A CROSS-SECTIONAL SEROLOGICAL SURVEY OF SHELTER CATS AND DOGS IN AUSTRALIA TO ASSESS THE PREVALENCE OF EXPOSURE TO, AND INFECTION

WITH DIROFILARIA IMMITIS Dr Sarah Mitchell Veterinary Operations Manager Companion Animals Pfizer Animal Health 38-42 Wharf Road West Ryde NSW 2114 Background Heartworm in dogs is a well known disease, having been first reported in 1626, in Italy. Heartworm in cats was not reported until 1921 in Brazil, and awareness of this disease has grown significantly in recent years. In most areas within Australia it has been widely accepted for many years that dogs should be on year-round heartworm prevention, and this has resulted in an anecdotal significant decrease in the incidence of heartworm cases seen in veterinary practice each year. Dogs are the preferred direct host for Dirofilaria immitis (D. immitis) and mosquitoes are the indirect host. There are several species of mosquitoes in Australia that are competent heartworm vectors (Russell & Geary 1996). Dog versus cat There are significant differences between canine and feline heartworm disease due to the difference in the host-parasite relationships between D. immitis and dogs and cats. The relationship between D. immitis and cats is regarded as a partially adapted hostparasite relationship (Nelson et al. 2007). Cats are innately more resistant to infection with D. immitis than are dogs, and are more likely to mount an immune response to the infection in an attempt to eliminate the parasite. Canine heartworm disease is associated with infection with adult worms in the cardiopulmonary system of dogs and historically research in feline heartworm disease has concentrated on investigating disease in cats associated with infection with adult D. immitis. However, due to recognition of the different host-parasite relationships and varying clinical signs between D.immitis in dogs and D. immitis in cats, more recent research has set out to define feline heartworm disease as a separate entity from canine heartworm disease. Through this research, pulmonary disease has been observed in cats associated with the immature stages of D. immitis, which are present in the pulmonary tissues (Dillon et al. 2007; Donahoe et al. 1976; Holmes et al. 1992). This clinical respiratory syndrome occurs due to infection with the immature stages of D. immitis, has become known as heartworm-associated respiratory disease (HARD), and is regarded as an emerging issue in feline medicine (Levy 2008). Although there are significant differences between the host-parasite relationship in dogs and cats with D. immitis, feline infection is likely to occur anywhere the parasite is found in dogs. Studies on dogs and cats necropsied at shelters have shown that adult worms can usually be found in cats at 5% to 20% of the rate that they are found in dogs in the same location (Ryan and Newcomb 1995).

Mitchell, Sarah, A cross-sectional serological survey of shelter cats in Australia to assess the prevalence of exposure to, and infection with Dirofilaria immitis (2011). Proceedings of the AVA Annual Conference, Canberra 2012 Page A1.5.1

Due to the relative resistance of cats to D. immitis, diagnosis is not as straightforward as it is in the dog where adult infections often involving many worms can be readily identified through simple in-house antigen tests. Antigen tests for D. immitis detect the antigen produced by the adult female heartworm and also from dying adult male and female worms. Thus because adult worms often fail to develop in the cat, and when they do, the population of adult worms is smaller and the likelihood of single infections is greater, the utility of antigen tests as a diagnostic tool for detection of feline heartworm is limited. If, however, the result is positive, it does confirm the presence of adult heartworm infection (Nelson et al. 2007). Microfilarial detection tests in dogs can have issues with specificity as other parasites can cause microfilaraemia. These tests are not very useful in cats due to cats rarely being microfilaraemic, and when microfilaraemia does occur, it tends to be transient. Because the significance of the immature stages of D. immitis in the pathophysiology of heartworm disease in the cat is now better understood and appreciated, antibody tests are increasingly used in the diagnostic processes for feline heartworm. These serological tests detect antibodies produced by the cat to heartworm larvae, as early as 8 weeks post infection, thereby confirming the presence of larval heartworm infection. There are limitations of antibody tests in that cats may spontaneously clear the infection with larval heartworm and the cat may have cleared the infection by the time the test is performed. This means that the test may not necessarily indicate the current presence of infection with immature D. immitis, just that the cat has been previously exposed to or infected with the larval stages of D. immitis, and moreover antibody tests cannot confirm disease or clinical sign causality. What antibody tests can be seen to do, is to confirm that the cat that tests positive is at risk of heartworm infection and/or HARD. Environmental and human influences Changing landscapes, climate and demographics impact the spread of infectious diseases of both humans and animals, especially vector-borne diseases. In other parts of the world, these factors have contributed to the expansion of the range of heartworm. Australia is also being impacted by these factors. The past 18-24 months have seen increases in cases of mosquito-borne diseases such as dengue fever, Kunjin, Murray Valley encephalitis, Barmah Forest virus infection and Ross River fever. It is possible that recent climatic and financial conditions have opened opportunities for the spread of heartworm in Australia: Increased rainfall and warm summers throughout the eastern states mean mosquito populations can thrive and support the heartworm lifecycle, and the global financial crisis in which the importance of pet health preventatives would have waned in many families has likely meant that more dogs and cats have missed doses of heartworm prevention. Recent peer-reviewed literature on the prevalence of heartworm disease in dogs and cats in Australia however is lacking, with no readily accessible peer-reviewed publications on either canine or feline heartworm in the past ten years.

Mitchell, Sarah, A cross-sectional serological survey of shelter cats in Australia to assess the prevalence of exposure to, and infection with Dirofilaria immitis (2011). Proceedings of the AVA Annual Conference, Canberra 2012 Page A1.5.2

Recent research Pfizer Animal Health (PAH) has recently invested in research to determine the experiences, attitudes, practices of Australian veterinarians related to heartworm in cats and dogs, testing and use of preventative therapies. In addition, PAH in collaboration with Animal Welfare League shelters in New South Wales, Queensland and South Australia have recently conducted a study to determine the prevalence in dogs and cats housed at these shelters. A postal survey was conducted in 2010, involving 100 veterinarians from all states in Australia. Information from the survey showed that: 97% of veterinarians recommend heartworm prevention for dogs, but only 43% recommended heartworm prevention for cats 55% of respondents have seen cases of canine heartworm in the past 10 years 8% of respondents have seen cases of feline heartworm in the past 10 years 100% test for canine heartworm using antigen tests, but only in a limited number of scenarios compared to what is regarded as best practice and recommended by the American Heartworm Society (Graham et al 2012) Only 27% test cats for heartworm

An Australian prevalence study, approved by Westmead Hospital Animal Ethics Committee, was conducted in 2011. In this study over 700 dogs and 500 cats, older than 18-24 months, entering or housed at the shelters were enrolled, examined, basic information collected and a blood sample taken. Blood samples taken from dogs were tested for canine heartworm antigen. Blood samples from cats were tested for heartworm antigen and antibody. Sample sizes were determined with EpiTools epidemiological calculators (Sergeant 2009) using data on the dog and cats populations in the shelter, sensitivity and specificity data for the tests to be used and estimated prevalences for the populations. For the cats, parallel sensitivity and specificity values were calculated involving both antigen and antibody test validity data. Results and conclusions of the study are not available at the time of writing these proceedings, but will be discussed at the conference. Conclusions Heartworm prevention is widely used, year-round in Australia, which has helped to decrease the number of heartworm cases seen. This decrease in cases can result in lack of awareness, and coupled with financial pressures may potentially lead to complacency with heartworm prevention. In the current environment of climate change, urbanisation and increased travel, and with suboptimal heartworm surveillance, this scenario may create an opportunity for reemergence of heartworm disease.

Mitchell, Sarah, A cross-sectional serological survey of shelter cats in Australia to assess the prevalence of exposure to, and infection with Dirofilaria immitis (2011). Proceedings of the AVA Annual Conference, Canberra 2012 Page A1.5.3

References Dillon, A.R., Blagburn, B.L., Tillson, D.M., Brawner, W.R., Welles, B., Johnson, B., Spenser, B., Kaltenboeck, B. and Rynders, P.E. 2007. 'Immature Heartworm Infection Produces Pulmonary Parenchymal, Airway and Vascular Disease in Cats'. ACVIM 2007. Donahoe, J.M.R., Kneller, S.K. and Lewis, R.E. 1976. 'Hematologic and Radiographic Changes in Cats After Inoculation with Infective Larvae of Dirofilaria immitis.' Journal of the American Veterinary Medical Association, vol. 168, pp. 413-7. Graham, W., Rubin, S., Boechk, A., Buzhardt, L. F., Jones, S., Miller, M., Payne, P., Rehm, P., Smith-Blackmore, M., Stannard,. R., Nelson, C. T., Atkins, C., Carithers, D., McCall, J., von Simson, C. (2012) Current Canine Guidelines for the Diagnosis, Prevention, and Management of Heartworm (Dirofilaria immitis) Infection in Dogs. http://www.heartwormsociety.org/veterinary-resources/canine-guidelines.html [15 January 2012]. Holmes, R.A., Clark, J.M., Casey, H.W., Henk, W. and Plue, R.E. 1992. 'Histopathologic and Radiographic Studies of the Development of Heartworm Pulmonary Vascular Disease in Experimentally Infected Cats'. Heartworm Symposium 1992, American Heartworm Society. Levy, J.K. 2008. 'Heartworm Disease Without Heartworms'. AAFP Spring 2008 Meeting, American Association of Feline Practitioners. Nelson, C.T., Seward, R.W., McCall, J.W., Rubin, S.B., Buzhardt, L.F., Graham, W., Longhofer, S.L., Guerrero, J., Robertson-Plouch, C., Paul, A. and Carithers, D. 2007. 'American Heartworm Society: 2007 Guidelines for the Diagnosis, Prevention and Management of Heartworm (Dirofilaria immitis) Infection in Cats.' http://www.heartwormsociety.org/veterinary-resources/feline-guidelines.html. [12 January 2009] Russell, R. C. and Geary, M. E. (1996) The Influence of microfilarial density of dog heartworm Dirofilaria immitis on infection rate and survival of Aedes notoscriptus and Culex annulirostris from Australia. Medical and Veterinary Entomology 10: 29-34. Ryan, W.G. and Newcomb, K.M. 1995. 'Prevalence of Feline Heartworm Disease - A Global Review'. Heartworm Symposium 1995, American Heartworm Society. Sergeant, E.S.G. 2009. Epitools epidemiological calculators. AusVet Animal Health Services and Australian Biosecurity Cooperative Research Centre for Emerging Infectious Disease. Available at: http://epitools.ausvet.com.au.

Mitchell, Sarah, A cross-sectional serological survey of shelter cats in Australia to assess the prevalence of exposure to, and infection with Dirofilaria immitis (2011). Proceedings of the AVA Annual Conference, Canberra 2012 Page A1.5.4