Malaria update 2012 Gunnar Holmgren, Infectious diseases clinic, Ryhov Hospital, Jönköping, Sweden May 2012 About 3.3 billion people are at risk of malaria globally, 350 million have the parasite in their body and 250 million will be ill with malaria in 2011 with almost 1 million dying as a result. Some new research suggests that 1.24 million died of malaria in 2010 with 85% of the deaths in Sub-Saharan Africa. This is the most important parasitic disease in the world. The majority of countries in tropical Sub-Saharan Africa (SSA) have a reduction of their potential income by between 1-1.3% per year due malaria and that over a 15 year period this loss of income amounts to almost 20% of their potential earnings. Put in another way probably 20% of their poverty is due to malaria. In the worst affected countries malaria accounts for 40% of public health expenditure, 30-50% of in-patient admissions and up to 60% of out-patient clinic visits. In 1997 it was estimated at the Global Malaria Conference in Hydrabad that annual deaths from malaria were between 2.5 million and 3.5 million. Cause Malaria is spread by the bite of the female Anopheles mosquito (45 species are important vectors with A. gambiae being the most effective) in which the parasite goes through a complicated life-cycle ending up in the salivary glands. A bite from a mosquito infected with such parasites is accompanied by the injection of saliva to stop the blood from clotting. With the saliva are injected parasites at the sporozoite stage where they can spread within 20 - 45 minutes through the blood stream of man and reach the hepatocytes of the liver. They bore into these hepatocytes and start asexual reproduction into schizonts and finally discharge merozoites into the blood stream. In Plasmodium falciparum (the main malaria parasite in Tropical Africa) the liver stage lasts 5.5 days. In P. malariae it lasts 15 days. The merozoites bore into red cells and form a vacuole with the invaginated red cell membrane. About 36 hours (54 hrs. in P. malariae) after invasion repeated nuclear division forms a schizont or better termed a meront and finally the growing parasite fills the red cell and is packed with merozoites. It then bursts and 6-36 merozoites are released to invade new red blood cells. The infection expands logarithmically at around 10-fold per cycle. The release of several substances at this bursting stage brings about the symptoms of malaria which include fever, headache, pain in muscles, nausea and vomiting. Finally, if untreated, so many red blood cells are damaged that the person becomes anaemic due to lack of intact blood cells. The spleen which is the dumping ground of broken down red cells may become enlarged and tender. The person may even become jaundiced as the break down products of haemoglobin finally overwhelm the liver's capacity to deal with them and the inflammatory effect of malaria on the liver cells function. After a series of asexual cycles in P. falciparum a sub-population of the parasite develops into sexual forms (gametocytes) with the process taking about 7-10 days. This is the stage that is then taken up by the female anopheles. The parasite then goes through the complex process within the mosquito which brings us back to our starting point as sporozoites in the salivary glands. The effect of malaria on nutrition Many episodes of malaria coming close together will decrease the intake of nutrients since the child that is acutely ill with the disease has nausea and vomiting, and may have a poor appetite even after the acute illness is over. The malaria medicines may also affect the appetite. The high fever increases catabolism and thus increases the need for nutrients. The effect of nutrition on malaria Paradoxically severe malnutrition has a relative protective effect against the severe complications of malaria. Children with kwashiorkor virtually never develop cerebral malaria until they have recovered from their malnutrition; they may then suddenly become sick with severe malaria. Maybe the parasite becomes as malnourished as the human host in severe malnutrition. Replication is slower with less severe complications of malaria. Conversely, most children with cerebral malaria are in the well-nourished group. It may be that iron deficiency anaemia also provides a limited protection against severe malaria, and iron supplements may aggravate the disease. Parasite types There are 4 main types of parasites: Plasmodium falciparum, P. malariae, P. ovale, and P. vivax. (P. knowlesi is found on Borneo and parts of peninsular Malaysia) Plasmodium falciparum is virtually the only life-threatening type and is also the dominant type in most parts of tropical Africa. In Zambia this type accounts for 95% of all malaria. In many parts of Asia the malaria threat is smaller than in Africa and P. vivax is dominant in many countries (India, Pakistan, Sri Lanka etc.) Only P. vivax and P. ovale can have a long-term liver stage where the parasite can hide for several years and flare-up again. This stage needs special treatment with primaquine to clear the parasite.
Disease situations There are four situations of malaria threat that are so different from each other that it is as if four different diseases are being described: 1. High transmission with an extremely effective vector (Anopheles gambiae) of P. falciparum. Here malaria will almost certainly be the main cause of morbidity and mortality in children under 5 and the main cause of illness amongst adults. Chloroquine treatment is virtually always useless. Impregnated mosquito nets and more effective treatment are the only hope in the community. 2. Low transmission of P. falciparum. Here the disease may occasionally come as epidemics (e.g. during El Nino-associated climate changes) that are a threat to the life and health of both adults and children with massive outbreaks of severe anaemia and cerebral malaria during the outbreaks but little impact otherwise. In an outbreak, there may even be discussion about short-term prophylaxis for children and pregnant women, as well as impregnated mosquito nets and early effective treatment. 3. Moderate transmission of P. vivax and ovale. Here both prophylaxis and treatment with chloroquine are usually effective but follow-up treatment of the liver stage with primaquine is still needed. Environmental changes such as draining marshlands and separating human from animal dwellings often make a big impact. 4. Low transmission of P. vivax, ovale and malariae. Here malaria is an exotic disease without much impact at the community level. Symptoms There are 4 main symptoms: headache, muscular pain, fever + rigors (shaking of the body due to the shivering attack), nausea + vomiting. These are usually in the absence of symptoms of respiratory infection such as sore throat, runny nose and coughing. There could be an almost identical presentation with influenza but here there are virtually always respiratory symptoms. Almost always after two days there will be some period completely without fever and the other symptoms. Signs There are 4 main signs on examining a person with malaria: raised temperature, anaemia, enlarged spleen, jaundice. However all four may be absent although the absence of fever throughout is rare. Intermittent fever is the usual picture with a normal temperature at times. The latter three signs are less usual in the first attack of malaria. Diagnosis Blood films with both thin and thick films are still the main-stay of diagnosis but these need to be accurately made, stained and assessed by an experienced technician to be useful. There are 4 new methods that are all expensive and mainly useful in looking for falciparum malaria: ParaSight-F, PCR, QBC malaria test, and HRP-2 (Malaquick) Severe complications
There are 4 main severe life-threatening complications all with P. falciparum and are due to inflammatory processes resulting from the release of cytokines when the red blood cells burst. Also the parasite causes red blood cells to become more sticky and to form aggregations including rosettes which slow down or stop circulation in the microcirculation to essential organs. This starts in the venules.: 1. cerebral malaria, 2. severe anaemia, 3. renal failure and blackwater fever (passing very dark urine because of a massive break-down of red blood cells with the release of free haemoglobin into the blood at a level that the kidneys cannot control and hence a leak of haemoglobin with its very dark colour in the urine), 4. pulmonary oedema (water in the lungs which become stiff and dangerously ineffective). Assessment There are 4 important clinical assessments to follow the course of severe malaria: daily (or twice daily) parasite counts, fluid input/output assessment, daily weighing for assessing fluid balance, measurement of conscious level using e.g. Glasgow or Blantyre coma score. Rapid treatment There are 4 rapidly-acting groups of drugs for treating acute malaria: 1. chloroquine or amodiaquine for P. vivax, malariae and ovale 2. artemether combination therapy (ACT) for malaria due to P. falciparum 3. quinine or quinidine for severe malaria due to P. falciparum, 4. mefloquine for malaria due to P. falciparum (Lariam®). Halofantrine has been very useful for P. falciparum treatment but now potentially life-threatening cardiac side-effects have been described which limit its value. A closely related drug, lumefantrine, without the cardiac side effects is now the commonest linked drug to artmesinine in ACT Dual therapy In an area with chloroquine resistant P. falciparum malaria, which means virtually all of Africa, (resistance to chloroquine with the other 3 types of malaria is still unusual) the strongly recommended treatment is “dual therapy.” Here are 4 examples: *1. Artemether + lumefantrine (Coartem®). Because of the Roll-Back-Malaria programme this is now widely available and is the best. In Europe it is marketed as Riamet® 2. Atovaquone + proguanil (Malarone®) is a rapidly acting anti-malarial which is good for early P. falciparum but very expensive. 3. Amodiaquine (Camoquin®) + Fansidar®. Cheap and well tested in Uganda but is likely to show resistance developing.
4. A new low-cost dual therapy was about to be launched but had to be dropped because of potential problems in those with G6PD deficiency: chloproguanil + dapsone (LapDap®) which was to have been combined with artmether. If single treatment with amodiaquine is used a follow-up treatment will almost certainly be needed with doxycycline. For severe malaria choose artemether i.m. or artesunate i.v. or quinine i.v. as these are the three that have been shown to give the quickest response. A short course with one of these until there is good improvement should be followed by oral Coartem® or doxycycline to clear the last parasites (in the early stages oral Coartem® may be possible). Some countries have suppositories with Artenam available where i.v. treatment is not possible. Follow-up treatment There are 4 follow-up slow-acting treatments when there are resistance problems: doxycycline, clindamycin (can be used in pregnancy), Fansidar® or cotrimoxazole. Chloroquine resistance There are 4 levels of resistance to chloroquine: no resistance;R1: good clinical effect and the blood slide becomes negative but the symptoms come back without new infection within 28 days; R2: same as R1 but blood slide never becomes negative; R3: no improvement either clinically or on parasite counts. Prophylaxis There are 4 main ways of avoiding malaria in Africa: 1. Avoid mosquito bites: e.g. use of mosquito nets impregnated with permethrin or deltamethrin over beds (97% effective in semi-immunes). Combine this with local insect repellants e.g. DEET. Put self-closing doors on all outside doors and mosquito nets on all windows. 2. Prophylaxis with an effective drug e.g. in East and Central Africa with mefloquine, or malarone or doxycycline(all 90% effective). Less effective: proguanil + chloroquine (75% effective); 3. Spray all houses within 1 km radius of an institution with insecticide regularly; such programmes of Residual indoor spraying (IRS) are mainly relevant in high density living areas. 4. Drain all puddles, marsh areas, remove tyres, cans and all rain-water accumulating items. Dosages in P. falciparum malaria treatment in Africa First choice when available Artemether-lumefantrine (Coartem®) For adults: 4 tablets as a single initial dose, 4 tablets again after 8 hours and then 4 tablets twice daily (morning and evening) for the following two days (total course of 24 tablets). For children: 5-15 kg: one tablet as an initial dose, one tablet again after 8 hours and then one tablet twice daily
(morning and evening) for the following two days (total course of 6 tablets); 15-25 kg: two tablets as an initial dose, two tablets again after 8 hours and then two tablets twice daily (morning and evening) for the following two days (total course of 12 tablets); 25-35 kg: three tablets as an initial dose, three tablets again after 8 hours and then three tablets twice daily (morning and evening) for the following two days (total course of 18 tablets). Above 35 kg as for adults. Second choice when available Atovaquone 1000 mg daily plus proguanil 400mg (Malorone®) daily by mouth after fatty meal for 3 days. In children 11-20 kg the dosage of atovaquone/proguanil is 250mg/100mg; 21-30 kg 500mg/200mg; 31-40 kg 750mg/300mg and above 40kg adult dose. Third choice when the above are not available: Amodiaquine 600mg daily by mouth for 2 days then 300 mg on 3rd day (total of 25mg/kg over three days) combined with Fansidar ® 3 tablets single dose. This is usually adequate treatment but if single treatment with amodiaquine is used and fever or symptoms come back after this add doxycycline 200 mg daily by mouth for 7-10 days Dosages in P. vivax, P. ovale and P. malariae First choice when chloroquine resistance is rare or at a low level of resistance. Start with: Full chloroquine course 10mg base/kg PO stat; then 5mg/kg at 12, 24 and 36 hrs in semi-immunes. Follow up in P. vivax and P. ovale with primaquine 0.25 - 0.5 mg/kg once daily with food for 14 days (in adults usual dose is 15-30 mg daily). Dosages in Severe malaria First choice when available Artemether (artemisinine derivative) 160 mg (children 1.6 mg/kg) i.m. twice a day for 3-7 days (an i.v. preparation of artenam is available in Sweden but rare in Africa) or if the person is not vomiting an alternative is oral Artemether + lumefantrine as above or artesunate 4 mg/kg daily for 3 days then doxycycline 200 mg orally daily for 7 days. Artenam by suppository is an excellent alternative but not yet widely available. Second choice (more likely to be available) Quinine dihydrochloride 20 mg salt/kg of body weight (loading dose) by infusion in 5% dextrose saline (5-10 ml/kg related to hydration level) over 4 hours or by i.m. injection. Eight to twelve hours later give a maintenance dose of quinine 10 mg salt/kg in dextrose saline over 4 hours. Repeat this dose every 8-12 hours until oral therapy is possible. If available then go over to Artemether + lumefantrine as above. Otherwise Oral therapy: quinine 10mg salt/kg orally thrice daily for 3 days then doxycycline 200 mg daily PO for 7 days, Alternative: mefloquine 15mg base/kg orally as first dose followed by one further dose of 10mg/kg 8 hrs later but more side-effects than above.
Conclusion There is undoubtedly an important improvement in malaria research, prevention including indoor residual spraying and impregnated bed-nets, better diagnosis including both microscopy and the rapid tests as well as Dual treatment with Coartem all resulting from the Roll-Back-Malaria interventions and the financing from the Global Fund and many others. Much discussion has taken place about whether eradication is possible and there is still no consensus about the right strategy in the most affected countries. Some feel that in this setting more lives will be saved if the focus is on containment rather than on eradication. It seems likely that until a highly effective and affordable vaccine is widely available, the idea of eradication of malaria in these areas is still a dream. In the field there are some precautions where Roll-Back-Malaria programmes have not yet been introduced and only microscopy is available for diagnosis. Make sure that the diagnosis is certain. In some laboratories in Africa many blood slides are falsely reported* as positive (especially if it is known by the laboratory assistant that the slide came from an expatriate). In some laboratories even if the slide is found negative, this is inevitably reported as “scanty malaria parasites seen” just so that the lab assistant is covered in case he missed a rare parasite. This can lead to many unnecessary treatments with all their side-effects * In one study in Tanzania blood slides were first assessed in the local hospital laboratory and then the same slide was taken to a malaria reference laboratory in Sweden and assessed by a very experienced expert. The results were very interesting. When the slide was reported as: + for malaria parasites there were 50% false positives. ++ for malaria parasites there were 10% false positives +++ for malaria parasites there were 3% false positives Only when the report stated ++++ for malaria parasites were all the slides truly malaria. Overall there were 3% false negatives. Of course such mistakes are understandable when the microscope is old, the stain reagents are old, the light source is poor and the laboratory assistant is poorly motivated (his salary may not have been paid for 6 months) or inadequately trained.
Proc. Interamer. Soc. Trop. Hort. 47:61-64. Vegetables/Hortalizas - October 2003 Qualidade e Conservação Pós-Colheita de Melão Gália Híbrido 'Arava' Submetido a Aplicações Pré- Colheita de Quelato de Cálcio e Boro Fábio del Monte Cocozza e Adimilson Bosco Chitarra, Universidade Federal de Lavras, Caixa Postal 37, 37200-000 Lavras, MG, Brasil, email@example.com Everardo Ferre
Proceedings of 12th ISMAS Symposium cum RSP-7 Workshop on Mass Spectrometry LC-MS/MS Studies on Identification and Characterization of Hydrolytic Products of Atorvastatin Ravi P. Shah, Vijay Kumar and Saranjit Singh * Department of Pharmaceutical Analysis National Institute of Pharmaceutical Education and Research (NIPER) Sector 67, S.A.S. Nagar 160062, Punjab