In vitro anthelmintic activity assessment of six medicinal plant aqueous extracts against donkey strongyles.

The lack of anthelmintic products licensed for donkeys and the rising number of small donkey milk farms in the countries of Western Europe and Italy have led to an increased interest in the study of reliable and safe plant-derived treatment alternatives. In this study, the aqueous extracts of Achillea millefolium L. (flowers), Artemisia absinthium L. (aerial parts), Centaurium erythraea Rafn. (flowers), Gentiana asclepiadea L. (rhizomes and roots), Inula helenium L. (rhizomes and roots) and Tanacetum vulgare L. (aerial parts), have been tested in vitro for their potential ovicidal and larvicidal activity against donkey nematodes. An egg-hatching assay (EHA) and larval development assay (LDA) were performed for the in vitro study, and median lethal concentration (LC-50) values for both EHA and LDA were calculated using probit analysis. All tested plant extracts showed strong anthelmintic activity against strongyle eggs and larvae at concentrations ranging between 125 and 1.95 mg/ml, except for C. erythraea, which exhibited very little or no effect at all at the tested concentrations. A strong ovicidal effect was observed in A. absinthium, with an LC-50 value of 0.486 mg/ml (95% confidence interval (CI) 0.21-1.09). Gentiana asclepiadea showed high efficacy against strongyle larvae, with an LC-50 value of 0.041 mg/ml (95% CI 0.01-0.16). The most significant (P < 0.01) anthelmintic activity was exhibited by I. helenium, with an LC-50 value of 0.041 mg/ml (95% CI 0.01-0.16) for EHA and 0.41 mg/ml (95% CI 0.27-0.62) for LDA. The results proved the anthelmintic efficacy of the tested plant extracts, highlighting the need for further research into plant bioactive molecules both in vitro and in vivo.

Anthelmintic effect of Cymbopogon citratus essential oil and its nanoemulsion on sheep gastrointestinal nematodes.

The anthelmintic resistance stimulated the search for strategies for controlling gastrointestinal nematodes, including the use of free essential oils or its nanoemulsion. This study evaluated the anthelmintic efficacy of Cymbopogon citratus essential oil (CcEO) and C. citratus essential oil nanoemulsion (CcEOn). Pysicochemical analyses were performed. The in vitro effect was determined using the egg hatch test (EHT) on Haemonchus contortus and in vivo effect was evaluated in sheep infected with gastrointestinal nematodes. The animals were treated with CcEO (500 mg/kg) or CcEOn (450 mg/kg) for the fecal egg count (FEC) and the determination of worm burden. The main component of CcEO was citral. The CcEO content in the nanoemulsion was 20% (v/v), and the mean particle size was 248 nm. In EHT, CcEO and CcEOn (1.25 mg/mL) inhibited larval hatching by 98.4 and 97.1%, respectively. Three animals treated with CcEO died whereas in the group treated with CcEOn one animal died. The FEC and total worm burden of the treated groups did not differ from the negative control (p>0.05). The CcEOn showed efficacy only on H. contortus (p<0.05). In conclusion, nanoencapsulation reduced toxicity and increased efficacy on H. contortus.

Anthelmintic effect of Cymbopogon citratus essential oil and its nanoemulsion on sheep gastrointestinal nematodes / Efeito anti-helmíntico do óleo essencial de Cymbopogon citratus e sua nanoemulsão sobre nematoides gastrintestinais de ovinos

Abstract The anthelmintic resistance stimulated the search for strategies for controlling gastrointestinal nematodes, including the use of free essential oils or its nanoemulsion. This study evaluated the anthelmintic efficacy of Cymbopogon citratus essential oil (CcEO) and C. citratus essential oil nanoemulsion (CcEOn). Pysicochemical analyses were performed. The in vitro effect was determined using the egg hatch test (EHT) on Haemonchus contortus and in vivo effect was evaluated in sheep infected with gastrointestinal nematodes. The animals were treated with CcEO (500 mg/kg) or CcEOn (450 mg/kg) for the fecal egg count (FEC) and the determination of worm burden. The main component of CcEO was citral. The CcEO content in the nanoemulsion was 20% (v/v), and the mean particle size was 248 nm. In EHT, CcEO and CcEOn (1.25 mg/mL) inhibited larval hatching by 98.4 and 97.1%, respectively. Three animals treated with CcEO died whereas in the group treated with CcEOn one animal died. The FEC and total worm burden of the treated groups did not differ from the negative control (p>0.05). The CcEOn showed efficacy only on H. contortus (p<0.05). In conclusion, nanoencapsulation reduced toxicity and increased efficacy on H. contortus.

Resumo A resistência anti-helmíntica estimulou a busca por estratégias de controle de nematoides gastrintestinais, incluindo óleos essenciais livres ou em nanoemulsão. Este estudo avaliou a eficácia anti-helmíntica do óleo essencial de Cymbopogon citratus (OECc) e da nanoemulsão do óleo essencial de C. citratus (nOECc). Análises físico-químicas foram realizadas. O efeito in vitro foi determinado no teste de eclosão de ovos (TEO) sobre Haemonchus contortus e o efeito in vivo foi avaliado em ovinos infectados com nematoides gastrintestinais. Os animais foram tratados com OECc (500 mg/kg) ou nOECc (450 mg/kg) para determinação do número de ovos por grama de fezes (OPG) e carga parasitária. O principal constituinte do OECc foi citral. O conteúdo de OECc na nanoemulsão foi 20% e o tamanho médio de partícula foi 248 nm. No TEO, OECc e nOECc (1,25 mg/mL) inibiram 98,4 e 97,1% da eclosão larvar, respectivamente. Três animais tratados com o OECc morreram, enquanto um animal do grupo tratado com a nOECc morreu. O OPG e a carga parasitária total dos grupos tratados não diferiram do controle negativo (p>0.05). A nOECc apresentou eficácia somente sobre H. contortus (p<0.05). Como conclusão, o nanoencapsulamento reduziu a toxicidade e aumentou a eficácia sobre H. contortus.

Screening of the 'Open Scaffolds' collection from Compounds Australia identifies a new chemical entity with anthelmintic activities against different developmental stages of the barber's pole worm and other parasitic nematodes.

The discovery and development of novel anthelmintic classes is essential to sustain the control of socioeconomically important parasitic worms of humans and animals. With the aim of offering novel, lead-like scaffolds for drug discovery, Compounds Australia released the 'Open Scaffolds' collection containing 33,999 compounds, with extensive information available on the physicochemical properties of these chemicals. In the present study, we screened 14,464 prioritised compounds from the 'Open Scaffolds' collection against the exsheathed third-stage larvae (xL3s) of Haemonchus contortus using recently developed whole-organism screening assays. We identified a hit compound, called SN00797439, which was shown to reproducibly reduce xL3 motility by ≥ 70%; this compound induced a characteristic, "coiled" xL3 phenotype (IC50 = 3.46-5.93 µM), inhibited motility of fourth-stage larvae (L4s; IC50 = 0.31-12.5 µM) and caused considerable cuticular damage to L4s in vitro. When tested on other parasitic nematodes in vitro, SN00797439 was shown to inhibit (IC50 = 3-50 µM) adults of Ancylostoma ceylanicum (hookworm) and first-stage larvae of Trichuris muris (whipworm) and eventually kill (>90%) these stages. Furthermore, this compound completely inhibited the motility of female and male adults of Brugia malayi (50-100 µM) as well as microfilariae of both B. malayi and Dirofilaria immitis (heartworm). Overall, these results show that SN00797439 acts against genetically (evolutionarily) distant parasitic nematodes i.e. H. contortus and A. ceylanicum [strongyloids] vs. B. malayi and D. immitis [filarioids] vs. T. muris [enoplid], and, thus, might offer a novel, lead-like scaffold for the development of a relatively broad-spectrum anthelmintic. Our future work will focus on assessing the activity of SN00797439 against other pathogens that cause neglected tropical diseases, optimising analogs with improved biological activities and characterising their targets.

A temporal assessment of nematode community structure and diversity in the rhizosphere of cisgenic Phytophthora infestans-resistant potatoes.

BACKGROUND: Nematodes play a key role in soil processes with alterations in the nematode community structure having the potential to considerably influence ecosystem functioning. As a result fluctuations in nematode diversity and/or community structure can be gauged as a 'barometer' of a soil's functional biodiversity. However, a deficit exists in regards to baseline knowledge and on the impact of specific GM crops on soil nematode populations and in particular in regard to the impact of GM potatoes on the diversity of nematode populations in the rhizosphere. The goal of this project was to begin to address this knowledge gap in regards to a GM potato line, cisgenically engineered for resistance to Phytophthora infestans (responsible organism of the Irish potato famine causing late blight disease). For this, a 3 year (2013, 2014, 2015) field experimental study was completed, containing two conventional genotypes (cvs. Desiree and Sarpo Mira) and a cisgenic genotype (cv. Desiree + Rpi-vnt1). Each potato genotype was treated with different disease management strategies (weekly chemical applications and corresponding no spray control). Hence affording the opportunity to investigate the temporal impact of potato genotype, disease management strategy (and their interaction) on the potato rhizosphere nematode community. RESULTS: Nematode structure and diversity were measured through established indices, accounts and taxonomy with factors recording a significant effect limited to the climatic conditions across the three seasons of the study and chemical applications associated with the selected disease management strategy. Based on the metrics studied, the cultivation of the cisgenic potato genotype exerted no significant effect (P > 0.05) on nematode community diversity or structure. The disease management treatments led to a reduction of specific trophic groups (e.g. Predacious c-p = 4), which of interest appeared to be counteracted by a potato genotype with vigorous growth phenotype e.g. cv. Sarpo Mira. The fluctuating climates led to disparate conditions, with enrichment conditions (bacterial feeding c-p = 1) dominating during the wet seasons of 2014 and 2015 versus the dry season of 2013 which induced an environmental stress (functional guild c-p = 2) on nematode communities. CONCLUSIONS: Overall the functional guild indices in comparison to other indices or absolutes values, delivered the most accurate quantitative measurement with which to determine the occurrence of a specific disturbance relative to the cultivation of the studied cisgenic P. infestans-resistant potatoes.

Interactions Between Nutrition and Infections With Haemonchus contortus and Related Gastrointestinal Nematodes in Small Ruminants.

Interactions between host nutrition and feeding behaviour are central to understanding the pathophysiological consequences of infections of the digestive tract with parasitic nematodes. The manipulation of host nutrition provides useful options to control gastrointestinal nematodes as a component of an integrated strategy. Focussed mainly on the Haemonchus contortus infection model in small ruminants, this chapter (1) illustrates the relationship between quantitative (macro- and micro-nutrients) and qualitative (plant secondary metabolites) aspects of host nutrition and nematode infection, and (2) shows how basic studies aimed at addressing some generic questions can help to provide solutions, despite the considerable diversity of epidemiological situations and breeding systems.

Atividade antagonista do óleo essencial de Lippia alba (Mill. ) N. E. Brown (Verbenaceae) sobre Meloidogyne incognita (Kofoid & White) Chitwood / Antagonist activity of the essential oil Lippia alba (Mill. ) N. E. Brown (Verbenaceae) on Meloidogyne incognita (Kofoid & White) Chitwood

RESUMO O controle de fitonematóides é uma tarefa difícil. A alta infestação de nematoides no solo obriga os produtores a usarem doses mais elevadas de nematicidas, ou ainda, a aumentarem a freqüência das aplicações, geralmente no solo, ocasionando maior potencial de dano ao homem e ao ambiente. Além disso, nematicidas convencionais vêm sofrendo grandes restrições de uso em muitos países. Desse modo, a busca de novas medidas de controle de fitonematóides é uma prioridade da agricultura sustentável. Desta forma, objetivou-se estudar o efeito de óleos essenciais de Lippia Alba, na mortalidade de juvenis pré-parasitas do segundo estádio de Meloidogyne incognita. O ensaio foi montado em delineamento inteiramente casualizado, com 4 repetições, em esquema fatorial 3×4 (3 quimiotipos x 4 concentrações) empregando-se três diferentes quimiotipos de L. Alba (I, II, III) nas concentrações de 0, 100, 500 e 1000 ppm, coletados em diferentes horas do dia (7, 9, 13, 16 e 19h). Os dados coletados (% de mortalidade) foram submetidos à análise de variância e os valores médios comparados por meio do teste de Tukey. Foi possível concluir que Lippia alba contém, em seu óleo essencial, compostos com efeitos significativos na mortalidade de juvenis de segundo estádio (J2) de M. incognita. Dessa forma, é importante destacar que o acentuado efeito nematicida do óleo essencial de L. alba sugere a possibilidade de seu uso no controle de M. incognita.

ABSTRACT The control of nematodes is difficult. The high infestation of nematodes in the soil requires the producers use higher doses of nematicides, increasing the frequency of applications, usually in soil, causing negative effects to humans and the environment. Furthermore, conventional nematicides have suffered great use restrictions in many countries. Thus, the search for new measures to control nematodes is a priority of sustainable agriculture. Thus, the objective of this study was available the effect of essential oils from Lippia Alba against Meloidogyne incognita. The experiment was conducted in a completely randomized design with four replications in a factorial 3x4 (3 chemotypes and 4 concentrations) using three different chemotypes of L. Alba (I, II, III) at concentrations of 0, 100, 500 and 1000 ppm, collected in different times of day (7 AM, 9 AM, 1 PM, 4 PM e 7 PM). The data collected (% mortality) were subjected to analysis of variance and mean values were compared using the Tukey test. It was able to conclude that L. alba contains in its essential oil, compounds with significant effects on mortality of M. incognita. Thus, it is important to note that the greatest nematicide effect of essential oil of L. alba suggests the possibility of its use to control M. incognita.

[Composition and Density of Soil Fauna in the Region with Enhanced Radioactivity Level (Komi Republic, Vodnyi)].

Studies on the influence of high levels of radiation on soil fauna were carried out in 2012 in the territory formed as a result of the activity of the enterprise for extraction and production of radium from reservoir water and waste of uranium ore from 1931 to 1956. At present the local radioactive pollution in this area is caused by the presence of heavy natural radionuclides 226Ra, 238U and products of their disintegration in soils. The oppression of soil invertebrate.fauna in pine forests and meadows with high levels of radionuclides and heavy metals is revealed. Also shown is the decrease in the number and density of different taxonomic groups of invertebrates, reduction of the diversity and spectrum of trophic groups and vital forms in the area with a high content of radionuclides in soil. Our results are in agreement with the results obtained by the similar studies showing negative influence of high-level ionizing radiation on soil fauna.

[Characteristics of soil nematode community of different agricultural areas in Jiangsu Province, China].

This paper investigated the genus diversity of soil nematodes of different agricultural areas in Jiangsu Province, analyzed the relationship between soil nematodes and soil environmental factors, and discussed the roles of soil nematodes as biological indicators of soil health. The results showed that, a total of 41 nematode genera were found in all six agricultural areas, belonging to 19 families, 7 orders, 2 classes. The numbers and community compositions of nematodes were obviously influenced by soil texture, fertilization and tillage practices. In all six agricultural areas, the numbers of nematodes in coastal agricultural area (400 individuals per 100 g dry soil) were significantly larger than that in Xuhuai, Ningzhenyang, and riverside agricultural areas. While the smallest number of nematodes was found in Yanjiang agricultural area (232 individuals per 100 g dry soil), which might be due to the differences in soil texture, annual rainfall and annual air temperature and other factors. The dominant genera of nematodes were similar in the adjacent agricultural areas. Correlation analysis showed that there was a significant positive correlation between the number of soil nematodes and levels of soil nutrients (soil organic matter, total nitrogen, available nitrogen, available potassium and available phosphorus). Redundancy analysis (RDA) indicated the total nitrogen, available potassium and pH obviously affected some soil nematode genera. The analysis of spatial distribution characteristics of soil nematode community in farmland of Jiangsu Province could provide data for health assessment of agricultural ecosystems.

Biostimulation as an attractive technique to reduce phenanthrene toxicity for meiofauna and bacteria in lagoon sediment.

A microcosm experiment was setup to examine (1) the effect of phenanthrene contamination on meiofauna and bacteria communities and (2) the effects of different bioremediation strategies on phenanthrene degradation and on the community structure of free-living marine nematodes. Sediments from Bizerte lagoon were contaminated with (100 mg kg(-1)) phenanthrene and effects were examined after 20 days. Biostimulation (addition of nitrogen and phosphorus fertilizer or mineral salt medium) and bioaugmentation (inoculation of a hydrocarbonoclastic bacterium) were used as bioremediation treatments. Bacterial biomass was estimated using flow cytometry. Meiofauna was counted and identified at the higher taxon level using a stereomicroscope. Nematodes, comprising approximately two thirds of total meiofauna abundance, were identified to genus or species. Phenanthrene contamination had a severe impact on bacteria and meiofauna abundances with a strong decrease of nematodes with a complete disappearance of polychaetes and copepods. Bioremediation counter balanced the toxic effects of phenanthrene since meiofauna and bacteria abundances were significantly higher (p < 0.01) than those observed in phenanthrene contamination. Up to 98 % of phenanthrene removal was observed. In response to phenanthrene contamination, the nematode species had different behavior: Daptonema fallax was eliminated in contaminated microcosms, suggesting that it is an intolerant species to phenanthrene; Neochromadora peocilosoma, Spirinia parasitifera, and Odontophora n. sp., which significantly (p < 0.05) increased in contaminated microcosms, could be considered as "opportunistic" species to phenanthrene whereas Anticoma acuminata and Calomicrolaimus honestus increased in the treatment combining biostimulation and bioaugmentation. Phenanthrene had a significant effect on meiofaunal and bacterial abundances (p < 0.05), with a strong reduction of density and change in the nematode communities. Biostimulation using mineral salt medium strongly enhanced phenanthrene removal, leading to a decrease of its toxicity. This finding opens exciting axes for the future use of biostimulation to reduce toxic effects of PAHs for meiofauna and bacteria in lagoon sediment.

Soluções de alho (Allium sativum L. ) no controle de nematódeos gastrintestinais em bovinos jovens da raça Holandesa / Garlic (Allium sativum L. ) solutions in control of gastrointestinal nematodes in young Holstein cattle

Para avaliar o efeito da atividade anti-helmíntica do alho suplementado, foram utilizadas 24 bezerras e novilhas da raça Holandesa, naturalmente infectadas. As soluções foram preparadas triturando-se o alho, (50%), mais água ou álcool 92º, (50%), administrando-se oralmente aos animais. Os tratamentos (T) foram constituídos pelo grupo controle negativo (T1); extrato alcoólico de alho a 60g e 120g/100Kg de peso vivo (T2) e (T3); extrato aquoso de alho a 60 e 120g/100kg de peso vivo (T4) e (T5); e o grupo controle positivo com albendazol a 10% (T6). Os tratamentos fitoterápicos foram repetidos a cada 14 dias, caso a infecção fosse superior a 400 ovos por grama de fezes (OPG). A técnica de coprocultura quantitativa e qualitativa foi empregada para avaliar a eficácia anti-helmíntica dos tratamentos. Foram observadas diferenças entre os grupos controle e tratados para OPG e na porcentagem relativa de larvas infectantes e desenvolvimento larval. O uso das soluções de alho demonstrou controle parcial de nematódeos gastrintestinais.

Twenty four Holstein calves and heifers naturally infected were used to evaluate the effect of garlic supply on helminth control. The solutions consisted to triturated garlic, (50%), plus water or alcohol 92º, (50%), orally-administrable on the animals. Treatments (T) consisted of a negative control (T1), garlic at 60 (T2) and 120g on alcoholic extract (T3); 60 (T4) and 120g on aqueous extract/100kg of life weight (T5) and the positive control with albendazol at 10% (T6). The phytotherapic treatments were repeated after 14th day if the infection was superior to 400 eggs per gram of faeces (EPG). The quantitative and qualitative coproculture technique was performed to evaluate the anthelmintic efficacy of the treatments. Differences between treated and control groups were observed in EPG and in relative percentage of infective larvae and the larval development. Partial control was found with the garlic solutions on gastrointestinal nematodes.

Application and commercialization of nematodes.

While nematodes are most commonly known for their negative impact on plants, animals, and humans, there are a number of species which are commercially explored. This review highlights some of the most important success stories for the application of nematodes. They are used as bioindicators in ecological and toxicity studies, as model organisms for elucidating fundamental biological questions and for high throughput screening of drugs. Besides these indirect uses, direct applications include the use of Beddingia siricidicola against a major forest pest and the commercialization of Steinernema, Heterorhabditis, and Phasmarhabditis as biological pest control products. New directions for the commercialization of nematodes are the use as living food, specifically loaded with essential nutrients for various fish and shrimp larvae. Even human parasites or closely related species have been successfully used for curing autoimmune disorders and are currently in the process of being developed as drugs. With the striving development of life sciences, we are likely to see more applications for nematodes in the future. A prerequisite is that we continue to explore the vast number of yet undiscovered nematode species.

Australian plants show anthelmintic activity toward equine cyathostomins in vitro.

Anthelmintic resistance in gastrointestinal parasites of horses is an increasing problem, particularly in cyathostomins, and there is a need to find alternative means for the control of these parasites. We screened crude extracts from 37 species of Australian native plants for their anthelmintic activity in vitro against cyathostomin larvae (development from egg to third larval stage), with the aim of identifying those species that may be suitable for incorporation into sustainable parasite management programs. Water extracts from seven species, namely Acacia baileyana, Acacia melanoxylon, Acacia podalyriifolia, Alectryon oleifolius, Duboisia hopwoodii, Eucalyptus gomphocephala and Santalum spicatum completely inhibited larval development (100% inhibition compared to the control), while another 10 species caused 90% inhibition at the initial screening concentration of 1400 µg of extractable solids/mL. The seven most potent extracts produced IC50 values (concentration of extract which resulted in a 50% inhibition of development) in the range 30.9-196 µg/mL. Fourteen extracts were incubated with polyvinylpolypyrrolidone (PVPP) before the assays, which removed the anthelmintic activity from 12 of these extracts, indicating that tannins were likely to be the bioactive compound responsible for the effect, while in two species, i.e. A. melanoxylon and D. hopwoodii, compounds other than tannins were likely to be responsible for their anthelmintic action. Our results suggest that a number of Australian native plants have significant anthelmintic activity against cyathostomin larval development in vitro. There is potential for these plants to be used as part of sustainable parasite control programs in horses, although more research is needed to identify the compounds responsible for the anthelmintic effects and confirm their activity in vivo.

Preliminary study on responses of marine nematode community to crude oil contamination in intertidal zone of Bathing Beach, Dalian.

This study investigated the responses of marine nematodes to crude oil contamination in polluted and relatively uncontaminated sites in Dalian Xingang, China, 40 days after an oil spill. Samples were taken at different tide levels on the beach and at different positions along the beach. We present the results of a comparison of nematode assemblages from undisturbed sediment from the Xiajiahezi Bathing Beach with those from sediment from the Xinghai Bathing Beach contaminated with crude oil. A total of 1666 nematodes from 26 genera were found in this study. Results showed significant differences in nematode assemblages between samples from undisturbed controls and those from the polluted area. Nematode abundance, number of species, diversity and species richness decreased significantly with increasing levels of crude oil contamination. Fifteen genera were eliminated and seemed to be composed of species intolerant to crude oil contamination; only the abundance of Marylynnia sp. increased slightly.

Challenges to grow oilseed rape Brassica napus in sugar beet rotations.

The study was carried out in 1989-1991 and repeated in 2003-2006 to compare life cycle and dynamics of Heterodera schachtii Schm. on sugar beet, oilseed rape, fodder radish and to work out recommendations on how to decrease the risk of yield reduction while it grows in sugar-beet rotations. Research was carried out in plot experiment in natural conditions. Nematode community on rape, fodder radish and sugar beet was analyzed. Data of nematode community showed that composition of nematode species was very similar. Heterodera shachtii were dominated species with rape and sugar beet. All tested Brassica crops are susceptible to H. schachtii. However there is significant difference in population dynamics. The highest total number of brown cysts, eggs and juveniles of all ages was observed in winter rape. H.schachtii developed two generations on sugar beet and one generation on mustard. The voluntary seed germination after harvest contributes to increasing H. schachtii population. Therefore it is necessary to destroy oilseed rape voluntary chemically or physically. This operation should be done in about 2-4 weeks. The exact time can be calculated using the temperature- based model. Growing regular fodder radish and mustard as the trap crops can significantly reduce population of H. schachtii. The time of sowing is not earlier than August 20th. While estimating the time of destruction of trap crops it should be taken into consideration that H. schachtii can complete life cycle without foliage.

Segregation and recombination of a multipartite mitochondrial DNA in populations of the potato cyst nematode Globodera pallida.

The discovery that the potato cyst nematode Globodera pallida has a multipartite mitochondrial DNA (mtDNA) composed, at least in part, of six small circular mtDNAs (scmtDNAs) raised a number of questions concerning the population-level processes that might act on such a complex genome. Here we report our observations on the distribution of some scmtDNAs among a sample of European and South American G. pallida populations. The occurrence of sequence variants of scmtDNA IV in population P4A from South America, and that particular sequence variants are common to the individuals within a single cyst, is described. Evidence for recombination of sequence variants of scmtDNA IV in P4A is also reported. The mosaic structure of P4A scmtDNA IV sequences was revealed using several detection methods and recombination breakpoints were independently detected by maximum likelihood and Bayesian MCMC methods.

A northward colonisation of the Andes by the potato cyst nematode during geological times suggests multiple host-shifts from wild to cultivated potatoes.

The cyst nematode Globodera pallida is a major pest of potato in South America where this specialist parasite is native. To investigate its phylogeography, we have genotyped individuals from 42 Peruvian populations using mitochondrial and nuclear molecular markers. A clear south-to-north phylogeographical pattern was revealed with five well-supported clades. The clade containing the southern populations is genetically more diverse and forms the most basal branch. The large divergence among cytochrome b haplotypes suggests that they diverged before human domestication of potato. As the nematodes studied have been sampled on cultivated potato, multiple host-shifts from wild to cultivated potatoes must have occurred independently in each clade. We hypothesise that this south-to-north pattern took place during the uplift of the Andes beginning 20 My ago and following the same direction. To our knowledge, this is the first study of a plant parasite sampled on cultivated plants revealing an ancient phylogeographical pattern.

Improving resilience against natural gastrointestinal nematode infections in browsing kids during the dry season in tropical Mexico.

The objective was to determine the effect of supplementary feeding on the resilience and resistance of Criollo kids against natural gastrointestinal nematode (GIN) infections, when browsing native vegetation during the dry season in tropical Mexico. Thirty-three two-month-old Criollo kids, raised nematode free, were included at weaning in a 20-week trial. The kids were placed into four groups. Two groups of eight kids were offered 100g/day soybean and sorghum meal (26%:74% respectively fresh basis) (treated/supplemented (T-S) and infected/supplemented (I-S)). Two groups remained with no supplement for the duration of the trial (infected/non-supplemented (I-NS) (n=9) and treated/non-supplemented (T-NS) (n=8)). Kids in groups T-S and T-NS were drenched with 0.2mg of moxidectin/kg body weight orally (Cydectin, Fort Dodge) every 28 days. Groups I-S and I-NS were naturally infected with GIN. The animals browsed native vegetation for an average of 7h/day together with a herd of 120 naturally infected adult goats. Cumulative live weight gain (CLWG), packed cell volume (PCV), haemoglobin (Hb), total plasma protein and plasma albumin were recorded every 14 days as measurements of resilience. Resistance parameters (faecal egg counts (FEC) and peripheral eosinophil counts (PEC)) were also measured. Bulk faecal cultures were made for each group every 28 days. Every month a new pair of initially worm-free tracer kids assessed the infectivity of the vegetation browsed by the animals. Tracer kids and faecal cultures showed that kids faced low mixed infections (Haemonchus contortus, Trichostrongylus colubriformis and Oesophagostomum columbianum). Under conditions of scarce vegetation, such as those in the present study, supplemented groups (I-S and T-S) had higher growth rates compared to the non-supplemented groups independently of the control of GIN infection with anthelmintic (AH) treatment (P<0.001). Supplementary feeding did not affect FEC or PEC. In the absence of supplementation, lack of AH treatment may lead to outbreaks of clinical nematodosis. The supplementary feeding was economically feasible.

Occurrence of plant parasitic nematodes (Tylenchina) in sugar beet fields in Fars province, Iran.

On a survey of plant parasitic nematode fauna, belonging to the suborder Tylenchina, about 110 soil and root samples were collected from different sugar beet field in fars Province in Iran. The samples were washed and the nematodes were extracted by Centrifugal Flotation Technique according to the modified method by De Grisse, 1969. They were fixed and transferred to glycerine, then permanent slides were mounted. Morphological and morphometrical characters of collected species were carefully studied and the characters were compared with the original descriptions and differences were discussed. In this study 18 species belonging to 11 different genera of subordes Tylenchina were identified as follows. Boleodorus thylactus, Ditylenchus destructor, D. dipsaci Geocenamus brevidens, G. rugosus, Helicotylenchus digonicus, H. pseudorobustus, H. vulgaris, Heterodera filipjevi, H. schachtii, Pratylenchus neglectus, P. thornei, Psilenchus hilarulus, P. hilarus, Stictylus mucronatus, Tylenchorhynchus ventrosinatus, Tylenchus davainei and Zygotylenchus guevarai. The species of the genera Ditylenchus, Heterodera, Pratylenchus and Psilenchus were comparatively more distributed than the others. This investigation revealed that sugar beet cyst nematode (Heterodera schachti) was found already in most sugar beet fields in this Province. Fathabod, Marvdasht and Tasuj-kwar were the most infested areas. Cysts collected from 100 gr. Of soil samples were 120 to 121 in number and the eggs and second stage larvae in each gr. of soil were 104.1 to 104.5 respectively.

Gene pool similarities of potato cyst nematode populations assessed by AFLP analysis.

AFLP was used to characterize 24 potato cyst nematode populations. This novel DNA fingerprinting technique enabled the identification of 987 marker loci by screening only 12 primer combinations. Data on presence or absence polymorphisms and data on the intensities of corresponding DNA fragments were collected. Separate analysis of both data sets revealed similar dendrograms for the nine G. rostochiensis populations included in this study. Both dendrograms consisted of two groups containing three and five related populations, respectively. One population differed from either of these groups. Each group represented a different pathotype as defined by Kort et al. (J. Kort, H. Ross, H. J. Rumpenhorst, and A. R. Stone, Nematologica 23:333-339, 1977). Previously, a similar arrangement was found after analysis of the genetic variation using random amplified polymorphic DNA (RAPD) (R. T. Folkertsma, J. N. A. M. Rouppe van der Voort, M. P. E. van Gent-Pelzer, K. E. de Groot, W. J. van den Bos, A. Schots, J. Bakker, and F. J. Gommers, Phytopathology 84:807-811, 1994). For the 15 G. pallida populations analyzed, complex AFLP patterns were obtained and therefore only qualitative AFLP data were used. Incongruities were observed between clustering on the basis of AFLP data and classical pathotyping. This strongly confirms earlier findings obtained with RAPDs, because the AFLP markers used in this study outnumbered the population characteristics revealed by RAPDs by a factor of five. To arrive at a reliable pathotype designation of potato cyst nematode populations molecular data and virulence characteristics should be integrated. Possible causes for the difference in distribution of polymorphisms among g. rostochiensis and G. pallida populations are discussed.