Glutathione-S-transferase: an important diagnostic antigen of liver amphistome Gigantocotyle explanatum, infecting the Indian water buffalo.

The foodborne trematodiases pose a significant health problem to the animals as well as the human population living in close proximities with the livestock and are still considered as the neglected tropical diseases by the World Health Organisation. The digenetic trematode, Gigantocotyle explanatum infecting the liver of Indian water buffalo, Bubalus bubalis, has been identified as one of the most common helminth parasite responsible for the disease, amphistomosis, in livestock. Despite huge abattoir prevalence, the epidemiological data and the actual economic losses incurred due to this parasite alone are yet to be established probably due to the limitations of routinely used diagnostic tests. The gold standard for the confirmation of such infections under field conditions is still the fecal egg count (FEC). However, the poor sensitivity and cumbersome nature of these tests necessitates the development of a more sensitive, reliable and easy to perform workflow/method. Immunological diagnosis of helminthic infections is still considered as an alternative to the FEC. Therefore, efforts have been made to utilize glutathione-S-transferase (GST), a vitally significant molecule of the adult G. explanatum, for the serodiagnosis of amphistomosis under both laboratory and field conditions. The GST antigen was first affinity purified from the somatic extract of the adult worms since its highest level was recorded in the somatic extracts followed by eggs and the excretory/secretory products. A five-fold affinity purified native GST antigen of about 25 kDa was found to be highly immunogenic as evident from high titre (1:25,600) of the polyclonal antibodies raised in the rabbits. The immunoblotting results revealed differential presence of GST in the adult worms, their eggs and excretory/secretory products. The immunolocalization studies revealed that the vitelline glands are the major source of GST in liver amphistome. Further, we were able to successfully screen animals naturally infected with G. explanatum using anti GST polyclonal antibodies in dot blot assay. High levels of both circulating GST antigen and anti GST antibodies were detected in the serum of the animals naturally infected with G. explanatum, while no cross reactivity was observed with the tropical liver fluke, F. gigantica which often infects the buffalo liver concurrently. The findings of the present study indicate that GST could be used as an important antigen for the diagnosis of G. explanatum infection in Indian water buffaloes.

A novel Kunitz protein with proposed dual function from Eudiplozoon nipponicum (Monogenea) impairs haemostasis and action of complement in vitro.

Serine peptidases are involved in many physiological processes including digestion, haemostasis and complement cascade. Parasites regulate activities of host serine peptidases to their own benefit, employing various inhibitors, many of which belong to the Kunitz-type protein family. In this study, we confirmed the presence of potential anticoagulants in protein extracts of the haematophagous monogenean Eudiplozoon nipponicum which parasitizes the common carp. We then focused on a Kunitz protein (EnKT1) discovered in the E. nipponicum transcriptome, which structurally resembles textilinin-1, an antihemorrhagic snake venom factor from Pseudonaja textilis. The protein was recombinantly expressed, purified and biochemically characterised. The recombinant EnKT1 did inhibit in vitro activity of Factor Xa of the coagulation cascade, but exhibited a higher activity against plasmin and plasma kallikrein, which participate in fibrinolysis, production of kinins, and complement activation. Anti-coagulation properties of EnKT1 based on the inhibition of Factor Xa were confirmed by thromboelastography, but no effect on fibrinolysis was observed. Moreover, we discovered that EnKT1 significantly impairs the function of fish complement, possibly by inhibiting plasmin or Factor Xa which can act as a C3 and C5 convertase. We localised Enkt1 transcripts and protein within haematin digestive cells of the parasite by RNA in situ hybridisation and immunohistochemistry, respectively. Based on these results, we suggest that the secretory Kunitz protein of E. nipponicum has a dual function. In particular, it impairs both haemostasis and complement activation in vitro, and thus might facilitate digestion of a host's blood and protect a parasite's gastrodermis from damage by the complement. This study presents, to our knowledge, the first characterisation of a Kunitz protein from monogeneans and the first example of a parasite Kunitz inhibitor that impairs the function of the complement.

Effects on haematological and serum biochemical parameters of Pangasianodon hypophthalmus to an experimental infection of Thaparocleidus sp. (Monogenea: dactylogyridae).

Monogenea (gill parasite) is a major problem in aquaculture that reduces the growth of cultured fish and adversely affects the economy. The present study was performed to evaluate the impact of various degrees of Thaparocleidus sp. (dactylogyrids, monogenean) infestation on haematological and serum biochemical parameters of Pangasianodon hypophthalmus. A standard cohabitation study, following complete randomized design in triplicate, was conducted to obtain low, moderate and high degrees of infestation in P. hypophthalmus along with the control (uninfested) group. Blood and serum were studied for haematological (total erythrocyte count (TEC), haemoglobin (Hb), packed cell volume (PCV), total leucocyte count (TLC) and indices viz. mean cell volume (MCV), mean cell haemoglobin (MCH), mean cell haemoglobin concentration (MCHC) and erythrocyte osmolarity brittleness (EOB)) and serum biochemical parameters (serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), lactate, total bilirubin and creatinine. Significant (p < 0.05) increase in TEC, Hb, TLC, EOB, SGOT, SGPT, LDH, lactate, bilirubin, and creatinine were noticed in moderate to high monogenean-infested group in comparison to the control group. However, significant (p < 0.05) reduction in MCH, and MCV and no difference (P > 0.05) in PCV were noticed in high degree parasitized group in comparison to the control group. The results of altered haematological and serum biochemical parameters in various degrees of monogenean-infested groups signify the density dependent physiological responses and changes in cells of the blood. The data of serum enzymes in the present study would be valuable for assessing the health status of the host and facilitate as a potential biomarker in relation to various degrees of monogenean infestation.

Clinical, hematological, biochemical, and ultrasonographic aspects of Platynosomum sp. (Trematoda: Dicrocoeliidae) infection of captive Callithrix penicillata.

Trematodes from the genus Platynosomum have been found to infect Neotropical primates in captivity, but little is known about their pathogeny in such hosts. This study evaluated the physiological effects of natural infection by the liver-dwelling trematode Platynosomum sp. in ten males and ten females of Callithrix penicillata kept in captivity at the Primate Center of the University of Brasília. The marmosets were examined twice, 6 months apart. The following parameters were analyzed: complete blood count, bleeding time, serum total protein, albumin, and the liver enzymes AST and ALT, and both a stool analysis and a liver ultrasonic evaluation were performed. We were able to characterize a group of abnormalities associated with this trematode infection which were mainly derived from the hepatitis caused by it: coagulation disorders, abnormal red blood cells, hypoalbuminemia, and abnormal levels of liver-linked serum enzymes. Eosinophilia and thrombocytopenia were also commonly seen. All of the aforementioned abnormalities were in good accord with typical effects of trematodes on liver parenchyma. We suggest that this set of abnormalities is characteristic of the infection of C. penicillata with Platynosomum sp., and should be among the most prominent aspects that the veterinary surgeon considers when suspecting such an infection. We also suggest that these clinical signs and abnormalities will be similar in other liver-dwelling trematode-infected primate species.

Experimental Infections of Bluegill with the Trematode Ribeiroia ondatrae (Digenea: Cathaemasiidae): Histopathology and Hematological Response.

Infections by the digenetic trematode, Ribeiroia ondatrae, cause severe limb malformations in many North American amphibians. Ribeiroia ondatrae also infects fishes as second intermediate hosts, but less is known about the pathology and immune responses initiated in infected fish, even though reports of infected fish date back to early 1900s. To this end, we experimentally exposed juvenile Bluegills Lepomis macrochirus to three doses of R. ondatrae cercariae and monitored the pathology, parasite infection success, and humoral responses over 648 h. All exposed fish became infected with metacercariae, and the average infection load increased with exposure dose. Histologically, infection was associated with acute hemorrhages in the lateral line and local dermis at 36 h, followed by progressive granulomatous inflammation that led to the destruction of encysted metacercariae. Correspondingly, over the course of 648 h we observed an 85% decline in average infection load among hosts, reflecting the host's clearance of the parasite. Infection was not associated with changes in fish growth or survival, but did correlate with leukocytosis and neutrophilia in circulating host blood. Understanding the physiological responses of R. ondatrae in Bluegill will help to clarify the ecological effects of this parasite and provide a foundation for subsequent comparisons into its effects on behavior, individual health, and population dynamics of Bluegill.

Antibody response of definitive hosts against antigens of two life stages of the neuropathogenic schistosome Trichobilharzia regenti.

BACKGROUND: The nasal avian schistosome Trichobilharzia regenti spends part of its intravertebrate period of life within the central nervous system. Migration of the parasites can be accompanied by neuromotor disorders or paralysis in natural definitive hosts (ducks) and even in laboratory mammals. Cercariae are also able to penetrate human skin and induce cercarial dermatitis. While the cellular and antibody responses against cercariae and migrating schistosomula have been investigated in mice, little is known about immune reactions in birds. This study first describes the dynamics of antibody response in infected ducks and identifies frequently recognized antigens that may serve as diagnostic markers of infection by T. regenti. METHODS: Groups of 35 domestic ducks and 10 mallards were exposed to different doses of T. regenti cercariae. Sera were collected at predefined time intervals and tested by ELISA for the presence of specific anti-cercarial IgY and IgM. Antigens recognized by the antibodies were identified on Western blots of cercariae and schistosomula. The applicability in immunodiagnostics was statistically evaluated by expression of specificity and sensitivity values for individual antigens. RESULTS: In ELISA, the levels of anti-cercarial IgM peaked on day 15 pi. Increased production of IgY associated with the later phases of infection was observed in most individuals around 20 dpi and culminated 30 dpi. The time course of antibody response did not differ among experimental groups, variations were only observed in the levels of specific IgY which depended rather on the age of ducks at the time of infection than on the infectious dose. On Western blots, 40 cercarial and 7 schistosomular antigens were recognized by IgY from infected ducks. Among them, 4 cercarial antigens of 50, 47, 32 and 19 kDa provided the most sensitive and specific reactions. CONCLUSIONS: Antigens of cercariae and schistosomula elicited distinct antibody response in ducks, which correlated positively with the age of animals at the time of infection. Several antigens originating in cercariae and fewer in schistosomula were recognized by IgY with diverse sensitivity and specificity; only a few seemed to be common to both stages. Four of them were considered as the most promising candidates for immunodiagnostics.

Plasma autoantibodies against heat shock protein 70, enolase 1 and ribonuclease/angiogenin inhibitor 1 as potential biomarkers for cholangiocarcinoma.

The diagnosis of cholangiocarcinoma (CCA) is often challenging, leading to poor prognosis. CCA arises via chronic inflammation which may be associated with autoantibodies production. This study aims to identify IgG antibodies directed at self-proteins and tumor-associated antigens. Proteins derived from immortalized cholangiocyte cell line (MMNK1) and CCA cell lines (M055, M214 and M139) were separated using 2-dimensional electrophoresis and incubated with pooled plasma of patients with CCA and non-neoplastic controls by immunoblotting. Twenty five immunoreactive spots against all cell lines-derived proteins were observed on stained gels and studied by LC-MS/MS. Among these, heat shock protein 70 (HSP70), enolase 1 (ENO1) and ribonuclease/angiogenin inhibitor 1 (RNH1) obtained the highest matching scores and were thus selected for further validation. Western blot revealed immunoreactivity against HSP70 and RNH1 in the majority of CCA cases and weakly in healthy individuals. Further, ELISA showed that plasma HSP70 autoantibody level in CCA was significantly capable to discriminate CCA from healthy individuals with an area under the receiver operating characteristic curve of 0.9158 (cut-off 0.2630, 93.55% sensitivity and 73.91% specificity). Plasma levels of IgG autoantibodies against HSP70 were correlated with progression from healthy individuals to cholangitis to CCA (r = 0.679, P<0.001). In addition, circulating ENO1 and RNH1 autoantibodies levels were also significantly higher in cholangitis and CCA compared to healthy controls (P<0.05). Moreover, the combinations of HSP70, ENO1 or RNH1 autoantibodies positivity rates improved specificity to over 78%. In conclusion, plasma IgG autoantibodies against HSP70, ENO1 and RNH1 may represent new diagnostic markers for CCA.

Transcriptional immune response of cage-cultured Pacific bluefin tuna during infection by two Cardicola blood fluke species.

Infections by two blood fluke species, Cardicola orientalis and Cardicola opisthorchis, currently present the greatest disease concern for the sea-cage culture of Pacific bluefin tuna (PBT) - a species of high global economic importance and ecological concern. In this study, we aimed to rapidly, quantitatively, and differentially identify infections by these two parasite species in cultured PBT as well as identify potential host immune responses. Using real-time qPCR, we were successful in quantitatively detecting parasite-specific DNA from within host blood, gill, and heart tissues; positively identifying parasitic infections 44 days earlier than microscopy methods previously employed. Both gill and heart became heavily infected by both parasite species in PBT within two months of sea-cage culture, which was only mitigated by the administration of anthelmintic praziquantel. Nevertheless, fish were observed to mount an organ specific transcriptive immune response during infection that mirrored the relative quantity of pathogenic load. In heart, significant (3-6 fold) increases in IgM, MHC2, TCRß, and IL-8 transcription was observed in infected fish relative to uninfected controls; whereas in the gills only IgM transcription was observed to be induced (11 fold) by infection. Interestingly, the relative quantity of IgM transcription was highly correlated to the relative abundance of C. orientalis but not C. opisthorchis DNA in the gill samples, even though this organ showed high prevalence of DNA from both parasite species. Taken together, these findings indicate that although ineffective at combating infection during primary exposure, a cellular immune response is mounted in PBT as a potential rejoinder to future Cardicola exposure, particularly against C. orientalis. Although future investigation into antibody effectiveness will be needed, this work provides valuable preliminary insight into host responsiveness to Cardicola infection as well as additional support for the need of anthelmintic treatment following primary parasite exposure during PBT culture.

Pro-inflammatory cytokines in the serum of silver catfish (Rhamdia quelen) naturally infected by Clinostomum complanatum: a preliminary study.

Clinostomum complanatum is a zoonotic parasite of birds, and its larval form (metacercariae) may cause disease in fish. The immune response of fish infected by C. complanatum is unknown, and therefore the objective of this study was to evaluate serum levels of pro-inflammatory cytokines in silver catfish (Rhamdia quelen) infected by this parasite. The analysis showed that fish infected with metacercariae presented significantly (P < 0.05) higher levels of tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1), and interleukin 6 (IL-6) when compared to those not infected. These data suggest that C. complanatum larvae activate the host immune response.

[Morphological features of cellular responses to different rates of trematode: Quinqueserialis quinqueserialis (Trematoda: Notocotilidae) invasion in muskrat (Ondatra zibethicus)].

The results of investigation of leukocyte morphology and leukocyte contents of blood and caecum depending on the trematode Quinqueserialis quinqueserialis invasion rate in muskrats from natural population are given. At low trematode invasion rates, there was observed systemic activation of lymphopoiesis and neutrophil granulocytopoiesis with a decrease in the monocyte-miacrophage response in caecum (trematodelocalization organ). At the same time, under high invasion rates, there was detected induction of T cell suppressor activity and the absence of a granulocyte response in the tissues under study. Intensification of B lymphocyte blast transformation in caecum tissues as well as the appearance of blast cells in the blood of infected muskrats was observed.

Evaluation of Gastrothylax crumenifer antigenic preparation in serodiagnosis of paramphistomiasis in sheep.

An evaluation of Gastrothylax crumenifer crude antigen preparation viz., Somatic Antigen (SAg), Excretory Secretory Antigen (ESAg) and Egg Antigen (EAg) in serodiagnosis of disease was undertaken. Test sera samples were obtained from 30 Paramphistomiasis Positive and 30 Gastrothylax free sheep slaughtered at Hazratbal Kashmir. The referral antigenic preparation were evaluated against Paramphistomiasis positive sera, via., control negative sera, using double immunodiffusion test (DID), (IEP) Immunoelectrophoretic assay and ELISA. The performance of referral antigens, as assessed from percent sensitivity and specificity, revealed an increasing trend from DID (Double immunodiffusion-An immunological technique used in the detection, identification and quantification of antibodies and antigens) to IEP (immunoelectrophoresis-A general name for a number of biochemical methods for separation and characterization of proteins based on electrophoresis and reaction with antibodies), followed by ELISA, detecting higher number of sheep positive for paramphistomiasis. In ELISA the ESAg and SAg were evaluated as most reactive antigens with no significant difference and EAg was the least antigenic. In IEP, EAg had the higher sensitivity (60%) and analogous specificity of SAg and ESAg. The formation of the preceptin lines in the proximity to EAg containing wells (cathode end) in IEP was suggestive of higher molecular weight of G. crumenifer specific protein molecules with slower rate of migration. Purification and characterization of G. crumenifer and identification of specific antigenic molecules, particularly in EAg has been suggested for qualitative improvement of diagnostic value of the antigens in the tests used here in.

Hematological parameters of Hoplias malabaricus (Characiformes: Erythrinidae) parasitized by Monogenea in lagoons in Pirassununga, Brazil / Parâmetros hematológicos de Hoplias malabaricus (Characiformes: Erythrinidae) parasitados por monogenea em lagoas de Pirassununga, Brasil

Hematology is an important pathological and diagnostic tool. This paper describes the hematological parameters of 76 specimens of Hoplias malabaricus infested with monogenean parasites, which were collected from two lagoons. The hematological parameters included: erythrocyte count (Er), microhematocrit (Hct), hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC). The blood parameters showed no significant changes associated with the infestation. Water temperature was found to be negatively correlated with MCV and Hct, with values of rs= −0.52, p<0.0001 and rs= −0.48, p<0.0001, respectively. The mean Relative Condition Factor was Kn=1.01, indicating good health conditions of the fish in these lagoons. No correlation was found between the monogenean infestation and the blood parameters, or between the environmental factors and the monogeneans.

A hematologia é uma importante ferramenta para patologia e diagnóstico. O presente estudo teve como objetivo descrever os parâmetros hematológicos de 76 espécimes de Hoplias malabaricus, coletados em duas lagoas, associados ao parasitismo por monogenético. Os parâmetros hematológicos foram: contagem de eritrócitos (Er), micro hematócrito (Hct), hemoglobina (Hb), volume corpuscular médio (VCM), hemoglobina corpuscular (HCM) e a concentração da hemoglobina cospuscular média (CHCM). Observou-se que não ocorreram mudanças significativas nos parâmetros sanguíneos relacionados com o parasitismo. Houve uma correlação negativa entre a temperatura da água o VCM e Hct, com valores de rs = −0,52, p<0,0001 e rs = −0,48, p<0,0001, respectivamente. O valor médio entre o fator de condição relativo foi de Kn = 1,01, indicando boas condições e os peixes estavam saudáveis nas lagoas estudadas. Não houve relação entre a infestação de monogeneas e os parâmetros sanguíneos, ou entre os fatores ambientais e as monogeneas.

Hematological parameters of Hoplias malabaricus (Characiformes: Erythrinidae) parasitized by Monogenea in lagoons in Pirassununga, Brazil.

Hematology is an important pathological and diagnostic tool. This paper describes the hematological parameters of 76 specimens of Hoplias malabaricus infested with monogenean parasites, which were collected from two lagoons. The hematological parameters included: erythrocyte count (Er), microhematocrit (Hct), hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC). The blood parameters showed no significant changes associated with the infestation. Water temperature was found to be negatively correlated with MCV and Hct, with values of rs= -0.52, p<0.0001 and rs= -0.48, p<0.0001, respectively. The mean Relative Condition Factor was Kn=1.01, indicating good health conditions of the fish in these lagoons. No correlation was found between the monogenean infestation and the blood parameters, or between the environmental factors and the monogeneans.

A new aporocotylid (Digenea) species from blood vascular system of gag grouper, Mycteroperca microlepis (Perciformes: Serranidae), off Alabama, with an emendation of Pearsonellum Overstreet & Køie, 1989.

Pearsonellum lemusi n. sp. (Digenea: Aporocotylidae) infects the blood vascular system of the gag grouper, Mycteroperca microlepis (Perciformes: Serranidae), in the north central Gulf of Mexico, approximately 80 km south of Dauphin Island, Alabama (29°34'09″N, 88°22'16″W). The new species can be most easily differentiated from its only congeners Pearsonellum corventum Overstreet and Køie, 1989 (type species) and Pearsonellum pygmaeus Nolan and Cribb, 2004 , both of which infect Australian serranids, by the combination of having a large adult body (3,237 × 570 µm), a cecal intersection comprising an elongated medial channel, anterior ceca >10% of total body length, ovary narrower than testis, and pre-ovarian uterus not looping between testis and ovary. The embryonated eggs of the new species infect gill epithelium, are spheroid, and measure 25-30 µm in diameter. Sympatric Gulf of Mexico serranids were negative for aporocotylid infections: coney, Cephalopholis fulva (n  =  1); Nassau grouper, Epinephelus striatus (3); red grouper, Epinephelus morio (32); yellowedge grouper, Epinephelus flavolimbatus (1); rock hind, Epinephelus adscensionis (1); red hind, Epinephelus guttatus (2); Warsaw grouper, Epinephelus nigritus (3); graysby, Cephalopholis cruentata (1); black grouper, Mycteroperca bonaci (1), and tattler, Serranus phoebe (2). The new species is the first aporocotylid described from a serranid outside of the southwestern Pacific Ocean. The diagnosis of Pearsonellum Overstreet and Køie, 1989 is herein emended to include anterior sucker having concentric rows of spines anterior to mouth, pharynx absent, esophagus length <1/2 total body length, vas deferens connecting with cirrus sac anteromedially, ovary occupying posterior 1/4-1/3 of body, primary vitelline duct dextral, and oviducal seminal receptacle extending posteriad in parallel with lateral body margin, not transverse nor constricted anteriorly or posteriorly by sharp bends or kinks.

Influence of ß-glucan on innate immunity and resistance of Lutjanus guttatus to an experimental infection of dactylogyrid monogeneans.

The present study investigated the effect of ß-1,3/1,6-glucan on growth, haematology, innate immunity and resistance against dactylogyrids on the spotted rose snapper (Lutjanus guttatus). Fish were fed during 5 weeks with commercial diet (control group) and same diet supplemented with three levels of ß-glucans (0·05%, 0·1% and 0·5%/kg feed). The results showed that at concentrations of 0·05% and 0·1%, fish growth was enhanced, and in weeks 2 and 4, an increase in cellular responses such as percentage of monocytes, neutrophils, respiratory burst activity and nitric oxide activity was observed. In diet with 0·5%ß-glucans, changes were registered at the end of the experiment. At week 2, 0·05%ß-glucans showed the best response to most of the analysed parameters. In a second trial, diet with 0·05%ß-glucans was chosen to prove its effect on the resistance of infected fish with monogeneans. The results showed that fish reduce significantly the number of dactylogyrids, and parameters such as WBC, percentage of lymphocytes, neutrophils and eosinophils were increased. In addition, WBC and percentage of thrombocytes increased in the control (+). The ß-glucans have the potential to be use in diet formulations of the spotted rose snapper and to limit the adverse effects of dactylogyrids.

Hematological characteristics of the spotted rose snapper Lutjanus guttatus (Steindachner, 1869) healthy and naturally infected by dactylogyrid monogeneans.

The aim of the present study was to obtain a basic knowledge of the hematology in order to determine changes in blood parameters of the spotted rose snapper Lutjanus guttatus. The morphological features of blood cells were described according to the observations made by light microscopy of Wright-Giemsa-stained blood films. The reference intervals and the mean value were determined for each hematological parameter evaluated in healthy fish and data were compared to those of naturally infected, with dactylogyrid monogeneans fish. Infected fish showed a prevalence of 100% and a mean intensity of 246.6 parasites per fish. Mean values of HCT, WBC, thrombocytes percentage and eosinophils percentage were significantly higher (P<0.05) in the infected fish. In addition, lymphocytes percentage and total protein were significantly lower (P<0.05) in the infected fish compared to healthy fish. Only total WBC count, lymphocytes percentage and eosinophils percentage in infected fish were outside reference interval. The hematology of the spotted rose snapper of this study might serve as a basis for future studies and diagnosis. Changes observed in blood parameters in infected fish suggest that the immune system of L. guttatus was affected by the presence of the parasites.

Influence of Calicophoron microbothrium amphistomosis on the biochemical and blood cell counts of cattle.

Sixteen Tuli steers aged 1 year were subdivided into four equal groups (I-IV) and infected with Calicophoron microbothrium metacercariae. Group I received a low dose (LD) of 5000 metacercariae, group II a medium dose (MD) of 15,000 metacercariae, group III a high dose (HD) of 25,000 metacercariae while group IV was the non-infected control (C) group. The experimental animals were monitored daily for clinical signs while ethylene diamine tetraacetic acid (EDTA) blood and serum samples were collected every 7 days until day 28 post-infection, when sample collection was terminated. Samples were processed for full blood count, eosinophils and blood biochemical values for calcium, magnesium, phosphorus, total protein and albumin. Moderate to severe diarrhoea developed in the MD and HD groups at day 21 post-infection. The diarrhoea coincided with a significant decrease (P < 0.05) in total plasma protein, calcium and phosphorus levels, particularly in the MD group. Similarly, a significant decrease (P < 0.05) in the packed cell volume (PCV), the haemoglobin (Hb) and red blood cell (RBC) levels occurred in the MD and HD groups from day 21 post-infection, while a significant increase (P < 0.05) in the circulating eosinophils occurred between 7 and 21 days post-infection in the LD and the HD groups.

[Increased accumulation of some trace elements in peripheral blood and bile of patients infested with Opisthorchis felineus (Rivolta, 1884) and Metorchis bilis (Braun, 1890)].

The invasion with Opisthorchis felineus (Rivolta, 1884) is known to be common in the Ob River region, West Siberia. These trematodes parasitize biliary tract of devinitive host (man or some species of animals). Other opisthorchiid species occurring in West Siberia, Metorchis bilis (Braun, 1890), has also been recorded recently as human parasite. Life cycles of both these trematodes include fish-eating mammal hosts. Eggs of O. felineus and M. bilis are very similar morphologically and can hardly be indentified. Chronic invasion with the helminthes is found to be a cause of disbolism of trace elements or high accumulation of some essential and toxic elements in the organism of definitive host. The aim of the present study was to determine concentrations of some essential and toxic elements in samples of peripheral blood and bile obtained from patients infested with Opisthorchis and/or Metorchis using instrumental neuron-activation technique. At first, all patients with microscopically confirmed opisthorchiasis (by microscopic examination of faeces and bile for the helminth eggs) were examined with serological method (ELISA) for specific anti-Opisthorchis and anti-Metorchis antibodies. Among 139 examined patients, 56.1% had specific antibodies against both Opisthorchis and Metorchis, 41.7% showed anti-Opisthorchis antibodies only, and 2.1% turn out to be seropositive for anti-Metorchis antibodies only. Of 31 elements detected in the samples of peripheral blood and bile, the concentration of nine essential and toxic elements (mercury, chromium, cesium, rubidium, lanthanum, bromine, selenium, zinc, and cobalt) in the patients with the mixt-infection of Opisthorchis and Metorchis (78 individuals) and with the Opisthorchis mono-infection (58 individuals) were significantly higher than those in healthy uninfested individuals. Among these elements, mercury and chromium showed the highest concentrations in infested patients. We suggest that the significant increase in concentrations of some toxic elements in blood and bile of patients may be associated with increased bioaccumulation of these elements in environment, especially in the local river water and fish. Moreover, concentrations of such elements as chromium, mercury, cesium, lanthanum, selenium, and zinc in blood and/or bile of the patients with mixt-infection of Opisthorchis and Metorchis were significantly higher than those in the patients with mono-infection of Opisthorchis. Accumulation of these elements in the organisms of patients was positively correlated with the number of parasites and the duration of helminth infection.

[Haptoglobin, serum amyloid-a and ceruloplasmin concentrations in goats with mixed helminth infection]. / Miks Helmint Enfeksiyonlu Keçilerde Haptoglobin, Serum Amiloid-A ve Seruloplazmin Konsantrasyonlari.

In this study, the concentrations of haptoglobin, serum amyloid A and ceruloplasmin were measured in goats with mixed gastrointestinal infections of nematodes and liver trematodes. Twelve patients who were diagnosed as having mixed helminth infection after detection of Trichuris spp., Trichostrongylidae spp. and Fasciola spp. and six healthy controls participated in the study. The concentrations of the acute phase proteins (haptoglobin p< or =0.001; serum amyloid-A p< or =0.05) were significantly higher in the goats with mixed gastrointestinal infection with nematodes and liver trematodes than the control goats. As a result of this study, we determined that the concentrations of haptoglobin and serum amyloid-A increased in goats with mixed helminth infection but further studies of the clinical importance of those increases in relation to prognosis and monitoring therapy of helminthic infections in goats should be made.