How to diagnosis influenza in pets
One of the common questions accompanying the onslaught of calls I've taken today is "How do you diagnose influenza in pets?"
Clinical signs, such as sneezing, coughing, fever and lethargy, are not useful for diagnosis. Influenza can produce highly variable disease, ranging from almost none to very severe - so you can't look at an animal and say it has influenza just based on the clinical signs. We don't know much about H1N1 influenza in different animal species (including pets), but this type of influenza can probably cause a wide range of disease in animals as well (at least in those it can infect).
The presence of someone in the household with influenza should get you thinking about flu in a sick pet, but it is far from diagnostic. Many, many people have influenza, but very few pets do. There are many other diseases that can produce signs similar to influenza in pets. The health of people in the household is an important thing to know, but we can't jump to conclusions based on the household history alone.
Laboratory testing is required for the diagnosis of influenza, and there are a few options:
- PCR testing of nasopharyngeal (throat) or nasal swabs, or fluid collected from the trachea: This molecular test detects influenza virus RNA. This is the fastest test and it is most sensitive when samples are taken early in disease.
- Serology: This involves testing blood for antibodies against influenza. Two samples are taken 10-14 days apart. If the antibody level rises 4-fold or greater, that is indicative of influenza infection. This is considered the most reliable method of diagnosis but takes time.
- Virus isolation from nasopharyngeal or nasal swabs, or tracheal fluid: Samples are inoculated into eggs to try to grow the virus. This can take quite a while and isolation of the virus can be difficult.
"Family protection" screening of healthy pets
As public awareness of zoonotic diseases increases, some new marketing opportunities are created. One is offering screening of pets for "protection" of the family. One company, Healthgene, offers a Family Protection Program that involves screening pets for selected zoonotic pathogens. Unfortunately, it involves the use of non-validated PCR tests for pathogens for which screening of healthy animals is not considered useful. They make various unsupported, illogical and sometimes downright incorrect statements such as "If, by chance, any positive results should occur, not only should the animal be treated immediately, but the client and anyone having contact with the animal should also notify their family physician." Despite the fact that the tests themselves are of questionable quality and the results are essentially useless, I'm sure they are developing a market.
Also, it's pretty concerning that this company misspells the names of various microoganisms for which they test on their website - they even misspell "protection" in one of their Family Protection Plan info sheets! Inability to spell isn't necessarily linked to inability to test properly, but it shows a lack of care and attention to detail that raises further red flags in my mind.
Zoonotic diseases ARE a concern. We need to pay attention to them and try to reduce the risks of transmission to people. Screening healthy animals is almost never a component of this. Save your money when it comes to "routine" screening of healthy animals - talking to your vet about potential problems and washing your hands are much better ways to reduce the risk of infection.
Severe diarrhea outbreak in Florida dogs
Tuttle Animal Medical Center in Florida has reported six dogs with severe bloody diarrhea, vomiting and fever, over the past month. The affected dogs were from the same general area in Sarasota County, and all but one died within 24 hours. However, care must be taken when interpreting information such as this. Apparently, most dogs were owned by people with limited finances, so it's hard to say whether they would have died if aggressive (and expensive) treatment could have been provided. A disease like parvovirus is highly fatal without treatment, but survival rates are excellent if proper treatment is provided.
Initial testing of these dogs to identify the causative agent has been unsuccessful, including a rapid in-clinic test for parvovirus. Because of limited finances, complete diagnostic testing has not been performed, and it's likely that only a very limited range of possible causes have been investigated. That's a problem with a user-pay system such as this. There's no incentive for owners of dead pets to pay for further testing that could help understand the problem and/or help other peoples' pets.
Various experts have been consulted, but it sounds like there is minimal material (e.g. saved fecal samples) to use for further testing. Trying to make a diagnosis based on clinical signs and basic laboratory data collected by the clinic during standard work-up and treatment is essentially impossible. Veterinary infectious disease expert Dr. Cynda Crawford told VIN News Service by e-mail last Wednesday "There is very little case material to work with, so am struggling with meaningful diagnostic approaches,...Everything is basically speculation at this point."
Florida's Division of Animal Industry is apparently "monitoring the situation." At this point, there's nothing that can really be done, but hopefully assistance with testing will be provided should further cases be identified. One official from this agency speculated that E. coli O157 could be the cause. That seems pretty unlikely. This bacterium can cause disease in dogs but it's quite rare. Six apparently unrelated cases due to such a rare cause seems pretty unlikely, although it shouldn't be dismissed.
A general tenet of medicine is "common things occur commonly." Situations like this are most often unusual presentations of a common disease (e.g. parvovirus) rather than a new disease. More aggressive diagnostic testing for known causes of disease, along with additional testing to try to identify new pathogens is needed if further cases are identified. Sometimes apparent outbreaks like this go away on their own without any intervention or diagnosis. Only time will tell whether this is a small local event or the "tip of the iceberg."
MRSA testing and false advertising
I received a flyer from Zoologix, a company that offers various (typically unvalidated and unproven) PCR tests for animals. The flyer headline was "Pets can carry MRSA - but testing can help."
Testing in certain situations is useful, but this is almost always limited to diagnosis of animals with active infections (i.e. they're sick). PCR is not a good way to make such a diagnosis, because the test doesn't tell you anything about the bacterium's susceptibility to other antibiotics. Screening of pets just to determine whether or not they carry MRSA is rarely needed, and currently there is no evidence that PCR is a reasonable test for this.
There are no validated PCR tests for MRSA in animals. We looked at using a human test in horses and it failed miserably. There are validated tests for use in people, and they are quite good: they accurately identify MRSA and differentiate it from other methicillin-resistant staphylococci and from methicillin-susceptible S. aureus. That's critical, because you have to know what a positive test really means.
I called the company and asked what the test actually detects. They said it detects the mecA gene, the gene that confers methicillin-resistance to staphylococci such as S. aureus. However, this gene can be present in other staphylococci that can be found in many healthy dogs and cats (10-30% in some studies). It does not actually detect MRSA and a large percentage of samples that give positive results will be false positives. The tests that are used in humans are specifically designed to look at two things in combination: whether S. aureus is present and whether it has the mecA gene (methicillin-resistance). This is the right approach because it excludes all those other false positives. Detecting mecA alone is completely useless. It's interesting that the flyer states "PCR testing is fast, effective and accurately differentiates MRSA from other bacteria - even other Staph strains." Based on what the company told me over the phone, with regard to the test they're advertising, that's a blatant lie.
This is an example of a combination of bad science and bad ethics. This company has no business marketing this test. It's false advertising, because the test isn't an MRSA test. Their justification for using it is similarly weak. Anyone thinking about using this test should run away quickly! The issues with this test (and others) should also be considered when deciding whether to use this company for any tests.
More (and accurate) information about MRSA can be found on the Worms & Germs Resources page.
Testing for ringworm with toothbrushes
If your veterinarian suspects your pet may have ringworm, there are several different ways he or she may test for the causative fungus (a dermatophyte) on your animal's fur and skin. Some of these techniques are more useful than others in different situations.
- Wood's lamp: A Wood's lamp is simply a special ultraviolet light. Approximately half of all Microsporum canis strains (the most common species of dermatophyte that causes ringworm in cats and dogs) will fluoresce blue-green under such a light. This type of testing is obviously very easy to perform. However, other debris in an animal’s hair coat may fluoresce as well, and other species of fungus that cause ringworm do not fluoresce, so this test is not useful by itself in most cases.
- Microscopy: Sometimes ringworm fungus can be seen on hair shafts from an infected pet when examined under a microscope. However, it is easy to confuse other debris and structures for dermatophytes. Also, not every hair on an infected animal will carry the fungus, so it's possible to miss the infected hairs altogether with this test.
Fungal culture: The best way to diagnose ringworm is to culture the fungus from the infected individual (person or animal). In animals, one of the best ways to collect a sample for culture is to comb over all the fur and skin with a new toothbrush, and then try to grow dermatophytes from the toothbrush. This allows the fur from all over the animal to tested, rather than just one little clump of fur plucked from one area. It can also make it easier to get a sample from the face and paws of cats, which is where these animals often carry the fungus. Although fungal culture is the best way to diagnose ringworm, remember that fungal culture takes much longer than bacterial culture – instead of days, it may take up to three weeks to grow some dermatophytes.
It's also important to remember that dogs, and more often cats, may carry dermatophytes on their fur even when they look healthy. A positive fungal culture from an animal with skin disease, particularly a cat, does not necessarily rule out other diagnoses, so your veterinarian may still recommend other tests as well. However, any animal with ringworm should be treated to prevent spreading the infection to other animals and people.
More information about ringworm is now available on the Worms&Germs Resources page, and in our archives.
Molecular Diagnostic Testing: Pros and Cons
An important step in diagnosing infectious diseases and determining the optimum approach to treatment and management is rapid and accurate diagnostic testing. Many different testing methods are used, particularly bacterial culture (at least for bacterial diseases). Molecular testing has revolutionized the field of microbiology, and is making inroads into the field of diagnostic testing. Polymerase chain reaction (PCR) testing is a very powerful tool that can be used to detect DNA or RNA from specific microorganisms. This technique can be very useful, but it can also be easily misused or misinterpreted.
The potential PROS of molecular diagnostic testing include:
- Rapid turnaround time: Testing can take as little as a few hours versus a few days for other tests like bacterial culture.
- Sensitivity: Organisms that are difficult or impossible to grow in a lab can be detected, and they can often be detected at lower levels than with other diagnostic methods.
The potential CONS of molecular diagnostic testing include:
- Sample contamination: This is a common concern with highly sensitive molecular tests - even a minute amount of contamination in the sample can cause a false positive result.
- Test inhibition: Samples from complex biological sites (e.g. stool) can contain substances that interfere with the many complex molecular reactions upon which the tests rely. Without good (and proven) methods to prepare the sample, this can result in a false negative result.
- Biologically irrelevant results: Some bacteria that cause disease are also commonly found as part of the normal microflora in healthy animals - simply finding it does not tell you that it is necessarily relevant to the problem. For example, Clostridium difficile can be found in the intestine of approximately 10% of healthy dogs and cats (or more, in some situations), but the diagnosis of C. difficile diarrhea requires detection of the bacterial toxins in stool samples, not just the bacterium itself. A molecular test that simply identifies the presence of C. difficile, even if it identifies strains that possess the genes to produce toxins, tells you nothing about whether the bacterium was actually producing toxins in the animal.
- Lack of validation: This is a common problem with many (if not most) molecular tests. Some companies, especially those that just run molecular tests, offer a huge array of completely unvalidated and sometimes illogical tests. It is also important to remember that tests must be validated for each species in which they are used - a test that works well in people will not necessarily work on a sample from a horse or a dog.
Molecular testing can be useful in some situations. If you are unsure, here are some things to ask the lab:
- Do they have a validated test that provides relevant results? If they don't have good data (ideally published data) that their test is useful, accurate and reproducible, I'd avoid it.
- Do they have a quality control program, which includes running positive and negative control samples with each test batch?
Finally, as with any test that we use in veterinary (or human) medicine, it's important to evaluate all results in the context of what is happening with the animal - treat the patient, not the test result.
