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Trematode infections caused by Fasciolidae and Paramphistomidae remain widespread in livestock, resulting in substantial economic losses. The two distinct fluke families are difficult to distinguish morphologically, and molecular identification provides the most reliable means of accurate diagnosis. In Ecuador, however, molecular data on these parasites are scarce. In this study, we collected trematodes from cattle rumen and bile ducts, molecularly identified them, and assessed their phylogenetic relationship to Fasciola hepatica to determine their introduction pathways into South America. Genomic DNA was extracted, and PCR was used to amplify the ITS2 (~500 bp) and COXI (~266 bp) regions; all amplicons were Sanger-sequenced. Phylogenetic trees for both markers were constructed using a Maximum Likelihood approach with 1000 bootstrap replicates in CIPRES v3.3. The rumen fluke exhibited 99% ITS2 and COXI similarity to an Indian Cotylophoron cotylophorum strain, while the bile-duct fluke showed 99% ITS2 and 100% COXI similarity to F. hepatica isolates from Australia and Nigeria, respectively. Distinct single-nucleotide polymorphisms (SNPs) in the ITS2 chromatograms suggest a diploid genome structure in both trematode species. This is the first report of C. cotylophorum in Ecuador, and its presence may be linked to the late 19th-century introduction of Zebu cattle (Bos taurus indicus) from India.

Understanding changes in the host-parasite relationship due to habitat fragmentation is necessary for better management and conservation of endangered species in fragmented landscapes. Pathogens and parasites can pose severe threat to species in restricted environments such as forest fragments where there is increased contact of wildlife with human and livestock populations. Environmental stress and reduced nutritional level in forest fragments can influence parasite infection and intensity on the native species. In this study, we examine the impact of habitat fragmentation on the prevalence of gastrointestinal parasites in lion-tailed macaques in a fragmented rainforest in Western Ghats. The prevalence of different gastrointestinal parasites was estimated from 91 fecal samples collected from 9 lion-tailed macaque groups in nine forest fragments. The parasites were identified up to genus level on the basis of the morphology and coloration of the egg, larva and cyst. The covariates included forest fragment area, group size and the presence/absence of human settlements and livestock in proximity. We used a linear regression model to identify the covariates that significantly influenced the prevalence of different parasite taxa. Nine gastrointestinal parasite taxa were detected in lion-tailed macaque groups. The groups near human settlements had greater prevalence and number of taxa, and these variables also had significant positive correlations with group size. We found that these parameters were also greater in groups near human settlements after controlling for group size. Livestock were present in all five fragments that had human settlements in proximity. The present study suggests that high prevalence and species richness of gastrointestinal parasites in lion-tailed macaque groups are directly related to habitat fragmentation, high anthropogenic activities and high host density. The parasite load partially explains the reason for the decline in immature survival and birth rate in small and isolated rainforest fragments in Anamalai Hills.

Excretory and secretory antigen of adult Cotylophoron cotylophorum and Gastrothylax crumenifer was prepared and characterized by SDS-PAGE. Adult paramphistomes were collected in PBS from rumen of sheep slaughtered at Perambur and Villivakkam slaughter houses, Chennai, Tamil Nadu. Excretory/Secretory (E/S) antigens of G. crumenifer and C. cotylophorum were prepared by incubating in PBS at 28.3 °C overnight in a shaking incubator and centrifuged. The culture supernatant was used as antigen. The protein concentration of E/S antigens of C. cotylophorum and G. crumenifer ranged from 0.4 to 7.1 mg/ml and 3.3 to 5.9 mg/ml, respectively. SDS-PAGE analysis of E/S antigen of C. cotylophorum revealed two polypeptide bands from the range of 32 and 40 kDa. Whereas, SDS-PAGE analysis of E/S antigen of G. crumenifer revealed only one polypeptide band of 35 kDa. Excretory/secretory antigen may be used for serodiagnosis of paramphistomosis infection in sheep.

Introduction: In the present study, the in vitro effect of methanolic extract of leaves of E. acaulis (EaME) on the key enzymes, namely pyruvate kinase (PK), phosphoenolpyruvate carboxykinase (PEPCK) and lactate dehydrogenase (LDH) of Cotylophoron cotylophorum have been investigated. Furthermore, the EaME was subjected to preliminary structural analysis using gas GC-MS. Materials and Methods: Adult live C. cotylophorum was collected from the rumen of slaughtered sheep and maintained in Hedon-Fleig solution. The flukes were incubated in five different sub-lethal concentrations of EaME for 2, 4, and 8 h. Control flukes without the plant extract were also maintained simultaneously. The enzymes were assayed in the sample following the standard protocols and were expressed as n moles mg protein−1. The effect of plant extract on the enzyme activity was calculated as percentage inhibition or acceleration. The structural elucidation of components of EaME was done using a quadrupole mass spectrometer and electron ionization method. Results: EaME significantly inhibited the PEPCK and PK activities of C. cotylophorum, and the inhibition was dose- and time-dependent. Interestingly, EaME accelerated the LDH activity catalyzing the reduction of pyruvate. The GC-MS analysis of EaME showed the presence of seven major compounds. Conclusion: The present study clearly revealed that the anthelmintic potential of Elytraria acaulis in impeding the energy-generating mechanism of C. cotylophoroum and hence, could be a potential source of natural anthelmintic.

•Vaccinating mice with recombinant proFgCatL1H (rproFgCatL1H) exhibited protection at 62.7–66.1%.•Anti-rproFgCatL1H antisera reacted specifically with rproFgCatL1H and other cathepsin L isotypes of F. gigantica, but not with antigens of other parasites.•Vaccination with rproFgCatL1H elicited a high level of IgG1, indicating the Th2-dominated response.•The level of IgG1 was strongly correlated with the number of worm recovery. Fasciola gigantica cathepsin L1H (FgCatL1H) is one of the major cathepsin L released by juveniles of F. gigantica to aid in the invasion of host's tissues. Due to its high sequence similarity with other cathepsin L (CatL) isoforms of late stage F. gigantica, it was considered to be a good vaccine candidate that can block all CatL-mediated protease activities and affect juveniles as well as adult parasites. In this study, recombinant proFgCatL1H protein expressed in yeast, Pichia pastoris, system was mixed with Freund's adjuvants and used to subcutaneously immunize mice that were later challenged with metacercariae of F. gigantica. The percentage of worm protection in the rproFgCatL1H-vaccinated mice compared to the non-immunized and adjuvant control mice were approximately 62.7% and 66.1%, respectively. Anti-rproFgCatL1H antisera collected from vaccinated mice reacted specifically with rproFgCatL1H and other cathepsin L isoforms of F. gigantica, but the antibodies did not cross react with antigens from other trematode and nematode parasites, including Eurytrema pancreaticum, Opisthorchis viverrini, Fischoederius cobboldi, Cotylophoron cotylophorum, Gigantocotyle explanatum, Paramphistomum cervi, and Setaria labiato-papillosa. The levels of IgG1 and IgG2a in mouse sera increased significantly at two weeks after immunization and were highest during the sixth to eighth weeks after immunization. The IgG1 level was higher than IgG2a at all periods of immunization, implicating the dominance of the Th2 response. The levels of IgG1 and IgG2a in the immune sera were shown to be strongly correlated with the numbers of worm recovery, and the correlation coefficient was higher for IgG1. The levels of serum aspartate aminotransferase and alanine transaminase were significantly lower in the sera of rproFgCatL1H-vaccinated mice than in the infected control mice indicating a lower degree of liver damage. This study demonstrated a high potential of FgCatL1H vaccine, and its efficacy is currently being studied in the larger economic animals.

A cross-sectional study was carried out to determine the prevalence of gastrointestinal (GI) nematodes and flukes (Fasciola and amphistomes) infection in communally grazed traditional cattle, zero-grazed small-scale dairy cattle and intensively grazed large-scale dairy cattle through examination of helminth eggs in faeces. Results indicated that the type of management, especially the grazing habit, has a significant influence on the prevalence and intensity of GI nematodes and flukes. The prevalence of GI nematodes in traditional, large-scale dairy and small-scale dairy cattle was 67%, 44.4% and 37%, respectively, with the highest faecal egg counts in calves. The overall prevalence of Fasciola gigantica in traditional, large-scale dairy and small-scale dairy cattle was 63.8%, 46.2% and 28.4%, respectively. The prevalence of amphistomes was 81.9%, 55.5% and 41.1% in traditional, large-scale dairy and small-scale dairy cattle, respectively. The high prevalence of flukes in the traditional system was attributed to communal grazing and watering management practices. Stomach flukes recovered in examined cattle at the abattoir were Calicophoron microbothrium and Cotylophoron jacksoni. About 42.1% of infected animals had both Fasciola and amphistomes. The prevalence of both GI nematodes and flukes varied greatly among villages and farms. The prevalence of both Fasciola and amphistomes was higher in adults (58.5%, 75.2%) than in yearlings (36.5%, 51.5%) or calves (24.9%, 47.2%). The variation in the prevalence of both GI nematodes and flukes among management and age groups within systems can be used as an entry point towards rational use of anthelmintics for each management system. More studies on seasonal transmission pattern of all these parasites are required in order to design rational, economic and locally sustainable parasite control programmes.

Objective: Paramphistomosis (stomach fluke disease) is a parasitic infection caused by digenetic trematodes and is considered to be one of the most important parasitic diseases affecting livestock worldwide. This disease is widely prevalent in India, and the highest incidence is reported during monsoon and post-monsoon months. In the present study, in vitro effect of aqueous extract of pods of Acacia concinna (AcP E) on the morphology and the histology of the digenetic trematode Cotylophoron cotylophorum have been investigated. Materials and Methods: The in vitro effect of Ac PE on the morphology and the histology of a digenetic trematode C. cotylophorum have been examined using scanning electron microscopy (SEM), transmission electron microscopy (TEM) and light microscopic techniques. Results: The SEM micrograph of treated flukes showed the appearance of few blebs near the oral region and rupture of the dorsal surface of the tegument. The light and TEM observations revealed significant deleterious changes in the internal organization of the fluke. Severe injury to the tegument due to bleb formation, detachment of tubercles, and vacuolization of the subtegumental region was observed. Nuclear indentation, cytoplasmic autolysis, and mitochondrial abnormalities were the other prominent observations. Conclusion: The results of the present study convincingly showed that Ac PE is an effective anthelmintic causing detrimental effect to C. cotylophorum and appears to be a potent phytotherapeutic agent to control paramphistomosis.

La paranfistomosis es una infección ocasionada por trematodes pertenecientes al suborden Paramphistomata, algunas de estas especies son parásitos del rúmen y retículo de rumiantes. Uno de los trematodos más fre- cuentes en todo el mundo es el Cotylophoron que afecta a rumiantes domésticos y silvestres, especialmente a vacas, ovejas, cabras y búfalos especialmente en regiones tropicales y subtropicales. Se realizó la evaluación parasitaria a 61 ejemplares de ganado bovino aparentemente sanos (42 hembras adultos y 19 menores de un año), manejados al pastoreo, en un fundo ganadero ubicado en la cuenca del río Nanay –Iquitos. Las heces fueron obtenidas mediante palpación rectal y procesada mediante el método Dennis modificado. Dos vacas fueron sacrificadas para obtener parásitos adultos. Los resultados indican la presencia de Cotylophoron cotylo- phorum (Fischoeder, 1901) Stiles & goldeberger, 1910 en los animales estudiados y es un nuevo registro para el Perú, encontrando porcentajes de infección de 62 y 53% en adultos y terneros respectivamente. El rango de la carga parasitaria (huevo/gramo de heces) en adultos fue 1—44 y 1—55 en animales menores de un año. El estudio revela la existencia del trematodo en un grupo de ganado cruzado manejados al pastoreo, aunque se desconoce los efectos negativos que podría estar causando en la reducida ganadería amazónica.

Entre los Paramphistomidae (Trematoda: Digenea) hay especies que parasitan el sistema digestivo de rumiantes disminuyendo su producción láctea y cárnica. En Colombia se ha registrado la paramphistomosis bovina en: 1) la región lechera altoandina de Antioquia donde se describe a Cotylophoron cotylophorum como el agente causal; y 2) en bovinos de producción cárnica de las Sabanas en el cálido piedemonte llanero y en los Llanos Orientales nacionales, donde se señala a varias especies de paramfistómidos, de los cuales falta la descripción que respalde el estatus taxonómico específico asignado en las publicaciones. Por lo tanto, el hallazgo de digeneos en bovinos sacrificados en el beneficiadero de Guamal, Meta, (Colombia) constituyó una oportunidad para iniciar con la descripción de los Paramphistomidae de esta región del país. Entre 2006 y 2008 se extrajeron 715 trematodos adultos del rumen de 32 bovinos provenientes de los departamentos del Meta y del Guaviare. El análisis morfológico muestra las siguientes características de la familia Paramphistomidae: el canal de Laurer atraviesa la vesícula excretora y se abre debajo del poro excretor; acetábulo pequeño del tipo Cotylophoron, la faringe tipo Calicophoron; ventosa genital sin esfínter; ausencia de bulbo esofágico y las vitelarias laterales y dorsales confluyendo en el extremo posterior del cuerpo, debajo del acetábulo, como en Cotylophoron panamensis. Con este estudio se ratifica la paramfistomosis bovina en el departamento del Meta y se registra por primera vez a Cotylophoron panamensis en bovinos del departamento del Guaviare, ampliando su rango de distribución geográficaen Colombia.

La paranfistomosis es una infección ocasionada por trematodes pertenecientes al suborden Paramphistomata, algunas de estas especies son parásitos del rúmen y retículo de rumiantes. Uno de los trematodos más frecuentes en todo el mundo es el Cotylophoron que afecta a rumiantes domésticos y silvestres, especialmente a vacas, ovejas, cabras y búfalos especialmente en regiones tropicales y subtropicales. Se realizó la evaluación parasitaria a 61 ejemplares de ganado bovino aparentemente sanos (42 hembras adultos y 19 menores de un año), manejados al pastoreo, en un fundo ganadero ubicado en la cuenca del río Nanay--Iquitos. Las heces fueron obtenidas mediante palpación rectal y procesada mediante el método Dennis modificado. Dos vacas fueron sacrificadas para obtener parásitos adultos. Los resultados indican la presencia de Cotylophoron cotylophorum (Fischoeder, 1901) Stiles & goldeberger, 1910 en los animales estudiados y es un nuevo registro para el Perú, encontrando porcentajes de infección de 62 y 53% en adultos y terneros respectivamente. El rango de la carga parasitaria (huevo/gramo de heces) en adultos fue 1-44 y 1-55 en animales menores de un año. El estudio revela la existencia del trematodo en un grupo de ganado cruzado manejados al pastoreo, aunque se desconoce los efectos negativos que podría estar causando en la reducida ganadería amazónica.