Effect of phenol and pesticides on the physicochemical properties of the hemolymph in fresh-water gastropod mollusks (Gastropoda, Pulmonata) infected by trematode parthenitae

The effect of solutions of phenol (100, 250, 400 mg/l), copper sulfate, carbophos and bazudine (0.01, 0.1, 1.0 mg/l) on physical and chemical properties of haemolymph of Lymnaea stagnalis (L.) and Planorbarius corneus (L.) infected with parthenitae of trematodes was studied. Haemolymph viscosity decreases and its active reaction shifts to weakly acid one. Haemolymph density decreases only in solutions of carbophos and bazudine. With heavy infection the concentration of haemoglobin in haemolymph of P. corneus does not change under the effect of phenol but the total volume of haemolymph reduces considerably.     

The effect of ammonium nitrate on the residual nitrogen content in the hemolymph of the pulmonate snail Planorbarius purpura (Mollusca: Pulmonata: Bulinidae) normally and in trematode invasion

The effect of the ammonium nitrate (concentrations 2000, 4000, 6000 mg/l) on the content of urea, urea nitrogen and residual nitrogen in the haemolymph of normal Planorbarius purpura and in trematode-infected individuals. It is shown, that stimulation of metabolic processes is more expressed in uninfected specimens, while the trematode-infected ones, the protective process is strained. When the toxin concentration reaches 6000 mg/l, the trematode infected specimens enter the phase of depression with 3.73% hyponitrogen level.     

Effect of trematoda infection and different concentrations of surface active agents onto physical and chemical characteristics of haemolymph of Planorbarius corneus (Mollusca: Pulmonata)

Effect of trematod infection and different concentrations of detergent (20, 50, 80 mg/dm3) onto physical and chemical characteristics of haemolymph of Planorbarius corneus (content of Hb, density, active reaction, amount of Hb per unit of total mass and soft mass of the body) have been investigated. It was noted that all changes of named characteristics caused by detergent were more pronounced in infected mollusks.     

Changes in the sulfhydryl group content in the hemolymph of freshwater gastropods infected with trematode parthenitae and larvae

A study was conducted on the changes in sulphhydryl groups in the haemolymph of Lymnaea stagnalis infected with partenites of Opisthioglyphe ranae, Planorbis corneus infected with partenites of Cotylurus cornutus and females of Viviparus viviparus infected with partenites and larvae of Echinoparyphium petrowi. At the infection in the haemolymph of hosts were recorded statystically reliable changes in the protein sulphhydryl groups which are blocked by toxines excreted by the parasites. The reduction rate in the content of these substances depends on the localization of the parasites and infection intensity of the hosts.     

The effect of the trematode invasion and accumulation of heavy metals onto the pond snail (Mollusca: Gastropoda: Lymnaeidae)

The cumulating of heavy metals (Cu, Cd, Pb, Zn) by the pond snail Lymnaea stagnalis from river and pond populations in a norm and under the trematode infection was examined. The copper and cadmium were accumulated by all specimens, mainly in the mantle and leg, in less content--in the hepatopancreas, and in the least content--in the shell and haemolymph. The lead was accumulated most intensively by the river specimens in the shell and mantle, while in the pond specimens it was accumulated in the mantle and leg. The greatest content of zink was found in the mantle, leg, and hepatopancreas. According to the ratio of cumulation the heavy metals by the river and pond populations the following series of ions were obtained respectively: Pb > Zn > Cu > Cd and Zn > Pb > Cu > Cd. The difference between uninfected specimens and one infected with trematodes was recorded in the ratio of cumulated heavy metals, in the ratio of benthic biological accumulation, and also based on bending force between metals compared in pairs. An intensity of differences was directly proportional to the intensity of the trematode infection.     

Entomopathogenic fungal infection (Deuteromycotina: Hyphomycetes) affects water relations of the german cockroach,Blattella germanica (L.) (Dictyoptera: Blattellidae)

• 1.1. The total body water, lipid content, and cuticular permeability of fungus infected and uninfected German cockroaches, Blattella germanica, were examined.
• 2.2. Infected adult cockroaches weighed less and had significantly more body water than did uninfected specimens of the same size.
• 3.3. Uninfected medium-size nymphs weighed significantly more than infected nymphs, but there was no difference in body size between infected and uninfected small nymphs.
• 4.4. Cuticular permeability and lipid content of infected and uninfected cockroaches was not significantly different.
• 5.5. Sequestering of water by the fungal cells is discussed as a possible factor in the pathology of this fungal parasite.

     

The effect of trematode invasion and chromium sulphate on the crude protein content in the haemolymph of Viviparus viviparus (Mollusca: Gastropoda: Pectinibranchia)

The combined effect of the trematode infection (Echinoparyphium sp.) and various concentrations of chromium sulphate (0.01, 1, 100 mg/l) onto the crude protein content in the haemolymph of the mollusc Viviparus viviparus was investigated. The normal contents of the crude protein is 0.7-1.22%. In adult specimens its concentration is 20-80% higher than in molluscs of junior age group. Sex difference by this index, which is higher in females, begins to manifest in four year old individuals only. The contents of crude protein in the haemolymph of pregnant females is 1.5 times higher than in latent ones. Under the low intensity of the trematode infection, the contents of crude protein in the haemolymph remains normal, while in the case of heavy infection, it decrease 100 times or lower. In the cases of 0.01-1 mg/l concentrations of chromium sulphate, the content of protein in the haemolymph of V. viviparus decreases, while in the case of 100 mg/l it increases abruptly, in comparison to the norm. The trematode infection intensifies these processes.     

Immune responses of the argasid tick Ornithodoros moubata moubata induced by infection with the filarial worm Acanthocheilonema viteae

Investigations were undertaken to determine whether the tick Ornithodoros moubata moubata mounted a detectable immune response to primary and secondary infections with Acanthocheilonema viteae. Uninfected control tick survival rate was 70%, but only 45% in the primary infection group. Post-secondary infection survival rate (82%) was comparable to controls, indicating that these selected ticks had some protective advantage. Mean A. viteae infective larvae recovery from ticks with secondary infections was 31.4% lower than expected, suggesting the development of immunity. SDS-PAGE of haemolymph for proteins induced post-primary infection yielded a stronger signal at 45 kDa than controls, which was further elevated post-secondary infection. Proteins at 48, 22 and 16 to 18 kDa were detected in haemolymph from infected ticks but not seen from controls. The direct effect of haemolymph on microfilarial viability was examined using a novel in vitro assay; in these preliminary trials no differences were observed in parasite viability when exposed to haemolymph from infected or uninfected groups of ticks.     

Development of a monoclonal antibody specific to yellow head virus (YHV) from Penaeus monodon

A monoclonal antibody specific to yellow head virus (YHV) was produced from a mouse immunized with gill extracts prepared from laboratory-reared Penaeus monodon dually infected with YHV and white spot syndrome virus (WSSV). One clone designated V3-2B specifically bound to native and SDS-treated viral specific antigens. Immunocytochemical studies of infected gills revealed viral specific immunoreactivities in the cytoplasm of gill tissue and in haemocytes. No antibody binding was observed in gills from non-infected shrimp. In addition, immunocytochemical examination of tissues from shrimp experimentally infected with YHV gave a positive reaction, while tissues from uninfected control shrimp or shrimp experimentally infected with WSSV did not. Western blot analysis indicated that the antibody reacted with a protein of approximately 135 kD that was present only in shrimp infected with YHV. In dot-blot indirect immunoperoxidase assays, the antibody was able to detect viral associated antigen in diluted haemolymph up to 1:50 dilution and in an ammonium sulfate precipitate of haemolymph up to 1:1000 dilution. The results suggested that this antibody might be useful for development of effective diagnostic techniques for both heavy and mild YHV infections in shrimp.     

Haematological alterations in Rattus norvegicus (Wistar) experimentally infected with Echinostoma paraensei (Trematoda: Echinostomatidae)

Laboratory rats (Rattus norvegicus) were infected with Echinostoma paraensei (Trematoda: Echinostomatidae). The rodents received 150 metacercariae each and blood samples were collected weekly until the fifth week of infection. The blood samples were analyzed for determination of haematocrit, total red blood cells with their dimensions, haemoglobin and haematimetric index (mean corpuscular volume, MCV; mean corpuscular haemoglobin, MCH; and mean corpuscular haemoglobin concentration, MCHC) and platelets. Red blood cells, haematocrit and haemoglobin in the first week had significantly lower levels than those of uninfected (control) rats, suggesting the development of normocytic and normocromic anaemia with anisocytic alteration. The number of eosinophils did not increase significantly among the groups. We concluded that E. paraensei produces haematological alterations in R. norvegicus, causing regenerative anaemia. This system can therefore be a useful model to study the direct and indirect effects of gastrointestinal infections.     

菜青虫感染玫烟色拟青霉后血淋巴蛋白质含量及几种保护酶活力的变化

对玫烟色拟青霉Paecilomyces fumosoroseus侵染3、4龄菜青虫Pieris rapae后,其血淋巴中蛋白质含量、体内保护酶及谷胱甘肽S-转移酶活力进行了研究。结果表明,感病的3、4龄菜青虫血淋巴中蛋白质含量明显低于同期未感染的幼虫;感病虫体内超氧化物歧化酶、过氧化氢酶和过氧化物酶的活力发生了不同程度的改变,其中对3龄菜青虫体内酶活力的影响比4龄幼虫大。此外被侵染的3、4龄菜青虫体内谷胱甘肽S-转移酶活力在感病前期显著高于同期未感染的菜青虫,而在感病后期明显低于同期未感染的菜青虫。     

Influence of anions and protons on the Adair coefficients of haemoglobins A and Cowtown (His HC3(146) beta----Leu)

We have measured the contribution of the alkaline Bohr effect of the C-terminal histidine residues of the beta-chains of haemoglobin A by comparing haemoglobin A with haemoglobin Cowtown in which those histidine residues are replaced by leucine. Oxygenation of a stripped 2.5 mM (haem) solution of haemoglobin A yielded 0.19 H+/haem, while oxygenation of a similar solution of haemoglobin Cowtown produced no change of pH. Oxygen equilibria measured at 60 microM-haem in 0.1 M-Hepes buffer gave an alkaline Bohr effect of -0.21 H+/haem for haemoglobin A and only -0.01 H+/haem for haemoglobin Cowtown, even though its Hill's coefficient was greater than 2 throughout the pH range studied. These results prove that the chloride-independent part of the alkaline Bohr effect is due to the C-terminal histidine residues of the beta-chains. Oxygen equilibria measured in 0.095 M-bis-Tris buffers with minimal chloride or with 0.1 M-chloride showed the contribution of those histidine residues to the alkaline Bohr effect to be about 0.2 H+/haem, independent of chloride concentration. Determination of the individual Adair coefficients in the three different buffers indicated that pH and chloride tend to have their greatest effects at the second or third steps of oxygenation when the change of quaternary structure is most likely to occur; between pH 7 and 9, the fourth Adair coefficient is only very slightly affected by pH and not significantly by chloride.     

Shell geometric morphometrics in Biomphalaria glabrata (Mollusca: Planorbidae) uninfected and infected with Schistosoma mansoni

Freshwater planorbid mollusks belonging to the genus Biomphalaria act as intermediate hosts for Schistosoma mansoni,the etiological agent of human intestinal schistosomiasis,in the Neotropical Region.Identification ofBiomphalaria spp.are carried out based on morphological characters,and the Schistosoma infection are determined by the presence of cercariae (verified through microscope preparation and mounting).Recently,the geometric morphometrics has proven to be a useful tool for determining shape differences in disease vectors arthropods.Due to this,we used geometric morphometrics to determine Biomphalaria glabrata shell differences (shape and size)between uninfected and infected specimens.We digitalized 12 anatomical points over the shell left side (from umbilicus to the last whorl) by combining type Ⅰ and Ⅱ landmarks and sliding semilandmarks;the coordinates were aligned by generalized Procrustes analysis.Principal component analyses were implemented for examining main variation axes,and discriminant analysis for testing group membership significance.We found significant separation between infected and uninfected shell conformation.All specimens were 100％correctly classified.The main differences occur in the peristome.The Kruskal-Wallis test finds significant differences in shell isometric size among infected and uninfected specimens.These findings correspond to other studies of traditional morphometrics,that infected snails showed the reduction in shell size in contrast to those uninfected specimens.     

Larval trematodes (Digenea) of planorbid snails (Gastropoda: Pulmonata) in Central Europe: a survey of species and key to their identification

A survey of the larval stages (cercariae and metacercariae) of trematodes (Digenea) found in planorbid snails in Central Europe (Austria, Czech Republic, south-east Germany, Hungary, Poland and the Slovak Republic) is presented based on a study of 7,628 snails of 12 species examined between 1998-2006. A total of 34 trematode larval stages, comprising cercariae of 28 species and metacercariae of seven species (one species occurred both as cercaria and metacercaria) of nine families were found in 898 (11.5%) snails of eight species. The dominant cercariae were those belonging to the Rubenstrema exasperatum (Rudolphi, 1819)/Neoglyphe locellus (Kossack, 1910) species complex, Tylodelphys excavata (Rudolphi, 1803) and Echinostoma spiniferum (La Valette, 1855) sensu Nasincová (1992), all from Planorbarius corneus (Linnaeus). Almost the same spectrum of cercariae of the families Echinostomatidae, Plagiorchiidae and Omphalometridae was found in the present study as in previous reports; however, a considerably lower spectrum of cercariae of the families Diplostomidae and Strigeidae was recorded. The most frequent metacercariae were those of Echinoparyphium aconiatum Dietz, 1909, Neoglyphe locellus and Moliniella anceps (Molin, 1859), all occurring mainly in P. corneus. The most heavily infected snail species was P. corneus, followed by Planorbis planorbis (Linnaeus) and Segmentina nitida (Müller). The widest spectrum of trematode species was found in P. planorbis and P. corneus. Forty-two cercariae identified to the species level belonging to 15 families, plus an additional 43 taxa recorded under generic or provisional names, were reported from 11 species of planorbids in previous studies carried out in Central Europe. However, the actual number of trematode species occurring in the planorbid snails is probably much lower, because many, if not most, larval stages reported under provisional names or unidentified to the species level may be conspecific with identified adult forms. A key to the cercariae and metacercariae recorded from planorbids in Central Europe, together with illustrations of those species encountered most frequently in the field, is provided to facilitate identification.     

Effect of different concentrations of heavy metal ions on the normal physical and chemical characteristics of the hemolymph of Palanorbarius purpura (Mollusca: Bulinidae) in the norm and during a trematode infection

Short-term simultaneous effect of high concentrations (LC25, LC50, LC75) of heavy metal ions. (Cu2+, Zn2+, Cd2+ and Pb2+) and infection with trematode partenites Echinoparyphium aconiatum onto haemolymph of mollusk has been investigated. It was noted that low doses of toxicant (2.5 and 10 maximum admitted concentrations) have variable effect. In infected molluscs the concentration of haemoglobin decreases, while in intact ones it increases. In relation to this index, it was found, that the ion Cu2+ is highly toxic, Zn2+ and Cd(2+)--toxic, Pb(2+)--moderately toxic.     

Evidence of intracolony transmission of Thelohania solenopsae (Microsporidia: Thelohaniidae) in red imported fire ants (Hymenoptera: Formicidae) and the first report of spores from pupae.

Red imported fire ant, Solenopsis invicta, colonies were infected horizontally by introducing live brood (mainly larvae and pupae) infected with Thelohania solenopsae. Live, infected brood introduced into uninfected colonies were adopted and raised to adulthood instead of being executed by the recipient colony. Introductions of infected larvae with uninfected pupae, which eclose into adult worker caste fire ants, resulted in an 80% infection rate of the inoculated colonies. Infections from introductions of infected pupae with uninfected larvae resulted in a 37.5% infection of inoculated colonies. Infections were also detected in 11.6 and 3.7% of the adult worker caste ants that eclosed from uninfected large larvae and pupae, respectively, that were held with infected adult workers. Microscopic examination of infected brood revealed sporoblasts and large numbers of spores of T. solenopsae in S. invicta pupae.     

High-performance thin-layer chromatographic analysis of lutein and beta-carotene in Cerithidia californica (Gastropoda) infected with two species of larval trematodes

High-performance thin-layer chromatography (HPTLC) analysis was done on lutein and beta-carotene in the digestive gland-gonad complex (DGG) and whole body of uninfected Cerithidia californica snails and those infected with the larval trematodes Mesostephanus appendiculatis or Euhaplorichis californiensis. HPTLC of the DGG extract on C-18 reversed-phase plates developed in petroleum ether-acetonitrile-methanol (1:2:2) mobile phase showed 2 identifiable pigment zones; the least polar zone had a retention factor (Rf) of 0.07, identical to a beta-carotene standard, and the more polar zone had an Rf of 0.41, identical to a lutein standard. Densitometric scanning of the pigment zones in sample versus standard chromatograms showed that the weight percent of lutein in the uninfected DGGs (3.4x10(-3)%) was significantly greater (P<0.05) than that of DGGs infected with either M. appendiculatis (0.35x10(-3)%) or E. californiensis (0.82x10(-3)%). Changes in beta-carotene in the infected DGGs were insignificant compared to the uninfected controls. However, the beta-carotene content of whole snails was significantly reduced (P<0.05) by infection with either trematode.     

Some aspects of the infection of bib, Trisopterus luscus (L.), by the parasitic copepod Lernaeocera lusci (Basset-Smith, 1896) in Portuguese waters

Winter samples totalling 2308 specimens of Trisopterus luscus (total length range: 13–36 cm) caught in northern Portugal were examined for infections with Lernaeocera lusci. Infection prevalence was of 42%, and infection intensity was 1.7 copepods per infected fish. Individual infection varied between 1 and 9 copepods per infected specimen; 30.6% of the fish had bilateral parasitosis. There was a significant correlation (P<0.01) between fish length and prevalence. Length-weight relationships of uninfected and infected fish were not significantly different (P > 0.05) and thus provide evidence that L. lusci has no effect on the condition of T. luscus in all the examined 1–cm size classes.     

棉铃虫感染中华卵索线虫后血淋巴游离氨基酸含量的变化

中华卵索线虫Ovomermis sinensis感染棉铃虫Helicoverpa armigera 1天后,棉铃虫幼虫血淋巴中游离氨基酸总量大幅度下降,各种游离氨基酸的含量也是如此变化。感染2～4天后的棉铃虫血淋巴中游离氨基酸总量变化不大,各种游离氨基酸的含量有升有降。感染5～6天后的棉铃虫血淋巴中游离氨基酸总量和各种氨基酸的含量都急剧上升。研究表明,中华卵索线虫寄生棉铃虫时,棉铃虫血淋巴中游离氨基酸含量的变化朝着有利于线虫生长发育的方向进行。     

Poisoning by copper sulfate of Lymnaea stagnalis infected with trematode parthenitae

The effect of different concentrations of copper sulphate on the content level of carotinoid pigments in the haemolymph of Lymnaea stagnalis infected with parthenitae of trematodes was studied. The content level of carotinoid pigments in haemolymph of molluscs steadily decreases with an increasing concentration of the toxicant. The mortality of experimental animals under such conditions rises considerably, the mortality of infected animals being much higher than that of non-infected ones.