Strategies for describing myxozoan pathogens, dreadful fish diseases in aquaculture.

Myxozoans are obligate endoparasites, cosmopolitan in distribution with both vertebrate and invertebrate hosts. Their myxospores consist of shell valves, polar capsules with coiled polar tubules that are extrudible, and infective amoeboid germs. Myxozoan parasites are most abundant, and due to their increasing number in recent years, they can pose an emerging threat to the fish industry worldwide. Hence, the immediate need is to devise a strategy to understand and detect parasites and parasitism. They may proliferate to different organs with the advancement of infection. This all warrants the development/devising of strategies and results of integrative studies in order to identify these dreadful parasites and resolve taxonomic issues. Different methods whether classical methods including gross morphology or advanced methods such as electron microscopy (SEM, TEM, STEM), Confocal laser scanning microscopy (CLSM), histopathological studies, site preference, host and tissue specificity, a molecular approach using new markers can be clubbed for identification because these parasites are hidden and are difficult to recognize. This group was earlier classified only on the basis of myxospores morphology, but due to the high structural variability of this group advanced methods and approaches have to be implied which can minimize the problems in assigning new species.

Evaluation of anthelmintic antimicrobial and antioxidant activity of Chenopodium album.

The objective of this study was to evaluate the anthelmintic, antimicrobial and antioxidant activities of Chenopodium album against gastrointestinal nematodes of sheep and some pathogenic microbes. A worm motility inhibition assay was used for in vitro study, and a faecal egg count reduction assay was used for an in vivo study. Various concentrations ranging from 100 to 500 µg/ml of the extract were subjected to antimicrobial screening by disc diffusion method against four selected bacterial (Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas multocida and Escherichia coli) and two fungal (Aspergillus flavus and Candida albicans) strains in order to estimate the medicinal potential of the herb. DPPH (1,1-diphenyl-2-picrylhydrazyl), riboflavin photo-oxidation, deoxyribose, lipid peroxidation assays were used for antioxidant activity. The extracts exhibited dose- and time-dependent anthelmintic effects on the Haemonchus contortus as compared to levamisole. The extract showed maximum inhibitory effect against S. aureus (28 ± 0.14 mm), while as mild inhibitory effect was observed against E. coli among the selected microbial strains. The effect produced by the different extract concentrations was comparable with the standard antibacterial agent streptomycin sulphate and antifungal agent nystatin, which were used as effective positive control in the study. The antioxidant activity showed that the extracts exhibited scavenging effect in concentration-dependent manner on superoxide anion radicals and hydroxyl radicals leading to the conclusion that the plant has broad spectrum anthelmintic, antimicrobial and antioxidant activities and could be a potential alternative for treating various diseases.

First record of myxozoan parasites from fresh water fishes of Jammu and Kashmir and their pathogenecity.

The present study was aimed to isolate myxozoan parasites infecting the native carp Schizothorax esocinus (Heckel) and pond reared carp Labeo rohita (Ham.) collected from fresh water ecosystems of Jammu and Kashmir from July 2013 to June 2014. Various organs such as eyes, gills, scales, fins, intestines, visceral organs and pericardial cavity of as many as 100 live specimens were examined. Results revealed that 14% of the examined L. rohita were infected with Myxobolus rocatlae (Ham.) and 6% of S. esocinus (Heckel) were infected with M. kashmirensis sp. nov. Clinical signs revealed mucous laden gills while the parasitological observation under stereozoom binocular microscope revealed multiple minute plasmodia on the gills. On rupturing these plasmodia with sharp needle 100-500 myxospores were liberated. The identification to species level was done on the basis of morphological and morphometric attributes of the myxospores. This is the first record of myxozoan parasites in fishes from the state of Jammu and Kashmir, India. Histological observations of infected gills revealed plasmodia of two types (LV4 intralamellar vascular type) in M. rocatlae and (LV3 intralamellar vascular type) in M. kasmirensis causing necrosis, hyperplasia, hypertrophy and vacuolization of the epithelial and vascular endothelium.

Myxobolus chushi n. sp. (Myxozoa:Myxosporea) parasitizing Schizothorax niger (Heckel), a native cyprinid fish from Wullar Lake in Kashmir Himalayas.

In the study, a new species, Myxobolus chushi n. sp. infecting gills of wild specimens of Schizothorax niger (Heckel) inhabiting Wullar Lake in Kashmir Himalayas, (J&K) India has been described based on morphology of the myxospore and using partial 18S rDNA sequencing. Pathological changes in the gills have been studied with the help of histological sections stained with Luna's method. Twenty fish specimens were examined, out of which four had oval, white plasmodia in gills measuring 2.0×0.5mm. The myxospores were spherical to ovoidal in shape with slightly attenuated posterior end, measuring 11.17±0.23 (10.60-11.40)µm in length and 9.14±0.06 (8.80-9.20)µm in width, having a prominent pore at the anterior end. The polar capsules were pyriform in shape, measuring 4.25±0.15 (4.00-4.40)µm in length and 2.38±0.27 (2.00-2.65)µm in width having polar filaments forming coils up to 5 in number. Parietal folds 9 in number present on the posterior part of the shell. The intensity of infection was recorded to be moderate as indicated by gill plasmodial index (GPI=2). The plasmodium was located in the vascular network occupying whole of the gill lamella therefore typed as intralamellar vascular type, LV3. Analysis of 18S small subunit (SSU) rDNA sequence of the isolate demonstrated 90% homogeneity with M. sp. KLT-2014 infecting scales of Labeo rohita from Myanmar and 89% with M. dermiscalis infecting scales of Labeo rohita from India.

Breaking the resistance of Escherichia coli: Antimicrobial activity of Berberis lycium Royle.

The antimicrobial activity of root bark of Berberis lycium and its principal component berberine was tested against a panel of microbial strains using agar well diffusion test and further analyzed using micro-broth dilution method. Preliminary analysis, on the basis of zone of Inhibition (ZOI) showed that the methanolic extract of B. lycium was highly effective against Escherichia coli (ZOI 41 ± 1 mm). Among the bacterial strains E. coli was found to be most susceptible and among fungi Candida albicans was the most susceptible for berberine as well as the crude methanolic extract of the plant. Methanolic extract of the plant was more effective for E. coli (MIC 1.7 ± 1.18; MBC 2.4 ± 1.18) than berberine (MIC 3.5 ± 0.57) (p < 0.05), whereas berberine was more effective than crude extracts for C. albicans. In addition, E. coli showed the development of resistant colonies after 72 h when tested with berberine but the development of such colonies was not observed with the methanolic extract of the plant. This could be due to the presence of resistance breaking molecules in the crude methanolic extract of B. lycium. Also the MIC index of crude methanolic extract was 1.39 for E. coli, which showed the mode of action to be bactericidal. HPLC analysis revealed the presence of berberine at highest concentration in methanolic extract of the plant, followed by aqueous extract. Potentiation of this berberine by resistance breaking molecules in the crude extract could be a possible explanation for its strong effectiveness.

A field survey on the status of gastrointestinal helminth parasites in hangul (Cervus elaphus hanglu) in Dachigam National Park of Kashmir.

One year crossectional survey was carried out to determine and describe the prevalence and intensity of gastrointestinal parasite infections in hangul (Cervus elaphus hanglu) in Dachigam National Park of Kashmir through faecal examinations. Out of 153 faecal samples examined, 82 (53.59 %) were found infected with GIT helminthes. In present study seven helminth species were found, including five nematode [Haemonchus contortus (55.39 %), Trichuris ovis (39.75 %), Dictyocaulus viviparus (28.4.00 %), Oesophogostomum circumcincta (13.7 %) and Chabertia ovina (4.02 %)] one trematode [Fasciola hepatica (17.3 %)] and one cestode species [Moneizia expansa (6.05 %)]. Based on the severity of infection 81.7 % of hangul positive samples were severely infected (epg > 1,500), 8.3 % heavily infected (epg = 1,100-1,500), 3.8 % moderately infected (epg = 800-1,000) and 7.2 % mildly infected (epg = 500). Season, sex and age were the factors that influenced the epidemiological prevalence of GIT helminths in hangul in the present study. The maximum helminth infection was observed in summer season and lowest in winter (P = 0.003). Lower age groups were more infected than adult animals (P > 0.05). Prevalence was higher in males than females (P > 0.05). The present study will initially be of great significance to add to existing knowledge of the epidemiology of GIT helminth of hangul which is the pioneering study on this animal in the valley and the findings will be quite helpful to devise the appropriate control and prophylactic strategies for GIT helminthiasis of hangul in the Dachigam national park.

Population analysis of the genera buildup on some commercially important vegetable crops grown in Kashmir Valley.

In order to list the genera buildup on Brassica oleracea (Cauliflower), Capsicum frustscens (Chillies), Spinacia oleracea (Spinach) and Phaseolus vulgaris (Farash bean) a general survey was carried in the summer season of 2013. On examining the soil samples as well as root samples of Cauliflower Boleodorus, Psilenchus, Helicotylenchus, Merlinius, Aglenchus and Filenchus were encountered with their absolute density of 18, 31, 34.5, 35, 35 and 31.5 %. On Chillies, Tylenchus, Helicotylenchus, Psilenchus, Pratylenchus and Hirschmaniella with their absolute frequency of 86, 96.6, 100, 100 and 96.6 % respectively were collected. Among these five genera recorded on Chillies the Relative prominence value of 16 % is lowest for Psilenchus and 26.7 % for Helicotylenchus. Merlinius, Psilenchus, Aglenchus, and Boleodorus were encountered on examining the soil and root samples of Spinach during the period of study. Boleodorus was met with the highest percentage of frequency of 90 % as well as highest proportion of absolute density of 37 % and lowest value of 24 % in case of Merlinius and Psilenchus. Soil samples as well as root samples of Farash bean, Aglenchus, Tylenchus, Hirschmaniella, Helicotylenchus and Psilenchus were encountered with their absolute density of 27, 19.5, 33, 33 and 35.4 %. The absolute frequency of 100 % was found in Tylenchus with the lowest frequency of 70 % in Helicotylenchus. Among the genera recorded on Farash bean relative prominence value happened to be highest in Hirschmaniella with the value as 23.6 % and lowest on Tylenchus with the value of 14.2 %.

A report on the incidence of Trichodina heterodentata from fishes of Jammu, J&K India.

The present study was aimed at screening for the presence of protozoan's among Cyprinid fishes collected from various fish ponds and farms in Jammu division of Jammu and Kashmir (J and K) state. Out of 75 fishes collected from local water bodies of Jammu division, only 35, (49.6 %) were infested with Trichodina. Trichodina infestations were studied in the period from November to February. In light infestation Trichodina was usually present on gills, fins and skin of apparently healthy fish. Clinical signs of Trichodiniasis appeared on fish with heavy infections and in presence of one or more stress factors including, rough handling during transportation from ponds, over crowdedness, malnutrition, high concentration of free ammonia and low oxygen concentration. Clinical signs of Trichodiniasis in fish such as sluggish movement, loss of appetite, black colouration, necrosis and ulcer on different parts of the body, detached scales and excessive accumulation of mucous in gill pouches were also observed.

First record of protozoan parasites in cyprinid fish, Schizothorax niger Heckel, 1838 from Dal lake in Kashmir Himalayas with study on their pathogenesis.

Trichodina heterodentata Duncan, 1977 and Ichthyophthirius multifiliis Fouquet, 1876 obtained from gills during a parasitological survey conducted for the protozoan parasitic fauna of Schizothorax niger a snow trout in Dal Lake, Kashmir, India during the period October 2013 and March 2015. Thirty out of 180 fish were found infected with protozoan parasites. During the study of their pathogenecity the most common deteriorating signs observed in gill tissue were necrosis, hypertrophy, hyperplasia and fusion of secondary lamellae. Prevalence of infection was found to be 16.66%. This is the first record of the protozoan fauna of the schizothoracines from Kashmir valley, India.

Synergistic approach for treatment of chicken coccidiosis using berberine--A plant natural product.

Despite the advent of anticoccidial drugs and vaccines, coccidiosis continues to result in substantial economic losses to the poultry industry. Berberine, a natural alkaloid is well known in studies involving synergistic approaches, thereby reducing the dosage of principal drugs. Therefore, a study was designed to see whether a synergistic anticoccidial effect could be obtained between amprolium and berberine, in vivo using broiler chicken. Anticoccidial activity was measured in comparison to the reference drug amprolium on the basis of oocyst output reduction, mean weight gain and feed conversion ratio. Oocyst output was measured using Mc-Masters counting technique. Different combinations of berberine and amprolium were tested and out of which 1:1 ratio was the most effective for controlling these parasites. Oral gavaging of 100(50 + 50) mg/kg body weight of 1:1 ratio of amprolium and berberine caused the equivalent reduction in number of oocysts (38.85 ± 9.61) one day prior to that of standard drug amprolium (49.95 ± 16.65) as well as pure berberine (44.4 ± 9.61) used in the study. Weight gain of birds was also highest in the synergistic group (1547.43 ± 12.86) among all the infected groups. Besides feed conversion ratio in the synergistic group was also better (1.387 ± 0.026). The results of this study proved the effectiveness of both amprolium and berberine and revealed synergism between amprolium and berberine against coccidian oocysts, confirmed by significant reduction in the number of coccidian oocysts shed in the feces, leading to better weight gain and improved feed conversion ratio. The study deep-rooted the synergistic potential of berberine, a natural bioactive compound for controlling a protozoan parasite and the results of this study corroborate with its use for treatment of severe diarrhoea, amoebiasis and intestinal infections.

[A comparative analysis of various antigenic proteins found in Haemonchus contortus--a review].

Many innovative researches on the development and introduction of recombinant vaccines against many economically important parasites were carried out in the 20th century. Research continues to hold promise with the development of immunological and molecular approaches for control of these parasites and in this regard it has already been seen that blood-sucking parasites such as Haemonchus contortus and Ostertagia ostertagi are susceptible to control by vaccines containing "novel" or "concealed" antigens. Haemonchus contortus is primarily pathogenic to sheep and its blood-feeding behaviour causes effects ranging from mild anaemia to mortality in young animals. Current means of control which are dependent on repeated treatment with anthelmintics are responsible for the increasing drug resistance of this parasite. Together with the growing concern of residual chemicals in the environment and food chain, this has led to attempts to better understand the biology of the parasite with an aim to develop alternate means of control, including the development of molecular vaccines. More problematic and also important is the formulation and delivery strategy to induce expulsion of this parasite, using vaccines containing recombinant "conventional" antigens. Tremendous progress has been made in the last decade in identifying several antigens from Haemonchus contortus which in their native form stimulate useful levels of protective immunity. Vaccines have been developed against H. contortus using 'novel' gut antigens from the parasite, but variable responsiveness of the host sheep has resulted in varying degrees of protection which are stimulated by these vaccines. Computer models have also been used to simulate vaccine efficacy in worm control and have yielded good results. This review will try to summarise the protective efficacy and also the molecular properties of principal candidate antigens which are expressed by this parasite. The review will try to cover the aspirations, current success, limitations and problems faced by researchers in the control of this economically important parasite.

Protein profiling of Haemonchus contortus found in sheep of Kashmir valley.

Economic losses due to helminth parasites in sheep throughout the world are considerable. Haemonchus contortus is a blood sucking intestinal helminth that lives in the abomasum of small ruminants worldwide. This parasite can be devastating to producers as it causes decreased production levels due to clinical signs such as anaemia, edema and death. For isolation of the proteins of the parasite, a well defined methodology was adopted. The abomasae of sheep in which this parasite resides were collected from abattoirs of various districts and were then carried to laboratory for screening. In case of collection sites falling in far areas, the organs were screened on spot. The parasites were collected in normal saline, washed and stored in 0.05 M PBS with pH of 7.4 at 0 °C. After refrigeration, frozen nematodes were thawed, homogenized and centrifuged at 1,000-15,000 rpm for 15 min. The supernatant was thus collected as a protein mixture and stored at -20 °C. Protein concentration of the samples was estimated by Lowry method. The samples were then analyzed through PAGE and then through SDS-PAGE. Protein estimation of the samples was estimated to be 4.2 mg/ml. The processed parasite samples were then subjected to PAGE and SDS-PAGE to determine the presence of the proteins. It showed high concentration of proteins in its whole protein profile. The proteins were seen as continuous bands intermixing with each other in PAGE analysis. The present study revealed two bands of molecular weights-55 and 33 kDa in PAGE analysis. The proteins when analyzed through SDS-PAGE were mostly found in the range of 25-70 kDa. The SDS-PAGE analysis showed four prominent bands. These bands were of the molecular weights of 66, 40, 33 and 26 kDa. The present work was a challenging one since only a single study was conducted in this region on this aspect and thus obviously was a big task to peep into the field where scanty input was available.

Does alteration in biodiversity really affect disease outcome? - A debate is brewing.

How changes in biodiversity alter the transmission of infectious diseases is presently under debate. Epidemiologists and ecologists have put a lot of effort to understand the mechanism behind biodiversity-disease relationship. Two important mechanisms, i.e. dilution and amplification theories have in some manner made it clear that biodiversity and disease outcome have an intimate relationship. The dilution effect theory seems to answer some overarching questions, but paucity of information about many disease systems is a real obstacle for its acceptance. Also, there is hardly any agreement on host population threshold and critical community size vis-à-vis wild life diseases. We suggest a multidimensional approach whereby the same disease system needs to be studied in different ecological zones and then the effect of biodiversity on disease outcome needs to be ascertained. Nonetheless, caution is to be taken while jumping to any conclusion as biodiversity-disease relationship is a multifactorial process.

In vivo anticoccidial activity of berberine [18, 5,6-dihydro-9,10-dimethoxybenzo(g)-1,3-benzodioxolo(5,6-a) quinolizinium]--an isoquinoline alkaloid present in the root bark of Berberis lycium.

Coccidiosis, caused by various Eimeria species, is a major parasitic disease in chicken. However the increasing resistance of these parasites to currently used anticoccidial drugs has stimulated the search for new methods of control. As part of this effort we investigated the root bark of Berberis lycium (barberry) as a potential source of compounds with anticoccidial activity. In the present study anticoccidial activity of different solvent extracts of the root bark of B. lycium and berberine was evaluated in vivo using broiler chicken. Results of the study demonstrated equipotent efficacy of pure berberine in comparison to that of standard drug amprolium on the basis of reduction in coccidian oocyst output, body weight gain of chicken and feed conversion ratio. Among the extracts crude methanolic extract showed highest anticoccidial activity tested at 300 mg/kg body weight which could be due to the presence of alcohol-soluble active ingredients in root bark of B. lycium. Toxicological studies revealed that B. lycium extracts as well as berberine were not lethal up to dosage of 2000 mg/kg body weight. LD(50) was not determined as mortalities were not recorded in any of the five groups of chicken. From the present study it can be concluded that root bark of B. lycium has the immense potential to contribute to the control of coccidian parasites of chicken. Our results corroborate the use of berberine for treatment of severe diarrhoea, amoebiasis and intestinal infections and could justify its use in folk medicine for treatment of haemorrhagic dysentery.

Trophic status and helminth infracommunities of fish populations in Kashmir Himalayan lakes.

The present study considers the influence of the trophic status of three Kashmir Himalayan lakes on the patterns of helminth infracommunities in populations of three species of fish during 2006 to 2008. Data were collected from three lakes of differing trophic status in the Kashmir Himalayas, namely Anchar, a hyper(eu)trophic lake; Dal, a eutrophic lake; and Manasbal, a meso(eu)trophic lake. Three species of fish examined included the native fish Schizothorax niger Heckel and two exotic species--Carassius carassius (Linnaeus) and Cyprinus carpio Linnaeus. The analysis of data showed a clear habitat effect on the abundance pattern of helminth species, thus revealing lake-specific differences in parasite infracommunities of both S. niger and C. carassius. Helminth infracommunity richness was the highest in host populations from the Anchar lake compared to other two lakes. Low values in the Manasbal lake emphasize the low diversity of their helminth infracommunities. On the other hand, there was no observed pattern of community structure in the case of C. carpio in the three lake sites. However due to bias in sampling there was no distinct effect of fish body size on parasite infracommunity structure, although the present results do show that fish parasite data can be meaningful in diagnosing changes in the trophic condition of eutrophic lakes.

Seasonal changes in infrapopulations of Diplozoon kashmirensis Kaw, 1950 (Monogenea: Diplozoidae) along a eutrophic gradient.

Seasonal population dynamics of the monogenean, Diplozoon kashmirensis Kaw, on the gills of two cyprinid fish species, Schizothorax niger Heckel and Carassius carassius (Linnaeus), was investigated in three limnologically distinct trophic habitats located along the flood plain of River Jhelum in Kashmir from June 2006 to May 2008. The parasite infrapopulations exhibited a marked seasonal regime in infestation pattern as the infection indices increased to a much higher plateau during summer season at all the lakes, while the lows were recorded in winter. The heterogeneity in infection pattern indicates that water temperature is an important determinant of the seasonality of infrapopulations at all the localities. Furthermore, the results of our work clearly indicate that the parasite infrapopulations increased proportionally with eutrophication level and, as such, the highly eutrophic habitat, Anchar Lake, was significantly more favourable for parasite infrapopulations than the less eutrophic ones. However, the lakes presented no significant interlake differences in water temperature. Therefore, we could argue that interlake differences in the infestation pattern of parasite can be safely attributed to respective water quality in the lakes rather than water temperature. We propose that infrapopulations of the diplozoid studied herein do respond to differences in water quality of lakes and, thus, could qualify as simple and reliable indicator species in short-term comparative assays by lake managers.

Species spectrum, diversity profile and infection indices of helminth parasite fauna of Chirruh snowtrout, Schizothorax esocinus (Heckel) in lake ecosystems of Kashmir Himalayas-Do similarity and host-parasite associations arise?

In order to assess the species richness and diversity profile of helminth parasite fauna in an endemic fish, an investigation was carried out in two urban and two rural lakes of Kashmir. Overall nine species of helminth parasites were observed in four lakes. Of these three were autogenic and six were allogenic. Heteroxenous parasite species were more in number than monoxenous species. Results showed significant differences in heteroxenous / monoxenous ratio between different lakes. Core species (Prevalence > 20) were only found in hypertrophic lake (Anchar Lake). Overall, majority of helminth species were either secondary or satellite species. Prevalence of some helminth parasites showed significant differences in different lakes. In addition mean intensity showed significant differences between autogenic and allogenic parasites (P < 0.05). Principle Component Analysis based on prevalence showed that Anchar Lake was strongly associated with most of helminth parasites. Diversity indices showed significant variation between different lakes. Maximum helminth species per host was in Anchar Lake. Finally we concluded that helminth parasite fauna showed significant differences in species richness and infection indices between different lakes. Diversity profile was higher in Anchar Lake in comparison to other three lakes. The results clearly show that environmental features of lake ecosystems have got an impact on distribution pattern of helminth parasites in S. esocinus. We suggest comparative parasitological study should be taken between different species of fish in order to have a clear picture regarding the species composition of helminth species in this region. Also we need to characterize the species spectrum of parasitic worms in fish of freshwater bodies of this region as well as other similar type of climatic zones because parasite fauna is an integral part of the inventory of biodiversity and as possible regulators of host populations in aquatic ecosystems.

Helminth communities of fish as ecological indicators of lake health.

This paper deals largely with the dynamics and changes in the helminth parasite communities of fish along the trophic gradient of lakes. The use of parasitological community data as a bioindicator of environmental health underlines the need to study parasite communities at comparable localities with known pollution levels. The comparison of the conditions in different habitats might be helpful to differentiate between normal fluctuations in ambient conditions and pollution-mediated effects. Therefore, the present study was designed to examine the community structure of parasites in snow trout (Schizothorax niger Heckel) inhabiting 3 lakes of contrasting trophic status in Kashmir. The idea of selecting the lakes, namely Anchar (strongly hypereutrophic), Dal (eutrophic) and Manasbal (mesotrophic) for this study was intentional as they depict different trophic gradients and exhibit the desirable pattern which was a prerequisite for this study. The findings presented in this article suggest an apparent lake-wise gradient in community structure, as the increase in trematode and cestode infections in Anchar was markedly greater, to levels clearly distinguishable from those in the other two water bodies. We conclude that human-induced eutrophication of lakes modifies the parasite community at component level and community-level studies on parasites may provide information on health status of lakes.

In vitro and in vivo anthelmintic activity of Euphorbia helioscopia L.

The anthelmintic potentials of the aqueous and methanol extracts of Euphorbia helioscopia were investigated. In folklore medicine, E. helioscopia (Euphorbiaceae) is used in the treatment of various gastrointestinal ailments and diseases. A worm motility inhibition (WMI) assay and egg hatch assay (EHA) were used for in vitro assessment, and a faecal egg count reduction (FECR) assay was used for an in vivo study. The in vitro study revealed anthelmintic effects of crude methanolic extracts of E. helioscopia on live Haemonchus contortus worms as evident from their paralysis and/or death at 8h after exposure, different concentrations (12.5 mg ml(-1), 25 mg ml(-1) and 50 mg ml(-1)) of aqueous and methanolic extracts were used against H. contortus which exhibited dose-dependent anthelmintic effects on H. contortus. Aqueous and methanolic extracts of aerial parts (stem, leaves and flowers) of E. helioscopia were found to have very low percent inhibitory egg hatching as compared to levamisole. It is concluded that the entire plant of E. helioscopia possesses significant anthelmintic activity and could be a potential alternative for treating cases of helminth infections in ruminants.

Infection level of monogenean gill parasite, Diplozoon kashmirensis (Monogenea, Polyopisthocotylea) in the Crucian Carp, Carassius carassius from lake ecosystems of an altered water quality: What factors do have an impact on the Diplozoon infection?

Monogeneans are of great interest to the ecologists because of their simple life cycle and are considered as one of the important and sensitive parasites to any changes in water quality. The monogenean gill parasite, Diplozoon kashmirensis of the Carassius carassius was examined with the aim to evaluate the infection level and the factors influencing the infection. Our results showed that highest prevalence (34.22%) of D. kashmirensis was in the lake having high trophic status and least prevalence (10.90%) in the lake having least trophic status. Infection levels were significantly higher at the basins/sites having deteriorated water quality in comparison to the basins/sites having better water quality. The combined effect of pollutants and eutrophication showed decrease in intensity of D. kashmirensis in one of the polluted/hypertrophied site in the hypertrophic lake (Anchar Lake), while as synergistic effect (i.e. increase) on prevalence and intensity was observed in one of the polluted/hypertrophied site in the eutrophic lake (Dal Lake). Sex was not an important factor influencing the D. kashmirensis in three lakes. The study showed that condition factor was significantly higher in the hypertrophic lake and in an uninfected fish. In addition microhabitat influenced the Diplozoon infection. Most preferable site for the D. kashmirensis was middle sector of the gill. Furthermore, the gill parasite showed significantly positive correlation (r(p)=0.6, P<0.05) with water temperature in the hypertrophic lake, thus showing the impact of water temperature on this parasite. It was concluded that intra lake differences, season, condition factor, microhabitat and water temperature influenced the infection of D. kashmirensis. It seems from the data that eutrophic and hypertrophic habitats were favourable for the D. kashmirensis. The D. kashmirensis showed both antagonistic and synergistic response to the combined effect of pollution and eutrophication.