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Ectopic location of adult worms and first-stage larvae of Angiostrongylus vasorum in an infected dog
Veterinary Parasitology 121 (2004) 293–296
Ectopic location of adult worms and first-stage larvae of Angiostrongylus vasorum in an infected dog S.D. Oliveira-Júnior, J.M.P. Barçante∗ , T.A. Barçante, V.M. Ribeiro, W.S. Lima Laboratório de Helmintologia Veterinária, Departamento de Parasitologia, Universidade Federal de Minas Gerais, ICB, Caixa Postal: 486, CEP 31270-901, Minas Gerias, Brazil Received 30 July 2003; received in revised form 4 February 2004; accepted 22 February 2004
Abstract A mongrel dog with angiostrongylosis died suddenly and was submitted to necropsy. Post-mortem findings showed adult worms in ectopic locations. This is the first report of Angiostrongylus vasorum adult worms being found within the pericardial sac and in the lumen of the bladder of an infected dog. A total of 587 A. vasorum first-stage larvae were recovered from different organs and tissues after concentration using Baermann apparatus. © 2004 Elsevier B.V. All rights reserved. Keywords: Angiostrongylus vasorum; Ectopic location; Adult worms; First-stage larvae
1. Introduction Angiostrongylus vasorum (Baillet, 1866) Kamensky (1905) is a protostrongylid nematode that inhabits the right ventricle of the heart, pulmonary artery and its branches in domestic dogs and wild canids (Cury et al., 2002). The life cycle evolves first-stage larvae (L1) that hatch into the alveoli; where the sexual reproduction and oviposture take place. The L1 pass up the bronchial tree and along digestive tract before being eliminated to the environment with the host feces. The intermediate host, snails and slugs, either terrestrial or aquatic become infected through invasion or ingestion of L1. Larvae invade mollusks tissues, where they undergo first and second molts, reaching the infective third-stage larvae (L3) (Guilhon and Afghahi, 1969; Rosen et al., 1970). Infection of definitive host results from ingestion of infected snail or slugs which contain infective L3 ∗ Corresponding author. Tel.: +55-31-34992975; fax: +55-31-34992970. E-mail address: [email protected] (J.M.P. Barçante).
0304-4017/$ – see front matter © 2004 Elsevier B.V. All rights reserved. doi:10.1016/j.vetpar.2004.02.018
294
S.D. Oliveira-J´unior et al. / Veterinary Parasitology 121 (2004) 293–296
(Guilhon and Cens, 1973). Paratenic hosts, such as frogs, lizards, mice and rats may also be ingested by definitive host (Bolt et al., 1993). The L3 invade mesenteric lymph nodes where they undergo third and fourth molts. Young adults nematode migrate via lymphatic or hepatic portal vessels to the right side of the heart and pulmonary artery, where they develop to sexual maturity (Patteson et al., 1993). Infection of the dog frequently leads to pneumonia, loss of racing performance, coughing and anemia (Jones et al., 1980). Severely infected dogs may develop cardiac insufficiency, pulmonary fibrosis followed by weight loss, hemorrhagic diatheses and death (Dood, 1973).
2. Case history A mongrel adult dog with angiostrongyliasis died suddenly. The diagnosis was based on clinical signs and presence of first-stage larvae on the fecal exams. The recovery of A. vasorum-L1 was performed using the modified Baermann apparatus (Barçante et al., 2003). This apparatus consisted of a funnel directly connected to a hemolysis tube through a latex hose. Dog fecal samples (50–100 g) were wrapped in surgical gauze and placed on the top of the funnel filled with tap-water at 41 ◦ C. After 12 h, the sediment was recovered from the bottom of the hemolysis tube and washed three times with phosphate-buffered saline (PBS) (136 mM NaCl, 2 mM KCl, 8 mM Na2 HPO4 , 10 mM KH2 PO4 , pH 7.4) by centrifugation at 250×g for 5 min, 20 ◦ C. The active and inactive larvae were counted under a stereomicroscope and identified as A. vasorum-L1 (Lima et al., 1985; Bessa et al., 1996). In the clinical examination the dog presented signs of weight loss, generalized dermatitis, pale mucous membrane, bilateral lung rattles and cardiac arrhythmia. At necropsy congestion in submandibular, cervical and popliteal lymph nodes was observed. A yellowish fluid was observed in the thoracic cavity. The lungs contained hemorrhage areas and grayish nodes. A total of 96 adult worms was recovered from the major pulmonary artery and 114 adult worms were recovered from the branches of the pulmonary artery. A reddish fluid with amount increased above normal was observed within the pericardial sac, where a male adult worm measuring 12.36 mm in length and 0.26 mm in width was found. The right ventricular wall was flaccid and congestion was seen on the walls of both heart ventricles. A total of 104 adult parasites was found in the right ventricle. After examination of other organs and tissues a female adult worm was found in the lumen of the bladder. The worm was measuring 14.47 mm in length and 0.35 mm in width, was whitish and no eggs were seen in the uterus. Each organ was entire cut up, wrapped up in surgical gauze and placed on the top of the funnel of the modified Baermann apparatus (Barçante et al., 2003). At the same way multiple skin sections of ears, abdomen, members, ventral and dorsal thorax were submitted to modified Baermann apparatus. Alive and active A. vasorum first-stage larvae (L1) were recovered from the skin, diaphragm, liver, pancreas, brain and cerebellum. A total of 5 ml of peripheral blood and 1 ml of cerebrospinal fluid were centrifuged (250 × g for 5 min, 20 ◦ C) and the sediment was re-suspend in 5 ml of distillated water. The suspensions were centrifuged (250 × g for 5 min, 20 ◦ C) and the sediments were analyzed under a stereomicroscope. Larvae were also recovered on peripheral blood and cerebrospinal fluid. Results of the systemic distribution of L1 are presented in Table 1.
S.D. Oliveira-J´unior et al. / Veterinary Parasitology 121 (2004) 293–296
295
Table 1 Number of A. vasorum first-stage larvae recovered from different organs and tissues of a dog in chronic infection, using modified Baermann apparatus Organ or tissue
Number of recovered L1
Brain Cerebellum Diaphragm Liver Pancreas Skin CSF (1 ml) Peripheral blood (5 ml)
8 4 4 550 9 8 2 2
Total
587
L1: first-stage larvae; CSF: cerebrospinal fluid.
All of the helminths, adults and larvae, were identified as A. vasorum (Lima et al., 1985; Bessa et al., 1996).
3. Discussion The clinical, hematological and pathological changes associated with the canine angiostrongylosis have been extensively described (Prestwood et al., 1981; Perry et al., 1991; Patteson et al., 1993). The cardiac and pulmonary lesions described in this work were similar to those reported in the literature for canine angiostrongylosis (Cury and Lima, 1996a; Costa and Tafuri, 1997). For the first time it was showed that A. vasorum-L1 can be recovered alive and active in many different organs from an infected dog. Ectopic location of parasite L1 can cause complications at the levels of the capillaries and sinusoids (Costa and Tafuri, 1997). This is the first report of A. vasorum adult worms being found within the pericardium sac and in the lumen of the bladder of an infected dog. The ectopic location of adults and larvae is really an important fact in the pathogenesis of the infection by A. vasorum. The presence of adults and larvae in sites different from those described in the natural route of parasite in its life cycle generally leads to animal death or lesions that can change the normal organ functions. Death of infected animals due to ectopic location of A. vasorum adult worms the kidney and femoral artery was previously described (Cury and Lima, 1995, 1996b). Although the knowledge of the ectopic location importance in the pathogenesis of canine angiostrongylosis the mechanisms, which determine the changed of a normal route to an erratic site still need to be studied.
References Barçante, J.M.P., Barçante, T.A., Dias, S.R.C., Vieira, L.Q., Lima, W.S., Negrão-Corrˆea, D., 2003. A method to obtain axenic Angiostrongylus vasorum first-stage larvae from dog feces. Parasitol. Res. 89, 89–93.
296
S.D. Oliveira-J´unior et al. / Veterinary Parasitology 121 (2004) 293–296
Bessa, E.C., Lopes, C.W.G., Pinheiro, N.L., Araujo, J.L.B., 1996. S´ıtios de desenvolvimento de Angiostrongylus vasorum (Baillet, 1866) Kamensky, 1905 (Nematoda: Protostrongylidae) em Subulina octona (Bruguière, 1789) (MOLUSCA: Subulinidae) em condições de laboratório. Bol. ICB 47, 13–19. Bolt, G., Monrad, J., Frandsen, F., Henrikesen, P., Dietz, H.H., 1993. The common frog (Rana temporaria) as a pontencial paratenic and intermediate host for Angiostrongylus vasorum. Parasitol. Res. 79, 428–430. Costa, J.O., Tafuri, W.L.E., 1997. Estudo anátomo-patológico de cães infectados experimentalmente pelo Angiostrongylus vasorum (Baillet, 1866) Kamensky 1905. Arq. Bras. Med. Vet. Zootech. 49, 389–407. Cury, M.C., Lima, W.S., 1995. Ocorrˆecia de Angiostrongylus vasorum no rim de um cão experimentalmente infectado. Arq. Bras. Med. Vet. Zootech. 47, 593–595. Cury, M.C., Lima, W.S., 1996a. Aspectos cl´ınicos de cães infectados experimentalmente com Angiostrongylus vasorum. Arq. Bras. Vet. Zootech. 48, 27–34. Cury, M.C., Lima, W.S., 1996b. Rupture of femoral artery in a dog infected with Angiostrongylus vasorum. Vet. Parasitol. 65, 313–315. Cury, M.C., Guimarães, M.P., Lima, W.S., Vitor, R.W.A., 2002. Western blot analyses of the humoral response of dogs experimentally infected with Angiostrongylus vasorum (Baillet, 1866). Vet. Parasitol. 106, 83–87. Dood, K., 1973. Angiostrongylus vasorum (Baillet, 1866) infestation in a greyhound kennels. Vet. Rec. 92, 195. Guilhon, J., Afghahi, A., 1969. Larval development of Angiostrongylus vasorum (Baillet, 1866) in the body of various species of terrestrial mollusks. CR Acad. Sci. Paris 268, 434–436. Guilhon, J., Cens, B., 1973. Angiostrongylus vasorum (Baillet, 1866) Etude biologique et morfologique. Ann. Parasitol. 48, 567–596. Jones, G.W., Neal, C., Turner, G.R.J., 1980. Angiostrongylus vasorum infection in dogs in Cornwall. Vet. Rec. 26, 83. Lima, W.S., Costa, H.M.A., Guimarães, M.P., Leite, A.C.R., 1985. Angiostrongylus vasorum (Baillet, 1866) Nematoda: Prothostrongylidae em cães de Minas Gerais, Brasil. Mem. Inst. Oswaldo Cruz 80, 233–235. Patteson, M.W., Gibbs, C., Wotton, P.R., Day, M.J., 1993. Angiostrongylus vasorum infection in seven dogs. Vet. Rec. 133, 565–570. Perry, A.W., Herting, R., Kennedy, M.J., 1991. Angiostrongylosis with disseminated larval infection associated with signs of ocular and nervous disease in an imported dog. Can. Vet. J. 32, 430–431. Prestwood, A.K., Greene, C.E., Mahaffey, E.A., Burgess, D.E., 1981. Experimental Canine Angiostrongylosis. I. Pathologic manifestation. J. Am. Anim. Hosp. Assoc. 17, 491–497. Rosen, L., Ash, L.R., Wallace, G.D., 1970. Life history of the canine lungworm Angiostrongylus vasorum (Baillet, 1866). Am. J. Vet. Res. 31, 131–143.
Ectopic location of adult worms and first-stage larvae of Angiostrongylus vasorum in an infected dog S.D. Oliveira-Júnior, J.M.P. Barçante∗ , T.A. Barçante, V.M. Ribeiro, W.S. Lima Laboratório de Helmintologia Veterinária, Departamento de Parasitologia, Universidade Federal de Minas Gerais, ICB, Caixa Postal: 486, CEP 31270-901, Minas Gerias, Brazil Received 30 July 2003; received in revised form 4 February 2004; accepted 22 February 2004
Abstract A mongrel dog with angiostrongylosis died suddenly and was submitted to necropsy. Post-mortem findings showed adult worms in ectopic locations. This is the first report of Angiostrongylus vasorum adult worms being found within the pericardial sac and in the lumen of the bladder of an infected dog. A total of 587 A. vasorum first-stage larvae were recovered from different organs and tissues after concentration using Baermann apparatus. © 2004 Elsevier B.V. All rights reserved. Keywords: Angiostrongylus vasorum; Ectopic location; Adult worms; First-stage larvae
1. Introduction Angiostrongylus vasorum (Baillet, 1866) Kamensky (1905) is a protostrongylid nematode that inhabits the right ventricle of the heart, pulmonary artery and its branches in domestic dogs and wild canids (Cury et al., 2002). The life cycle evolves first-stage larvae (L1) that hatch into the alveoli; where the sexual reproduction and oviposture take place. The L1 pass up the bronchial tree and along digestive tract before being eliminated to the environment with the host feces. The intermediate host, snails and slugs, either terrestrial or aquatic become infected through invasion or ingestion of L1. Larvae invade mollusks tissues, where they undergo first and second molts, reaching the infective third-stage larvae (L3) (Guilhon and Afghahi, 1969; Rosen et al., 1970). Infection of definitive host results from ingestion of infected snail or slugs which contain infective L3 ∗ Corresponding author. Tel.: +55-31-34992975; fax: +55-31-34992970. E-mail address: [email protected] (J.M.P. Barçante).
0304-4017/$ – see front matter © 2004 Elsevier B.V. All rights reserved. doi:10.1016/j.vetpar.2004.02.018
294
S.D. Oliveira-J´unior et al. / Veterinary Parasitology 121 (2004) 293–296
(Guilhon and Cens, 1973). Paratenic hosts, such as frogs, lizards, mice and rats may also be ingested by definitive host (Bolt et al., 1993). The L3 invade mesenteric lymph nodes where they undergo third and fourth molts. Young adults nematode migrate via lymphatic or hepatic portal vessels to the right side of the heart and pulmonary artery, where they develop to sexual maturity (Patteson et al., 1993). Infection of the dog frequently leads to pneumonia, loss of racing performance, coughing and anemia (Jones et al., 1980). Severely infected dogs may develop cardiac insufficiency, pulmonary fibrosis followed by weight loss, hemorrhagic diatheses and death (Dood, 1973).
2. Case history A mongrel adult dog with angiostrongyliasis died suddenly. The diagnosis was based on clinical signs and presence of first-stage larvae on the fecal exams. The recovery of A. vasorum-L1 was performed using the modified Baermann apparatus (Barçante et al., 2003). This apparatus consisted of a funnel directly connected to a hemolysis tube through a latex hose. Dog fecal samples (50–100 g) were wrapped in surgical gauze and placed on the top of the funnel filled with tap-water at 41 ◦ C. After 12 h, the sediment was recovered from the bottom of the hemolysis tube and washed three times with phosphate-buffered saline (PBS) (136 mM NaCl, 2 mM KCl, 8 mM Na2 HPO4 , 10 mM KH2 PO4 , pH 7.4) by centrifugation at 250×g for 5 min, 20 ◦ C. The active and inactive larvae were counted under a stereomicroscope and identified as A. vasorum-L1 (Lima et al., 1985; Bessa et al., 1996). In the clinical examination the dog presented signs of weight loss, generalized dermatitis, pale mucous membrane, bilateral lung rattles and cardiac arrhythmia. At necropsy congestion in submandibular, cervical and popliteal lymph nodes was observed. A yellowish fluid was observed in the thoracic cavity. The lungs contained hemorrhage areas and grayish nodes. A total of 96 adult worms was recovered from the major pulmonary artery and 114 adult worms were recovered from the branches of the pulmonary artery. A reddish fluid with amount increased above normal was observed within the pericardial sac, where a male adult worm measuring 12.36 mm in length and 0.26 mm in width was found. The right ventricular wall was flaccid and congestion was seen on the walls of both heart ventricles. A total of 104 adult parasites was found in the right ventricle. After examination of other organs and tissues a female adult worm was found in the lumen of the bladder. The worm was measuring 14.47 mm in length and 0.35 mm in width, was whitish and no eggs were seen in the uterus. Each organ was entire cut up, wrapped up in surgical gauze and placed on the top of the funnel of the modified Baermann apparatus (Barçante et al., 2003). At the same way multiple skin sections of ears, abdomen, members, ventral and dorsal thorax were submitted to modified Baermann apparatus. Alive and active A. vasorum first-stage larvae (L1) were recovered from the skin, diaphragm, liver, pancreas, brain and cerebellum. A total of 5 ml of peripheral blood and 1 ml of cerebrospinal fluid were centrifuged (250 × g for 5 min, 20 ◦ C) and the sediment was re-suspend in 5 ml of distillated water. The suspensions were centrifuged (250 × g for 5 min, 20 ◦ C) and the sediments were analyzed under a stereomicroscope. Larvae were also recovered on peripheral blood and cerebrospinal fluid. Results of the systemic distribution of L1 are presented in Table 1.
S.D. Oliveira-J´unior et al. / Veterinary Parasitology 121 (2004) 293–296
295
Table 1 Number of A. vasorum first-stage larvae recovered from different organs and tissues of a dog in chronic infection, using modified Baermann apparatus Organ or tissue
Number of recovered L1
Brain Cerebellum Diaphragm Liver Pancreas Skin CSF (1 ml) Peripheral blood (5 ml)
8 4 4 550 9 8 2 2
Total
587
L1: first-stage larvae; CSF: cerebrospinal fluid.
All of the helminths, adults and larvae, were identified as A. vasorum (Lima et al., 1985; Bessa et al., 1996).
3. Discussion The clinical, hematological and pathological changes associated with the canine angiostrongylosis have been extensively described (Prestwood et al., 1981; Perry et al., 1991; Patteson et al., 1993). The cardiac and pulmonary lesions described in this work were similar to those reported in the literature for canine angiostrongylosis (Cury and Lima, 1996a; Costa and Tafuri, 1997). For the first time it was showed that A. vasorum-L1 can be recovered alive and active in many different organs from an infected dog. Ectopic location of parasite L1 can cause complications at the levels of the capillaries and sinusoids (Costa and Tafuri, 1997). This is the first report of A. vasorum adult worms being found within the pericardium sac and in the lumen of the bladder of an infected dog. The ectopic location of adults and larvae is really an important fact in the pathogenesis of the infection by A. vasorum. The presence of adults and larvae in sites different from those described in the natural route of parasite in its life cycle generally leads to animal death or lesions that can change the normal organ functions. Death of infected animals due to ectopic location of A. vasorum adult worms the kidney and femoral artery was previously described (Cury and Lima, 1995, 1996b). Although the knowledge of the ectopic location importance in the pathogenesis of canine angiostrongylosis the mechanisms, which determine the changed of a normal route to an erratic site still need to be studied.
References Barçante, J.M.P., Barçante, T.A., Dias, S.R.C., Vieira, L.Q., Lima, W.S., Negrão-Corrˆea, D., 2003. A method to obtain axenic Angiostrongylus vasorum first-stage larvae from dog feces. Parasitol. Res. 89, 89–93.
296
S.D. Oliveira-J´unior et al. / Veterinary Parasitology 121 (2004) 293–296
Bessa, E.C., Lopes, C.W.G., Pinheiro, N.L., Araujo, J.L.B., 1996. S´ıtios de desenvolvimento de Angiostrongylus vasorum (Baillet, 1866) Kamensky, 1905 (Nematoda: Protostrongylidae) em Subulina octona (Bruguière, 1789) (MOLUSCA: Subulinidae) em condições de laboratório. Bol. ICB 47, 13–19. Bolt, G., Monrad, J., Frandsen, F., Henrikesen, P., Dietz, H.H., 1993. The common frog (Rana temporaria) as a pontencial paratenic and intermediate host for Angiostrongylus vasorum. Parasitol. Res. 79, 428–430. Costa, J.O., Tafuri, W.L.E., 1997. Estudo anátomo-patológico de cães infectados experimentalmente pelo Angiostrongylus vasorum (Baillet, 1866) Kamensky 1905. Arq. Bras. Med. Vet. Zootech. 49, 389–407. Cury, M.C., Lima, W.S., 1995. Ocorrˆecia de Angiostrongylus vasorum no rim de um cão experimentalmente infectado. Arq. Bras. Med. Vet. Zootech. 47, 593–595. Cury, M.C., Lima, W.S., 1996a. Aspectos cl´ınicos de cães infectados experimentalmente com Angiostrongylus vasorum. Arq. Bras. Vet. Zootech. 48, 27–34. Cury, M.C., Lima, W.S., 1996b. Rupture of femoral artery in a dog infected with Angiostrongylus vasorum. Vet. Parasitol. 65, 313–315. Cury, M.C., Guimarães, M.P., Lima, W.S., Vitor, R.W.A., 2002. Western blot analyses of the humoral response of dogs experimentally infected with Angiostrongylus vasorum (Baillet, 1866). Vet. Parasitol. 106, 83–87. Dood, K., 1973. Angiostrongylus vasorum (Baillet, 1866) infestation in a greyhound kennels. Vet. Rec. 92, 195. Guilhon, J., Afghahi, A., 1969. Larval development of Angiostrongylus vasorum (Baillet, 1866) in the body of various species of terrestrial mollusks. CR Acad. Sci. Paris 268, 434–436. Guilhon, J., Cens, B., 1973. Angiostrongylus vasorum (Baillet, 1866) Etude biologique et morfologique. Ann. Parasitol. 48, 567–596. Jones, G.W., Neal, C., Turner, G.R.J., 1980. Angiostrongylus vasorum infection in dogs in Cornwall. Vet. Rec. 26, 83. Lima, W.S., Costa, H.M.A., Guimarães, M.P., Leite, A.C.R., 1985. Angiostrongylus vasorum (Baillet, 1866) Nematoda: Prothostrongylidae em cães de Minas Gerais, Brasil. Mem. Inst. Oswaldo Cruz 80, 233–235. Patteson, M.W., Gibbs, C., Wotton, P.R., Day, M.J., 1993. Angiostrongylus vasorum infection in seven dogs. Vet. Rec. 133, 565–570. Perry, A.W., Herting, R., Kennedy, M.J., 1991. Angiostrongylosis with disseminated larval infection associated with signs of ocular and nervous disease in an imported dog. Can. Vet. J. 32, 430–431. Prestwood, A.K., Greene, C.E., Mahaffey, E.A., Burgess, D.E., 1981. Experimental Canine Angiostrongylosis. I. Pathologic manifestation. J. Am. Anim. Hosp. Assoc. 17, 491–497. Rosen, L., Ash, L.R., Wallace, G.D., 1970. Life history of the canine lungworm Angiostrongylus vasorum (Baillet, 1866). Am. J. Vet. Res. 31, 131–143.