphology and Gram-staining character can be examined microscopically. Biochemical tests on pure bacterial cultures and then serotyping are used for identification purposes. Different complex media, in tubes or plates, wiU give a rapid identification by a combination of characteristics. The determination of the O factor, and in special cases also of the Vi antigen as weU as the H antigen, are performed by direct sude agglutination tests using specific O, Vi or H antisera. Screening is faciUtated by the avaUabiuty of antisera directed against several factors, and this can be pursued further by the use of monovalent antisera. Reference laboratories are often required to confirm the identity of an isolate and give information as to its exact serovar, biotype, phage type, antibiotype and plasmid profile. Serological tests Blood samples should not contain anti-coagulant and should be coUected from several cUnicaUy affected and convalescent animals. Second samples may be taken if an evaluation of the antibody response is required. In sporadic cases caused by ubiquitous serovars, screening for the causal serovar by agglutination tests can only be done by specialised laboratories which have an assortment of reference antigens. In practice, a preliminary bacteriological diagnosis is recommended, otherwise the identity of the probable causal serovar can be sought by using a restricted number of antigens selected according to the results of bacteriological tests conducted previously. Precipitating antibody or complement fixation tests do not present real advantages compared with the detection of agglutinating antibodies. Agglutinating antigens may be dense suspensions of either live or killed bacteria. The use of live antigens cannot be recommended, however, because they present a hazard to those performing the tests, and in fact it is preferable to use separate O and H antigens, involving treatments which inactivate the strain. Bacterial suspensions which contain only O or H antigen (sometimes Vi antigen) factors will allow separate screening for O and H agglutinins. The H antigen consists of a dense suspension of highly motile salmonellae which have been cultured in a nutrient broth and then treated with 0. If the strain used is bi-phasic, H antiserum corresponding to the phase to be suppressed is added to the agar. Slow microplate agglutination with stained antigens facilitates the test and reading of results. Monovalent antisera can be used to determine any O or H agglutination of antigen suspensions diluted in isotonic saline. Two techniques are available to associate a sample under examination with the two antigen types. The antigen is dispensed into a tube containing diluted serum which is then incubated at 37?C, or at 50?C (for 24 h for the O antigens and 4 h for the H antigens), or which is centrifuged to give a more rapid result. Serological examinations are generally conducted on a representative sample of a population and not on a single animal. An interpretation of the results will depend on a knowledge of the humoral antibody response to infection of the animal species involved. Whatever test is used, the results obtained should be evaluated after taking into account the techniques used and the clinical and epidemiological findings. A further disadvantage is that agglutinating antibodies may not be demonstrable until about three weeks after infection. Some live vaccines have been developed more recently, a few being approved by some national control authorities. This diversity of vaccines means that only general recommendations can be made in this chapter. Seed management a) Characteristics For dead or hving vaccines, the bacterial strain (or each bacterial strain) should be identified by historical records and characterised by stable phenotypic and/or genetic markers. Living vaccinal strains should be marked by stable characters allowing distinction from wild strains. Markers and attenuation of virulence should be stable, and preferably obtained by two independent mutations. A primary seed lot for each bacterial strain used in a vaccine preparation should be used during the entire period of use of the vaccine. Culture may be on solid medium, in Roux flasks, or in liquid medium, in which case large-scale fermentation equipment may be used. StabiUty and non-reversion to virulence after serial passages in susceptible species should be shown. Ui) Efficacy Laboratory experiments and field trials must be used to show that the vaccine is effective. The laboratory experiments consist of vaccination-chaUenge tests in the target species, at the recommended dose and age. Manufacture Vaccine must be made in suitable clean rooms with access restricted to approved persons. Personnel must be provided with protective clothing in production areas and in animal rooms. Culture seed-lots are prepared from the primary seed-lot, and the vaccine must be less than four passages removed from the seed-lot, i. The vaccine may be prepared by inoculation of a suitable medium, such as nutrient broth, with a fresh culture and incubation at 37?C for 24 hours, with or without aeration. Alternatively the organisms may be grown on and harvested from a soud medium, such as nutrient agar. The time of inactivation of dead vaccines should be at least three times that taken to reduce the viable number to an undetectable level. Preservatives, excipient for lyophilisation, or stabiliser for multidose containers or other substances added to or combined with a vaccinal preparation must have no deleterious effect on the immunising potency of the product. A standard freeze-drying process should be developed for a particular apparatus, loss of viabiUty of about 50% being considered as a starting point; the bacterial concentration shaU be adjusted to aUow for loss of viabiUty during the freeze-drying process. Batch control a) Sterility Tests for sterility and freedom from contamination of biological materials may be found in the chapter on General Information. Each batch should be tested in the target species at the recommended age and route, using at least twice the field dose. The results of vaccination need to take into account the flock immunity developing after a natural infection. The salmonellae of all sub-species are pathogenic for man and many animal species, but a few serovars are adapted to a host species. In the case of those of sub-species I, which are the most frequent, the clinical signs, post-mortem findings and epidemiological patterns may vary according to the serovar involved. Identification of the agent: the considerations applying to salmonella in general (see the previous chapter) apply. At present the preferred method for monitoring poultry breeding flocks and hatcheries for salmonella is by means of bacteriological examination of samples obtained from these establishments. Samples should be taken at specified regular intervals and examined in a laboratory authorised for that purpose by the veterinary authorities. Rappaport-Vasiliadis medium is widely used for the detection of salmonella in samples from poultry flocks and hatcheries. Serological tests: the considerations applying to salmonella in general (see the previous chapter) apply. Earlier proposals for the nomenclature of Salmonella species are described in refs. However, because these organisms are treated as serovars rather than species in the nomenclature of Le Minor et al. Many serovars of salmonella have been shown to cause human food poisoning, although they may not be pathogenic in the animal from which the food was derived. Recently this has been associated with poultry meat and eggs, although these organisms are also isolated from other livestock. Sampling for salmonella the frequency of sampling and the type of samples obtained will depend largely on the clinical signs, epidemiological data, laboratory facilities available and the objectives. Individual samples for bacteriological tests are taken as aseptically as possible and before any antibiotic treatment has commenced. Monitoring of poultry breeding flocks and hatcheries for salmonella: general considerations At present the preferred method for monitoring poultry breeding flocks and hatcheries for salmonella is by means of bacteriological examination of samples obtained from these establishments. Samples for bacteriological monitoring are obtained in the case of rearing flocks from the premises in which the birds are housed, or in the case of adult laying birds either from the premises in which the birds are housed or from the hatchery to which the hatching eggs from that flock are consigned. The samples to be taken are: a) On the premises in which birds are housed fresh faeces (each sample at least one gram), dead or culled birds or in the case of day old birds the chick box liners. Additionally it is recommended that environmental samples are also taken in both the premises and the hatchery at a similar frequency. Where the laying flock is sampled only on the premises, environmental sampling of the hatchery is required. The total number of samples to be taken on each occasion is shown below and is based on the random statistical sample required to give a probability of 95% to detect one positive sample given that infection is present in the population at a level of 5% or greater. Number of birds in Number of samples to be taken the flock on each occasion 25-29 20 30-39 25 40-49 30 50-59 35 60-89 40 90-199 50 200-499 55 500 or more 60 Up to five samples may be pooled together for bacteriological culture. All samples should be selected at random to represent the house or in the case of samples taken at the hatchery to represent the hatching eggs from that poultry flock. The following minimum frequency of sampling is recommended: a) Rearing flocks: At day old and three weeks prior to moving to laying accomodation. Where birds are moved from the rearing premises other than direct to laying accomodation a further sample should be taken three weeks S. All samples should be fully marked and identified as to the date of sampling and the flock to which the samples relate. Samples should be stored in a refrigerator at between 1?C and 4?C until they are dispatched to the laboratory (for not more than five days). All samples should be examined in a laboratory authorised for that purpose by the Veterinary Authorities. Bacteriological methods for culture of salmonella from poultry flocks and hatcheries As described in the previous chapter (Chapter 44 Salmonellosis: Section 1: Identification of the agent), many methods are available for the culture of salmonella. A practical method which is in wide use, using Rappaport-VasUiaids medium for the detection of salmonella in chick box liners, cloacal swabs, composite faeces samples, carcasses and environmental samples, is described below. Samples submitted for testing for the presence of salmonella should be examined on consecutive days. Samples should be stored in a refrigerator between 1?C and 4?C until required for examination. Day 5 a) the incubated plates and composite media or equivalent shall be examined and the findings recorded, discarding cultures which are obviously not salmonella. Sude serological tests shall be performed using salmonella polyvalent O (Groups A-S) and polyvalent H (phase 1 and 2) agglutinating sera on selected suspect colonies collected from the blood agar or MacConkey plates. Serological tests See Section 2: Serological tests, in the previous chapter (Chapter 44 Salmonellosis). The disease is caused by one of the unconventional slow agents, the biochemical nature of which remains unknown. The natural infection is commonly passed from ewe to lamb before parturition and during the period to weaning. Infection can also pass to unrelated animals, especially when lambing occurs in confined areas. The incubation time between primary infection and clinical disease is nearly always longer than one year and may sometimes exceed the commercial life-span of the sheep. Animals that never develop clinical signs can still be a source of infection to others. Identification of the agent: No definitive diagnostic test for scrapie infection is available. Scrapie can however be transmitted to laboratory rodents by injecting them with infected tissue, but the variable efficiency of primary isolations coupled with incubation times of 1-2 years preclude this as a routine diagnostic procedure. Clinical diagnosis is supported by pathological diagnosis of spongiform encephalopathy. This is characterised by the histological demonstration of bilateral and usually symmetrical spongiform change of neuropil and neuronal vacuolation in the grey matter of the brain stem and, often, spinal cord. The detection of specific scrapie-associated fibrils in extracts of diseased brain may become a useful, additional criterion for scrapie diagnosis. Serological tests: Scrapie infection is not known to elicit any specific immune response and there is no basis for establishing a diagnosis by detecting specific antibodies. For example, an affected sheep may either lead or trail behind a flock that is being driven (1, 7, 10). These changes progress to more definite signs that are of 2 kinds: a) Pruritus: There is often an extensive loss of wool from the flanks and hindquarters as the result of scraping or rubbing against objects and by Scrapie (B32) 425 nibbling. The compulsive nature of this behaviour can often lead to the development of skin lesions. Evidence of self-inflicted lesions may also be found on the head, ears, forelegs and base of the tail. Affected sheep often tremble and exhibit a nibbling reflex when rubbed over the rump, or even when rubbing themselves. Some sheep exhibit a high-stepping or trotting action of the forelegs, but more generally there is ataxia of the hind limbs, with much stonbling if the animal is obliged to move. When at rest, an affected sheep may stand awkwardly while supporting its hindquarters against a fence, or it may be seen to sway drowsily and even lose its balance. Characteristically, clinical signs are progressive and can lead to incoordination and an inability to stand. The clinical disease has a very variable course and may last for a week or up to several months, but invariably ends in death. The general bodily condition often remains good for most of the period of clinical disease, although a significant loss of weight may occur just before death in many but not all cases. This variation is probably due to differences between breeds of sheep, between strains of the agent and from environmental factors. It is not uncommon for affected animals with pruritus to show no incoordination, and vice-versa.
If you have questions or don?t understand what has been said alcohol and erectile dysfunction statistics purchase discount levitra professional, ask for clarifcation erectile dysfunction co.za buy cheap levitra professional 20 mg line. These fears often include fear for the well being of your family erectile dysfunction doctors in san fernando valley order genuine levitra professional online, fear of discomfort erectile dysfunction pills dischem order levitra professional online now, fear of what your future holds and fear of dying erectile dysfunction pills at walmart 20 mg levitra professional visa. Although a transplant unit is a place of hope erectile dysfunction medication free trial discount levitra professional 20 mg online, it is important to realize that patients can die during the course of transplantation erectile dysfunction treatment in kuala lumpur discount 20mg levitra professional. Sometimes family members and patients may not talk about their fears of death as way of trying to protect one another erectile dysfunction new drug levitra professional 20 mg low price, but these fears are very real and sharing them with family and staff often helps. Some patients may be angry at a higher power; others direct their anger at their family or the health care team. Loved ones may feel angry at the patient for being sick and disrupting their lives. Although anger is a normal reaction to stress, it is important to fnd a healthy outlet (for example, talking to someone) to relieve the tension. Caregivers may feel guilty for feeling angry or frustrated about their lives being disrupted or may feel helpless that they are unable to do more for the patient. These feelings are perfectly normal human emotions and coping with them may be diffcult. You and your loved ones should not hesitate to use the experience, understanding and support of the transplant team and unit staff to help you deal with these emotions. Many patients fnd support groups helpful in coping with the emotions and stress of transplant. A group is held weekly on the adult transplant unit for patients and their families. One option for your plan is CarePages that offers free, private web sites and patient blogs. The University of Michigan Health System has partnered with CarePages to make it easier for you and your family to communicate. You will fnd a link to the CarePages private site as well as instructions for using the program at There are many items and amenities provided at no charge to you on the transplant units. The transplant team encourages you to bring personal items to make the hospital room feel more at home. Items from home can also provide distraction and often help decrease stress and boredom. Many patients have found it diffcult to read or concentrate during the hospitalization. Check with your nurse coordinator or social worker if you have questions about items not on this list. This handbook 26 A Patient and Family Guide to Blood & Marrow Transplant 2013 University of Michigan Comprehensive Cancer Center Items to consider bringing: Pajamas, sweats, or loose ftting, comfortable street clothing to change daily (Tops that open in the front are preferable to pullovers; need loose collars for chest central lines. These units specialize in the care of blood and marrow stem cell and cord blood transplant patients. Although the entire Mott Hospital is Hepa fltered, keeping the door to your room closed is clinically necessary. Each room has a small closet, a small bedside stand with 3 drawers, and an overthe-bed table. You will also have your own refrigerator in your room it does not have a freezer component. This allows you access to television programming, a movie channel, a music system, the internet, and computer 29 2013 University of Michigan Comprehensive Cancer Center A Patient and Family Guide to Blood & Marrow Transplant games and comes with a keyboard at no charge. Ideas and Amenities for Your Stay Most of our patients prefer to wear their own clothing during their stay. If needed, we have free laundry facilities in the family lounge room 7-241 with free detergent. Exercise is very important in maintaining your stamina and preventing complications. In addition to physical therapy giving you exercises to complete daily and walking the halls, we have a family ftness center in room 7-231 with a treadmill, bike, and elliptical machines available. We are working to try to get patients a designated time of the day that they could use it too, but not yet. The Pediatric unit has activity areas that are designed for age appropriate activities for your child by our Child Life Department. There are a small activity room, 7-359, a large activity room, 7-411, and a teen activity room 7-418 flled with fun activities. We encourage you and your family/caregivers to use your time to get your needed rest. There is a family room located behind the hospital elevator, room 7-201 for larger family groups. This is where the kitchen facilities are located as well as the laundry facilities. The Pediatric unit also has a family lounge with kitchen facilities located on the unit in room 7-609. Lodging arrangements for visitors are available through the Patient and Visitor Accommodations Program. There are 3 stations, one in the lobby of Mott Hospital, one at the connector between the Taubman Center and Parking lot (P3) and Mott Hospital, and one on level 3 at the end of the connector from the P4 parking structure. There is also screening on the 7th foor when they arrive if they have missed the other stations. In general, children may visit if they are healthy and have not been exposed to contagious diseases (such as chicken pox, measles, colds, or fu) within the previous 48 hours. Because this is a transplant unit where patients are at risk when exposed to any illness, we ask that visitors and family members wait to visit until they are well. Separate family shower facilities are located in the hallway to 7 East room 7-357T. Family members will need to take all shower supplies with them as there are no supplies in the shower room. Family caregivers may have meal trays delivered to the room for a small fee that will be added to the hospital bill. Guest meal trays are ordered the same way patient meals are ordered, by calling the room service call center. Telephone Service There is no charge for local calls using the patient telephones. Standard telephone jacks are used in all rooms and personal phones, cell phones, answering machines or fax machines may be used. Patient Mail When you are in the hospital, you can receive mail at the following address. It is important to include your exact room number and hospital unit in the mailing address to ensure the mail gets to you. Also, write the word patient in the lower left-hand corner to facilitate quick delivery of your letter or card. Patient Name Mott Children & Womens Hospital Bone Marrow Transplant Unit Patient Unit 7Mott / Room # 1540 E. This will help your family and friends keep track of your progress through your transplant and allow you to determine the amount and content of information being released through your journey. Plants/Flowers As a transplant patient, you will be susceptible to infections until your white blood cell count and immune system recover. During this time, fresh-cut fowers and plants are not permitted because they may carry a large number of germs in their water and soil. Meals Your meals are ordered by you through the room service call center 23663 between the hours of 6:30am and 8pm. There will be some restrictions in your diet while you are hospitalized in order to reduce the chance for infection. A dietician will meet with you to review these dietary guidelines, restrictions, and options. Food is permitted from home if prepared following our guidelines and you will have your own refrigerator in your room. There is also a Nourishment room located on each unit, Adult, room 7-209, and Pediatrics, rooms 7-443 and 7-527, available for your use. Bringing Food to Transplant Patients Transplant patients are on a special diet throughout the transplant admission. Transplant patients may eat food brought in from the outside if the following guidelines are met: 33 2013 University of Michigan Comprehensive Cancer Center A Patient and Family Guide to Blood & Marrow Transplant Food Preparation Guidelines: 1. Always wash your hands and the surface the food will touch before handling food with soap and water. If you are handling raw meat you must wash your hands with soap and water after handling the meat and before touching another food substance. If a plate, cutting board, or utensil was used for pre-cooked food, do not use again for cooked food. Fruits with a thick peel (such as oranges, bananas) or those that can be eaten once peeled (such as apples) are allowed to be eaten fresh. Portion food into individual servings in tightly sealed containers and cool in the refrigerator or freezer prior to transporting to the hospital. This leaves food in the danger zone temperature of 40140 degrees which is prime temperature for bacteria to grow. Avoid fountain pop/soda, milkshakes, frosties, frozen yogurt or soft serve ice cream from bulk serving machines. Bakery items with cream and custard fllings as well as baked custard, are only allowed if they have been refrigerated, or are fresh out of the oven. Food should be re-heated in the microwave until hot (steaming) and promptly eaten. Smoking the University of Michigan Health System has a no smoking policy for the entire complex including all buildings, parking structures, and grounds. Tobacco contains fungus spores that can lead to a severe pneumonia in transplant patients. This conditioning phase may start before admission, may last one to ten days, and typically fnishes one or two days before the infusion of the stem cell product (blood, marrow, or cord blood). The conditioning you receive has been very carefully planned as part of a treatment protocol, a precisely timed and organized approach to the treatment of your disease. The protocol outlines the treatment (chemotherapy, immunotherapy, and/or radiation therapy) you need as well as blood tests, x-rays, and other procedures. The protocol was written by a doctor who is an expert in the feld of cancer and transplant. Before your admission, you will be asked to sign an informed consent that reviews your treatment protocol. This document assures that you have been given important information, including the risks and benefts about your treatment and the transplant process. Reduced Intensity Conditioning: In some cases for patients receiving allogeneic transplants, your doctor may elect to give you a less intensive or reduced intensity conditioning regimen. In such cases, the transplant doctor may not feel that your body can tolerate the full dose of chemotherapy that may normally be given to patients for transplant. In a reduced intensity transplant, chemotherapy is given to suppress your immune system, thereby allowing the new stem cells to take hold and start growing. As the chemotherapy is less intensive, the complications you may experience in the frst 2-4 weeks after transplant, such as mouth sores, infections, kidney or liver problems, are often much less. However, since a lower dose of chemotherapy is used, transplants do not immediately kill as many cancer cells as full dose treatments. Thus, patients undergoing reduced intensity transplants should be in remission or near complete remission. Your transplant doctor will be able to tell you if a reduced intensity transplant is possible for you. Conditioning: Chemotherapy Chemotherapy is an important part of the protocol and will consist of one or more different types of medications, depending on the type of disease you have and the type of transplant you are receiving. The chemotherapy is given on a specifc schedule that is thought best for eliminating your cancer cells and/ or bone marrow cells. You will be given a calendar indicating what drugs you will be given on each day of your protocol and how you should expect to feel. The doses of chemotherapy used in transplant are often much higher than those you may have received in prior treatments. To kill cancer cells in your body it is often necessary to use the highest doses possible. Therefore you may experience more severe and different side effects than the ones you may have had in the past. Unfortunately, chemotherapy also damage normal fast-growing cells such as those in your mouth, throat, bowels, skin, hair, and bone marrow. You may experience nausea and vomiting, mouth and 38 A Patient and Family Guide to Blood & Marrow Transplant 2013 University of Michigan Comprehensive Cancer Center throat sores, diarrhea, a rash or change in the color of your skin, and hair loss. Good oral hygiene including frequent mouth care is required of all transplant patients to help limit the number of mouth or throat sores. If you develop diarrhea, we will identify the cause and treat to correct accordingly. In most cases, the hair loss is temporary and should grow back within a few months. Chemotherapy may also affect organs of the body such as the kidneys, liver, heart, and lungs. Although severe side effects are infrequent, they can progress and cause more serious problems. We will monitor your kidney and liver function daily through blood tests and listen to your heart and lungs daily to check for changes. If you notice any changes, it is important for you to notify your nurse immediately. Conditioning: Immunotherapy Immunotherapy is most frequently used in patients who are receiving transplants for a non-cancer related diagnosis. When it is used as part of the conditioning regimen, it is usually given intravenously for several days prior to transplant and may also be given after transplant. The main goal of immunotherapy is to suppress the immune system and prevent the rejection of the new donor cells. For some of these drugs, test doses are given frst to determine if you are allergic or can tolerate the drug. Conditioning: Radiation Therapy Radiation therapy (or irradiation) is a part of some transplant regimens. Radiation is given by a machine that sends high speed x-rays (Photons) into your body. The type of radiation you may receive will be discussed with you by your transplant doctor and the radiation oncology team. If radiation is planned, you will meet with the radiation oncology team before your transplant admission. The team consists of the radiation oncology doctor (who is in charge of your treatments), the radiation technologist (who administers the treatments), and the radiation nurse (who will monitor you for side effects). Depending on the type of radiation you receive, you may have a planning appointment call simulation. Simulation is composed of having you on lay on an x-ray table, making an immobilization device if needed, taking x-rays if necessary to prepare blocks (used to block tissue that will not be treated such as lungs, liver) and placing markings (permanent marker or tattoos) for lining up purposes on the treatment machine. The special machines in this department require the temperature to be cold, therefore, you may want to wear a robe and bring an extra blanket. You should take off all metal jewelry, safety pins, clips, etc) before your appointment. During this time you will be asked to remain in a certain position (lying down, standing, or sitting). You will be alone in the treatment room but the technologist can hear you, talk to you, and see you on a video monitor. Initially you may experience 40 A Patient and Family Guide to Blood & Marrow Transplant 2013 University of Michigan Comprehensive Cancer Center nausea, vomiting, and diarrhea. After you complete the radiation treatments you may develop other side effects such as red, dry skin which may look like a suntan; dry mouth which occurs from the effect of the radiation on the salivary glands; parotitis (swelling of the parotid glands in front of your ears); infertility; lung changes and cataracts (see complications section for more information). Your doctors and nurses will be monitoring you very closely for these side effects. If you have questions regarding your treatments, ask a member or your transplant or radiation team. Stem Cell Infusion Transplant After your conditioning treatment, you are ready to receive your stem cell infusion or transplant. If you are receiving stem cells from a donor (an allogeneic transplant), the infusion is given soon after the cells have been collected and processed. If you receive your own stem cells (an autologous transplant) or cord blood stem cells, a blood bank technologist will bring the frozen product to your room. The most common side effect is its bad taste (fsh, garlic, cold creamed corn, V8 juice). This taste can be very strong and cause nausea and 41 2013 University of Michigan Comprehensive Cancer Center A Patient and Family Guide to Blood & Marrow Transplant vomiting. This contributes to the bad taste/ breath as well as having this smell linger in your room for two to three days especially if you have a lot of bags of stem cells. This is from any red blood cells that were in the product that did not survive the freezing/ thawing process. Recovery Within 7-10 days of receiving your conditioning therapy, your blood counts will decrease and remain low until the new bone marrow cells begin to grow.
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Terrorists would be most likely to acquire radioactive material from the open market, as many radioactive isotopes are used in commercial practices such as communication technology and medical procedures. Furthermore, regulatory procedures to track global shipments of radiological materials used in commercial applications are not as consistent or widespread as those in place to regulate the transfer of nuclear material. A real concern is that terrorists with a high level of technical expertise may be able to use radiological material to contaminate a target without using an explosive device. Some isotopes can be dissolved and sprayed, while some can be vaporized, or even 148 burned. These delivery methods would require special technical skills and expertise in order to carry out an effective terrorist attack. Another concern is the fact that so many alQaeda affiliates are willing to die in the process of carrying out an attack. For this reason, if the group is able to procure a large amount of radioactive material, it may be less concerned about the technical expertise required to conduct a sophisticated attack; instead, they may opt to sacrifice a life in order to deliver a lethal dose of radiological material to a target. Radiological material that is ingested or inhaled is the most deadly, and this can only really occur in close proximity to a highly radioactive source. Such an attack would certainly instill fear and anxiety and would probably have serious economic consequences. Additionally, an agent disseminated using an explosive device could result in significant structural damage. The use of a chemical or biological agent, albeit crude, has the potential to cause fear and panic disproportionate to the actual effect of the agent. However, while they may succeed in terrorizing a population and causing massive economic disruption, it is unlikely that such agents would be capable of causing mass casualties. Indeed, the large majority of al-Qaeda literature dealing with operational methods outline the use of conventional explosives and guerrilla warfare. These accounts are not totally speculative; as Nikolai Patrushev, Chief of the Russian Federal Security Service, told journalists on August 19, 2005, Terrorists are striving to gain access to biological, nuclear and chemical arms. One of the main concerns is funding: Russian officials have reported that expected financial contributions from foreign countries have not been 151 able to meet the security needs of several important facilities. Others believe that a preferable path to nuclear acquisition for the group would be to obtain an off-the-shelf device. Although unlikely, even if such scenarios come to fruition, significant obstacles to terrorist acquisition and use of such devices remain. Reports stating that Osama bin Laden purchased biological or chemical weapons are largely speculative, as are reports indicating that bin Laden was able to acquire materials for chemical weapons from the 154 former Soviet Union during the mid-1990s. Even so, it is important to keep the threat in perspective and not attribute to these terrorists any capabilities that they still do not possess. It can be argued that the most dangerous aspect of the al-Qaeda network is its ability to recruit and replenish its ranks with young jihadis who are willing to die for their cause around the globe. As for the prospect of nuclear terrorism, however, exhaustive efforts should be made to prevent the al-Qaeda network from acquiring nuclear devices or fissile materials that can be used to build such weapons. Besides the technical difficulties inherent in the manufacture of nuclear devices, the acquisition of fissile materials remains the greatest obstacle to nuclear warhead assembly. When considering the specter of nuclear weapons in the hands of al-Qaeda, there simply is no acceptable margin of error. While the killing potential of these agents is theoretically high given modest technical and delivery means, it likely pales in comparison to the real threat of conventional weapons. In the near future, the al-Qaeda network is likely to continue to use conventional weapons and atypical weapons in creative ways. Esposito, Unholy War: Terror in the Name of Islam (Oxford: Oxford University Press, 2002), pp. National Memorial Institute for the Prevention of Terrorism, Terrorism Knowledge Database, Terrorist Group Pro? Federation of American Scientists, Intelligent Resource Program, Al-Qai?da-The Base, B/ Staten, Dirty Bomb Plot Thwarted Says Attorney General, Emergency Net News, June 10, 2002; Elaine Sciolino, Terror Verdict Due Today for a Frenchman on Trial in Morocco, New York Times, Sept. Pamela Hess, Al Qaida May Have Chemical Weapons, United Press International, Aug. Hala Jaber and Nicholas Rufford, M15 Foils Poison-Gas Attack on the Tube, Sunday Times (London), Nov. Says: Suspected Al Qaeda Operative Held as Enemy Combatant, Washington Post, June 11, 2002. Muhammad Wajdi Qandyl, Searching for Weapons of Mass Destruction and Al-Qa?ida, Al-Akhbar (Cairo), Jan. Toby Harnden, Rogue Scientists Gave bin Laden Nuclear Secrets, Daily Telegraph (London), Dec. Alleges Pakistani Businessman Urged al Qaeda to Acquire Nuclear Weapons, Miami Herald, Feb. Max Delaney, Under Attack al-Qaeda Makes Nuclear Claim, Moscow News, March 3, 2004. Martin Arostegui, Terrorism in Morocco Deeper Than Imagined, United Press International, June 7, 2003; Frenchman on Trial in Morocco Over Suicide Bombings, Agence France Presse, Aug. Glasser and Kamran Khan, Pakistan Continues Probe of Nuclear Scientists, Washington Post, Nov. Jeffrey Bale, Anjali Bhattacharjee, Eric Croddy, and Richard Pilch, Ricin Found in London: An al-Qa?ida Connection? Duncan Campbell, Vikram Dodd, Richard Norton-Taylor, and Rosie Cowan, Police Killer Gets 17 Years for Poison Plot, the Guardian (London), April 14, 2003. Schanzer, citing Terror Handbook Found at Ansar al-Islam Camp: Report, Associated Foreign Press, April 9, 2003. Schanzer, citing Isma?il Zayir, Ansar al-Islam Group Accuses Talabani of Spreading Rumors About its Cooperation with al-Qa?ida, Al-Hayat, Aug. Says it Found Qaeda Lab Being Built to Produce Anthrax, New York Times, March 23, 2002. Alan Culluson and Andrew Higgins, Computer in Kabul Holds Chilling Memos, Wall Street Journal, Dec. Marc Sageman, Understanding Terror Networks (Philadelphia: University of Pennsylvania Press, 2004), p. Anonymous, Imperial Hubris: Why the West is Losing the War on Terror (Dulles: Brassey?s, 2004), p. Stephen Ulph, For Mujahideen, Bomb-Making Information Readily Accessible Online, Jamestown, Aug. Raymond Zilinskas, Director of the Chemical and Biological Weapons Nonproliferation Program at the Center for Nonproliferation Studies, Interview by author, Monterey, California, Aug. Abu al-Usood al-Faqir, Instances of Radiation Pollution from 1945A1987, al-Farouq Website. Eric Croddy, Chemical and Biological Warfare: A Comprehensive Survey for the Concerned Citizen (New York: Copernicus, 2002), p. John Mintz, Technical Hurdles Separate Terrorists From Biowarfare, Washington Post, Dec. Congress, Senate, Committee on Governmental Affairs, Permanent Subcommittee on Investigations (Minority Staff), Staff Statement, Hearings on Global Proliferation of Weapons of Mass Destruction: A Case Study on the Aum Shinrikyo, Oct. Zimmerman and Cheryl Loeb, Dirty Bombs: the Threat Revealed, Defense Horizons, Center for Technology and National Security Policy, National Defense University, Washington, D. Joby Warrick, Soviet Germ Factories Pose New Threat: Once Mined for Pathogens in Bioweapons Program, Labs Lack Security, Washington Post, Aug. Dafna Linzer, Nuclear Capabilities May Elude Terrorists, Experts Say, Washington Post, Dec. Biosecure was commissioned to produce the background documents and initial drafts of the report documents. The Symposium was attended individual experts, have reviewed developments in: by 72 participants from 30 countries. Technological barriers to acquiring and background papers and during the Trends using a biological weapon have been signifcantly Symposium around which this report is structured: eroded since the Seventh Review Conference. Biosciences are developing at an unprecedented there had been no further novel developments rate. Summaries of these sections are provided below this increases the likelihood of such and each is considered in detail in each chapter in developments in the foreseeable future. Biotechnology is increasingly important around unprecedented rate the world as a manufacturing technology and has the rate and scale of progress in the life sciences therefore become a potential target for biological and biotechnology continues to grow rapidly. It should be explored whether is an increasing difusion of knowledge around the there are any risks not already captured by existing world and more interconnection between knowledge treaties and laws of weapons that cause damage to hubs, many of them virtual. Laboratories operate in equipment supplies, or material associated with the more diverse geographic locations and across diferent bio-economy. Moving from concept to application is becoming ever simpler, unlocking further potential for progress. It could achieve this by devising an efective, already been partially implemented in both developed on-going, and suitably resourced mechanism to: and developing countries. Develop specifc questions that can be answered pressing need for more comprehensive sets of baseline through an on-going review of developments in and reference pathogen data. Outsourcing of key production steps has reduced the need for dedicated vaccine production the global ability to detect and treat disease has infrastructure. The meeting noted that one positive outcome of Single-use equipment and modular production developments since the Seventh Review Conference technologies shorten turn-around times. A more is that our collective capacity to combat disease has distributed production base in industry reduces the markedly improved, regardless of whether the outbreak distance a product has to travel to its point of use. However, regulatory and liability issues associated with Whilst it would require remaining logistic, economic and diagnostics, drugs and vaccines in health emergencies technical barriers to be surmounted, the conference continue to limit potential for progress and this is an noted that it should now be possible to assemble issue that should be addressed. Such a system could scale from local needs through the scientifc advances reported at the conference to international responses. A structure that enabled could also facilitate almost every step of a biological data such as pathogen sequences to be shared more weapons programme and technological barriers to efectively and efciently would facilitate a rapid an acquiring and using a biological weapon have been efective response. As expertise and know-how conspicuously eroded since the Seventh Review matures, opportunities for technological leapfrogging Conference. Developing countries can then access Both novel and traditional approaches continue to opportunities and capabilities in the feld that match, if ofer opportunities for acquiring an agent from nature. The sometimes-formidable challenges associated with the synthesis of existing agents and the development of novel agents have been overcome by using gene transfer and other biosynthetic engineering approaches. The Biological and Toxin Weapons Convention 8 Modifcation of biological agents enables them to be more easily optimised for use in a biological weapon. Developments in scale-up and production technologies have changed production signatures. Although these trends diminish the need for stockpiling, the proliferation of such capabilities, such as freeze-drying capacity, has actually reduced the space required to store biological weapons. It is also now easier to deliver a biological agent given advances in areas such as nanoparticles and sophisticated modelling of dispersal patterns using the techniques of aerobiology. Making use of them for prohibited purposes would probably currently require the resources of a state but this situation may change in the future, reinforcing the need for on-going eforts to review relevant developments in science and technology. No concrete examples of such developments were identifed either through the literature reviews or in workshop discussions. However, a number of potential future scenarios were Biotechnology is an increasingly important global identifed that do give cause for concern. Degrading manufacturing where, for example, the mechanisms of action of infrastructure and capability has long been a tool of war, weapons are not clearly chemical or biological, where insurgency and armed confict. The bio-economy itself components are signifcantly diferent from existing is therefore a potential target for biological weapons. States Parties could proactively consider the implications of these scenarios before they can be the conference also noted an increased need realised, thereby providing a window of opportunity to for education and outreach to promote the aims develop appropriate responses and actions. Further consideration of progress in this area may be needed prior to the 9th Review Conference and this reinforces the importance of a fexible process for ongoing reviews (see Section 2). Each round, twenty fellows skills and a sustaining network to help them guide participate in two in-residence workshops during a socially responsible future for synthetic biology. This requires an understanding of how developments in the life sciences and biotechnology might impact the treaty. Such a process might also usefully consider criteria for identifying relevant or signifcant developments. The Biological and Toxin Weapons Convention 12 3 the global ability to deal with disease Since the Seventh Review Conference, developments in the understanding of disease have increased capacity to deal with disease, regardless of whether the outbreak is naturally occurring or the result of a malevolent act. This includes structural between pathogenicity, transmissibility and drug and genetic analysis, production or isolation of resistance identifcation of mutations that can confer pathogens with increased transmissibility or host range additional functional characteristics without degrading and improved tools to investigate these. This includes Other factors structural and genetic analysis, investigation of other Improved insights into other factors that impact factors which efect virulence (infection location, pathogenicity, such as physical location in the host acclimatisation) and improved tools to investigate these. Increased virulence Improved understanding of interactions between Pathogens with increased virulence have been genetic elements also plays an important role in the produced: production and action of toxins, for example, gene interaction mapping of Ricin toxin. The Biological and Toxin Weapons Convention 18 Structural analysis Drug development Improved techniques for structural analysis: Better tools for drug development are available. More comprehensive baseline data is needed to assist in comparisons and establish a norm against which to compare unusual events. The Biological and Toxin Weapons Convention 19 We are increasingly able to diferentiate between 3. These have been improved by increased discipline of microbial forensics can help establish sensitivity and ability to detect a wider range of agents attribution if a malevolent deployment is suspected using a wider range of samples and to detect functional (see below). This has been and record environmental signals in the gut163 achieved through the use of nanoparticles;172, 177 and probiotics for disease detection. This improves our Advances in mass spectrometry have contributed to understanding of disease progression, enables our understandings of disease, facilitated detection earlier detection of disease, diferentiates between of disease outbreaks as they occur, and identifcation diferent infections, and facilitates triaging of of the agent responsible. N?Faly Magassouba, Infectious and available since the Seventh Review Conference, for Tropical Diseases Department, National Hospital example paper-based microscopes costing less than Donka, Conakry, Guinea. Research into clinical waste management has A variety of diferent technologies and approaches revealed that fragments of genetic material survive have been used for bioforensics, including: standard cleaning and decontamination techniques. These advances have increased the amount of these technologies and approaches have increased benchmark and background data available for our capacity for investigating the origins of a given diagnostics, enabled a wider set of samples to be pathogen by: used in diagnostics, decreased time for confrming. Similarly, cheap, paper based microscopes resistance;245 and have been developed with diferent levels of resolution. The Biological and Toxin Weapons Convention 26 There has also been progress in developing highOnline laboratories and facilities now ofer all of throughput approaches to diagnostics, for example these services at a single site, as an organism design the use of Luminex microarray hybridization to identify support service. However, bioinformatics capabilities remain a Synthetic biology is reducing the time needed to major challenge in this area. Examples example, a lack of quality assurance in toxin include: antibody-based drugs; novel drugs for diseases diagnostics has been demonstrated internationally. A number of infectious agents reduces the time required to of new avenues for designing vaccines have been develop vaccines, drugs and other counter-measures. Outsourcing of key production steps has reduced the need for dedicated vaccine production infrastructure. It is increasingly simpler, faster and cheaper to industrialize production processes. Single-use equipment and modular production technologies shorten turn-around times. A more distributed production base in industry reduces the distance a product has to travel to its point of use. However, regulatory and liability issues associated with diagnostics, drugs and vaccines in health emergencies continue to limit potential for progress and this is an issue that should be addressed. High-throughput platforms and big-data approaches continue to reveal a wealth of potential new targets and candidates for drugs and vaccines. The design, testing and optimisation processes have been streamlined by the digitalization of biology supported by better computational technologies, improved capacity for rational design, integration of synthetic biology approaches, more sophisticated modelling tools, enhanced synthesis technologies and a wider array of platform technologies (see Box 3). The Biological and Toxin Weapons Convention 27 Box 3: An example of how the digitization of biology accelerates vaccine development. Examples since the Seventh Computational and bioinformatics approaches have Review Conference include, a protocol for testing the improved design strategies, including: ability of a specifc protein to rescue knock-out strains. For example, research has techniques to engineer microbes to sense and demonstrated the value of administering both eradicate pathogens;284, 285 antibodies and antibiotics in treating anthrax. Progress has been made in developing antibodythis included the use of: based drugs, against: Smaller facilities using more compact equipment Notable progress has been made in the development increases the range of potential sites and reduces of more efective and stable vaccines, such as through logistical challenges, in some cases also ofering cost the use of virus-like particles, for example: benefts. In other cases, improvements in efcacy that elicit broader and more potent immunity than and efciency, such as those associated with automation traditional infuenza vaccines;312, 319 and miniaturization, may incur increased costs. There have been improvements in vaccine expression, in particular through insect cell Existing approaches to increase impact and efcacy line and suspended cell cultures and the use of bulk continue to be developed, for example: 311 production material. Post-production purifcation steps have been improved or simplifed, as a result of the New or improved vaccines have been designed and growing regulatory attention paid to ensuring viral developed to prevent: 313 removal or inactivation. Examples since the Seventh disposable, aseptic, genderless, universal connectors, Review Conference include the use of suspended and non-proprietary flms and materials of construction cultures in vaccine production329, 330 and industrial use 343 for its bags, connectors, and tubing.
Diseases
Pillay syndrome
Facial clefting corpus callosum agenesis
SCAD deficiency
Hyperchylomicronemia
Angiofollicular lymph hyperplasia
Ectrodactyly ectrodermal dysplasia
Myopathy with lysis of myofibrils
Vitamn B12 responsive methylmalonicaciduria
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If the inoculum contains large numbers of spores, a solid layer of growth will cover the plate. Difficulties can occur when the spore suspensions contain other spore-forming bacteria which may completely overgrow the culture. When growth has occurred, the addition of a drop of sulfanilic acid-alpha naphthyl reagent produces a red colour if any nitrate has been reduced to nitrite. Most aerobic bacteria break down the peroxide to water and oxygen, producing a bubbly foam, but B. External appearance of affected comb: cappings with moist and sunken appearance, some punctured with larval remains. The classical method (Sturtevant, 1932) is to dilute the honey 1:5 with water and centrifuge at high speed to deposit the spores. Honey (25 ml) from each sample is diluted with 25 ml of water to permit easier handling. The tubes are submerged in running water for 18 hours or in a water bath with 3-4 water changes in the same period. Serological tests Apart from the fluorescent antibody technique described above for detection of the organism there are no avaUable serological tests. Reproduced with the kind permission of the author and Ehrenwirth-Verlag, Munich (Federal RepubUc of Germany). The identification of its presence by the observation of signs of disease in the field is unreliable. The most usual and obvious sign is the death of larvae shortly before they are due to be sealed in their cells, but this may be caused for reasons other than European foulbrood. Most infected colonies display few visible signs which then often quickly abate spontaneously before the end of each active season. Infection remains enzootic within individual colonies because of mechanical contamination of the honeycombs by the durable organism. Identification of the agent: the examination of suitable preparations of larval remains by high-power microscopy for the presence of numerous lanceolate cocci is adequate for most practical purposes, especially when it is done by experienced individuals. The only certain means of making a diagnosis is by isolating and identifying the causative organism. This can be differentiated quite readily from all other bacteria associated with bees by its fastidious cultural requirements. The isolated bacterium can be identified and differentiated by means of simple tube agglutination tests. The disease is caused by Melissococcus pluton and occurs mostly during the early summer when colonies are growing quickly. Most sick larvae become displaced from their coiled position in the bottom of their cells before they die. Many are quickly detected and removed by nurse bees, leaving empty cells scattered randomly among the remaining brood. Infected larvae that escape detection by adult bees and then die, first become flaccid and turn a light yellow colour becoming increasingly brown, at the same time melting into a semi-liquid mass. They then dry out and form a dark brown European foulbrood (B76) 695 scale, and can easily be removed from the cells. Severely affected brood may have a very stale or sour odour, but often there is no smell. Signs of disease usually disappear spontaneously from infected colonies before the end of the active season but are likely to return in subsequent years (1, 5). Identification of the agent Before any decomposition occurs, diseased larvae or those that have recently died can be dissected on a microscope shde by pinching the cuticle about the centre of the body with 2 pahs of forceps which are then pulled apart. The mid-gut contents are left exposed on the shde, still within the gelatinous, transparent peritrophic membrane. This is partially or almost completely filled with bacteria which are easily seen as opaque chalk-white clumps. The contents of the mid-guts of healthy larvae, which are less easily dissected, have a golden-brown colour. The mid-guts of healthy larvae that contain much light-coloured pollen may resemble those that are filled with bacteria. For a bacteriological investigation a loopful of a dilute aqueous suspension of the mid-gut contents is transferred to a clean microscope shde and mixed with a 2 loopful of 5% aqueous nigrosin. This is spread over a few cm, dried gently over a flame, and examined directly by high-power microscopy. Similar preparations made from aqueous suspensions of whole dead or decomposing larvae are likely to appear more confusing, especiaUy when secondary organisms predominate. It can be cultivated on a medium (expressed in g/1 or ml/1) comprised of yeast extract or certain peptones (2) 10; cysteine or cystine, 0. The medium is preferably autoclaved in 100 ml lots in screw capped bottles at 116?C for 20 minutes and poured into petri plates immediately before use. These plates are streaked with dUute aqueous suspensions of diseased larvae, or ideaUy, of diseased larval mid-guts. The latter can be prepared beforehand by allowing them to dry on a shde which may then be kept, for years if necessary, at 4?C, or -20?C. This treatment also eliminates most secondary organisms after a few weeks without affecting the viabiuty of M. This organism is isolated most efficiently by inoculating decimal dUutions of the aqueous suspension into agar which has been maintained molten at 45?C and which is then poured into plates. This bacterium is somewhat pleomorphic in vitro, often appearing in rod-like forms. Alternatively the cultures can be suspended in an medium of 10% sucrose, 5% yeast extract and 0. A number of other bacteria are frequently associated with and may be confused with M. Its incidence in healthy bees is very low in winter and early spring but it increases in summer. It sometimes resembles streptococci when grown in certain media, and has been confused with M. However, its cultural characteristics closely resemble those of Corynebacterium pyogenes (6) and it multiplies poorly in the form of thin rods, under the conditions necessary for the cultivation of M. It is reponsible for the sour smell sometimes encountered with European foulbrood. It forms small transparent colonies within 24 hours and is a facultative anaerobe. It multiplies on a variety of the more common media with or without carbohydrates or C02. When it is not diluted out it produces sufficient acid to prevent the multiplication of M. In bee colonies it multiplies only in the decomposing remains of larvae, and then its spores often predominate over all other bacteria even to their apparent exclusion. It forms very resistant spores and becomes well established in bee colonies with enzootic European foulbrood. It produces a spreading growth of transparent colonies, some of which are motile and move in arcs over the surface of the agar. Cultures have the characteristic stale odour that is associated with European foulbrood when the bacillus is present. Both rods and spores are larger than those o/Bacillus larvae (see American foulbrood). Assays are made by agglutination tests in tubes containing suspensions of bacteria equivalent to 0. Acknowledgments Illustrations by Karl Weiss, extracted from Bienen-Pathologie, 1984. Reproduced with the kind permission of the author and Ehrenwhth-Verlag, Munich (Federal Republic of Germany). The parasite hyphae invade the posterior region of the mid-gut giving rise to large numbers of spores within a short time. The parasite is ubiquitous and occurs in greatest numbers in the spring when there is an increase in the brood. Spores can persist for up to 2 years in faecal droppings and up to 1 year in honey and in bee carcasses. Identification of the agent: In the acute form, brown faecal marks are seen on the comb with sick or dead bees in the vicinity of the hive. In milder cases, there may be no special signs, perhaps only poor colonies with large quantities of brood and few adult bees. Microscopical examinations of homogenates of the abdominal contents of affected bees will reveal the oval spores of Nosema, about 5-7 im by 3-4 un with a dark edge. The appearance of Nosema infestations can be confused with yeast cells, fungal spores, fat and calciferous bodies or cysts of Malpighamoeba mellificae. Only positive identifications by observation of typical spores in the mid-gut or faeces can be regarded as significant. The extent of infestation is determined by counting the spores in 10 areas of a slide and calculating the average number of spores per area. Infection occurs by the ingestion of the spores in the feed or after the grooming of the body hairs of the bee. Primary bodies pass down the hyphae and enter the epithelial cell, where they reproduce. Infected bees are unable to fly and have been shown to be infected with up to 500 million spores. The parasite is ubiquitous and multiplies at a specific rate throughout the year, with maximum numbers occurring during the spring, which coincides with the increase in the brood. In winter spores are rarely to be found, or are perhaps only found in bees which are heavily infested. The development stages necessary for hibernation are difficult to find microscopically because of their very small size. Any inherent natural defense by a bee colony against a heavy infestation with the parasite depends on the colony size as well as on the prevailing weather conditions during the early part of the autumn of the previous year. If these conditions are unfavourable, the overall life expectancy of the colony is reduced due to hormonal imbalance. This may lead to the premature death of bees during the winter at a time when they are responsible for the brood. In a typical case of a colony being depleted on account of a Nosema infestation, the queen can be observed surrounded by a few bees, confusedly attending to brood that is already sealed. In faecal droppings they may persist for up to two years, and in honey or in the carcasses of dead bees they may last for one year. Fumes from a solution of acetic acid of at least 60% will inactivate any spores, depending on the concentration, within a few hours; higher concentrations are even more effective and will kill spores within a few minutes. Disinfection can be carried out by putting acetic acid solution into bowls or onto sponges which can soak up the liquid. Following disinfection after an outbreak, all combs should be well ventilated for at least 14 days prior to use. Identification of the agent In acute forms of infestation, especially in early spring, brown faecal marks will be noted on the comb. At the entrance of the hive both sick and dead bees will be seen, and during winter there will be an increased mortality rate in the colony. In milder infestations there may be no special signs except for colonies of weak appearance with large numbers of brood but only a few adult bees. To diagnose a Nosema infestation, a number of bees are killed by compression of the thorax. The posterior pair of abdominal segments are removed with forceps to reveal the mid-gut, complete with the malpighian tubules, the small intestine and rectum. The mid-gut is normally brown but following a Nosema infestation becomes white and very fragile. Bee nosematosis (B77) 701 It is advisable to distinguish between a Nosema infestation and that of Malpighamoeba mellificae. In the former infection there is seldom an indication of dysentery, whereas in the latter there may be a sulphur-yeUow diarrhoea with a distinct odour. Three drops of the suspension are dropped onto a shde under a covershp and examined microscopically at x 400 magnification. Nosema spores are not to be confused with yeast cells, fungal spores, fat and cale bodies, nor with cysts of Malpighamoeba mellificae. As a variation of this method, a relatively fresh preparation of mid-gut may be examined. The intestine is ground up and a sample put directly onto a shde, diluted with a httle water, covered with a covershp and examined microscopically. The degree of infestation is determined by counting the spores in 10 known areas of a shde and calculating the average number of spores per unit area. These results may be evaluated as: Number of spores per unit area < 20 spores Weak infection 20 100 spores Increased infection > 100 spores Strong infection. A laboratory method for the simultaneous detection of Nosema spores and Amoeba cysts consists of the individual examination of the colonies (30-60 bees per colony). A suspension of the abdomens of dead bees is prepared by grinding with 5-10 ml water; the amount of water depends on the number and condition of the bees. One drop of the suspension is placed on a shde and examined microscopically at x 200 magnification. The results are assessed as <20 spores per area as negative; > 20 spores per area, positive. Nosema spores, pollen and parts of the malpighian tubules pass through the 100 um filter, but are collected on the 40 um filter. Only cysts inside the malpighian tubules can be taken as a positive result, because Amoeba cysts are often confused with fungal spores and yeast cells. It penetrates the intersegmental skin between the abdominal sclera of adult bees to ingest haemolymph. The number of parasites steadily increases with increasing brood activity and the growth of the bee population, especially in late summer and autumn, when clinical signs of infestation can first be recognised. The life span of the mites depends upon temperature and humidity, but in practice it can be said to last from 5 to 12 days. Identification of the agent: the clinical signs of varroasis can only be recognised at a late stage of infestation so that diagnosis entails the examination of bee debris. The earliest and most precise diagnosis can be made only after an application of a medication that obliges the mites to leave the bees. Larger amounts of debris, especially that produced in winter, can be examined using a flotation procedure. However, this method is less accurate due to the unequal distribution of mites and the usually small sample sizes. The mite inserts itself between the abdominal sclera in adult bees (6) where it penetrates the intersegmental skin in order to ingest haemolymph. The mites usually reproduce slowly at the beginning of the year, but their number increases with the bee population. The lifespan of mites on larval or adult bees depends upon temperature and humidity. In heavuy infested bee colonies clinical signs of varraosis can often first be seen in the latter end of the year (7), but heavy infestations are usuaUy reached 3-4 years after the primary invasion. This is due to an increased susceptibility to acute paralysis virus, as well as to the infection of wounds and loss of haemolymph (1, 2). Graph of populations of bees and mites over 1 year: Brood numbers (solid line); mite numbers (broken line). Right: bee with heavy infestation of mites, showing newly emerged but crippled bee. Diagram of Varroa jacobsoni Oudemans (female) a) dorsal aspect Note the flat shell-like b) antenor aspect (back and 4pairs of legs c) ventral aspect) d) Compare the bee louse (Braula coeca, female). Identification of the agent An easy method of diagnosis of varroasis is by the examination of the debris generated by bees themselves. Unless this insert is covered with such a gauze, or else smeared with grease, the bees will dispose of the mites outside the hive. The debris produced within a few days in late summer usuaUy contains Uttie other than visible mites (5, 8). An insert is placed in the hive as before but a medication is used to cause the mites to fall off the bees, so that after a given time, numbers of mites may be observed on the floor insert. Large amounts of debris can be examined in the laboratory using a flotation procedure (3). Debris is first dried for 24 hours, then flooded with industrial alcohol and stirred continuously for 10-20 minutes. In a second method, preferably 18-day-old drones and 13-day-old worker bees are selected. The brood, which in this case can have been stored at -20?C, is partially thawed and the ceU cappings removed. The sieves are rinsed thoroughly several times until the mites are located exclusively on the finest sieve. These sievings are emptied out onto a light-coloured surface and examined for mites (9). In a third method, about 200-250 bees are removed from open brood combs during the summer when the mite infestation will be heaviest. The bees are killed in a special container, covered either with petroleum spirit or alcohol, and the whole stirred for 10 minutes. The mites are then separated from the bees by means of a sieve having a mesh size of approximately 2-3 mm (9).
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