Utility of thoracic ultrasonography in a rapid diagnosis of angiostrongylosis in young dogs presenting with respiratory distress.

The nematode, Angiostrongylus vasorum is a nematode that lives in the pulmonary arteries of canids and has an obligate gastropod intermediate host. It can cause various clinical signs. with the two most common clinical scenarios consisting of acute respiratory distress and haemorrhagic diathesis, either separately or together. Younger dogs (< 2 years) are overrepresented, and dogs often show pulmonary granulomata (radiographically and pathologically). Thoracic ultrasonography offers a safe, rapid, commonly available, non-invasive means of assessing the lungs. We prospectively examined the utility of thoracic ultrasonography in the diagnosis of angiostrongylosis in 26 client-owned dogs <2 years old, presenting with respiratory distress. We identified small hypoechoic subpleural nodules in 15/26 dogs; 14 of these were subsequently confirmed to have angiostrongylosis by faecal Baermann concentration test, A. vasorum antigen testing or both. The remaining 11 dogs without subpleural nodules had negative faecal analysis and A. vasorum antigen testing and diagnosed with other respiratory diseases. This resulted in a sensitivity of 100% and a specificity of 92% for the detection of angiostrongylosis by thoracic ultrasonography in young dogs presenting with respiratory distress. Our results suggest that thoracic ultrasonography might offer a safe, rapid, relatively accurate diagnostic test for diagnosis of angiostrongylosis in young adult dogs with respiratory distress living in endemic areas.

Echocardiographic Evaluation of the Right Atrial Area Index in Dogs with Pulmonary Hypertension.

BACKGROUND: Right atrial area (RAA) is a prognostic factor in human patients with pulmonary arterial hypertension (PAH). Reference intervals for RAA have been described in healthy dogs. OBJECTIVES: To evaluate RAA indexed to the body surface area in dogs with PAH as an indicator of right atrial size, PAH severity and right-sided congestive heart failure (R-CHF). ANIMALS: A total of 119 client-owned dogs, 48 dogs with PAH and 71 control dogs. METHODS: Prospective observational study. Pulmonary arterial hypertension was classified according to the tricuspid regurgitation pressure gradient (TRPG) as mild (36-50 mmHg), moderate (51-75 mmHg), or severe (>75 mmHg). The RAA index was calculated as the RAA divided by body surface area. RESULTS: The RAA index was higher in dogs with moderate PAH (13.3 cm2 /m2 ; range, 3.4-24.7 cm2 /m2 ) and severe PAH (12.1 cm2 /m2 ; range, 5.4-21.8 cm2 /m2 ) than in those with mild PAH (6.7 cm2 /m2 ; range, 4.8-10.7 cm2 /m2 ) or in controls (7.3 cm2 /m2 ; range, 4.2-10.2 cm2 /m2 ; P < 0.001). The RAA index was higher (P < 0.0001) in dogs with R-CHF (17.5 cm2 /m2 ; range, 12.7-24.7 cm2 /m2 ) compared to those without R-CHF (7.6 cm2 /m2 ; range, 4.4-19.4 cm2 /m2 ). The most accurate cutoff value of the RAA index to identify R-CHF was >12.3 cm2 /m2 (sensitivity, 100%; specificity, 89.5%). In dogs with PAH, severity of tricuspid regurgitation (TR) was the only independent predictor of RAA index based on multivariate analysis (P < 0.02). CONCLUSIONS AND CLINICAL IMPORTANCE: The RAA index can be used to evaluate right atrial size in dogs and may be more effective than TRPG in predicting R-CHF in dogs with PAH. The severity of TR is the main determinant of the RAA index in dogs with PAH.

Feline heartworm disease: A'Rubik's-cube-like' diagnostic and therapeutic challenge.

Feline heartworm disease presents a unique diagnostic, therapeutic, and preventive challenge for veterinarians. Due to the elusive clinical nature and peculiar physiopathology of heartworm infection in cats, a multistep diagnostic process is mandatory. Clinical signs may be absent or atypical. At the present time there is no single ante mortem diagnostic test that can reach a high level of sensitivity for feline heartworm infection. The most efficient approach for the diagnosis of feline heartworm disease is based upon a synergic association of several tests: thoracic radiography and serum antibody tests for rising index of suspicion, and echocardiography and serum antigen tests for confirming the infection. Other tests should be considered of secondary importance, even if they can help to support the diagnosis. Treatment of feline heartworm disease is typically based on clinical signs, as adulticidal therapy is associated with a high rate of complications and cats frequently self-cure. Chemoprophylaxis, knowledge of the biology of the parasite, and a high index of suspicion seem to be the most important tools for combating feline heartworm disease.

Can heartworm prevalence in dogs be used as provisional data for assessing the prevalence of the infection in cats?

Cats are considered a susceptible host for Dirofilaria immitis; however, increased host resistance is reflected by relatively low adult worm burdens in natural and experimental infections; the prolonged prepatent period (8 months); the low level and short duration of microfilaremia; and the short life span of adult worms (2-3 years). From April to September 2006, 212 cats and 608 dogs, all exposed for at least one transmission season, were screened for D. immitis infection in a multi-center study in the Po River Valley in northern Italy. Cats were initially evaluated by antibody testing; positive subjects were followed up by antigen testing and echocardiography (and necropsy if death occurred). The prevalence in dogs was 29% by a modified Knott test and antigen testing compared with a prevalence of 4.7% in cats by an antibody test; six of these infections (2.8%) were confirmed by the follow-up evaluations. This field study demonstrated that the prevalence of heartworm infection in cats in this area is within the expected limits of 9-18% of the prevalence in dogs. Antibody testing likely underestimates the real prevalence of D. immitis infection in cats. These results also emphasize the importance of preventive treatment in cats.

Clinical evolution and radiographic findings of feline heartworm infection in asymptomatic cats.

Clinical manifestations of heartworm disease in cats are variable; most cats seem to tolerate the infection well for extended periods. Heartworm-infected cats may undergo spontaneous self-cure due to the natural death of parasites without any symptomatology, or they may suddenly show dramatic and acute symptoms. Sudden death in apparently healthy cats is not a rare event. Thoracic radiographs are important tool for the diagnosis of cardiopulmonary disease. However, thoracic abnormalities are often absent or transient and highly variable in heartworm-infected cats. Findings, such as enlargement of the peripheral branches of the pulmonary arteries, with a varying degree of pulmonary parenchymal disease and hyperinflation, are the most typical features consistent with infection. A field study was performed for cats referred to the Veterinary Hospital Città di Pavia from January 1998 to December 2001 for routine health examinations and procedures to evaluate the clinical evolution and radiographic findings of feline heartworm infection. Thirty-four asymptomatic cats diagnosed with feline heartworm infection by antibody and antigen tests together with an echocardiogram that allowed worm visualization were included in the follow-up study. Cats were routinely examined every 3 months from the time of heartworm diagnosis until the outcome (self-cure or death). Self-cure was defined as no positive serology for heartworm antigens and no visualization of worms by echocardiography. A final examination for antibodies was carried after 12 months as a final confirmation of self-cure. Twenty-eight cats (82.4%) self-cured; including 21 that showed no clinical signs of infection throughout the study. Six cats died. The most common clinical features observed were acute respiratory symptoms and sudden death. Infection lasted over 3 years in the majority of the cats enrolled in the study. Thoracic radiograph appearance was variable, and the most commonly observed findings were focal and diffuse pulmonary parenchymal abnormalities.

Changes in the levels of eicosanoids in cats naturally and experimentally infected with Dirofilaria immitis.

Feline heartworm (Dirofilaria immitis) infection is a severe, life-threatening disease. The eicosanoids are lipid mediators derived from the metabolism of the arachidonic acid, involved in the regulation of the immune response and of inflammatory reactions. In this study, naturally infected cats showed significant higher levels of prostaglandin E(2) (PGE2), thromboxane B(2) (TXB(2)) and leukotriene B(4) (LTB4) than uninfected cats. Changes in the levels of eicosanoids during the infection were observed in experimentally infected cats. PGE2 increased significantly during the first 60 days post-infection, then progressively decreased until day 180 post-infection. At this time, PGE2 values are still significantly higher than those observed before the infection. TxB2 and LTB4 increased progressively from the beginning of infection and reached their maximum levels 180 days post-infection. In experimentally infected, ivermectin-treated cats, 15 days after treatment (45 days after infection) both PGE2 and LTB4 levels were similar to those observed in experimentally infected, untreated cats. No significant differences of PGE2 levels were found before the infection and at the end of the experiment (165 days post-treatment, 195 days post-infection). Increased levels of LTB4 were found 15 days post-treatment, afterward they progressively decreased. These data show that D. immitis infection influences the production of intravascular eicosanoids in cats. The high levels of PGE2 observed in the early phase of infection could be related to the survival of the worms, while those of TxB2 and LTB4 detected at the end of the study could mediate the inflammatory reactions and thrombi formation during the feline dirofilariosis.

Aelurostrongylus abstrusus (cat lungworm) infection in five cats from Italy.

Infection by the cat lungworm Aelurostrongylus abstrusus is considered uncommon. Here, the authors report the clinical, diagnostic and therapeutic features of five infections recently observed in Italy. All cats were under 12 months of age. All except one cat had symptomatic infections, with cough, dyspnea, and weight loss with radiographic signs of broncopneumonia. All cats were eosinophilic. Larvae were present in fresh fecal smears and on flotation exam in all cats. Baermann larval recovery permitted definitive identification and, in one case, larvae per gram of feces (lpg) counts. One dose of ivermectin (400 microg/kg) was not effective in one cat, while one dose of selamectin (6 mg/kg) was effective in one of three cases and fenbendazole at 50 mg/kg given daily for 15 days was effective in four of four cases.

Heartworm disease in dogs: unusual clinical cases.

Two unusual cases of heartworm infection are described, including one of spontaneous rupture of cardiac chambers, most likely due to the progressive increase in pressure within the right ventricle caused by Dirofilaria immitis infection. Another case report describes persistent left cranial vena cava, which interfered with the surgical removal of heartworm via the left jugular vein.

Clinical, cytological and molecular evidence of Mesocestoides sp. infection in a dog from Italy.

A 12-year-old, 13 kg, mixed-breed male dog was referred for anorexia and depression. The dog showed discomfort on abdominal palpation. Abdominal ultrasound examination revealed multiple, small, round anechoic cystic structures. Cystic fluid obtained with fine needle aspiration contained several 2-4 mm white motile flecks. Microscopic examination of the fluid revealed numerous irregularly shaped organisms measuring several hundred microns to 3 mm, the morphology of which was suggestive of intact and fragmented acephalic metacestodes of the genus Mesocestoides sp. Molecular analysis confirmed that the peritoneal infection was caused by Mesocestoides sp.

Efficacy of long-term monthly administration of ivermectin on the progress of naturally acquired heartworm infections in dogs.

This study was designed to evaluate the efficacy of prolonged monthly ivermectin treatment against Dirofilaria immitis in client-owned dogs with naturally acquired infections and to clinically monitor the animal's response to the slow killing of heartworms, with death of the worms distributed over a period of up to 2 years. A total of 17 male and female dogs of different breeds and ages were used. Prior to treatment, all of the dogs tested positive for heartworm antigen (Ag) and all but two had microfilariae (mf). The dogs were randomly allocated to one group of seven dogs which received a commercial formulation of ivermectin (minimum, 6 mcg IVM/kg) plus pyrantel (minimum, 5 mg PP/kg) (Heartgard Plus Chewables, Merial, Ltd.), another group of seven dogs which received a commercial formulation of IVM (min, 6 mcg/kg) (Heartgard Chewables, Merial Ltd.), and a group of three dogs which served as an untreated controls. All dogs were evaluated prior to initiation of treatment and thereafter at 3- to 5-month-intervals for mf, Ag, and radiographic and echocardiographic findings. All of the 17 dogs, with the exception of two dogs in the IVM group, had circulating mf of D. immitis prior to the 1st monthly dose, and a few also had mf of Dirofilaria repens. After 4 monthly doses, only one dog in the IVM/PP group and two dogs in the IVM group had a patent heartworm infection, and no heartworm mf were seen in the 14 treated dogs thereafter. After 10 monthly doses, the number of Ag-positive dogs in both of the treated groups decreased gradually. Efficacy, based on the reduction in number of Ag-positive dogs, was similar for the IVM/PP and IVM groups, with overall efficacy scores for the 14 dogs of 21, 21, 43, and 71% after 10, 14, 19, and 24 monthly doses, respectively. Two of the seven dogs treated with IVM/PP, one of the seven treated with IVM, and two of the three untreated controls showed echocardiographic evidence of a parasitic burden prior to treatment, and all of these scores had decreased by the end of the study. Only one dog (IVM/PP group) had a cardiovascular pattern of heartworm disease by echocardiography prior to treatment, but this dog's score increased to two and the scores of two additional dogs increased from zero to two (IVM group) or three (IVM/PP group) by the end of the study. Only 1 (IVM/PP group) of the 17 dogs showed a pulmonary pattern of heartworm disease by radiography prior to treatment, but this dog's score increased to three by the end of the study. The radiographic scores of two additional dogs in the treated groups increased from zero to three (IVM/PP) or two (IVM) by the end of the study. Thus, monthly administration of IVM to dogs with clinical, radiographic or echocardiographic evidence of heartworm disease is ill-advised and such treatment of even the asymptomatic dog should be done only with much caution and frequent monitoring by the veterinarian.

Feline dirofilariosis: antibody response to antigenic fractions containing specific 20 to 30 kDa polypeptides from the adult Dirofilaria immitis somatic antigen.

Fractions from the adult somatic antigen (SA) Dirofilaria immitis complex, containing polypeptides from 20 to 30kDa, previously identified as molecular markers of feline dirofilariosis are isolated by sequential application of gel filtration and anion exchange chromatography. Indirect enzyme-linked immunosorbent assays, employing these fractions (20-26kDa/ELISAF1 and 30kDa/ELISAF7) show multivalent diagnostic capacities: they were able to detect pre-patent infections 2 months after infection, infections in clinical phase, and the fall of antibodies after the worms were removed from the heart, or the application of a ivermectin treatment. The results obtained by the two tests correlated well, in spite of the fact that ELISAF1 was most useful to detect antibodies in sera from cats in the clinical phase, while ELISAF7 has more sensitivity for the early detection of the infections. Both ELISAs were useful in the detection of the decrease of antibodies after the worms were removed by surgery or pharmacological treatment.

Primary cardiac rhabdomyosarcoma in a cat.

A seven-year-old domestic shorthair (DSH) cat was presented with anorexia and dyspnea. Pleural-pericardial effusion was detected with thoracic radiographs and echocardiography. Echocardiography demonstrated a large, soft-tissue mass in the right ventricular wall, protruding both into the pericardial space and into the right ventricle. Postmortem examination findings included a large mass in the right ventricular wall and multiple smaller masses on the external surface of the left ventricle and on the internal surface of the pericardium. Results of the histopathological and immunohistochemical examinations of the masses were consistent with rhabdomyosarcoma. This is the first reported case of primary cardiac rhabdomyosarcoma in the cat.

IgG response against infective larvae of Dirofilaria immitis in experimentally infected cats.

Somatic antigens from third stage larvae of Dirofilaria immitis (SL3) were used to detect IgG response against heartworm infection in 8 experimentally infected cats. A moderate specific anti-SL3 IgG response was found one month post-infection. Afterwards, antibodies decreased reaching a basal level 4 months post-infection and remained at this level until the end of the study. 6 months post-infection. Western blot analysis showed specific recognition of polypeptides of 79, 73, 60, 52, 40 and 39 kDa by sera from infected cats 1 month post-infection, but not by sera taken prior to the infection. The low antigenicity of the SL3 antigen in the cat should allow the parasite to escape the host's immune response.

Efficacy of moxidectin for the prevention of adult heartworm (Dirofilaria immitis) infection in dogs.

The authors report the efficacy of orally administered moxidectin for the prevention of canine heartworm infection in two endemic areas in northern Italy. Two trials were conducted on a total of 257 dogs, including 137 treated with moxidectin (minimum dose of 3 mcg/kg body weight), 85 with ivermectin (minimum dose 6.6 mcg/kg b.w.) and 35 untreated controls. Results of testing for microfilariae and circulating adult female antigens were negative for all treated dogs at the end of both trials. No adverse reactions to moxidectin were observed. In the study areas, prevalence values for Dirofilaria immitis infection calculated on the basis of the untreated controls and testing dogs which had no preventive treatment in the previous transmission season ranged 23-65%. This study confirms the efficacy and safety of moxidectin in the prevention of adult heartworm infection in dogs.

Utility of adult antigens of Dirofilaria immitis for the early detection of dirofilariosis and for the evaluation of chemoprophylactic treatment in experimentally infected cats.

Early detection of feline dirofilariosis was achieved by ELISA testing using adult somatic (SA) and excretory/secretory (E/S) antigens. Sera from experimentally infected cats showed significantly higher IgG levels at 2 months post-infection (p.i.) when compared to sera obtained prior to infection. This difference was apparent at all serum dilutions (1:100 to 1:800) in ELISA(SA) and at all dilutions except 1:800 when tested with ELISA(E/S). Changes in specific anti-SA and anti-E/S IgG levels were found in infected cats treated with the macrolid parasiticide. ELISA(E/S) was able to detect a significant decrease in IgG levels in ivermectin treated cats at 3 months post-treatment (p.t.), compared to infected, untreated controls. However, testing with ELISA(SA) showed a significant decrease in antibody levels in treated cats only at 4 months after treatment. Sera from untreated cats recognized previously identified specific markers for feline dirofilariosis on SA (22, 26, 30 and 40 kDa) and on E/S (22 and 25 kDa) in Western blot analysis. Sera from treated cats, on the other hand, gave only weak bands or none at all, indicating a decrease in antibody concentration. These results suggest that serologic testing with ELISA is useful in the early detection of Dirofilaria immitis infection and, together with Western blot analysis, can assist in the evaluation of response to chemoprophylactic treatment of feline dirofilariosis, two aspects of prime importance in the prevention of this disease in cats.

Effects of tetracycline on the filarial worms Brugia pahangi and Dirofilaria immitis and their bacterial endosymbionts Wolbachia.

Wolbachia endosymbiotic bacteria have been shown to be widespread among filarial worms and could thus play some role in the biology of these nematodes. Indeed, tetracycline has been shown to inhibit both the development of adult worms from third-stage larvae and the development of the microfilaraemia in jirds infected with Brugia pahangi. The possibility that these effects are related to the bacteriostatic activity of tetracycline on Wolbachia symbionts should be considered. Here we show that tetracycline treatment is very effective in blocking embryo development in two filarial nematodes, B. pahangi and Dirofilaria immitis. Embryo degeneration was documented by TEM, while the inhibition of the transovarial transmission of Wolbachia was documented by PCR. Phylogenetic analysis on the ssrDNA sequence of the Wolbachia of B. pahangi confirms that the phylogeny of the bacterial endosymbionts is consistent with that of the host worms. The possibility that tetracycline inhibition of embryo development in B. pahangi and D. immitis is determined by cytoplasmic incompatibility is discussed.

Preliminary results on the effect of tetracycline on the embryogenesis and symbiotic bacteria (Wolbachia) of Dirofilaria immitis. An update and discussion.

The distribution and phylogeny of Wolbachia in filarial species suggests that these endosymbiotic bacteria may be important in the biology of their filarial hosts. An experiment to falsify this hypothesis would be to treat filarial worms with antibiotics which are active against intracellular bacteria. Indeed, it has already been shown that tetracycline treatment inhibits development in a model filarial species (Brugia pahangi) at different stages of the life cycle, in both mosquito and mammalian hosts. Here we discuss these previous data and present new results on the effect of tetracycline on the embryogenesis of the canine filaria Dirofilaria immitis.