Hepatobiliary Ascariasis in a Piglet.

PURPOSE: Ascariasis caused by the helminth Ascaris suum is the most common parasitosis of swine worldwide and it may involve all age categories of pigs. The present study reports an unusual localization of A. suum worms in the biliary system of a piglet slaughtered for human consumption. METHODS: The liver was subjected to ultrasound scan and pathological examination. The isolated worms were morphologically examined and the DNA was extracted for the molecular identification of the species involved. RESULTS: A total of 43 preadult nematodes were found within the gallbladder and the bile ducts. Parasites were morphologically identified as belonging to the genus Ascaris and molecularly as A. suum. At gross examination, the liver was moderately enlarged, with the bile ducts severely dilated. A chronic inflammatory infiltrate was noted, often centered around ectatic bile ducts (up to 5 mm in diameter), lined by hyperplastic epithelium and filled with sections of nematodes. The worm sections showed smooth cuticle, coelomyarian musculature, and an intestinal tract lined by columnar, uninucleated cells within a pseudocoelom. The ex vivo ultrasonographic examination of the liver allowed the visualization of several nematodes in the bile duct lumen and could be suggested for in vivo diagnosis. Unfortunately, the absence of the intestine did not allow to define the pathogenesis of the infection. CONCLUSION: Although, given the unusual nature of this finding, it is difficult to identify predisposing factors for this A. suum localization, it suggests that ascariasis should be considered in the differential diagnosis of pigs with hepatobiliary disease.

Preliminary evaluation of the protective effects of recombinant AMA1 and IMP1 against Eimeria stiedae infection in rabbits.

BACKGROUND: Eimeria stiedae parasitizes the bile duct, causing hepatic coccidiosis in rabbits. Coccidiosis control using anticoccidials led to drug resistance and residues; therefore, vaccines are required as an alternative control strategy. Apical membrane antigen 1 (AMA1) and immune mapped protein 1 (IMP1) are surface-located proteins that might contribute to host cell invasion, having potential as candidate vaccine antigens. METHODS: Herein, we cloned and expressed the E. stiedae EsAMA1 and EsIMP1 genes. The reactogenicity of recombinant AMA1 (rEsAMA1) and IMP1 (rEsIMP1) proteins were investigated using immunoblotting. For the vaccination-infection trial, rabbits were vaccinated with rEsAMA1 and rEsIMP1 (both 100 µg/rabbit) twice at 2-week intervals. After vaccination, various serum cytokines were measured. The protective effects of rEsAMA1 and rEsIMP1 against E. stiedae infection were assessed using several indicators. Sera were collected weekly to detect the specific antibody levels. RESULTS: Both rEsAMA1 and rEsIMP1 showed strong reactogenicity. Rabbits vaccinated with rEsAMA1 and rEsIMP1 displayed significantly increased serum IL-2 (F (4, 25) = 9.53, P = 0.000), IL-4 (F (4, 25) = 7.81, P = 0.000), IL-17 (F (4, 25) = 8.55, P = 0.000), and IFN-γ (F (4, 25) = 6.89, P = 0.001) levels; in the rEsIMP1 group, serum TGF-ß1 level was also elevated (F (4, 25) = 3.01, P = 0.037). After vaccination, the specific antibody levels increased and were maintained at a high level. The vaccination-infection trial showed that compared with the positive control groups, rabbits vaccinated with the recombinant proteins showed significantly reduced oocyst output (F (5, 54) = 187.87, P = 0.000), liver index (F (5, 54) = 37.52, P = 0.000), and feed conversion ratio; body weight gain was significantly improved (F (5, 54) = 28.82, P = 0.000). CONCLUSIONS: rEsAMA1 and rEsIMP1 could induce cellular and humoral immunity, protecting against E. stiedae infection. Thus, rEsAMA1 and rEsIMP1 are potential vaccine candidates against E. stiedae.

Feline platynosomiasis in Klang Valley, Malaysia: Diagnosis, pathology and management.

Platynosomum fastosum (synonym Platynosomum concinnum, Platynosomum illiciens ) is a hepatic fluke causing platynosomiasis or 'lizard poisoning' in cats. This disease is generally being underestimated by veterinary practitioners due to lack of awareness and difficulty in diagnosis although the severe cases of platynosomiasis could be fatal. This study was designed to detect the presence of cat liver fluke through faecal examination among cats kept in shelters, pet cats and stray cats around Klang Valley, Malaysia. The detection of liver flukes among stray cats was based on post-mortem examination. A total of 201 faecal samples were collected from eight shelters (n = 119) and five veterinary clinics (n = 82) in Klang Valley were subjected to simple floatation and formalin-ether sedimentation techniques for ova detection. P. fastosum ova were identified in three faecal samples obtained from shelters (2.52%) and three samples collected from veterinary clinics (3.66%) by faecal examination. A total of 51 stray cats were procured from Pest and Animal Control Unit of the respective municipality. The cats were euthanised and necropsied to collect liver samples and bile duct. The liver was then dissected for isolation and identification of the fluke and bile duct fluid were aspirated for detection of fluke ova. Twelve cats (23.5%) were positive for P. fastosum and ova were found in their bile. Macroscopically, affected cats showed mottled liver (33.3%), distended gall bladder with thick tenacious bile (66.7%) that microscopically exhibited hepatic steatosis (25.0%) and hepatitis (33.3%). The severity of parasite load was almost equally distributed between the positive cats with low (n = 5, 55.6%; > 125 adult fluke) and high parasite burden (n = 4, 44.4%; < 125 adult fluke) cats, respectively. This study revealed the detection of P. fastosum among pet, shelters and stray cats in Klang Valley, Malaysia.

Hepatic Capillaria hepatica (Bancroft, 1893) infection in cat (Felis catus)-histopathological findings and first report from Iran.

Capillaria hepatica (syn. Calodium hepaticum) is a globally distributed nematode with a high affinity to the liver of a wide range of mammalian hosts, including humans. Documented reports of the nematode in cats and associated histopathology are rare. Here, we describe a case of C. hepatica infection in a 5-year-old male stray cat from Iran. At post-car accident necropsy, all body parts appeared normal except for the liver, in which a few yellowish-white granulomatous nodules were observed through the capsule and in the organ. Histopathological examination of the tissue revealed a large number of clustered parasite eggs in the parenchyma. The barrel-shaped, un-embryonated eggs (55.19 × 28.37 µm), with inconspicuous caps at both ends, were covered with striated shells. The presence of ova in the liver tissue had resulted in the development of hepatic inflammation with hepatocellular necrosis associated with the development of multifocal granulomas. As predators of small rodents, the cats might have a significant role in the epidemiology of C. hepatica. Infection of hosts through ingestion of embryonated eggs in contaminated water, food, or soil is of major importance in the epidemiology of C. hepatica. Since the rare reports of feline infection have come mainly from accidental detection of the parasite, any hepatic disease presenting difficulties to find an etiological agent may virtually be associated with the infection with this little-known nematode.

Liver fluke infections by Amphimerus sp. (Digenea: Opisthorchiidae) in definitive and fish intermediate hosts in Manabí province, Ecuador.

Amphimerus sp. is a fluke that dwells in the biliary tracts of vertebrate definitive hosts including humans, domestic, and wild mammals in Latin America. Opisthorchiid liver infections are rarely studied in the Americas confirming its status as a neglected tropical disease. In Ecuador, small trematode eggs were reported in human cases from the province of Manabí in 1949, and recently, Amphimerus sp. adults were recovered from human and reservoir hosts in the province of Esmeraldas. Due to the lack of research on the infectious sources of Amphimerus sp. in the continent, we have developed a series of epidemiological studies with parasitological and molecular techniques to elucidate the endemicity of opisthorchiid fluke infections. We developed a cross-sectional study in three communities at Pedro Pablo Gómez parish in the province of Manabí, Ecuador. We examined a total of 176 fecal samples to detect opisthorchiid eggs, and four fish species to find opisthorchiid metacercariae. To study adult worms, we treated and purged seven patients in a family and dissected the livers of a dog and a cat infected. We observed morphological features of adults and metacercariae and used polymerase chain reaction with restricted fragment length polymorphism (PCR-RFLP) and DNA sequencing of a section of the ITS2 gene for identification. Small trematode eggs were detected in 63 (35.8%) out of 176 fecal samples of residents in the three study sites. Adult opisthorchiid flukes were recovered from human patients, a dog and a cat, and they were morphologically and molecularly identified as Amphimerus sp. Opisthorchiid metacercariae were also identified molecularly as Amphimerus sp. in four fish species, i.e., Rhoadsia altipinna, Bryconamericus bucay, Andinoacara rivulatus, and Piabucina aureoguttata. Metacercariae of the heterophyid Haplorchis pumilio were also found in the four fish species examined. This is the first study to confirm the current endemicity of Amphimerus sp. in Pedro Pablo Gómez, Manabí, Ecuador. The adult worms isolated here shared morphological characteristics with previous Amphimerus sp. descriptions and were molecularly similar to Amphimerus sp. described in the province of Esmeraldas. Moreover, this study is the first to document four fish species as infection sources of Amphimerus sp. detected via a molecular protocol targeting the metacercariae of the parasite. Fish species identified here should be targeted for public health campaigns to avoid further human liver-fluke infections by Amphimerus sp. or potential intestinal-fluke infections by H. pumilio or others.

Chronic whipworm infection exacerbates Schistosoma mansoni egg-induced hepatopathology in non-human primates.

BACKGROUND: Schistosomiasis continues to inflict significant morbidity and mortality in the tropical and subtropical regions of the world. The disease endemicity overlaps with the transmission of other parasitic diseases. Despite the ubiquity of polyparasitism in tropical regions, particularly in rural communities, little is known about the impact of multiple helminth infections on disease progression. In this pilot study, we describe the influence of chronic Trichuris trichiura infection on Schistosoma mansoni egg-induced hepatopathology in infected baboons. METHODS: Baboons with or without underlying whipworm infection were challenged with S. mansoni cercariae to establish schistosomiasis. Adult S. mansoni worms were recovered by perfusion and enumerated, hepatic granulomas were quantified via light microscopy, and transcriptional profiling of tissues were completed using RNA sequencing technologies. RESULTS: Co-infection with both S. mansoni and T. trichiura resulted in higher female schistosome worm burden and significantly larger liver granuloma sizes. Systems biology analyses of peripheral blood mononuclear cells (PBMC) revealed pathways associated with increased liver damage in co-infected baboons. CONCLUSIONS: Underlying chronic whipworm infection intensified schistosome egg-induced liver pathology in infected baboons. RNA-Seq analysis provided insight into pathways associated with increased liver damage, corroborating histological findings.

Therapeutic effect of Moringa oleifera and Thymus vulgaris oils against hepatic coccidiosis in experimentally infected rabbits.

The present study was conducted to detect the therapeutic effect of Moringa oleifera and Thymus vulgaris oils on hepatic coccidiosis in experimentally infected rabbits. Also, immunomodulatory effect of the two oils was detected. Twenty-four Newzealand rabbits were used in this study and divided into 4 groups; healthy rabbits, experimentally infected rabbits with Eimeria stiedae oocysts, and two infected treated groups (one with moringa (200 mg/kg) and the other with thyme (500 mg/kg) oils). The results showed highly significant reduction in oocysts shedding (P<0.001 and P<0.05) in the two infected and treated rabbits than the infected non-treated rabbits in almost all days post infection (PI). Thyme oil was more potent and stopped oocysts shedding earlier at the day 34 PI compared to moringa oil at the day 41 PI. Microscopically, there was a damage in the oocysts shed by treated rabbits. Macroscopically, the livers of thyme oil treated rabbits showed more enhancement with protection percentage 75% than those treated with moringa oil in which protection percentage was 55%. The highest titer of antibodies was detected in moringa oil treated rabbits. It was concluded that both moringa and thyme oils had an anti-coccidial effect with thyme oil superiority. So, thyme oil could be useful as an alternative product for the control of rabbit coccidiosis.

Assessment of histological liver alterations in dogs naturally infected with Leishmania infantum.

BACKGROUND: The liver plays a central role in the development of canine visceral leishmaniasis. Studies of natural infection in animals and humans indicate a direct relationship between resolution of infection and the formation and maturation of granulomas in the liver. However, in contrast to other reports in the literature, the present study found no differences in the characteristics of hepatic granulomas that could be related to resistance or susceptibility to Leishmania. Here, we describe the hepatic alterations observed in dogs with differing clinical manifestations of visceral leishmaniasis in an endemic area in the state of Bahia, Brazil. METHODS: We examined 148 animals in an endemic area. The animals were clinically examined, and the infection was determined by ELISA, spleen aspirate culture and quantitative PCR. The animals were grouped into asymptomatic or symptomatic based on the number of signs of LV. The histological liver evaluation was performed in a blinded way. RESULTS: Our results indicated no association between the characteristics of granulomas and clinical presentation. We found an association between the intensity of this inflammatory response and parasite load in the animals' spleens. It is important to note that while hepatic alterations, such as portal and perivascular inflammation and the presence of larger amounts of granulomas, were linked with higher parasite loads, we found the inverse to be true with respect to intrasinusoidal lymphocytosis, the formation of intrasinusoidal inflammatory cell aggregates and Kupffer cell hypertrophy. CONCLUSIONS: Our findings suggest that the presence of mononuclear inflammatory cells inside the sinusoids is more important than that of organized granulomas in terms of the containment of parasitism by the host. We suggest that the presence of granulomas indicates the failure of a first line of defense mechanism in the control of parasite infection, which could be related to the presence of inflammatory cells and Kupffer cell hypertrophy inside the sinusoids. We further demonstrated that dogs with active Leishmania spp. infection present a higher frequency of inflammatory changes in the liver. In addition to being correlated with the severity of clinical manifestation, these hepatic alterations were also associated with changes in hematological and biochemical parameters.

In vitro anthelmintic effect of biologically synthesized silver nanoparticles on liver amphistome, Gigantocotyle explanatum.

In order to ensure global food security a rationale approach is required to control all those factors which directly or indirectly affect the food productivity. The neglected helminthic diseases alone are responsible for huge economic losses to the agrarian stakeholders. The problem is further compounded by the emerging drug resistance in flukes against the commonly used anthelmintics like triclabendazole. Therefore, the search for alternatives including the nano-based approaches has become a necessity to develop future control strategies. In the present study the effect of biologically synthesized silver nanoparticles (AgNPs) was investigated on an economically important amphistome parasite, Gigantocotyle explanatum, obtained from the infected liver of the Indian water buffaloes, Bubalus bubalis. In vitro treatment of the adult worms with different doses of AgNPs severely affected the worm motility and caused ROS mediated damages in the treated flukes. The antioxidant system and the detoxification ability of the worms appeared to be disrupted along with pronounced DNA damage in the treated worms as compared to the controls. Following the treatment of worms with different concentrations of AgNPs there was a significant (p < 0.05) increase in lipid peroxidation and protein carbonylation levels which are the key oxidative stress markers. The tegumental surface which is metabolically active, was severely damaged as evident from the loss of papillae, severe blebbing, shearing and erosion of the surface structures. Such topographical disruptions would facilitate the penetration of the nanoparticles deep within the tissues that might greatly reduce the invasive potential of the flukes as evident from the decreased motility. Taken together our findings suggest that the AgNPs posses great anthelmintic potential and could be further exploited for the development of anthelmintic formulations which may be tested in vivo.

Hepatic Dicrocoelium dendriticum infection in a miniature horse.

A miniature horse which died following humane seizure from an Ontario farm was emaciated with serous atrophy of fat. Autopsy revealed hepatic atrophy and moderate periportal fibrosis. Eggs and trematodes seen in the liver were identified as Dicrocoelium dendriticum. This appears to be the first reported case of infection of equids in North America with Dicrocoelium dendriticum.

Infection parDicrocoelium dendriticumhépatique chez un cheval miniature. Un cheval miniature qui est mort à la suite d'une saisie humanitaire dans une ferme de l'Ontario était émacié et avait une atrophie séreuse des tissus adipeux. L'autopsie a révélé une atrophie hépatique et une fibrose périportale modérée. Des oeufs et des trématodes observés dans le foie ont été identifiés comme Dicrocoelium dendriticum. Il semblerait qu'il s'agit du premier cas signalé d'infection des équidés par Dicrocoelium dendriticum en Amérique du Nord.(Traduit par Isabelle Vallières).

Infections by Hepatic Trematodes in Cats from Slaughterhouses in Vietnam.

Domestic cats are a final host for some hepatic trematodes, and cat meat is used for human consumption in several parts of northern Vietnam. Since there is no tradition for commercial cat rearing in the area, cats are purchased alive from other areas as well, i.e., southern Vietnam and neighboring countries, and brought to slaughterhouses. This uncontrolled trade in live cats could pose a risk for spread of various parasites, including hepatic trematodes. Hence, in this study, we investigated the infections by hepatic trematodes in cats from the slaughterhouses. Cat livers were sampled from 12 slaughterhouses. Clonorchis sinensis and Platynosomum fastosum were found in 14 of 78 necropsied cats of local origin, while more than half of the non-local cats (170 of 318) were infected by C. sinensis, P. fastosum, or Opisthorchis viverrini. The non-local cats had higher prevalence (odds ratio = 6.61, P < 0.01, 95% CI: 2.34-19.41) and intensity of infection (count ratio = 6.47, P < 0.01, 95% CI: 1.77-23.59) by C. sinensis than local cats. Prevalence of P. fastosum infection did not differ significantly between the 2 groups of cats. Opisthorchis viverrini was found at low prevalence (2.5%) in non-local cats. The presence of O. viverrini in cats sampled in northern Vietnam and the high prevalence and intensity of infection in cats of non-local origin suggest that cats are transported over great distances, and this poses a risk of spreading these trematodes.

Feline platynosomiasis: analysis of the association of infection levels with pathological and biochemical findings.

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.

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.

Molecular and phylogenetic analyses of the liver amphistome Explanatum explanatum (Creplin, 1847) Fukui, 1929 in ruminants from Bangladesh and Nepal based on nuclear ribosomal ITS2 and mitochondrial nad1 sequences.

Explanatum explanatum flukes, liver amphistomes of ruminants, cause significant economic loss in the livestock industry by inducing severe liver damage. A total of 66 flukes from 26 buffaloes and 7 cattle in four different geographic areas of Bangladesh and 20 flukes from 10 buffaloes in the Chitwan district of Nepal were subjected for analysis. The sequences (442 bp) of the second internal transcribed spacer (ITS2) of ribosomal DNA and the variable fragments (657 bp) of mitochondrial nicotinamide dehydrogenase subunit 1 (nad1) of E. explanatum flukes from Bangladesh and Nepal were analysed. The aim of this study was molecular characterization of the flukes and to elucidate their origin and biogeography. In the ITS2 region, two genotypes were detected among the flukes from Bangladesh, while flukes from Nepal were of only one genotype. Phylogenetic analyses inferred from the nad1 gene revealed that at least four divergent populations (groups I-IV) are distributed in Bangladesh, whereas two divergent populations were found to be distributed in Nepal. Fst values (pairwise fixation index) suggest that Bangladeshi and Nepalese populations of group I to IV are significantly different from each other; but within groups III and IV, the populations from Bangladesh and Nepal were genetically close. This divergence in the nad1 gene indicates that each lineage of E. explanatum from diverse geography was co-adapted during the multiple domestication events of ruminants. This study, for the first time, provides molecular characterization of E. explanatum in Bangladesh and Nepal, and may provide useful information for elucidating its origin and dispersal route in Asia.

Comparative Risk of Liver and Intestinal Fluke Infection from Either Wild-Caught or Cultured Fish in Vietnam.

A survey for fish-borne zoonotic trematode (FZT) metacercariae in fish from the Northern Mountain Region of Vietnam was conducted from May to August 2014. The major objective was to evaluate the impact of water body types on the prevalence of FZT. A total of 846 fish (31 species) were collected from three water body types, including aquaculture ponds, rivers, and reservoirs. A pepsin digestion method was used for the recovery of metacercariae from fish. Overall, 29 of the 31 fish species were infected with FZTs. Prevalence was 56.8% in river fish, 46.7% in pond fish, and 29.3% in reservoir fish. The prevalence of Clonorchis sinensis metacercariae was 10.6% in reservoir fish, and 2.6% in river fish; fish from ponds were uninfected. The infection intensity of C. sinensis was 29.7 metacercariae/reservoir fish and 2.1 metacercariae/river fish. The prevalence of C. sinensis was highest (25.9%) in Toxabramis houdemeri. The intestinal flukes Haplorchis pumilio, Haplorchis taichui, Haplorchis yokogawai, Centrocestus formosanus and Procerovum varium were recovered from 29 fish species, with an overall prevalence of 46.8% and a mean infection intensity of 23.6 metacercariae. The results indicate that a C. sinensis sylvatic cycle involving wild fish species is important in the epidemiology of liver fluke and that consumption of raw or inadequately prepared wild-caught fish is a the major risk factor for human Clonorchiasis in this region. Both wild-caught and cultured fish present a high risk for infection with intestinal flukes. The habitat requirements of the snail host may be the important determining factor in favoring a sylvatic cycle of C. sinensis. This information is relevant to the development of effective prevention and control strategies, and emphasizes the priority that must be given to education of the local communities on the risks of consuming raw or improperly prepared fish dishes.

A massive invasion of Parafasciolopsis fasciolaemorpha in elk (Alces alces) in Lublin Province, Poland.

UNLABELLED: Liver fluke Parafasciolopsis fasciolaemorpha is a parasite typical of elk (Alces alces). The present study describes the identification of a massive invasion of the liver fluke P. fasciolaemorpha in an elk in the Sobibór Landscape Park, Eastern Poland. In February 2016, samples of liver and faeces were collected from the elk during postmorte examination. A section of liver tissue and three grams of faeces were examined for the presence of flukes or fluke eggs by decantation. In total, 11,150 juvenile and mature flukes of P. fasciolaemorpha were found in the examined liver. Multiple cavities with distinct walls, filled with dark liquid, trematodes, fluke eggs and cellular detritus were seen in the cross section of the parenchyma. Additionally, some of the bile ducts were plugged with calcerous deposits. The three grams of examined faeces were found to contain 322 grey and golden-coloured eggs of P. fasciolaemorpha. The presence of such a massive infection of P. fasciolaemorpha in the examined elk has a significant impact on the health of the individual. We can assume P. fasciolaemorpha has a significant influence on the health and abundance of the entire elk population in Poland. KEY WORDS: Parafasciolopsis fasciolaemorpha, elk, Sobibór Landscape Park, Poland.

Prevalence of Platynosomum fastosum infection in free roaming cats in northeastern Brazil: Fluke burden and grading of lesions.

The objective of this paper was to investigate prevalence and parasite load of the trematode Platynosomum fastosum infection and the risk of developing cholangitis/cholangiohepatitis among infected domestic cats in a city of northeastern Brazil, and to characterize the influence of fluke burden in the formation of lesions in liver and gallbladder. A total of 141 cats was necropsied and examined for the presence of P. fastosum. The observed prevalence was 42.6% (60/141, 95% CI=34.2-50.9). Parasite load varied from 1 to 219 flukes per animal. Adult cats were more frequently infected than kittens (p<0.01). Risk of cholangitis was about three times higher in infected animals (RR=3.23, 95% CI=2.01-5.07, p<0.01). Centrolobular congestion, portal cholangitis and cholestasis was significantly more common in infected as compared to non-infected cats (p<0.01). Similarly, fibrosis, cholangitis and mucous gland hyperplasia were significantly more prevalent in the bile duct of infected animals (p<0.01). Mild fibrosis and cholecystitis were observed principally in infected cats. Hyperplastic precursor lesions of cholangiocarcinoma were observed, but no solid tumor was diagnosed. These results reveal high prevalence of P. fastosum in free roaming cats in northeastern Brazil, with increased risk of liver disease.

Disease surveillance of Atlantic herring: molecular characterization of hepatic coccidiosis and a morphological report of a novel intestinal coccidian.

Surveillance for pathogens of Atlantic herring, including viral hemorrhagic septicemia virus (VHSV), Ichthyophonus hoferi, and hepatic and intestinal coccidians, was conducted from 2012 to 2016 in the NW Atlantic Ocean, New Jersey, USA. Neither VHSV nor I. hoferi was detected in any sample. Goussia clupearum was found in the livers of 40 to 78% of adult herring in varying parasite loads; however, associated pathological changes were negligible. Phylogenetic analysis based on small subunit 18S rRNA gene sequences placed G. clupearum most closely with other extraintestinal liver coccidia from the genus Calyptospora, though the G. clupearum isolates had a unique nucleotide insertion between 604 and 729 bp that did not occur in any other coccidian species. G. clupearum oocysts from Atlantic and Pacific herring were morphologically similar, though differences occurred in oocyst dimensions. Comparison of G. clupearum genetic sequences from Atlantic and Pacific herring revealed 4 nucleotide substitutions and 2 gaps in a 1749 bp region, indicating some divergence in the geographically separate populations. Pacific G. clupearum oocysts were not directly infective, suggesting that a heteroxenous life cycle is likely. Intestinal coccidiosis was described for the first time from juvenile and adult Atlantic herring. A novel intestinal coccidian species was detected based on morphological characteristics of exogenously sporulated oocysts. A unique feature in these oocysts was the presence of 3 long (15.1 ± 5.1 µm, mean ±SD) spiny projections on both ends of the oocyst. The novel morphology of this coccidian led us to tentatively name this parasite G. echinata n. sp.

Molecular diagnosis of Eimeria stiedae in hepatic tissue of experimentally infected rabbits.

The early detection of Eimeria stiedae in the hepatic tissue of experimentally infected rabbits was investigated using molecular assay. Forty 6-week-old male New Zealand rabbits were divided into two groups. Group A (30 animals) was infected with 2.5 × 10(4) sporulated oocysts of E. stiedae per animal on Day 0 and Group B (10 animals) was used as the uninfected controls. Three animals from Group A and one from Group B were sacrificed at 0, 3, 6, 9, 12, 15, 18, 21, 24 and 27 days post infection (PI). Gross and microscopic post-mortem findings were recorded. Polymerase chain reaction (PCR) of the E. stiedae internal transcribed spacer 1 genomic region was conducted on blood, liver tissue, and feces from the Group A experimentally infected animals. Macroscopically, the liver showed irregular yellowish white nodules pathognomonic to E. stiedae infection beginning on Day 15 PI. Hepatomegaly and ascites were obvious from Day 21-24 PI. The presence of different E. stiedae schizonts and gametocytes in the histopathological sections of the biliary epithelium were evident on Day 15 PI. The E. stiedae PCR was first positive in liver tissues on Day 12 and in fecal samples on Day 18 PI, but the blood samples were negative. In conclusion, the PCR can be used for early diagnosis and control of E. stiedae schizonts before shedding of the oocysts in feces.