Parasitol Res DOI 10.1007/s00436-015-4763-x
ORIGINAL PAPER
New insights into diagnosis of Platynosomum fastosum (Trematoda: Dicrocoeliidae) in cats Rafael Antonio Nascimento Ramos 1 & Victor Fernando Santana Lima 1 & Maria Fernanda Melo Monteiro 1 & Marília de Andrade Santana 1 & Raphael Lepold 1 & Maria Aparecida da Gloria Faustino 1 & Laura Rinaldi 2 & Giuseppe Cringoli 2 & Leucio Câmara Alves 1
Received: 3 August 2015 / Accepted: 28 September 2015 # Springer-Verlag Berlin Heidelberg 2015
Abstract Platynosomum fastosum is a hepatic trematode which causes the so-called lizard poisoning in cats. This parasite is reported in tropical and sub-tropical areas infecting the liver, gallbladder, and bile ducts of cats. Despite its clinical importance, the diagnosis of P. fastosum has been poorly investigated so far. In this study, three cases of infection by P. fastosum in cats are reported. The FLOTAC technique was utilized for the first time in the detection of eggs of this parasite. Three cats (two males and one female), being one of them with a history of diarrhea and vomiting, were diagnosed positive for the presence of P. fastosum eggs through the FLOTAC technique. In conclusion, this study provides important and new insights into the diagnosis of P. fastosum in cats, expanding the geographical knowledge of this parasite in Brazil. Keywords Cat liver fluke . Lizard poisoning . FLOTAC
Introduction Over the last years, helminth infections in cats have gained the scientific interest of researchers and practitioners due to the severity of the clinical manifestations they cause in these animals (Deplazes et al. 2011). Among the main parasites, intestinal nematodes (e.g., Toxocara sp. and Ancylostoma sp.) are * Rafael Antonio Nascimento Ramos
[email protected] 1
Department of Veterinary Medicine, Federal Rural University of Pernambuco, Recife, Brazil
2
Department of Veterinary Medicine and Animal Productions, University of Napoli Federico II, Naples, Italy
well known and have been extensively studied due to the relevance in public health. These pathogens are widely distributed throughout the world and have been considered important causes of disease, especially in young cats (Bowman et al. 2010). However, little attention has been given to trematode species that parasitizes cats. For example, Platynosomum fastosum (Trematoda: Dicrocoeliidae) is a small hepatic trematode which causes the so-called lizard poisoning in cats (Catcott 1964). Infection by this parasite occurs via ingestion of intermediate (e.g., snails) or paratenic (e.g., lizards) hosts containing infective metacercariae (Foley 1994). Usually, this trematode is restricted to tropical and sub-tropical areas where intermediate hosts have suitable conditions to development (Basu and Charles 2014). In infected animals, the main sites of parasitism are the liver, gallbladder, and bile ducts (Maldonado 1945). Therefore, in cases of high parasitism, clinical signs characterized by hepatic insufficiency and hepatic encephalopathy may be the cause of death (Pimentel et al. 2005; Headley et al. 2012). Despite of its clinical importance, the diagnosis of the infection by P. fastosum and its clinical implications have been poorly investigated so far. Indeed, most of cases diagnosed are occasional findings obtained during necropsies (Carreira et al. 2008). For instance, in Brazil, few cases have been reported over the last years, and all of them have been detected through the presence of eggs of P. fastosum in feces or during the post-mortem examination (Table 1; adapted from Basu and Charles 2014). In fact, the great importance of platynosomiasis in the feline medicine contrasts with the scarcity of studies on the diagnosis and clinical presentation of infected animals. Until now, many gaps remain in the history of this parasite indicating that this infection may occur sporadically or has been underestimated over the time
Parasitol Res Table 1
Infections by Platynosomum sp. in cats from Brazil
Geographical location
Case reports and prevalence
Method of diagnosis
Reference
Rio de Janeiro
Case report: 4 cats
Necropsy and histology of liver
Santos et al. (1981)
São Paulo
0.46 % (01/215)
Willis-Mollay and centrifugal flotation in water-ether
Ogassawara et al. (1986)
São Paulo
1.07 % (02/187)
São Paulo
27.3 % (03/11)
Centrifugal flotation in water-ether and centrifugal sedimentation in sucrose Gennari et al. (1999) solution Necropsy Silva et al. (2001)
São Paulo
1.45 % (02/138)
Centrifugal flotation in sucrose solution
Minas Gerais
40 % (20/50)
Necropsy
Ragozo et al. (2002) Mundim et al. (2004)
Rio Grande do Sul
Case report: 3 cats
Abdominal ultrasonography, exploratory laparotomy, and liver biopsy.
Ilha et al. (2004)
Rio de Janeiro
33 % (24/72)
Abdominal ultrasonography and Faust technique
Salomão et al. (2005)
Bahia
Case report: 1 cat
Abdominal ultrasonography
Sampaio et al. (2006)
Rio de Janeiro
30.7 % (04/13)
Zinc sulfate centrifugal flotation technique
Souza-Dantas et al. (2007) Xavier et al. (2007)
São Paulo
Case report: 1 cat
Necropsy and histology of liver
São Paulo
Case report: 1 cat
Abdominal ultrasonography, liver biopsy, direct examination of biliary fluid Carreira et al. (2008)
Minas Gerais
Case report: 2 cats
Necropsy and histology of liver
Vieira et al. (2009)
Paraíba
0.9 % (03/327)
Necropsy and histology of liver
Andrade et al. (2012)
Rio Grande do Sul
Case report: 1 cat
Sedimentation
Michaelsen et al. (2012)
São Paulo
Case report: 1 cat
Abdominal ultrasonography, cholecystoduodenostomy, and biopsy of liver
Daniel et al. (2012)
Mato Grosso
26.03 % (38/146)
Necropsy
Ramos et al. (2013)
Rio de Janeiro
Case report: 8 cats
Abdominal ultrasonography and radiographic examination
Azevedo et al. (2013)
Bahia
Case report: 1 cat
Necropsy
Jesus et al. (2015)
Adapted from Basu and Charles (2014)
due to the inefficiency of the methods of diagnosis. Therefore, the aim of this study was to report for the first time the use of the FLOTAC technique in the detection of P. fastosum eggs in cats. In addition, cases of platynosomiasis in cats were diagnosed in an area never reported before.
Material and methods Sampling and animals Three mixed-breed cats (two males and one female) with different ages were used in this study. All animals (A1, A2, and A3) were patients at the Veterinary Teaching Hospital (VTH) of the Universidade Federal Rural de Pernambuco (UFRPE), Recife (08° 03′ 14″ S and 34° 52′ 52″ W), Brazil. Fresh feces were collected and the material obtained was transferred to plastic vials, labeled, and stored at 8 °C until laboratorial analysis. According to the owners, none of the animals have been dewormed in the last 6 months.
Laboratory procedures In order to avoid the diagnosis failure, each sample was analyzed using three different techniques: (1) Willis-Mollay flotation technique (Willis 1921); (2) spontaneous sedimentation of Hoffman, Pons, and Janer (Hoffman et al. 1934); and (3) FLOTAC technique (Cringoli et al. 2010). The FLOTAC was performed with two flotation solutions (FS2—saturated sodium chloride 1.200, and FS7—zinc sulphate 1.350). All methods were performed according to the instructions reported in the original protocols. In addition, a routine physical examination and serum biochemistry were performed in all animals.
Results Eggs of P. fastosum were detected in all samples analyzed but only when using the FLOTAC technique. The number of eggs per gram (EPG) of feces ranged from 7 to 385 per animal (mean=136 EPG). The P. fastosum eggs presented a mean length and width of 42.5 μm (±5.2) and 28.2 μm (±3.1),
Parasitol Res
respectively. Morphologically, they were golden brown (unstained) oval, thick shelled, and embryonated, with symmetrical ends containing an operculum at only one end (Fig. 1). Of the cats studied, one of them (A1) presented continuous episodes of diarrhea and vomiting, whereas the other two were asymptomatic (A2 and A3). The biochemical analysis revealed levels of urea (79.3 μg/dL), creatinine (1.9 μg/dL), and alkaline phosphatase (105 μg/dL) higher only in the A1. Interestingly, all animals lived in areas were the presence of lizards, snails, and frogs have been frequently reported. Moreover, according to the owner, the only animal with clinical manifestation (A1) hunts and eats routinely these small animals. It is important to note that one of them (A3) comes from the municipality of Salvador (12° 58′ 16″ S and 38° 30′ 39″ W), an endemic area for platynosomiasis, 4 months before of the diagnosis.
Discussion This study provides new insights into the copromicroscopic diagnosis of P. fastosum in cats which was diagnosed here for the first time using the FLOTAC technique. Infections by P. fastosum are occasionally detected during the necropsy (Carreira et al. 2008). Although, some different copromicroscopic methods have already been evaluated for the identification of platynosomiasis in cats, information is still limited (Souza-Dantas et al. 2007; Krecek et al. 2010). Indeed, it has been reported that the efficacy in the diagnosis of P. fastosum in cats may be influenced by several intrinsic characteristics such as the small size of eggs and the identification of immature eggs (Palumbo et al. 1976; Ferreira et al. 1999). It is known that a single adult fluke may
produce 10–100 eggs per day; therefore, the number of eggs released is an important factor which may influence the diagnosis (Palumbo et al. 1976). To our knowledge, until now, a Bgold standard^ test for the diagnosis of P. fastosum in cats has not been well established in the scientific literature. Distinct methods of feces analyses (e.g., centrifugal sedimentation and flotation) employed for the parasitological diagnosis of this infection exhibit different values of sensitivity (Leal et al. 2011; Basu and Charles 2014). Based on our findings, the FLOTAC technique herein proposed may be considered an important tool to diagnose P. fastosum eggs in feces of cats; in addition, data herein obtained may be extended to the diagnosis of platynosomiasis in other vertebrate host species. In Brazil, since 1981 when the first case of platynosomiasis in cats was reported, only six studies detected this trematode using copromicroscopic methods, but the FLOTAC technique was never mentioned. The other cases were based on the detection and identification during post-mortem examination (see Table 1). In fact, it is widely known that the FLOTAC technique is a reliable method for the diagnosis of several species of intestinal parasites of medical and veterinary concern (Cringoli et al. 2010; Lima et al. 2015). Indeed, the technique may be used for the simultaneous detection of different helminthes as well as intestinal protozoa of cats; this issue is of relevance because parasitic co-infections are very common in pets (Gaglio et al. 2008). In this study, only an animal (A1) showed unspecific clinical (i.e., diarrhea and vomiting) which could be related to any parasitic infection. In addition, some biochemical parameters such as urea, creatinine, and alkaline phosphatase were higher, being this latter change related to the hepatic function. Curiously, according to the owner, the A1 hunts and eats lizards, snails, and frogs which play an important role in the life cycle of this trematode. In conclusion, the FLOTAC technique is an efficient copromicroscopic technique that may be used to enhance the diagnosis of P. fastosum in cats. Additional studies with a large number of samples are needed in order to better evaluate the use of this tool in the diagnosis of platynosomiasis. Finally, this study provides important and new insights into the diagnosis of P. fastosum in cats, increasing the geographical knowledge of this parasite in Brazil. Compliance with ethical standards
Fig. 1 Egg of Platynosomum fastosum detected through FLOTAC technique. Note the presence of an operculum at only one end (Scale bar=20 μm)
Conflict of interest The authors declare that they have no competing interests.
Parasitol Res
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