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There are many diagnostic tests for "Cryptosporidium". They include microscopy, staining, and detection of antibodies. Microscopy can help identify oocysts in fecal matter. To increase the chance of finding the oocysts, the diagnostician should inspect at least 3 stool samples. There are several techniques to concentrate either the stool sample or the oocysts. The modified formalin-ethyl acetate (FEA) concentration method concentrates the stool. Both the modified zinc sulfate centrifugal flotation technique and the Sheather’s sugar flotation procedure can concentrate the oocysts by causing them to float. Another form of microscopy is fluorescent microscopy done by staining with auramine.
Other staining techniques include acid-fast staining, which will stain the oocysts red. One type of acid-fast stain is the Kinyoun stain. Giemsa staining can also be performed. Part of the small intestine can be stained with hematoxylin and eosin (H & E), which will show oocysts attached to the epithelial cells.
Detecting antigens is yet another way to diagnose the disease. This can be done with direct fluorescent antibody (DFA) techniques. It can also be achieved through indirect immunofluorescence assay. Enzyme-linked immunosorbent assay (ELISA) also detects antigens.
Polymerase chain reaction (PCR) is another way to diagnose cryptosporidiosis. It can even identify the specific species of "Cryptosporidium". If the patient is thought to have biliary cryptosporidiosis, then an appropriate diagnostic technique is ultrasonography. If that returns normal results, the next step would be to perform endoscopic retrograde cholangiopancreatography.
The diagnosis of balantidiasis can be an intricate process, partly because the related symptoms may or may not be present. However, the diagnosis of balantidiasis can be considered when a patient has diarrhea combined with a probable history of current exposure to amebiasis through travel, contact with infected persons, or anal intercourse. In addition, the diagnosis of balantidiasis can be made by microscopic examination of stool or tissue samples.
Diagnosis can be difficult due to the lack of recognizable oocysts in the feces. PCR-based DNA tests and acid-fast staining can help with identification. The infection is often treated with trimethaprine-sulfamethaxozol [Bactrim, co-trimoxazole], because traditional anti-protozoal drugs are not sufficient. To prevent transmission, food should be cooked thoroughly and drinking water from streams should be avoided while outdoors.
There is no vaccine to control "Cyclospora" infection in humans at present, but one is available for reduction of fetal losses in sheep.
Some conventional parasitological techniques (CPT) such as wet blood film, and stained blood smears are used because so far, the best identifier is looking at the blood of the potentially infected host. Other tissues can be looked at, but the gold standard is identifying trypanosomes in the blood. Before the infection becomes severe, it is difficult to catch as many times these cryptic infections are undetectable by direct microscopy. Since CPT is not very sensitive, it cannot be used as a sole method of diagnosis.
The Haematocrit Centrifugation Technique (HCT) is a much better alternative. Using HCT trypanosomes can be detected in the blood even in field conditions. Buffy coat can be used to increase detection. Detection with this method is approx 85 trypanosomes per millilitre.
Rather than using live animals as test subjects, Canada used serological tests such as complement fixation tests to detect trypanosomes, and have been very successful. Other tests used look at detecting antibodies generated by the host species against T.evansi antigens. This is done using the enzyme-linked immunosorbent assays (ELISA) method. Now polymerase chain reaction (PCR) and DNA probes are being used to detect Surra in animals.
The presence of "T. cruzi" is diagnostic of Chagas disease. It can be detected by microscopic examination of fresh anticoagulated blood, or its buffy coat, for motile parasites; or by preparation of thin and thick blood smears stained with Giemsa, for direct visualization of parasites. Microscopically, "T. cruzi" can be confused with "Trypanosoma rangeli", which is not known to be pathogenic in humans. Isolation of "T. cruzi" can occur by inoculation into mice, by culture in specialized media (for example, NNN, LIT); and by xenodiagnosis, where uninfected Reduviidae bugs are fed on the patient's blood, and their gut contents examined for parasites.
Various immunoassays for "T. cruzi" are available and can be used to distinguish among strains (zymodemes of "T.cruzi" with divergent pathogenicities). These tests include: detecting complement fixation, indirect hemagglutination, indirect fluorescence assays, radioimmunoassays, and ELISA. Alternatively, diagnosis and strain identification can be made using polymerase chain reaction (PCR).
Many treatment plants that take raw water from rivers, lakes, and reservoirs for public drinking water production use conventional filtration technologies. This involves a series of processes, including coagulation, flocculation, sedimentation, and filtration. Direct filtration, which is typically used to treat water with low particulate levels, includes coagulation and filtration, but not sedimentation. Other common filtration processes, including slow sand filters, diatomaceous earth filters and membranes will remove 99% of "Cryptosporidium". Membranes and bag and cartridge filters remove "Cryptosporidium" product-specifically.
While "Cryptosporidium" is highly resistant to chlorine disinfection, with high enough concentrations and contact time, "Cryptosporidium" will be inactivated by chlorine dioxide and ozone treatment. The required levels of chlorine generally preclude the use of chlorine disinfection as a reliable method to control "Cryptosporidium" in drinking water. Ultraviolet light treatment at relatively low doses will inactivate "Cryptosporidium". Water Research Foundation-funded research originally discovered UV's efficacy in inactivating "Cryptosporidium".
One of the largest challenges in identifying outbreaks is the ability to identify "Cryptosporidium" in the laboratory. Real-time monitoring technology is now able to detect "Cryptosporidium" with online systems, unlike the spot and batch testing methods used in the past.
The most reliable way to decontaminate drinking water that may be contaminated by "Cryptosporidium" is to boil it.
In the US the law requires doctors and labs to report cases of cryptosporidiosis to local or state health departments. These departments then report to the Center for Disease Control and Prevention. The best way to prevent getting and spreading cryptosporidiosis is to have good hygiene and sanitation. An example would be hand-washing. Prevention is through washing hands carefully after going to the bathroom or contacting stool, and before eating. People should avoid contact with animal feces. They should also avoid possibly contaminated food and water. In addition, people should refrain from engaging in sexual activities that can expose them to feces.
Standard water filtration may not be enough to eliminate "Cryptosporidium"; boiling for at least 1 minute (3 minutes above of altitude) will decontaminate it. Heating milk at 71.7 °C (161 °F) for 15 seconds pasteurizes it and can destroy the oocysts' ability to infect. Water can also be made safe by filtering with a filter with pore size not greater than 1 micrometre, or by filters that have been approved for “cyst removal” by NSF International National Sanitation Foundation. Bottled drinking water is less likely to contain "Cryptosporidium", especially if the water is from an underground source.
People with cryptosporidiosis should not swim in communal areas because the pathogen can reside in the anal and genital areas and be washed off. They should wait until at least two weeks after diarrhea stops before entering public water sources, since oocysts can still be shed for a while. Also, they should stay away from immunosuppressed people. Immunocompromised people should take care to protect themselves from water in lakes and streams. They should also stay away from animal stools and wash their hands after touching animals. To be safe, they should boil or filter their water. They should also wash and cook their vegetables.
The US CDC notes the recommendation of many public health departments to soak contaminated surfaces for 20 minutes with a 3% hydrogen peroxide (99% kill rate) and then rinse them thoroughly, with the caveat that no disinfectant is guaranteed to be completely effective against Cryptosporidium. However, hydrogen peroxide is more effective than standard bleach solutions.
One way to diagnose "C. felis" is by taking blood and performing a peripheral blood smear to look for the erythrocytic piroplasms. The erythrocytic piroplasms are usually shaped like signet rings and are 1 to 1.5 µm. Not all cats that are infected will have the piroplasms on their blood smear, especially if they are early in disease course. Another method of diagnosing infection in sick cats is to take needle aspirates of affected organs and find the schizonts inside mononuclear cells in the tissues; examination of tissue is also useful for the diagnosis after cats have died. Blood samples can be sent away for polymerase chain reaction (PCR) testing to confirm infection. Other diseases that might resemble cytauxzoonosis should be ruled out. A major rule-out for "C. felis" is "Mycoplasma haemofelis" (formerly known as "Haemobartonella felis"); clinical signs can be similar to cytauxzoonosis and the organism may be confused on the peripheral smear. Because it causes similar signs in outdoor cats during the spring and summer, tularemia is another disease the veterinarian may want to rule out.
Other laboratory tests are often abnormal in sick cats. The CBC of an infected cat often shows a pancytopenia, or a decrease in red blood cells, white blood cells, and platelets; in some cases there is not a decrease in all three values. Clotting tests may be prolonged. Increased liver enzymes are common, and electrolyte disturbances, hyperglycemia, and acid-base disturbances can also be observed.
Microscopic demonstration of the large typically shaped oocysts is the basis for diagnosis. Because the oocysts may be passed in small amounts and intermittently, repeated stool examinations and concentration procedures are recommended. If stool examinations are negative, examination of duodenal specimens by biopsy or string test (Enterotest) may be needed. The oocysts can be visualized on wet mounts by microscopy with bright-field, differential interference contrast (DIC), and epifluorescence. They can also be stained by modified acid-fast stain.
Typical laboratory analyses include:
- Microscopy
- Morphologic comparison with other intestinal parasites
- Bench aids for "Isospora"
Preventative measures require effective personal and community hygiene. Some specific safeguards include the following:
- Purification of drinking water.
- Proper handling of food.
- Careful disposal of human feces.
- Monitoring the contacts of balantidiasis patients.
One strategy for the prevention of infection transmission between cats and people is to better educate people on the behaviour that puts them at risk for becoming infected.
Those at the highest risk of contracting a disease from a cat are those with behaviors that include: being licked, sharing food, sharing kithchen utensils, kissing, and sleeping with a cat. The very young, the elderly and those who are immunocompromised increase their risk of becoming infected when sleeping with their cats (and dogs). The CDC recommends that cat owners not allow a cat to lick your face because it can result in disease transmission. If someone is licked on their face, mucous membranes or an open wound, the risk for infection is reduced if the area is immediately washed with soap and water. Maintaining the health of the animal by regular inspection for fleas and ticks, scheduling deworming medications along with veterinary exams will also reduce the risk of acquiring a feline zoonosis.
Recommendations for the prevention of ringworm transmission to people include:
- regularly vacuuming areas of the home that pets commonly visit helps to remove fur or flakes of skin
- washing the hands with soap and running water after playing with or petting your pet.
- wearing gloves and long sleeves when handling cats infected with.
- disinfect areas the pet has spent time in, including surfaces and bedding.
- the spores of this fungus can be killed with common disinfectants like chlorine bleach diluted 1:10 (1/4 cup in 1 gallon of water), benzalkonium chloride, or strong detergents.
- not handling cats with ringworm by those whose immune system is weak in any way (if you have HIV/AIDS, are undergoing cancer treatment, or are taking medications that suppress the immune system, for example).
- taking the cat to the veterinarian if ringworm infection is suspected.
The use of trypanotolerant breeds for livestock farming should be considered if the disease is widespread.
Fly control is another option but is difficult to implement.
The main approaches to controlling African trypanosomiasis are to reduce the reservoirs of infection and the presence of the tsetse fly. Screening of people at risk helps identify patients at an early stage. Diagnosis should be made as early as possible and before the advanced stage to avoid complicated, difficult and risky treatment procedures.
For basic diagnosis, specific helminths can be generally identified from the faeces, and their eggs microscopically examined and enumerated using fecal egg count method. However, there are certain limitations such as the inability to identify mixed infections, and on clinical practice, the technique is inaccurate and unreliable.
A novel effective method for egg analysis is the Kato-Katz technique. It is a highly accurate and rapid method for "A. lumbricoides" and "T. trichiura"; however not so much for hookworm, which could be due to fast degeneration of the rather delicate hookworm eggs.
The main methods of controlling surra has been chemotherapy, and chemoprophylaxis in animals.
There is currently no vaccine against Chagas disease. Prevention is generally focused on decreasing the numbers of the insect that spreads it ("Triatoma") and decreasing their contact with humans. This is done by using sprays and paints containing insecticides (synthetic pyrethroids), and improving housing and sanitary conditions in rural areas. For urban dwellers, spending vacations and camping out in the wilderness or sleeping at hostels or mud houses in endemic areas can be dangerous; a mosquito net is recommended. Some measures of vector control include:
- A yeast trap can be used for monitoring infestations of certain species of triatomine bugs ("Triatoma sordida", "Triatoma brasiliensis", "Triatoma pseudomaculata", and "Panstrongylus megistus").
- Promising results were gained with the treatment of vector habitats with the fungus "Beauveria bassiana".
- Targeting the symbionts of Triatominae through paratransgenesis can be done.
A number of potential vaccines are currently being tested. Vaccination with "Trypanosoma rangeli" has produced positive results in animal models. More recently, the potential of DNA vaccines for immunotherapy of acute and chronic Chagas disease is being tested by several research groups.
Blood transfusion was formerly the second-most common mode of transmission for Chagas disease, but the development and implementation of blood bank screening tests has dramatically reduced this risk in the 21st century. Blood donations in all endemic Latin American countries undergo Chagas screening, and testing is expanding in countries, such as France, Spain and the United States, that have significant or growing populations of immigrants from endemic areas. In Spain, donors are evaluated with a questionnaire to identify individuals at risk of Chagas exposure for screening tests.
The US FDA has approved two Chagas tests, including one approved in April 2010, and has published guidelines that recommend testing of all donated blood and tissue products. While these tests are not required in US, an estimated 75–90% of the blood supply is currently tested for Chagas, including all units collected by the American Red Cross, which accounts for 40% of the U.S. blood supply. The Chagas Biovigilance Network reports current incidents of Chagas-positive blood products in the United States, as reported by labs using the screening test approved by the FDA in 2007.
Parasitic infections can usually be treated with antiparasitic drugs.
Albendazole and mebendazole have been the treatments administered to entire populations to control hookworm infection. However, it is a costly option and both children and adults become reinfected within a few months after deparasitation occurs raising concerns because the treatment has to repeatedly be administered and drug resistance may occur.
Another medication administered to kill worm infections has been pyrantel pamoate. For some parasitic diseases, there is no treatment and, in the case of serious symptoms, medication intended to kill the parasite is administered, whereas, in other cases, symptom relief options are used. Recent papers have also proposed the use of viruses to treat infections caused by protozoa.
They are treated with antiprotozoal agents. Recent papers have also proposed the use of viruses to treat infections caused by protozoa.
The following diagnostic methods are not routinely available to patients. Researchers have reported that they are more reliable at detecting infection, and in some cases can provide the physician with information to help determine whether "Blastocystis" infection is the cause of the patient's symptoms:
Serum antibody testing: A 1993 research study performed by the NIH with United States patients suggested that it was possible to distinguish symptomatic and asymptomatic infection with "Blastocystis" using serum antibody testing. The study used blood samples to measure the patient's immune reaction to chemicals present on the surface of the "Blastocystis" cell. It found that patients diagnosed with symptomatic "Blastocystis" infection exhibited a much higher immune response than controls who had "Blastocystis" infection but no symptoms. The study was repeated in 2003 at Ain Shams University in Egypt with Egyptian patients with equivalent results.
Fecal antibody testing: A 2003 study at Ain Shams University in Egypt indicated that patients symptomatically infected could be distinguished with a fecal antibody test. The study compared patients diagnosed with symptomatic "Blastocystis" infection to controls who had "Blastocystis" infection but no symptoms. In the group with symptoms, IgA antibodies to "Blastocystis" were detected in fecal specimens that were not present in the healthy control group.
Stool culture: Culturing has been shown to be a more reliable method of identifying infection. In 2006, researchers reported the ability to distinguish between disease causing and non-disease causing isolates of "Blastocystis" using stool culture. "Blastocystis" cultured from patients who were sick and diagnosed with "Blastocystis" infection produced large, highly adhesive amoeboid forms in culture. These cells were absent in "Blastocystis" cultures from healthy controls. Subsequent genetic analysis showed the "Blastocystis" from healthy controls was genetically distinct from that found in patients with symptoms. Protozoal culture is unavailable in most countries due to the cost and lack of trained staff able to perform protozoal culture.
Genetic analysis of isolates: Researchers have used techniques which allow the DNA of "Blastocystis" to be isolated from fecal specimens. This method has been reported to be more reliable at detecting "Blastocystis" in symptomatic patients than stool culture. This method also allows the species group of "Blastocystis" to be identified. Research is continuing into which species groups are associated with symptomatic (see Genetics and Symptoms) blastocystosis.
Immuno-fluorescence (IFA) stain: An IFA stain causes "Blastocystis" cells to glow when viewed under a microscope, making the diagnostic method more reliable. IFA stains are in use for Giardia and Cryptosporidium for both diagnostic purposes and water quality testing. A 1991 paper from the NIH described the laboratory development of one such stain. However, no company currently offers this stain commercially.
Prevention and control measures to prevent soil-transmitted helminthiasis are the following: availability of clean water for personal and domestic uses, improved access to sanitation which includes the use of properly functioning and clean toilets by all community members, education on personal hygiene such as hand washing and hygienic and safe food preparation; eliminating the use of untreated human faeces as fertilizer.
Only specialized laboratories can adequately diagnose "Babesia" infection in humans, so "Babesia" infections are considered highly under-reported. It develops in patients who live in or travel to an endemic area or receive a contaminated blood transfusion within the preceding 9 weeks, so this aspect of the medical history is vital. Babesiosis may be suspected when a person with such an exposure history develops persistent fevers and hemolytic anemia. The definitive diagnostic test is the identification of parasites on a Giemsa-stained thin-film blood smear.
So-called "Maltese cross formations" on the blood film are diagnostic (pathognomonic) of babesiosis, since they are not seen in malaria, the primary differential diagnosis. Careful examination of multiple smears may be necessary, since "Babesia" may infect less than 1% of circulating red blood cells, thus be easily overlooked.
Serologic testing for antibodies against "Babesia" (both IgG and IgM) can detect low-level infection in cases with a high clinical suspicion, but negative blood film examinations. Serology is also useful for differentiating babesiosis from malaria in cases where people are at risk for both infections. Since detectable antibody responses require about a week after infection to develop, serologic testing may be falsely negative early in the disease course.
A polymerase chain reaction (PCR) test has been developed for the detection of "Babesia" from the peripheral blood. PCR may be at least as sensitive and specific as blood-film examination in diagnosing babesiosis, though it is also significantly more expensive. Most often, PCR testing is used in conjunction with blood film examination and possibly serologic testing.
Other laboratory findings include decreased numbers of red blood cells and platelets on complete blood count.
In animals, babesiosis is suspected by observation of clinical signs (hemoglobinuria and anemia) in animals in endemic areas. Diagnosis is confirmed by observation of merozoites on thin film blood smear examined at maximum magnification under oil using Romonovski stains (methylene blue and eosin). This is a routine part of the veterinary examination of dogs and ruminants in regions where babesiosis is endemic.
"Babesia canis" and "B. bigemina" are "large "Babesia" species" that form paired merozoites in the erythrocytes, commonly described as resembling "two pears hanging together", rather than the "Maltese cross" of the "small "Babesia" species". Their merozoites are around twice the size of small ones.
Cerebral babesiosis is suspected "in vivo" when neurological signs (often severe) are seen in cattle that are positive for "B. bovis" on blood smear, but this has yet to be proven scientifically. Outspoken red discoloration of the grey matter "post mortem" further strengthens suspicion of cerebral babesiosis. Diagnosis is confirmed "post mortem" by observation of "Babesia"-infected erythrocytes sludged in the cerebral cortical capillaries in a brain smear.
The gold standard for diagnosis is identification of trypanosomes in a patient sample by microscopic examination. Patient samples that can be used for diagnosis include chancre fluid, lymph node aspirates, blood, bone marrow, and, during the neurological stage, cerebrospinal fluid. Detection of trypanosome-specific antibodies can be used for diagnosis, but the sensitivity and specificity of these methods are too variable to be used alone for clinical diagnosis. Further, seroconversion occurs after the onset of clinical symptoms during a "T. b. rhodesiense" infection, so is of limited diagnostic use.
Trypanosomes can be detected from patient samples using two different preparations. A wet preparation can be used to look for the motile trypanosomes. Alternatively, a fixed (dried) smear can be stained using Giemsa's or Field's technique and examined under a microscope. Often, the parasite is in relatively low abundance in the sample, so techniques to concentrate the parasites can be used prior to microscopic examination. For blood samples, these include centrifugation followed by examination of the buffy coat; mini anion-exchange/centrifugation; and the quantitative buffy coat (QBC) technique. For other samples, such as spinal fluid, concentration techniques include centrifugation followed by examination of the sediment.
Three serological tests are also available for detection of the parasite: the micro-CATT, wb-CATT, and wb-LATEX. The first uses dried blood, while the other two use whole blood samples. A 2002 study found the wb-CATT to be the most efficient for diagnosis, while the wb-LATEX is a better exam for situations where greater sensitivity is required.
The gold standard for diagnosis is visualization of the amastigotes in splenic aspirate or bone marrow aspirate. This is a technically challenging procedure that is frequently unavailable in areas of the world where visceral leishmaniasis is endemic.
Serological testing is much more frequently used in areas where leishmaniasis is endemic. A 2014 Cochrane review evaluated different rapid diagnostic tests. One of them (the rK39 immunochromatographic test) gave correct, positive results in 92% of the people with visceral leishmaniasis and it gave correct, negative results in 92% of the people who did not have the disease. A second rapid test (called latex agglutination test) gave correct, positive results in 64% of the people with the disease and it gave correct, negative results in 93% of the people without the disease. Other types of tests have not been studied thoroughly enough to ascertain their efficacy.
The K39 dipstick test is easy to perform, and village health workers can be easily trained to use it. The kit may be stored at ambient temperature and no additional equipment needs to be carried to remote areas. The DAT anti-leishmania antigen test, standard within MSF, is much more cumbersome to use and appears not to have any advantages over the K39 test.
There are a number of problems with serological testing: in highly endemic areas, not everyone who becomes infected will actually develop clinical disease or require treatment. Indeed, up to 32% of the healthy population may test positive, but not require treatment. Conversely, because serological tests look for an immune response and not for the organism itself, the test does not become negative after the patient is cured, it cannot be used as a check for cure, or to check for re-infection or relapse. Likewise, patients with abnormal immune systems (e.g., HIV infection) will have false-negative tests.
Other tests being developed include detects erythrosalicylic acid.
"N. fowleri" can be grown in several kinds of liquid axenic media or on non-nutrient agar plates coated with bacteria. "Escherichia coli" can be used to overlay the non-nutrient agar plate and a drop of cerebrospinal fluid sediment is added to it. Plates are then incubated at 37 °C and checked daily for clearing of the agar in thin tracks, which indicate the trophozoites have fed on the bacteria. Detection in water is performed by centrifuging a water sample with "E. coli" added, then applying the pellet to a non-nutrient agar plate. After several days, the plate is microscopically inspected and "Naegleria" cysts are identified by their morphology. Final confirmation of the species' identity can be performed by various molecular or biochemical methods.
Confirmation of "Naegleria" presence can be done by a so-called flagellation test, where the organism is exposed to a hypotonic environment (distilled water). "Naegleria", in contrast to other amoebae, differentiates within two hours into the flagellate state.
Pathogenicity can be further confirmed by exposure to high temperature (42 °C): "Naegleria fowleri" is able to grow at this temperature, but the nonpathogenic "Naegleria gruberi" is not.
Avoiding food or water that may be contaminated with stool can help prevent the infection of "Cystoisospora" (Isosporiasis). Good hand-washing, and personal-hygiene practices should be used as well. One should wash their hands with soap and warm water after using the toilet, changing diapers, and before handling food (CDC.gov). It is also important to teach children the importance of washing their hands, and how to properly wash their hands.
Trypanosomiasis could, in future be prevented by genetically altering the tsetse fly. As the tsetse fly is the main vector of transmission, making the fly immune to the disease by altering its genome could be the main component in an effort to eradicate the disease. New technologies such as CRISPR allowing cheaper and easier genetic engineering could allow for such measures.