Efficacy and toxicity of praziquantel in helminth-infected barbel (Barbus barbus L.).

This study evaluated efficacy and toxicity of the pyrazinoisoquinoline anthelmintic praziquantel (PZQ) in barbel infected with metacercariae of Diplostomum spathaceum and adult Pomphorhynchus laevis, and assessed antioxidant biomarkers and the lipid peroxidation response in juvenile barbel post-treatment. The estimated 96-hr LC50 of PZQ was 28.6 mg/L. For evaluation of efficacy, barbel naturally infected with D. spathaceum were exposed to a 10 and 20 mg/L PZQ 4-day bath treatment. Both concentrations were 100% effective against D. spathaceum and significantly (p < .01) affected the activity of catalase, superoxide dismutase, glutathione reductase and glutathione-S-transferase as well as levels of reduced glutathione in liver and muscle. The efficacy of orally administered PZQ was assessed in adult barbel naturally infected with P. laevis. Fish were administered 10, 30 and 50 mg/kg of body weight and examined via gut dissection after 6 days. The 50 mg/kg dose significantly decreased the intensity of infection. Praziquantel is a feasible bath treatment for barbel infected with D. spathaceum and has potential for oral treatment of broodfish infected with P. laevis.

Anthelmintic activity of Rosmarinus officinalis against Dactylogyrus minutus (Monogenea) infections in Cyprinus carpio.

Monogenean parasites are important ectoparasites of fish, and are responsible for severe economic impacts in the aquaculture industry. They are usually treated with chemicals, but the chemicals can have harmful side effects in the fish and may pose threats to human health. Rosemary (Rosmarinus officinalis) is a common medicinal herb, with antimicrobial and antitumor properties. Here, we examined the anthelmintic activity of rosemary extract against the monogenean (Dactylogyrus minutus) in vitro and in vivo using bath treatment and oral administration. The in vitro experiments showed that parasite survival was affected by both rosemary extract concentration and the solvent (water and ethanol). Parasites were dead at 61.8±5.6 and 7.8±1.4min when exposed to 100 and 200g aqueous rosemary extract solution/L of water respectively. It took 166.7±48.2 and 5.4±1.01min to kill the parasites when exposed to 1 and 32g ethanol rosemary extract solution/L of water respectively. Moreover, pure component of rosemary extract obtained commercially used in in vitro experiments showed that 1,8-Cineole was the most toxic component of the main components tested. Parasite intensity and prevalence in fish exposed to 50 and 100g aqueous rosemary solution/L water for 30min were significantly lower than they were in controls (p<0.05). In oral treatment experiments, diets of Cyprinus carpio were supplemented with eight different concentrations of aqueous rosemary extract. The intensity of parasites was significantly less in fish fed for 30days with feed containing 60, 80 and 100ml aqueous extract/100g feed than in control (p<0.05). Together these results indicate that rosemary is a promising candidate for prevention and control of monogenean infection.

Praziquantel treatment for yellowtail kingfish (Seriola lalandi): dose and duration safety study.

Regulatory approval is being sought to use praziquantel (PZQ) to treat flukes infecting yellowtail kingfish (YTK), but accurate safety data were not available. We investigated the effect of increased doses or prolonged exposure of orally administered PZQ on YTK by assessing changes in haematological and biochemical characteristics, and mortality. Fish were intubated daily for 3 days with 0, 100, 300 and 500 mg PZQ kg(-1) BW day(-1) or once daily for 9 days at 0 and 100 mg PZQ kg(-1) BW day(-1). Blood was taken 24 h after the cessation of treatment. There was no significant difference between any of the haematological or biochemical indices in YTK treated with PZQ and controls, indicating that PZQ is safe for use at 100 mg PZQ kg(-1) BW day(-1) in YTK and that exposure to high doses or prolonged duration does not have negative effects on the YTK haematological or biochemical parameters we measured.

Application of small RNA technology for improved control of parasitic helminths.

Over the last decade microRNAs (miRNAs) and small interfering RNAs (siRNAs) have emerged as important regulators of post-transcriptional gene expression. miRNAs are short, non-coding RNAs that regulate a variety of processes including cancer, organ development and immune function. This class of small RNAs bind with partial complementarity to their target mRNA sequences, most often in the 3'UTR, to negatively regulate gene expression. In parasitic helminths, miRNAs are being increasingly studied for their potential roles in development and host-parasite interactions. The availability of genome data, combined with small RNA sequencing, has paved the way to profile miRNAs expressed at particular developmental stages for many parasitic helminths. While some miRNAs are conserved across species, others appear to be unique to specific parasites, suggesting important roles in adaptation and survival in the host environment. Some miRNAs are released from parasites, in exosomes or in protein complexes, and the potential effects of these on host immune function are being increasingly studied. In addition, release of miRNAs from schistosome and filarial parasites into host plasma can be exploited for the development of specific and sensitive diagnostic biomarkers of infection. Interfering with miRNA function, as well as silencing key components of the pathways they regulate, will progress our understanding of parasite development and provide a novel approach to therapeutic control. RNA interference (RNAi) by siRNAs has proven to be inconsistent in parasitic nematodes. However, the recent successes reported for schistosome and liver fluke RNAi, encourage further efforts to enhance delivery of RNA and improve in vitro culture systems and assays to monitor phenotypic effects in nematodes. These improvements are important for the establishment of reliable functional genomic platforms for novel drug and vaccine development. In this review we focus on the important roles of miRNAs and siRNAs in post-transcriptional gene regulation in veterinary parasitic helminths and the potential value of these in parasite diagnosis and control.

Immune modulation by helminth parasites of ruminants: implications for vaccine development and host immune competence.

Parasitic helminths reside in immunologically-exposed extracellular locations within their hosts, yet they are capable of surviving for extended periods. To enable this survival, these parasites have developed complex and multifaceted mechanisms to subvert or suppress host immunity. This review summarises current knowledge of immune modulation by helminth parasites of ruminants and the parasite-derived molecules involved in driving this modulation. Such immunomodulatory molecules have considerable promise as vaccine targets, as neutralisation of their function is predicted to enhance anti-parasite immunity and, as such, current knowledge in this area is presented herein. Furthermore, we summarise current evidence that, as well as affecting parasite-specific immunity, immune modulation by these parasites may also affect the ability of ruminant hosts to control concurrent diseases or mount effective responses to vaccination.

Risk factors and control of intestinal parasite infections in sled dogs in Poland.

Training and racing constitute serious challenges for working sled dogs. Attainment of the highest levels of stamina and speed are possible only by completely healthy dogs. Infections with nematodes as whipworm Trichuris sp. or hookworms Uncinaria/Ancylostoma can significantly reduce the fitness of working dogs leading to anemia or even to death. In the middle of the racing season, between December 2009 and April 2010, 108 individual fecal samples were collected from 25 sled dog kennels situated in different regions of Poland. Saturated salt flotation was performed for helminth egg detection. The immunofluorescent assay MeriFluor Cryptosporidium/Giardia and nested PCRs on 18S rRNA (Cryptosporidium spp.) and TPI gene (Giardia spp.) were carried out for detection of intestinal protozoa. Overall prevalence of 6 species of intestinal parasites was 68% in sled dogs (73/108). In 51 samples the eggs of a single species of helminth were detected (47%), two nematode species were detected in 13%, three species of nematodes were found in two dogs. The most prevalent helminths were the hookworms Uncinaria/Ancylostoma-identified in 36% of kennels, and in 34% of sled dogs. Toxocara eggs were detected in 36% of kennels, in 17% of dogs. Trichuris sp. eggs were found in 20% of kennels (5/25), in 13% of dogs. Cysts/oocysts of intestinal protozoa were detected in 31% of sled dogs. The most prevalent was Giardia spp. infection-in 54% of kennels [13/24], in 28% of dogs. Cryptosporidium spp. infections were identified in 37.5% of kennels [9/24], in 13% of dogs. Two sequenced Giardia isolates presented 100% homology with G. intestinalis Assemblage C isolate (AY228641.1), specific for dogs. A range of factors was shown to affect the prevalence of intestinal parasites in sled dogs. The highest prevalence of parasites was found among dogs from large kennels (housing >3 dogs), in dogs less than 2 years old, and in kennels, where prophylactic treatment was carried out 1-4 times a year. The present study has demonstrated a high prevalence of intestinal parasites in working sled dogs in Poland, including the zoonotic human pathogens Toxocara or Cryptosporidium.

Verminoses dos bovinos: percepção de pecuaristas em Minas Gerais, Brasil / Cattle helminthiasis: farmers perception in Minas Gerais, Brazil

O presente trabalho foi realizado com o objetivo de caracterizar o controle dos endoparasitos em rebanhos bovinos do Estado de Minas Gerais. Foram aplicados questionários em 1.304 propriedades rurais, dos quais 1.289 foram validados e utilizados no estudo. As propriedades localizavam-se em 555 municípios inseridos nas 66 microrregiões do Estado de Minas Gerais e foram selecionados por amostragem aleatória. Os entrevistados foram questionados quanto ao perfil sócioeconômico, características de produção e de manejo sanitário da população bovina de cada propriedade, além das atitudes tomadas para o combate dos endoparasitos. Os resultados demonstraram que as diversas práticas de controle das verminoses recomendadas para os rebanhos bovinos são desconhecidas pela maioria dos pecuaristas.

This study aimed to characterize the cattle endoparasites control in Minas Gerais State. Questionnaires were applied in 1,304 farms and 1,289 were validated and used in this study. The farms were located at 555 municipalities inserted in 66 Minas Gerais micro-regions and were selected by random sampling. The interviewees were asked about socioeconomic profile, production characteristics and cattle sanitary management of each farm, and attitudes to control the endoparasites. Results showed that the parasites control practices recomemended to cattle are unknown to most of the farmers.

Towards the implementation of the "basket of options" approach to helminth parasite control of livestock: emphasis on the tropics/subtropics.

The virtual reliance on anthelmintic drugs alone to control internal parasites of livestock is inappropriate and ultimately unsustainable. In the tropics and subtropics, widespread and high levels of anthelmintic resistance, particularly in nematode parasites of small ruminants, is rife. But more to the point, many farmers in these regions of the world are resource poor and cannot afford, or are reluctant to purchase drugs that may also be of dubious quality. As it is with any intervention, the benefits must outweigh the costs. This is not only in terms of conventional parameters such as reduced mortality and increasing productivity (meat, milk, fibre and traction power) of livestock, but also within the broad framework of helminths of veterinary/human importance, the aim should be a positive impact on reducing the threat of helminth zoonoses. However, understanding the issues involved and education of the end-users (farmers) is of fundamental importance, before any internal parasite control program should be promoted. Within the above context, we provide examples of how the "basket of options" approach could be adopted for the control of three quite disparate helminth problems in the tropics and subtropics, viz.: strongyle nematode infections of donkeys, the Taenia solium cysticercosis/taeniosis problem of pig and man and Haemonchus contortus infections in small ruminants. The "best practice" approaches can be defined as those "basket of options" that are practical, affordable, available and appropriate, whether to the commercial producer, or to the resource-poor farmer. Constraints that may restrict applying such options are accessibility to, and affordability of, suitable remedies and above all, the availability of information needed to make informed decisions in this regard.

Immunology of helminth infections, with special reference to immunopathology.

Immune responses resulting in immunity to infection or disease, share the same basic humoral and cellular mechanisms. While immunity to helminth infection has evolved to mediate rapid elimination of the parasite, the strategies evolved by the parasites themselves aim to delay this rejection process and ensure the survival and distribution of their progeny. Ineffective or incomplete immunity results in persistence of parasites or their products within the host tissues, inappropriate or chronic stimulation by parasite antigens, hyper-reactivity and tissue damage or immunopathology. A long standing classification by Gell and Coombs identifies four major types of hypersensitivity responses accounting for most of the immunopathogenesis, three of which are mediated by antibody and one, delayed type hypersensitivity (DTH), by T cells. This paper aims to give a short review of these four classical hypersensitivity reactions with particular reference to infections of large animals with helminth parasites. In addition, in view of the functionally different helper T cell subsets now identified, the existing DTH response is redefined as DTH Type 1 (Th-1 mediated) and two new classes of T cell-dependent DTH responses are proposed; DTH Type II, associated with the Th-2 type cytokines IL-4 and IL-5 and eosinophilic granuloma formation, and DTH Type III, associated with IL-4 and TGF-beta and fibrosis. Finally, some implications of immunopathology on parasite control strategies are discussed.

Field activity cost estimates for the first 3 years of the World Bank Loan Project for schistosomiasis control in China.

The World Bank Loan Project for schistosomiasis in China commenced field activities in 1992. In this paper, we describe disease control strategies for levels of different endemicity, and estimate unit costs and total expenditure of screening, treatment (cattle and humans) and snail control for 8 provinces where Schistosoma japonicum infection is endemic. Overall, we estimate that more than 21 million US dollars were spent on field activities during the first three years of the project. Mollusciciding (43% of the total expenditure) and screening (28% of the total) are estimated to have the most expensive field activities. However, despite the expense of screening, a simple model predicts that selective chemotherapy could have been cheaper than mass chemotherapy in areas where infection prevalence was higher than 15%, which was the threshold for mass chemotherapy intervention. It is concluded that considerable cost savings could be made in the future by narrowing the scope of snail control activities, redefining the threshold infection prevalence for mass chemotherapy, defining smaller administrative units, and developing rapid assessment tools.