Untersuchungen über den Befall des Rübenzystennematoden (Heterodera schachtii Schmidt) durch pilzliche Eiparasiten in zwei Zuckerrüben-Getreide-Fruchtfolgen

Abstract Investigations on fungal egg parasites of the sugar beet nematode ( Heterodera schachtii Schmidt) in two sugar beet-cereal-rotations
In two field trials with rotations of sugar beet-cereals-green manure the population dynamics of Heterodera schachtii was investigated as well as the frequency of its fungal egg parasites. During the rotation, highest rates of fungal parasitism (20–48%) were observed after years with sugar beet, independently of the nematode population density. In the interim parasitism decreased to about 10%. A highly significant correlation was found between nematode population density and the amount of parasitized eggs and larvae. The effect of resistant catch crops (oil radish'Pegletta and white mustard'Maxi') on fungal egg parasites was not different from the effect of Phacelia or fallow. There are indications, however, that a high nematode multiplication on a susceptible catch crop promotes antagonistic fungi and, in consequence, a nematode population decrease may occur in the following sugar beet crop.     

Tropical soil microflora of spice-based cropping systems as potential antagonists of root-knot nematodes

Suppression of plant parasitic nematodes with nematode predators, parasites or antagonists is an eco-friendly approach than the toxic chemicals. In a study, soil borne fungi from the rhizosphere of major spice crops were collected from diverse cropping systems prevailing in three southern states of India. A series of in vitro studies were conducted using 73 freshly collected fungal isolates and 76 isolates obtained from other sources. Out of this 67 isolates were not parasitic on females of root-knot nematodes whereas 115 isolates, though colonized the egg masses, did not show any signs of parasitism on nematode eggs. Fifty-nine isolates showed 50-90% inhibition in egg hatch. Pochonia chlamydospora, Verticillium lecanii, Paecilomyces lilacinus, and few isolates of Trichoderma spp. showed >25% parasitism on root-knot nematode eggs. The most promising isolates in this study were one isolate each of Aspergillus (F.45), Fusarium (F.47), and Penicillium (F.59); three each isolates of Trichoderma (F.3, F.52, and F.60) and Pochonia (F.30 and Vc.3) Verticillium (Vl); and two isolates of fungi that could not be identified (F.28 and F.62). Parasitism by Aspergillus tamarii, Aspergillus ustus, Drechslera sp., Humicola sp., and Scopulariopsis sp. on root-knot nematode eggs or females, reported in the present study, are new reports.     

Epidemiology of intestinal helminth parasites of dogs in Ibadan, Nigeria

An epidemiological study of gastrointestinal helminths of dogs (Canis familiaris) in two veterinary clinics in Ibadan, Nigeria, was conducted between January 2001 and December 2002. Faecal samples collected from 959 dogs were processed by modified Kato-Katz technique and then examined for helminth eggs. The results of the study showed that 237 (24.7%) of the dogs examined were infected with different types of helminths. The prevalences for the various helminth eggs observed were: Toxocara canis 9.0%, Ancylostoma spp. 17.9%, Toxascaris leonina 0.6%, Trichuris vulpis 0.5%, Uncinaria stenocephala 0.4% and Dipylidium caninum 0.2%. The faecal egg intensities, determined as mean egg count/gram of faeces ( +/- SEM), were: T. canis 462.0 +/- 100.5, Ancylostoma spp. 54.1 +/- 8.6, T. leonina 0.8 +/- 0.4, T. vulpis 0.1 +/- 0.0, U. stenocephala 1.0 +/- 0.7 and D. caninum 0.2 +/- 0.1. Host age was found to be a significant factor with respect to the prevalence and intensity of T. canis and Ancylostoma spp. There was no significant difference in the prevalence of intestinal helminth parasites between male (27.0%) and female (22.5%) dogs (P>0.05). The prevalence of helminth parasites was significantly higher (P < 0.05) in the local breed (African shepherd) (41.2%) than in Alsatian dogs (16.2%) or in other exotic breeds (21.0%). Single parasite infections (85.7%) were more common than mixed infections (3.5%).     

The specificity of nematode allergens in the diagnosis of human visceral larva migrans.

Extracts of nematodes have been used as skin test antigens in the diagnosis of nematode infections for many years. Ascaris lumbricoides and Toxocara canis are two nematodes commonly involved in human parasitism, the latter being associated with the clinical condition of Visceral Larva Migrans. In vitro and in vivo experiments reported in this paper confirm experimentally, as well as clinically, the existence of cross-reacting antigens between T. canis and Ascaris spp., and probably between T. canis and other nematodes. These cross-reactions compromise the usefulness of skin tests in the diagnosis of such parasitic infections.     

Prevalence of endoparasites in dog faecal deposits in Jordan.

Of 756 dog faecal deposits collected from the road side and public places in five Jordanian Governorates 466 (61.6%) harboured parasite ova, oocysts and/or larvae. Of these, 67.7%, 26.8%, 4.1% and 1.4% revealed one, two, three and four different types of parasite ova, oocysts or larvae in the same sample, respectively. Examination of the infected samples revealed that 44.1% contained taeniid ova, 19.8% Dipylidium caninum, 19% Toxocara canis, 8% Sarcocystis spp. oocysts, 5% hookworm larvae, 1.5% Diphyllobothrium latum, 0.1% Capillaria spp, and 0.1% Trichuri vulpis eggs.     

Fungal ovicidal activity on Toxocara canis eggs

BackgroundVisceral toxocariasis is a parasitic zoonosis caused by Toxocara canis. The prevalence of this parasite in dogs, soil contamination and the resistance of eggs increase human exposure to the disease. Moreover, the difficulties of the control measures justify the need for alternative ones.AimsThe objective of this study was to evaluate the in vitro ovicidal activity of fungi isolated from soils from public places in the city of Pelotas, Rio Grande do Sul, Brazil, on Toxocara canis.MethodsSamples of soil from ten localities were inoculated onto Petri dishes with 2% water–agar (WA) that contained antibiotics, and incubated at 25 °C/21 days. Isolated fungi were tested in vitro for ovicidal activity, with five replicates. One mL of an embryonated Toxocara canis egg suspension (10³ eggs) was poured over the fungal cultures after 10 days of growth. At intervals of 7, 14 and 21 days, 100 eggs were removed from each plaque and evaluated by optical microscopy.ResultsAcremonium, Aspergillus, Bipolaris, Fusarium, Gliocladium, Mucor and Trichoderma were isolated from the soil. A significant ovicidal type 3 effect was observed in Trichoderma, Fusarium solani complex and Acremonium. Those isolates from the genus Trichoderma showed their ovicidal effect on the 14th day of fungus–egg interaction. The other fungal genera tested showed a type 2 effect.ConclusionsThese results suggest that the use of Trichoderma and Fusarium solani complex in biological control of T. canis is promising; however, further studies should be performed.     

Fungal chitinases and their biological role in the antagonism onto nematode eggs. A review

Chitin, the most abundant aminopolysaccharide in nature, is a rigid and resistant structural component that contributes to the mechanical strength of chitin-containing organisms. Chemically, it is a linear cationic heteropolysaccharide composed of N-acetyl-D-glucosamine and D-glucosamine units. The enzymatic degradation of chitin is performed by a chitinolytic system with synergistic and consecutive action. Diverse organisms (containing chitin or not) produce a great variety of chitinolytic enzymes with different specificities and catalytic properties. Their physiological roles involve nutrition, parasitism, chitin recycling, morphogenesis, and/or defense. Microorganisms, as the main environmental chitin degraders, constitute a very important natural source of chitinolytic enzymes. Nowadays, the most used method for pest and plant diseases control is the utilization of chemical agents, causative of significant environmental pollution. Social concern has generated the search for alternative control systems (i.e., biological control), which contribute to the generation of sustainable agricultural development. Interactions among the different organisms are the natural bases of biological control. Interest in chitinolytic enzymes in the field of biological control has arisen due to their possible involvement in antagonistic activity against pathogenic chitin-containing organisms. The absence of chitin in plants and vertebrate animals allows the consideration of safe and selective “target” molecules for control of chitin-containing pathogenic organisms. Fungi show appropriate characteristics as potential biological control agents of insects, fungi, and nematodes due to the production of fungal enzymes with antagonistic action. The antagonistic interactions between fungi and plant nematode parasites are among the most studied experimental models because of the high economic relevance. Fungi which target nematodes are known as nematophagous fungi. The nematode egg is the only structural element where the presence of chitin has been demonstrated. In spite of being one of the most resistant biological structures, eggs are susceptible to being attacked by egg-parasitic fungi. A combination of physical and chemical phenomena result in their complete destruction. The contribution of fungal chitinases to the in vitro rupture of the eggshell confirms their role as a pathogenic factor. Chitinases have been produced by traditional fermentation methods, which have been improved by optimizing the culture conditions for industrial processes. Although wild-type microorganisms constitute an alternative source of chitinolytic enzymes, the advances in molecular biology are allowing the genetic transformation of fungi to obtain strains with high capability as biocontrol agents. Simultaneously, a better understanding of rhizosphere interactions, additional to the discovery of new molecular biology tools, will allow the choosing of better alternatives for the biological control of nematodes in order to achieve an integrated management of the soil ecosystem.     

Environmental contamination with Toxocara sp. eggs in public squares and parks from Santiago, Chile, 1999

Most of cases of visceral larva migrans syndrome are caused by Toxocara canis larvae. Man acquires the infection by accidental ingestion of ripe eggs of the helminth. In order to find out the frequency of the presence of Toxocara sp. eggs in dog fecal specimens from public places of Santiago city in 1999 july-september a study was carried out in 84 squares and 12 parks from 32 counties. Three fecal samples were collected in each of these places, making a total of 288. Each sample was processed according to a modified Telemann method and three smears of each were microscopically examined. Toxocara sp. eggs were found in 39 (13.5%) fecal specimens. In 28/84 (33.3%) squares and in 8/12 (66.7%) parks these eggs were detected. These results make necessary the control of dogs and cats populations with the aim of reduce the presence of animals without responsible owners. It should be advisable to prevent dogs and cats defecation in public areas grounds, trying to exclude these animals from recreation areas.     

Intestinal nematode infections in Turkish military dogs with special reference to Toxocara canis

The prevalence and potential zoonotic risk factors of intestinal nematodes of military working dogs, which are used for different military purposes, were assessed. Faecal samples from 352 defined-breed Turkish military dogs were investigated and 107 (30.4%) dogs were found to be infected with one or two nematode species. The following nematodes, with their respective prevalences, were diagnosed in the faecal samples: Toxascaris leonina (21.8%), Toxocara canis (13.3%), Trichuris vulpis (2.9%) and Uncinaria stenocephala (1.2%). Toxocara canis infections were more frequently seen in puppies (0-6 months old). The prevalence of T. canis was significantly higher in male than in female dogs and also higher in dogs which were exercised daily than in those without exercise. The highest prevalence was found in Belgian malinois breed dogs. Toxocara canis infections were not influenced by the floor type of the kennels (i.e. concrete or soil floor). There was no difference in the occurrence of T. canis infection when the last anthelmintic treatment was carried out less or more than 3 months prior to sampling. It is suggested that T. canis infected military dogs would be a threat not only for dog trainers but also for military personnel, notably during national and international operations.     

Initial stage of development and migratory behavior of Toxocara canis larvae in BALB/c mouse experimental model

In the present study, the initial developmental stage of Toxocara canis eggs and larvae, and number of recovered larvae from BALB/c mouse-infected organs are described. In vitro culture of T. canis detects the frequencies of interphasic, mitotic and embryonated eggs only within a 7-day period. Analysis by egg counting was carried out for 32 days. The results showed that at 7 days after cultivation, the frequency of larvae was 50.4% and that this frequency reached 52.8% in 32 days. In the experimental infection of BALB/c mice with T. canis, the number of recovered larvae statistically increased in the brain and liver, with doses of approximately 200 and 1000 eggs. After 7 days of infection, a larger number of larvae were obtained in the lung and liver, although a maximum amount was found in the brain after a 15- or 30-day post-infection period.     

Fungal parasitism of the cyst nematode Heterodera schachtii: infection and enzymatic activity

Abstract Fungal egg parasites isolated from eggs of the cyst nematode Heterodera avenae in Sweden were investigated with respect to their ability to infect cyst nematode eggs of H. schachtii in vitro. The infection was studied by interference phase contrast microscopy of whole cysts and of cryosections of cysts exposed to the fungi on agar plates.
Verticillium suchlasporium was the most effective parasite, infecting 53% of the nematode eggs, while V. chlamydosporium infected 12% of the eggs. The fungi Paecilomyces lilacinus, Cylindrocarpon destructans or Fusarium oxysporum did not parasitize nematode eggs; nor did Arthrobotrys oligospora , a nematode trapping fungus nor Penicillium viridicatum which served as a control fungus.
The ability of the fungi to infect eggs was correlated with their lytic enzyme activity. Fungi that readily infected eggs also showed chitinase activity and presence of proteolytic activity. The Verticillium species had an activity between 3.7 and 14.6 μmol N -acetyl-glucosamine per mg protein per hour (CU) while it was 4.5 CU or lower for P. lilacinus . Other isolates did not shown any chitinase activity.     

In-vitro Assays of Meloidogyne incognita and Heterodera glycines for Detection of Nematode-antagonistic Fungal Compounds

In-vitro methods were developed to test fungi for production of metabolites affecting nematode egg hatch and mobility of second-stage juveniles. Separate assays were developed for two nematodes: root-knot nematode (Meloidogyne incognita) and soybean cyst nematode (Heterodera glycines). For egg hatch to be successfully assayed, eggs must first be surface-disinfested to avoid the confounding effects of incidental microbial growth facilitated by the fungal culture medium. Sodium hypochlorite was more effective than chlorhexidine diacetate or formaldehyde solutions at surface-disinfesting soybean cyst nematode eggs from greenhouse cultures. Subsequent rinsing with sodium thiosulfate to remove residual chlorine from disinfested eggs did not improve either soybean cyst nematode hatch or juvenile mobility. Soybean cyst nematode hatch in all culture media was lower than in water. Sodium hypochlorite was also used to surface-disinfest root-knot nematode eggs. In contrast to soybean cyst nematode hatch, root-knot nematode hatch was higher in potato dextrose broth medium than in water. Broth of the fungus Fusarium equiseti inhibited root-knot nematode egg hatch and was investigated in more detail. Broth extract and its chemical fractions not only inhibited egg hatch but also immobilized second-stage juveniles that did hatch, confirming that the fungus secretes nematode-antagonistic metabolites.     

The nematode-trapping efficacy of two chlamydospore-forming fungi against Haemonchus contortus in sheep

An in vitro study was carried out to determine efficacy of Indian isolates of the nematode-trapping fungi Arthrobotrys musiformis and Duddingtonia flagrans to capture infective larvae of Haemonchus contortus. These fungi have previously been screened and selected for their survival in the gastrointestinal tract of sheep without losing growth and nematode capturing potential. Following the feeding of chlamydospores of these two fungi alone or in combination in sheep experimentally infected with Haemonchus contortus, coprocultures were set up to enumerate the infective third stage larvae. The number of larvae captured from faeces of fungus-fed sheep was significantly higher compared with fungus-unfed controls irrespective of the fungus used. The fungal combination produced no antagonistic effect and thus can be used as efficiently as the fungi alone in the biological control of animal parasitic nematodes.     

Variation in Efficacy of Isolates of the Fungus ARF Against the Soybean Cyst Nematode Heterodera glycines

An unnamed fungus, designated ARF, that parasitizes eggs and sedentary stages of cyst nematodes is a potential biological control agent of Heterodera glycines. The objectives of this study were to determine whether ARF isolates differ in their ability to suppress nematode numbers in soil and to compare the efficacy of ARF in heat-treated and native soil. The effectiveness of 11 ARF isolates was compared by introducing homogenized mycelium into heat-treated soil. Soybean seedlings were transplanted into pots containing fungus-infested soil and inoculated with H. glycines. After 30 or 60 days, the number of nematodes and the percentage of parasitized eggs were determined. Three isolates (907, 908, and TN14), which were previously reported to be weak egg parasites in vitro, consistently suppressed nematode numbers by 50% to 100%. Of the isolates previously reported to be aggressive egg parasites, four (903, BG2, MS3, and TN12) reduced nematode numbers by 56% to 69% in at least one experimental trial, but the other four had no effect on nematode numbers. When the efficacy of isolate TN14 was tested in heat-treated and native soil, nematode suppression was greater in the heat-treated soil in only one of two trials. In both soil treatments, nematode numbers were reduced by more than 60%. We conclude that virulence toward nematode eggs in vitro is a poor indicator of effectiveness of an ARF isolate in soil, and that the presence of soil microbes may reduce, but does not completely inhibit, activity of isolate TN14.     

Biocontrol: Fungi as Nematode Control Agents

The fungal antagonists of nematodes consist of a great variety of organisms belonging to widely divergent orders and families of fungi. They include the nematode-trapping fungi, endoparasitic fungi, parasites of nematode eggs and cysts, and fungi which produce metabolites toxic to nematodes. The diversity, adaptations, and distribution of nematode-destroying fungi and taxonomic problems encountered in their study are reviewed. The importance of nemato-phagous fungi in soil biology, with special emphasis on their relationship to populations of plant-parasitic nematodes, is considered. While predacious fungi have long been investigated as possible biocontrol agents and have often exhibited spectacular results in vitro, their performance in field studies has generated little enthusiasm among nematologists. To date no species has demonstrated control of any plant pest to a degree achieved with nematicides, but recent studies have provided a much clearer concept of possibilities and problems in the applied use of fungal antagonists. The discovery of new species, which appear to control certain pests effectively under specific conditions, holds out some promise that fungi may be utilized as alternatives to chemical control after a more thorough and expanded study of their biology and ecology.     

Complete mitochondrial genomes of Baylisascaris schroederi, Baylisascaris ailuri and Baylisascaris transfuga from giant panda, red panda and polar bear

Roundworms of the genus Baylisascaris are the most common parasitic nematodes of the intestinal tracts of wild mammals, and most of them have significant impacts in veterinary and public health. Mitochondrial (mt) genomes provide a foundation for studying epidemiology and ecology of these parasites and therefore may be used to assist in the control of Baylisascariasis. Here, we determined the complete sequences of mtDNAs for Baylisascaris schroederi, Baylisascaris ailuri and Baylisascaris transfuga, with 14,778 bp, 14,657 bp and 14,898 bp in size, respectively. Each mtDNA encodes 12 protein-coding genes, 22 transfer RNAs and 2 ribosomal RNAs, typical for other chromadorean nematodes. The gene arrangements for the three Baylisascaris species are the same as those of the Ascaridata species, but radically different from those of the Spirurida species. Phylogenetic analysis based on concatenated amino acid sequences of 12 protein-coding genes from nine nematode species indicated that the three Baylisascaris species are more closely related to Ascaris suum than to the three Toxocara species (Toxocara canis, Toxocara cati and Toxocara malaysiensis) and Anisakis simplex, and that B. ailuri is more closely related to B. transfuga than to B. schroeder. The determination of the complete mt genome sequences for these three Baylisascaris species (the first members of the genus Baylisascaris ever sequenced) is of importance in refining the phylogenetic relationships within the order Ascaridida, and provides new molecular data for population genetic, systematic, epidemiological and ecological studies of parasitic nematodes of socio-economic importance in wildlife.     

The epidemiology of Toxocara canis

Bred as hunter, companion and pet, the dog has a long and honourable association with man. Yet the domestic dog can host a wide range of parasites - many of which can also infect humans. One of these, the ascarid nematode Toxocara canis (Fig. 1), is of particular interest because of retinal damage that may result from larvae becoming trapped in the eye. At the Hospital for Tropical Diseases in London, about 20-30 patients with toxocariasis are treated annually. Widespread fouling of public parks, playgrounds and pedestrian areas with dog faeces - especially in large cities - is well -recognized as one of the main sources of Toxocara infection. Yet as Stephen Gillespie discusses here, epidemiological indicators vary widely and the risk of infection is often treated too lightly.     

Related factors to human toxocariasis in a rural community of Argentina

The objective of this study was to evaluate the relationship between toxocariasis frequency and demographic, environmental, sanitary variables, eosinophylia, and other intestinal parasites in a rural population of Argentina. Serological examination of 100 individuals was carried out by using ELISA technique for the detection of anti-toxocara antibodies. Eosinophiles in peripheral blood, presence of intestinal parasites, and demographic, environmental, and socio-cultural data were evaluated. Eighty-one feces samples of dogs belonging to the studied people were analyzed to detect eggs of Toxocara canis. Thirty of them were from 30 dogs and 51 were pools from dog feces. Samples of dirt from around the homes (n: 47) and from public park (n: 4) were taken. To determine the associations, the c(2) and Fisher tests were used. The seroprevalence was 23%. Eosinophilia in peripheral blood was detected in 86.95% seropositive individuals and in 37.66% seronegative individuals (p < 0.001, OR = 11.03). Of the 23 people with positive serology, 69.56% had at least one intestinal parasite. All individuals with positive serology had dogs in their homes. Among the dog owners there was a significant association between the presence of anti-toxocara antibodies and home flooding. Eggs of T. canis were detected in the feces of 5/81 dogs and three of these dogs belonged to individuals with positive serology. Eggs of Toxocara spp. were found in 41.17% of the dirt samples, eight of which came from the area surrounding the homes of individuals with positive serology (p = 0.032; OR = 4.36). Taking into account all the variables influencing the frequency of toxocariasis in this population, the implementation of Public Health programs specifically focused on anti-parasitic treatment of dogs is recommended.     

Formulation of a strategy for the application of Duddingtonia flagrans to control caprine parasitic gastroenteritis

Experiments on the influence of egg density and varying quantities of chlamydospores on the nematode-trapping ability of Duddingtonia flagrans, influence of D. flagrans on the larval translation of gastrointestinal nematodes, doses of chlamydospores required for the effective control of gastrointestinal nematodosis and the epidemiology of nematode parasites were conducted in goats, which generated baseline data required for strategic application of the biocontrol agent. The nematode-trapping ability of D. flagrans, measured by numerical enumeration of infective third-stage larvae developed in the faecal culture, revealed that the efficacy is dependent on both nematode egg and chlamydospore density. Pasture plot studies revealed that D. flagrans, if deposited at the same time as nematode eggs, prevents translation of third-stage larvae of caprine nematodes from the faecal pats onto the grass blades. Feeding of 1 x 106 chlamydospores per kg body weight and above to goats virtually eliminated larvae from the pasture. Application of as few as 1 x 104 and 1 x 105 chlamydospores per kg body weight had a profound impact on larval recovery. The effect persisted as long as the chlamydospores were fed. Monthly faecal worm egg counts of adult goats maintained under a semi-intensive management system on the Chhattisgarh plain and pasture larval burden revealed that June to August were the months of high risk for nematodosis. Haemonchus was the dominant species recorded throughout the year. The present data can best be utilized by formulating a strategic control measure when the larval challenge to the animal is maximum (June to August), so as to prevent establishment of patent infection. The observations reinforced the strategy to be adopted for nematode parasite control in goats by applying the biocontrol option at the onset of the monsoon.     

Decontamination by anaerobic stabilisation of the environment contaminated with enteronematode eggs Toxocara canis and Ascaris suum

Investigations were carried out under operating conditions of Field Composting Factory in Brezno (Slovak Republic) to determine the effect of anaerobic stabilization of organic wastes from public areas on the survival of model helminth Toxocara canis and Ascaris suum eggs. Due to anaerobic conditions, low temperature, low C:N ratio and changes in physical and chemical properties of organic waste, less than 64% of A. suum eggs remained viable after 150 days of stabilisation. The anaerobic stabilisation had a greater effect on the viability of T. canis eggs than on A. suum eggs. The infectivity of T. canis eggs was confirmed by a follow-up experiment in laboratory mice. A small number of T. canis larvae were found in their brain and muscles on day 28 after infection. The results refer to the risks of dissemination, survival and potential spread of endoparasitic developmental stages in the environment through organic wastes subjected to low temperature stabilisation.