Treatment of primate platynosomosis: A word of caution about the use of praziquantel in marmosets.

BACKGROUND: The successive reports of Platynosomum illiciens in Neotropical captive primates have increased interest in platynosomosis; however, its treatment is little known. METHODS: Callithrix penicillata (n = 10) naturally and chronically infected with P. illiciens were treated with praziquantel (25 mg/kg BW, three s.c. doses at 24 hours intervals), and coproparasitological tests performed over 67 days. The proportions of primates with a reduction in fecal egg counts (FEC) or negative results progressively increased after treatment, and at the last fecal tests, marmosets were negative. RESULTS AND CONCLUSIONS: Although all primates tolerated the initial days of study well, 40% (4/10) of them died between the 8th and 16th days after the onset of treatment. Clinical signs and necropsies indicated the occurrence of hepatic involvement, biliary obstruction, and cholangitis. Marmosets with a higher previous FEC were more likely to die after treatment. Use of praziquantel should be considered carefully on a case-by-case basis.

Platynosomum illiciens (Trematoda: Dicrocoeliidae) in a hybrid marmoset (Callithrix sp.) in the Municipality of Seropédica, RJ, Brazil - Case report / Platynosomum illiciens (Trematoda: Dicrocoeliidae) em um sagui híbrido (Callithrix sp.) no município de Seropédica, RJ, Brasil - Relato de caso

Abstract Platynosomum illiciens is a liver trematode encountered infecting mainly felids although it has also been reported in birds and in additional mammalian species, including non-human primates. The current study reports a natural P. illiciens infection primate of the genus Callithrix. The diagnosis was made using a combination of copro-parasitological techniques, morphological evaluation of adult specimens recovered from the liver during necropsy, and molecular analyses. Eggs were brown in color, oval, operculated, and contained a miracidium. Adult specimens recovered during necropsy were measured and showed dimensions compatible with P. illiciens. Molecular characterization of the trematode involved amplification by polymerase chain reaction (PCR), in combination with nucleotide sequencing, of an approximately 900 base pairs fragment corresponding to 18S-ITS1-5.8S ribosomal DNA. Sequenced amplicons showed 100% nucleotide identity with sequences deposited in the GenBank database as derived from specimens of P. illiciens recovered from cats in Malaysia and Brazil. It was concluded that the morphological and molecular analyses presented herein, confirmed the identification of the trematode recovered as P. illiciens.

Resumo Platynosomum illiciens é um trematódeo do fígado, encontrado principalmente em felinos, embora também tenha sido relatado em aves e outras espécies de mamíferos, incluindo primatas não humanos. Este estudo relata um caso de parasitismo natural por P. illiciens em um primata do gênero Callithrix. O diagnóstico foi feito com uma combinação de técnicas coproparasitológicas, avaliação morfológica de espécimes adultos recuperados do fígado durante a necropsia e análise molecular. Os ovos eram de cor marrom, ovais, operculados e continham um miracídio. Espécimes adultos recuperados durante a necropsia foram medidos e mostraram dimensões compatíveis com P. illiciens. A caracterização molecular do trematódeo envolveu a reação em cadeia da polimerase (PCR) e o sequenciamento de um fragmento de, aproximadamente, 900 pares de bases correspondentes ao DNA robosomal 18S-ITS1-5.8S, que apresentou 100% de identidade da sequência de nucleotídeos com as sequências do GenBank depositadas como derivadas de P. illiciens recuperados de gatos na Malásia e no Brasil, respectivamente. Concluiu-se pelas análises morfológicas e moleculares aqui apresentadas, que o trematódeo é da espécie P. illiciens.

Comparison of fecal analysis methods for the detection of Platynosomum fastosum in naturally infected cats.

OBJECTIVES: Diagnosis of Platynosomum fastosum Kossack 1910, a liver fluke of cats, can be challenging, with infections potentially not identified depending on the fecal analysis method used. The aim of this study was to compare three methods for identification of P fastosum eggs in feces. METHODS: Feces from 50 cats with unknown infection status from a location endemic for P fastosum were analyzed. The methods used included double centrifugation with Sheather's sugar flotation solution (specific gravity 1.28); the Mini-Parasep (Apacor) flotation kit with ZnSO4 (specific gravity 1.20); and the Mini-Parasep sedimentation kit with Triton X/ethyl acetate solution. RESULTS: Thirty-four cats were identified as positive for P fastosum. Of these cats, 33 were identified with the double centrifugation method, 16 with the Mini-Parasep flotation kit and 11 with the Mini-Parasep sedimentation kit. Using the total number of samples identified as positive by at least one method as the true positive, the sensitivity of the methods was 97.1% for double centrifugation, 47.1% for the Mini-Parasep flotation kit and 32.4% for the Mini-Parasep sedimentation kit. Other parasite eggs (eg, Ancylostoma tubaeforme, Trichuris species and Toxocara cati) also were detected more frequently with the double centrifugation method. CONCLUSIONS AND RELEVANCE: Of the three methods tested, double centrifugation with Sheather's sugar flotation solution was the most effective in identifying cats with P fastosum and other helminth eggs in their feces. The lower specific gravity of the flotation solution in the Mini-Parasep flotation kit likely influenced the results. A primary drawback of the Mini-Parasep sedimentation kit was the amount of sediment. While more sediment was examined than recommended in the manufacturer's instructions, this was still only a small portion of the total sediment. In addition, reading the slides took longer than with the other methods.

Feline platynosomiasis: analysis of the association of infection levels with pathological and biochemical findings / Platinossomiase felina: análise da associação dos níveis de infecção com achados patológicos e bioquímicos

Abstract Platynosomiasis is a common feline hepatic disease caused by Platynosomum fastosum (Trematoda - Dicrocoelidae), which is also known as ‘lizard poisoning’. Most reports of feline platynosomiasis show that this disease is sporadic and manifests with uncommon lesions; its pathogenicity is still not well understood. This study aimed to describe liver injuries and enzymatic changes associated with natural P. fastosum infection in 47 stray cats in an endemic area. Overall, 38.3% (18/47) of cats were parasitized, and 2,358 flukes (P. fastosum) were collected (131 – mean intensity of parasitism; 50.2 – mean abundance). The alanine transaminase (ALT) measure was significantly higher in parasitized animals, while alkaline phosphatase (ALP) showed no statistical difference between parasitized and non-parasitized animals. In infected animals, gross pathological lesions and microscopic liver injuries ranged from mild to severe, and were similar to those in previous descriptions of feline platynosomiasis. Nonetheless, the intensity of parasitism was not related to the severity of macroscopic or microscopic hepatic injuries. However, feline platynosomiasis should be considered in the differential diagnosis of feline liver disorders, as well as, in any program of helminth control, even if no clinical abnormalities are present.

Resumo Platinossomiase é uma doença hepática felina comum causada por Platynosomum fastosum (Trematoda - Dicrocoelidae), também é conhecida como “envenenamento por lagartixa”. A maioria dos relatos de platinossomiase felina mostra que esta doença é esporádica e se manifesta com lesões incomuns; sua patogenicidade ainda não é bem compreendida. Este estudo objetivou descrever as lesões no fígado e alterações enzimáticas associadas à infecção natural por P. fastosum em 47 gatos errantes em uma área endêmica. No total, 38,3% (18/47) dos gatos estavam parasitados, e 2.358 trematódeos (P. fastosum) foram coletados (131 – intensidade média de parasitismo; 50,2 – abundância média). A quantidade de alanina transaminase (ALT) foi significativamente maior nos animais parasitados, enquanto a fosfatase alcalina (ALP) não apresentou diferença estatística entre os animais parasitados e não parasitados. Nos animais infectados, lesões patológicas macroscópicas e microscópicas hepáticas variaram de leve a grave, e foram semelhantes a descrições anteriores de platinossomiase felina. No entanto, a intensidade do parasitismo não foi relacionada à gravidade das lesões hepáticas macroscópicas ou microscópicas. Contudo, a platinossomiase felina deve ser considerada no diagnóstico diferencial de distúrbios hepáticos em felinos, assim como, em qualquer programa de controle de helmintos, mesmo que nenhuma anormalidade clínica esteja presente.

Cattle naturally infected by Eurytrema coelomaticum: Relation between adenosine deaminase activity and zinc levels.

The enzyme adenosine deaminase (ADA) is critical for modulating the immune system, and in the presence of zinc, its activity is catalyzed. The aim of this study was to evaluate the ADA activity in pancreas of cattle naturally infected by Eurytrema coelomaticum in relation to the results of zinc levels, pathological findings and parasite load. For this study 51 slaughtered cattle were used. The animals were divided into two groups: Group A consisting of animals naturally infected by E. coelomaticum (n=33) and Group B of uninfected animals (n=18). Blood and pancreas were collected of each animal for analysis of zinc and ADA, respectively. Infected cattle showed a reduction on seric levels of zinc, and decreased ADA activity in the pancreas (P>0.05). A positive correlation between zinc levels and ADA activity was observed. Thus, high parasite load and severity of histopathologic lesions affect the ADA activity in pancreas, as well as the zinc levels in serum of infected animals (negative correlation between these variables). Therefore, we can conclude that cattle infected by E. coelomaticum have low ADA activity in pancreas, which can be directly related to zinc reduction, responsible for ADA activation and catalyzes.

NTPDase and 5'-nucleotidase as inflammatory markers in cattle naturally infected by Eurytrema coelomaticum.

The aim of this study was to evaluate seric NTPDase and 5'nucleotidase activities of cattle naturally infected by Eurytrema coelomanticum, as well as to correlate them to histopathological lesions in the pancreas and the degree of parasitism. Blood samples and pancreas of 51 bovines were collected on a slaughterhouse in Southern Brazil: 33 from cattle naturally infected by E. coelomanticum (the Group A), and 18 from uninfected animals (the Group B). Infected animals showed an average of 532 parasites per pancreas. In the pancreatic histology, ducts displayed hyperplasia, stenosis, proliferation of fibrous tissue, and interstitial inflammatory infiltration of lymphocytes. The serum from infected animals showed an increase in NTPDase activity when ATP was used as substrate (P<0.001). For the ADP substrate, there was no difference between groups regarding NTPDase activity (P=0.37), as well as 5'-nucleotidase activity (P=0.27). Correlating NTPDase activity (ATP substrate) with the degree of histopathological lesions (rho=0.66, P<0.001) and the parasitic load on the pancreas (rho=0.65, P<0.001), a positive correlation was observed. Similar results were found between the degree of histopathological lesions and NTPDase activity (ADP substrate; rho=0.29, P=0.03), and 5'nucleotidase activity (rho=0.35, P=0.01). Based on the results of NTPDase and 5'nucleotidase enzymes in cattle naturally infected by E. coleomanticum, it is possible to suggest that these enzymes are involved in the modulation of inflammation, and they can act as markers of inflammatory response.

Platynosomum fastosum (Digenea: Dicrocoeliidae) infection in a domestic cat in northeastern Brazil: high fluke burden and associated lesions / Infecção por Platynosomum fastosum (Digenea: Dicrocoeliidae) em gato doméstico no nordeste do Brasil: alta carga parasitária e lesões associadas

Introduction: Platynosomum fastosum is a dicrocoeliid fluke that infects birds and mammals, especially wild and domestic cats. Case report: A middleaged male domestic cat was euthanized and examined. Its clinical features had included apathy, emaciation, hepatomegaly and mild jaundice on the mucosae. At necropsy, we noticed enlarged liver and dilated gallbladder. After opening the gallbladder lengthwise, multiple whitish flattened ellipsoidal structures were observed and identified as P. fastosum. The larger parasites were found in gallbladder, with significant different size from cystic duct and intrahepatic biliary ducts (p<0.05). Histopathological analysis revealed fluke-induced suppurative cholecystitis and chronic cholangitis. Precursor lesions of cholangiocarcinoma were reported. Conclusion: This report was part of the first prevalence survey of P.fastosum in municipality of Maracanaú, state of Ceará, northeastern Brazil. (AU)

Introdução: Platynosomum fastosum é um trematódeo dicrocelídeo que infecta aves e mamíferos, especialmente felídeos domésticos e silvestres. Relato do caso: Um gato doméstico macho, apresentando apatia, emaciação, hepatomegalia e icterícia leve nas mucosas, foi submetido à eutanásia e subsequente necropsia. Ao exame macroscópico foi observada hepatomegalia e dilatação da vesícula biliar. Após abertura da vesícula, foram observadas múltiplas estruturas foliáceas esbranquiçadas, posteriormente identificadas como P. fastosum. Os maiores exemplares foram encontrados na vesícula, com tamanho estatisticamente diferente daqueles encontrados no ducto cístico e nos ductos biliares intrahepáticos (p<0.05). Análise microscópica revelou colecistite supurativa e colangite crônicas induzidas pela infecção parasitária. Lesões precursoras de colangiocarcinoma foram reportadas. Conclusão: este relato está inserido no primeiro estudo de prevalência de P. fastosumno município de Maracanaú, Ceará, nordeste do Brasil. (AU)

De novo assembly and characterization of the transcriptome of the pancreatic fluke Eurytrema pancreaticum (trematoda: Dicrocoeliidae) using Illumina paired-end sequencing.

Eurytrema pancreaticum is one of the most common trematodes living in the pancreatic and bile ducts of ruminants and also occasionally infects humans, causing eurytremiasis. In spite of its economic and medical importance, very little is known about the genomic resources of this parasite. Herein, we performed de novo sequencing, assembly and characterization of the transcriptome of adult E. pancreaticum. Approximately 36.4 million high-quality clean reads were obtained, and the length of the transcript contigs ranged from 66 to 19,968 nt with mean length of 479 nt and N50 length of 1094 nt, and then 23,573 unigenes were assembled. Of these unigenes, 15,353 (65.1%) were annotated by blast searches against the NCBI non-redundant protein database. Among these, 15,267 (64.8%), 2732 (11.6%) and 10,354 (43.9%) of the unigenes had significant similarity with proteins in the NR, NT and Swiss-Prot databases, respectively. 5510 (23.4%) and 4567 (19.4%) unigenes were assigned to GO and COG, respectively. 8886 (37.7%) unigenes were identified and mapped onto 254 pathways in the KEGG Pathway database. Furthermore, we found that 105 (1.18%) unigenes were related to pancreatic secretion and 61 (0.7%) to pancreatic cancer. The present study represents the first transcriptome of any members of the family Dicrocoeliidae, which has little genomic information available in the public databases. The novel transcriptome of E. pancreaticum should provide a useful resource for designing new strategies against pancreatic flukes and other trematodes of human and animal health significance.

Molecular characterization of Eurytrema coelomaticum in cattle from Paraná, Brazil / Caracterização molecular de Eurytrema coelomaticum em bovinos do estado do Paraná, Brasil

This study investigated the occurrence of Eurytrema spp. in cattle by analysis of the partial 18S rRNA gene sequence. Trematodes from 44 bovine pancreas were collected and classified based on typical morphological features. PCR assay and sequence analyses of amplified products confirmed that the trematodes classified as Eurytrema coelomaticum were phylogenetically distinct from those identified as E. pancreaticum. The results of this study represent the first molecular characterization of E. coelomaticum within the Americas, and provide an efficient method to differentiate digenean trematodes of domestic animals.

Este estudo avaliou a ocorrência de Eurytrema spp. em bovinos por meio da análise da sequência parcial do gene 18S rRNA. Trematódeos procedentes de 44 pâncreas bovinos foram coletados e classificados com base em aspectos morfológicos típicos. A técnica de PCR e a análise da sequência de nucleotídeos dos produtos amplificados confirmaram que os trematódeos classificados como Eurytrema coelomaticum eram filogeneticamente distintos daqueles identificados como E. pancreaticum. Os resultados deste estudo representam a primeira caracterização molecular de E. coelomaticum nas Américas e disponibiliza um método eficiente para diferenciar trematódeos digenéticos em animais domésticos.

Ocorrência de Platynosomum illiciens em felinos selvagens mantidos em cativeiro no estado da Bahia, Brasil / Occurrence of Platynosomum illiciens in enclosured wild cats in the state of Bahia, Brazil

This paper reports the occurrence of eggs of Platynosomum illiciens, found in the feces of three species of wild cats Herpailurus yagouaroundi, Puma concolor and Leopardus tigrinus from material collected at the enclosures of the Park Zoobotânico Getúlio Vargas in Salvador, Bahia, Brazil. It is the first record of this parasite in P. onca and L. tigrinus.

O presente trabalho relata a ocorrência de ovos de Platynosomum illiciens nas fezes de três espécies de felídeos silvestres Herpailurus yagouaroundi, Panthera onca e Leopardus tigrinus coletados nos recintos do Parque Zoobotânico Getúlio Vargas em Salvador, Bahia, Brasil. Sendo o primeiro registro deste parasito em P. onca e F. tigrinus.

Eurytrema procyonis and pancreatitis in a cat.

A young adult male domestic shorthair cat was presented for physical examination, routine vaccinations, and a fecal examination. Physical examination revealed no significant abnormalities. Eggs of the raccoon pancreatic fluke Eurytrema procyonis were detected by fecal flotation. Results of a complete blood count and serum biochemistry panel were normal. Abdominal sonography revealed an enlarged hypoechoic pancreas with a hyperechoic rim, and a distended and thickened pancreatic duct. Serum pancreatic lipase immunoreactivity (PLI) was increased. These findings supported the possibility of fluke-associated pancreatitis. Treatment with praziquantel/pyrantel/febantel was associated with resolution of sonographic abnormalities and normalization of PLI.

Aspectos quantitativos e morfométricos das infrapopulações de platynosomum illiciens (trematoda: dicrocoeliidae) em paroaria dominicana (passeriformes: emberizidae) do estado da Bahia, Brasil / Quantitative and morfometrical aspects in infrapopulations of platynosomum illiciens (trematoda: dicrocoeliidae) in paroaria dominicana (passeriformes: emberizidae) from the state of Bahia, Brazil

Durante o estudo dos metazoários parasitos de Paroaria dominicana Linnaeus, 1758 (Passeriformes), quatro infrapopulações de Platynosomum illiciens (Brown, 1901) (Trematoda) foram coletadas nos ductos hepáticos, com intensidades parasitárias de 10, 11, 18 e 36 espécimes respectivamente. Diferenças entre os valores morfométricos médios e o número total de ovos das infrapopulações foram estatisticamente testadas. O peso do fígado não se correlacionou ao peso das aves, ao comprimento do corpo das aves e à abundância parasitária. Foram detectadas correlações positivas entre o tamanho das infrapopulações e o número de ovos produzidos e entre o tamanho do corpo dos parasitos e o tamanho dos ovos. Correlações negativas foram observadas entre a intensidade parasitária e o peso do fígado; entre o tamanho das infrapopulações e o tamanho dos indivíduos dessas infrapopulações; entre o número e o tamanho dos ovos e entre o tamanho do corpo dos parasitos e o número de ovos. Os resultados indicaram que as infrapopulações maiores se estabeleceram em aves com fígados menores e que quanto maiores as infrapopulações foram observados menor tamanho corporal e menor tamanho dos ovos, os que se apresentaram em maior quantidade.

During the study of the metazoan parasites of Paroaria dominicana Linnaeus, 1758 (Passeriformes), four infrapopulations of Platynosomum illiciens (Brown, 1901) (Trematoda), parasitic in the liver ducts were collected, with intensity of infection 10, 11, 18 and 36 specimens respectively. Differences between the means of morphometrical values and the total number of eggs were statistically tested. The liver weigh were not significantly correlated with avian total weigh and length, and with the total parasitic abundance. Positive correlations were observed between the size of infrapopulations and the total number of eggs, and between parasite body size and the eggs. size. Negative correlations were observed between parasitic intensity and liver weigh; between the size of infrapopulations and the parasite body size; between total number and the size of eggs and between parasite body size and total number of eggs. These results suggests that host with smaller liver had greater infrapopulations. These greater infrapopulations showed smaller parasite body size and higher size and number of eggs.

Eurytrema pancreaticum: the in vitro effect of praziquantel and triclabendazole on the adult fluke.

The efficacy and tolerance of the 80 microg/ml praziquantel (PZQ) and 40 microg/ml triclabendazole (TCZ) against adult stage Eurytrema pancreaticum in vitro were investigated at 3, 12, and 15 h incubation. Motility of the flukes and histopathological changes were studied. Sudden paralysis and death were observed after exposed to PZQ as early as 3h incubation. In contrast, the TCZ treated flukes showed active mobility at all intervals. By light microscopic examination, severe damages in various organs such as tegument, muscle, and testes were observed early at 12h incubation of these drugs. PZQ caused more severe damage to flukes than TCZ. There were vigorous contraction of musculature, progressive shrinkage of circular and longitudinal muscles, vacuolization and disintegration of the tegument disrupting the worms' outer surface including detachment of spines in the PZQ treatment. The cells in testes were slightly increased in size and followed by degeneration leaving several hollow spaces. The uterus and vitelline glands remained unaffected. The direct observation of the fluke motility and light microscopic study highly suggested that PZQ was more effective than TCZ treatment for the eurytremiasis infection.

Quantitative hepatobiliary scintigraphy in normal cats and in cats with experimental cholangiohepatitis.

Quantitative hepatobiliary scintigraphy using 99mTc-mebrofenin was performed on eight normal cats and on the same cats after induction of experimental cholangiohepatitis by infection with the liver fluke Platynosomum concinnum. Hepatobiliary scintigraphy was performed 3 times at 10 weeks, 4 months and 6 months after infection. In addition, routine biochemical tests, hepatic ultrasound and ultrasound guided hepatic biopsy samples were obtained at the same time points, and the results compared with hepatobiliary scintigraphy. The normal hepatic extraction fraction was determined to be 85%, and the normal hepatic excretion half time (T 1/2) was 14 minutes. There was no significant change in scintigraphic parameters compared to pre-infection values at any time following infection with the liver fluke. No correlation between scintigraphic parameters and histologic scores was found; however, significant correlation was identified between parasite burden and histologic scores 6 months following infection. Despite the presence of severe multifocal histologic abnormalities, minimal clinical, biochemical and scintigraphic derangements were identified using this model of cholangiohepatitis. Based on this study, hepatobiliary scintigraphy appears to be an insensitive test for structural hepatobiliary abnormalities. The role of hepatobiliary scintigraphy in functional hepatobiliary abnormalities of the feline liver has not been determined.