Common Nematode Research Articles

Factors affecting the morphometric parameters of eggs of Trichocephalus suis Schrank, 1788, Ascaris suum Goeze, 1782 and Oesophagostomum dentatum Rudolphi, 1803 in the Republic of Crimea

The publication presents the results of the morphometric parameters of eggs of the most common intestinal nematodes of pigs in the Republic of Crimea that were measured using a computer program ImageJ. It was found that the sizes of eggs of Trichocephalus suis Schrank, 1788, Ascaris suum Goeze, 1782 and Oesophagostomum dentatum Rudolphi, 1803 differed significantly within the species in a fairly wide range. Also, the factors affecting the morphometric parameters of eggs of these nematodes were determined. Their changes depended significantly on the intensity of invasion, the season of the year and the stage of the biological cycle. Thus, it was found that with an increasing intensity of invasion the sizes of the eggs of these parasites decreased. Also, in the process of their maturation and development in the environment under the influence of favorable conditions of the time of year the sizes of the eggs of these species changed in different ways – they decreased in some species (A. suum, O. dentatum) and increased in the others (T. suis). Thus, the combined effect of certain specific factors in a particular area will contribute to a faster or slower maturation and development of the eggs of a particular type of helminth in the environment. Consequently, the period of the biological cycle of the same species of parasites in different areas will span a different interval of time. This issue was covered poorly in both domestic and foreign literature and will require further study and systematization.

RIOK-2 protein is essential for egg hatching in a common parasitic nematode

The atypical protein kinase RIOK-2 is a non-ribosomal factor essential for ribosome maturation in yeast and human cells; however, little is known about its physiological role in pathogens. Our earlier work examined the expression profile of a RIOK-2 gene (Ss-riok-2) in Strongyloides stercoralis – a prevalent nematode parasite of dogs and humans. Herein, we demonstrate that Ss-RIOK-2 encodes a catalytically active kinase, distributed primarily in the cytoplasm of intestinal and hypodermal cells in transgenic larvae. Its expression oscillates as the free-living L1s develop into infective L3s. Overexpression of a catalytically impaired Ss-RIOK-2-D228A mutant delayed the development of transgenic larvae, while ectopic expression of another dominant negative isoform with a mutation in the ATP-binding site (K123A) abrogated the process of egg hatching, which could be rescued by co-expressing a wild-type Ss-RIOK-2 but not by its Ss-RIOK-1 ortholog. Collectively, our findings show a critical and specific role of Ss-RIOK-2 during the development of a pathogenic roundworm, which can be exploited to develop anti-infectives.

Integrated evidence reveals a new species of Cosmocerca (Ascaridomorpha: Cosmocercoidea) from the Asiatic toad Bufo gargarizans Cantor (Amphibia: Anura).

Species of Cosmocerca Diesing, 1861 (Ascaridomorpha: Cosmocercoidea), are common nematode parasites of amphibians. In the present study, a new species of Cosmocerca, namely C. simile n. sp., was described using light and scanning electron microscopy, and sequencing different nuclear and mitochondrial genetic markers (i.e. small ribosomal DNA (18S), large ribosomal DNA (28S), internal transcribed spacer (ITS) and cytochrome c oxidase subunit 1 (cox1)). Cosmocerca simile n. sp. differs from its congeners based on body size, morphology and number of plectanes, relative length of spicules and gubernaculum and spicules to total body length and morphology and length of tail. Molecular analysis showed no nucleotide polymorphisms among different individuals of the new species regarding nuclear DNA. Very low intraspecific nucleotide variation (0.52-0.78%) was detected in cox1 mtDNA. In contrast, the level of interspecific nucleotide variation between C. simile n. sp. and its congeners were distinctly higher (2.74-18.1% in the partial ITS region and 10.2-13.5% in the partial cox1 region, respectively) than that of intraspecific variation. Phylogenetic analyses using maximum likelihood (ML) inference based on the partial ITS and cox1 sequence data both supported the new species to be a member of the genus Cosmocerca, and formed a sister relationship to C. japonica. The newly obtained genetic data are important for further studies of DNA-based taxonomy, population genetics and phylogenetics of the Cosmocercoidea.

Integrated management of root-knot nematode (Meloidogyne spp.) in Florida tomatoes combining host resistance and nematicides

Host resistance governed by the Mi gene is available in many commercial tomato cultivars, and is effective against the most common tropical root-knot nematode species (Meloidogyne incognita, M. javanica and M. arenaria). The resistance has some drawbacks like the lack of activity against other common root-knot nematode species like M. hapla and M. enterolobii, the widespread emergence of resistance-breaking races of M. incognita, and the reported heat sensitivity of the Mi gene. In Florida, many tomato fields often also harbour soilborne pathogens and weeds, such as nutsedge, together with root-knot nematodes. Therefore, most tomato fields in Florida are fumigated, and nematode-resistant tomato cultivars are rarely planted. Two field trials were conducted during fall 2017 and spring 2018 to evaluate the potential of Mi-tomato cultivars in Florida, with or without additional use of a soil fumigant and/or nematicide. Root-knot severity in both seasons was significantly reduced in cultivars having Mi (Sanibel, Skyway, and Tasti Lee + Mi), as compared to the cultivar without Mi (FL 47). Fumigation with chloropicrin at 225 kg/ha and nematicides (fluensulfone in fall and fluazaindolizine in spring) also significantly reduced nematode damage. This was mostly noted for the cv. FL 47, and mostly during the fall growing season, when early nematode pressure was higher. End-of-season root-knot nematode soil populations were significantly lower with Mi cultivars. Chloropicrin on the other hand tended to increase end-of-season root-knot soil populations. No breakdown of resistance due to high soil temperature (>30 °C) was observed. Soil fumigation significantly increased fruit yield, especially in fall (95% increase) and to a lesser extent in spring (14.5% increase), and mostly so for cv. FL 47 and cv. Sanibel. Overall, cv. Sanibel produced the highest fruit yields.

Evaluation of nematocidal effects of some medicinal plant extracts against root-knot nematodes (Meloidogyne incognita)

Hundreds of highly toxic chemical pesticides and their conventional unsafe formulations of varying toxicities are used extensively to control pests, diseases, and weeds to increase agricultural production. The use of bio or botanical pesticides, which have been found effective, safe, and eco-friendly, could possibly provide a viable solution. Thus, the development and production of environment-friendly botanical pesticides and their water-based formulations, to replace the highly toxic agro-chemicals and unsafe formulations, has gained significant importance towards developing appropriate strategies for crop protection. The present study was aimed to evaluate the nematocidal nature of the aqueous extracts of Allium sativum, Urtica dioica, Sophora mollis, Ephedra intermedia, and Tanacetum baltistanicum. For this purpose, the plant material was dried in shade and mechanically ground into a powder form. The methanolic extracts of each plant sample were obtained and further extracted into different organic and aqueous fractions. The polar organic and aqueous fractions were further subjected to in vitro studies against Meloidogyne incognita, a common root-knot nematode. The results revealed that the polar organic and aqueous extracts of all the tested plants showed excellent results with total mortality of 75-95% at the concentrations of 0.125-1% after 72 h of the treatment. These results can be exploited further for their efficacy against M. incognita through on-field applications. The nematocidal effect of tested extracts indicates that some polar oxygenated secondary metabolites with lipophilic properties may be responsible to damage the cytoplasmic membrane of the nematode cells by interfering with the enzyme protein structure through their functional groups.

Presence of Toxocara spp. and Other Zoonotic parasites Ova in Children's Playground in Karaman, Turkey

Human toxocariosis (HT) is a widespread and neglected parasitic disease around the world and it is caused by Toxocara canis and Toxocara cati, a common nematode found in dogs and cats. Childiren are caught to HT after ingestion of embriyonated Toxocara spp. eggs via contaminated materials such as soil, hair and etc. The aim of this study is to investigate Toxocara spp. and other zoonotic parasites in children’s playgrounds in Karaman province of Turkey. In total, 103 samples (68 sand soil, 26 soil and 9 stool) from 20 randomly selected children's playgrounds in May 2018 in Karaman province, were investigated. Samples were examined by flotation in saturated NaCl solution and parasite ova were diagnosed under the light microscope morphologically. Of the 20 screened playgorunds, 11 [55%, confidence interval (CI=33.6-75.2)]and 27 analyzed sample (26.2%, CI=18.4-35.2) were positive one or more parasite species. While Toxocara spp. eggs were the most common species in total (19.4%, CI=12.6-27.8), taeniid (Taenia spp., Echinococcus spp.) eggs and Ancylostoma spp. eggs were found in seven (6.8%, CI=2.97-12.7) and one (0.97%, CI=0.05-4.21) samples respectively. Also, one soil sample was found to be contaminated with both Toxocara and taeniid eggs. These results demonstrate that children’s playgrounds in Karaman may be a source for HT and other zoonotic infections. We advise to be fenced children’s playgrounds in order to prevent pet animal’s accessibility.

Toxocariasis of the Nervous System.

Toxocariasis is a helminthozoonosis caused by the infection of a human host by the larva of Toxocara spp., predominately involving Toxocara canis and Toxocara cati, which are common nematodes in dogs and cats, respectively. Human transmission occurs through contact with animals or by consumption of food contaminated with parasite's eggs. The purpose of this article is to review the current knowledge regarding human neurotoxocariasis. We conducted a systematic review of the existing literature concerning toxocariasis of the nervous system. Clinical spectrum of humantoxocariasisvaries widely from a subclinical course to significant organ morbidity. Clinical course depends on parasitic load, the migration route of the larvae and host response. Human neurotoxocariasis is a relatively rare entity yet associated with severe sequelae. Manifestations include meningitis (usually eosinophilic), encephalitis, myelitis, cerebellar vasculitis, space-occupying lesion, behavioral abnormalities, and optic neuritis. Even though valid diagnostic criteria are lacking, neurotoxocariasis should be suspected in patients with neurologic symptoms and cerebrospinal fluid (CSF) pleocytosis with eosinophilia, positive serology for anti-Toxocara antibodies, in serum and/or CSF, sterile CSF and clinical improvement after antihelminthic treatment. Neurotoxocariasis is treated by benzimidazole components, most frequently albendazole, corticosteroids, or diethylcarbamazine. Parasite larvae migrate through tissues and are able to reach the nervous system causing neurotoxocariasis. Its clinical spectrum varies and includes myelitis, meningoencephalitis, brain abscess, and vasculitis. Neurotoxocariasis should always be suspected in patients with neurologic symptoms accompanied by eosinophilia in blood and/or CSF. Early diagnosis and treatment could prevent long-term neurologic impairment.

Simplified Orange (Citrus spp.) Production Guide for Small-scale Farmers

Orange production provides both nutritional and financial benefits to farmers across Africa. However, these farmers do not realize the full benefits due to low yields caused by poor agronomic practices currently applied in the region. This guide, therefore, highlights key practices that farmers need to adopt for better yields. Farmers should always follow the best practices right from the selection of a variety to harvesting practices for high yield and better quality fruits to be achieved. Proper land preparation helps in early weed control and improves water infiltration and growth of roots. Weeds should always be kept below economic thresholds to ensure efficiencies in the use of water and nutrient. Like other plants, oranges require proper nutrition for growth and development of big fruits. Both manure and inorganic fertilizers should be applied depending on the availability and cost. Soil analysis helps in determining the rates of application. During production, farmers should scout for pests such as aphids, false codling moth, whiteflies, leaf miners, thrips, fruit fly and common spiral nematode and diseases like Pseudocercospora leaf and fruit spot, Phytophthora spp. and orange fruit scab which are common in the area. These pests cause significant yield losses if not timely controlled. When ready, harvesting of fruits should be careful and gentle without causing injuries.

Survey on Zoonotic Importance and Prevalence of Dirofilaria immitis Infection in Dogs of Tabriz, Iran

Background: Dirofilaria immitis is one of the most important parasites of the circulatory system of dogs that causes severe cardiovascular events in the animal and it is reported to be the most common nematode in humans and dogs from many parts of Iran. Methods: In order to investigate the presence of Dirofilaria immitis infection in Tabriz, Iran, a cross-sectional study was performed on 200 owned and stray dogs. The blood samples were analyzed using Knott method. The prevalence of D. immitis infection was reported to be 15% in owned dogs. Results: The highest levels of infection were reported in dogs less than one year of age and the lowest in dogs aged over 9 years. In a statistical survey, there was a significant relationship between infection and gender. Conclusion: According to the findings of this study, due to the presence of D. immitis infection in this region, the need for a more comprehensive examination and control of the infection in this region is emphasized.

Yellow and purple nutsedge and coffee senna as hosts of common plant nematodes in Florida.

Yellow (Cyperus esculentus) and purple (C. rotundus) nutsedges, and coffee senna (Senna occidentalis) are common weeds in the southern USA and each have been reported as alternative hosts for plant-parasitic nematodes. Our objective was to determine the host suitability of these weeds to plant-parasitic nematodes common in Florida agriculture and turfgrass systems. The root-knot nematode (RKN) species tested included Meloidogyne arenaria, M. enterolobii, M. floridensis, M. graminis, M. hapla, M. incognita, and M. javanica. The host status of sting nematode, Belonolaimus longicaudatus, was also evaluated, but only on the nutsedge species. All RKN species evaluated reproduced on both nutsedge species and had a reproductive factor greater than one, except for M. graminis on yellow nutsedge. However, only M. hapla, M. javanica, and M. graminis induced visual galls on yellow nutsedge and only M. graminis caused galling on purple nutsedge. Meloidogyne arenaria and M. graminis reproduced at a greater rate on purple nutsedge than on yellow nutsedge. Both nutsedge species were good hosts to B. longicaudatus. Coffee senna was a host to M. enterolobii, a poor host to M. incognita, and nonhost to the other RKN species evaluated.

Nematodes associated with terrestrial slugs in the Edmonton region of Alberta, Canada.

A survey of nematodes associated with terrestrial slugs was conducted in residential gardens, nurseries, greenhouses and agricultural sites located in and around Edmonton, Alberta, Canada. A total of 2406 slugs were collected from 82 sites. Slugs were decapitated and cadavers were incubated for two weeks, with emerging nematodes removed and processed for identification. Nematodes were identified using molecular sequence data for the 18S ribosomal DNA. Nematodes were recovered from 20 of the 82 sites surveyed, with 24.4% of the slugs infected with nematodes. A total of seven nematodes were identified to species level, including Caenorhabditis elegans, Panagrolaimus papillosus, Pellioditis typica, Pelodera pseudoteres, Rhabditella axei, Rhabditoides inermiformis and Phasmarhabditis californica. An additional four specimens were identified to genus level, including Oscheius sp. (9), Pristionchus sp., Rhabditis sp. and Rhabditophanes sp. (1). The two most common nematode species were C. elegans and P. pseudoteres. The facultative parasite, P. californica, was recovered from a single Arion rufus specimen, collected from a seasonal nursery. To our knowledge, this study represents the first survey of slug-associated nematodes in Canada.

Absence of mutations associated with resistance to benzimidazole in the beta-tubulin gene of Ascaris suum.

INTRODUCTION: Benzimidazoles are commonly used for the control of veterinary nematodes. Resistance to benzimidazoles has been associated with three single nucleotide polymorphisms in the β-tubulin gene of common nematodes. However, these mutations are infrequent in the genus Ascaris spp.METHODS: In order to determine mutations associated with benzimidazole resistance in Ascaris suum, worms were collected from slaughtered pigs and a partial region of the β-tubulin gene was sequenced.RESULTS: All parasites showed the wildtype genotype for codons 167, 198, and 200 of the β-tubulin gene.CONCLUSIONS: This is the first report of genetic sequences associated with benzimidazole resistance in A. suum.

Factors associated with the hatchability of Haemonchus species eggs and molecular identification of Haemonchus contortus

Haemonchus species is a common parasitic nematode infecting all ruminants and causing significant economic losses in developing countries. Accurate and reliable identification of various Haemonchus species and their molecular epidemiology is essential to formulate effective control strategies against Haemonchosis. The present study was carried out to identify larval stages of H. contortus in goat and sheep using morphological features and Polymerase chain reaction. A total of 400 abomasa were collected from different abattoirs for parasitological examination. Only female worms were stored for eggs isolation. The eggs were collected by macerating the whole female worms with adequate phosphate buffer saline (PBS) using a sterile mortar and pestle followed by filtering through a sieve. The collected eggs were counted by modified McMaster technique and incubated in different culture media such as phosphate buffered saline (PBS), normal saline (NS), tap water (TW) and distilled water (DW). The highest hatchability was obtained at PBS (57%) followed by NS (43%) and TW (35%) at fourth day of incubation at around neutral pH (7) and temperature 29°C. Distilled water, Acidic (pH<5) or Alkaline (pH>8) pH and high temperature (>35°C) suppressed the normal hatching rate. Different larval stages were morphologically identified according to their body length, tail and diameter in um, mouth cavity and esophageal structure. 5-7% blood sera or glucose enhanced the hatchability upto 58.33% and other growth enhancer showed no favourable effects. The amplification of 265 bp from ITS-2 of rDNA of H. contortus was successfully conducted to facilitate their molecular identification.

Seasonality and changing prevalence of common canine gastrointestinal nematodes in the USA

BackgroundThe three most commonly diagnosed gastrointestinal parasites of pet dogs within the USA are the whipworm, the hookworm, and the roundworm. The collection of large data sets from various sources throughout the industry have produced a number of publications on parasite prevalence in recent years. In this study, we look at data captured by the Companion Animal Parasite Council from 2012–2018, which includes 4.3–7.2 million annual fecal exams, to evaluate not only changes in prevalence, but also possible seasonal fluctuations of the three most common canine gastrointestinal parasites.MethodsAnnual and monthly data were collected from the CAPC parasite prevalence maps for canine roundworms, hookworms and whipworm. The map data were provided to CAPC by two large national reference laboratories. The data were evaluated for changes in prevalence on a monthly basis throughout each year as well as changes in prevalence from year to year from 2012–2018. Additionally, positive test results and total tests performed for each of the three parasites from 2012–2018 during individual months were totaled without using the year as a variable in order to evaluate the results for seasonality (i.e. all tests and positive results occurring in January, regardless of year, were totaled and analyzed).ResultsEvaluation of gastrointestinal nematode prevalence data from over 39 million fecal samples examined over a 7-year period revealed a subtle, yet significant, increasing prevalence for roundworms, an increasing prevalence for hookworms, and a slightly decreasing prevalence for whipworms. Seasonality was demonstrated for roundworms, hookworms, and to our knowledge, for the first time canine whipworms. Highest seasonal prevalence for roundworms, hookworms, and whipworms occurred during December–January, July–August, and January–February, respectively.ConclusionsEvaluation of monthly gastrointestinal parasite prevalence data from over 39 million fecal samples collected over a 7-year period revealed a slightly increasing prevalence for roundworms, an increasing prevalence for hookworms, and a slightly decreasing prevalence for whipworms. In addition to the annual changes in prevalence, seasonal prevalence was shown for the first time for whipworms. Prevalence of both whipworm and roundworm peaked in the winter, while prevalence of hookworm peaked in the late summer and early autumn.

Diversity and incidence of plant-parasitic nematodes associated with saffron (Crocus sativus L.) in Morocco and their relationship with soil physicochemical properties

Summary Saffron (Crocus sativus) fields in Morocco’s Taliouine and Taznakht regions were surveyed between January and April 2018 to study the diversity and incidence of plant-parasitic nematodes and assess the effects of soil physicochemical properties on the nematodes. Fourteen nematode genera were identified in soil and root samples collected from 66 saffron fields. The most common plant-parasitic nematodes in the Taliouine region were Pratylenchus spp. and Helicotylenchus spp. In the Taznakht region, the most common nematodes were Pratylenchus spp., Tylenchorhynchus spp. and Ditylenchus dipsaci. Nematodes, particularly Pratylenchus spp. and Ditylenchus spp., were abundant and frequent throughout the region. Several nematode genera were significantly associated with soil texture and mineral content, indicating that soil properties play an important role in plant-parasitic nematode communities. This description of plant-parasitic nematode assemblages associated with saffron fields in Morocco and their relationship with soil physicochemical properties provides a starting point from which appropriate nematode management strategies can be implemented.

First Report of the Root Lesion Nematode, Pratylenchus hippeastri, on Apple in South Africa

Root lesion nematodes (Pratylenchus spp.) are the most common plant-parasitic nematodes present in apple orchards in South Africa. During a survey conducted in 2017 and 2018 in the main apple production regions in South Africa, P. hippeastri was detected in 85% of samples that contained root lesion nematodes. Morphological and morphometric characters as well as DNA sequencing of the D2–D3 expansion segments of 28S rRNA confirmed the identity of the nematodes extracted from apple roots. Apple rootstocks supported reproduction of these nematodes with an average reproduction factor of 14, measured 90 days after inoculation. This study confirms the host status of apple (Malus domestica Borkh.) for P. hippeastri and is the first record of P. hippeastri in South Africa.

Prevalence and risk factors associated with swine gastrointestinal nematodes and coccidia in the semi-arid region of northeastern Brazil.

In northeastern Brazil, with has a predominantly dry climate, farmers seek alternative sources of income and livelihood by the rearing pigs. The gastrointestinal parasites that affect these animals represent an obstacle in the production and can cause significant economic loss. This study aimed to determine the prevalence and risk factors associated with gastrointestinal nematodes and Coccidia in swine herds in the Sousa microregion, Paraíba state, northeastern Brazil. This was a cross-sectional study, and the sampling was designed to determine the prevalence of endoparasite-positive farms and pigs. We randomly selected 51 farms and 187 pigs. Fecal samples were collected from each animal, and eggs per gram and oocysts per gram feces were recorded. The data collected in the epidemiological questionnaires were used to determine the possible risk factors associated with endoparasite-positive animal status. The prevalence of gastrointestinal nematodes and Coccidia in the pigs was 79.5% (149/187). Coccidia were the most prevalent parasite found, with 56.6% (106/187) of the pigs testing positive, followed by nematodes 22.9% (43/187). Strongylidae was the most common nematodes found (67.5%, 29/43), followed by Trichuris sp. (30.2%, 13/43) and Ascaris sp. (2.3%, 1/43). In 29 coprocultures, Strongylidae was identified: Oesophagostomum (82.2%, 25/29), Strongyloides (62.0%, 18/29), and Hyostrongylus (27.5%, 8/29). Mixed nematode and coccidial infection were observed in 72.4% (21/29) of the samples. Relevant risk factors were related to the type of management adopted by family farmers. Changes in management measures could improve the health profile of farms.

First Report of the Root-Lesion Nematode, Pratylenchus alleni, on Soybean in Wisconsin, U.S.A.

HomePlant DiseaseVol. 103, No. 8First Report of the Root-Lesion Nematode, Pratylenchus alleni, on Soybean in Wisconsin, U.S.A. PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of the Root-Lesion Nematode, Pratylenchus alleni, on Soybean in Wisconsin, U.S.A.K. Saikai and A. E. MacGuidwinK. SaikaiUniversity of Wisconsin–Madison, Department of Plant Pathology, Madison, WI 53706Search for more papers by this author and A. E. MacGuidwin†Corresponding author: A. E. MacGuidwin; E-mail Address: aem@plantpath.wisc.eduhttp://orcid.org/0000-0002-4807-8953University of Wisconsin–Madison, Department of Plant Pathology, Madison, WI 53706Search for more papers by this authorAffiliationsAuthors and Affiliations K. Saikai A. E. MacGuidwin † University of Wisconsin–Madison, Department of Plant Pathology, Madison, WI 53706 Published Online:13 Jun 2019https://doi.org/10.1094/PDIS-03-19-0501-PDNAboutSections ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Root-lesion nematodes (Pratylenchus spp.) are economically important nematode pathogens on a wide range of crops. In soybean fields in Wisconsin, it is the most common nematode pest present. In 2013, root-lesion nematodes were detected during routine diagnostics from a soil sample collected in a soybean field in Grant Co., WI. Numerous males were observed, and all life stages appeared smaller than what is typical for Pratylenchus penetrans in Wisconsin. Cultures were established by placing a single adult female in a Petri dish containing ‘IOChief’ sweet corn root explants growing on Gamborg’s B5 medium without auxins or cytokinin. After 3 months, infected root pieces in agar were transferred to new in vitro root cultures on Gamborg’s B5 medium to continue to increase the population. Nematodes were collected for morphological and molecular identification by incubating roots on a Baermann funnel for 48 h. Morphometric data were collected for 25 females and 25 males; measurements are presented as means (range). The body length of females was 482.1 μm (427.8 to 540.6 μm), with a maximum body width of 20.3 μm (18.4 to 22.6 μm). Values of a, b, and c ratios were 23.8 (21.8 to 26.3), 5.1 (4.5 to 6.0), and 20.1 (16.0 to 23.6), respectively. The height of the head was 2.3 μm (1.9 to 2.7 μm), with two lip annules. The stylet length was 14.1 μm (13.4 to 15.1 μm) with anteriorly flattened knobs. The lateral field was marked with four lines at midbody. The vulva percentage was 79% (77 to 81%). Females had a round spermatheca filled with sperm and a postuterine sac with a length of 13.3 μm (8.7 to 19.9 μm) that was slightly shorter than the body width at the vulva. The female tail was subcylindrical or slightly conical with a smooth tail tip and a tail length of 24.2 μm (20.8 to 31.7 μm) with 20 annules (16 to 25). Males were shorter with a body length of 418.0 μm (373.2 to 518.4 μm) and stylet length of 12.6 μm (11.5 to 14.0 μm). The spicules were arched, 16.5 μm long (12.8 to 19.4 μm), and the length of the gubernaculum was 4.0 μm (3.0 to 5.3 μm). The isolate was identified as Pratylenchus alleni based on morphological characters (Castillo and Vovlas 2007; Handoo and Golden 1989). Molecular characterization was based on 18S rDNA (Chizhov et al. 2006), the D2 to D3 expansion region of 28S rDNA (Subbotin et al. 2008), and cytochrome c oxidase subunit I (Derycke et al. 2010) from five specimens. The 28S sequence (GenBank accession no. MK037385) was 99% identical to P. alleni reported in Canada (Dauphinais et al. 2018) with a difference of two nucleotides. The 18S sequence (GenBank accession no. MK035844) was 98% similar to P. scribneri, but the COI sequence (GenBank accession no. MK045330) was only 84% similar to P. scribneri. There were no sequences for either loci for P. alleni available in GenBank. DNA samples of a Canadian isolate of P. alleni obtained from Agriculture and Agri-Food Canada, Saint-Jean-sur-Richelieu Research and Development Centre (Saint-Jean-sur-Richelieu, Canada) were 99 and 100% identical to the 18S and COI sequences of the Wisconsin isolate, respectively. Therefore, we identified the Wisconsin isolate as P. alleni based on the morphological and molecular data. Three soybean plants of cultivar ‘Corsoy’ were inoculated with 1,000 mixed-stage nematodes of the isolate and grown at 24°C with a 14-h photoperiod in pasteurized loamy sand soil. The reproductive factor (final population density/initial population density) 3 months after inoculation was 3.8, and many lesions were observed on roots. To our knowledge, this is the first report of P. alleni on soybean in Wisconsin. Further investigation about the economic impact of this species to soybean production and its distribution in Wisconsin are planned.The author(s) declare no conflict of interest.

Reliability of three common fecal egg counting techniques for detecting strongylid and ascarid infections in horses

The detection and quantification of nematode eggs using fecal egg count techniques have an irreplaceable role in equine parasitic control. The reliability, particularly precision and accuracy, of individual techniques have been described only for strongylid infections. The aim of this study was to compare three fecal egg count techniques used for the detection of the two most common equine nematode infections: strongylid and ascarid. The Simple McMaster, Concentration McMaster and Mini-FLOTAC techniques were tested on spiked fecal samples with various levels of egg concentration (50, 100, 200, 500, 1000 and 3000 eggs per gram) and naturally infected mixed strongylid-ascarid samples with 30 replicates. The Simple McMaster, Concentration McMaster and Mini-FLOTAC techniques had precision coefficients of variation of 44.33, 35.64 and 18.25% for the strongylid infection and 62.95, 35.71 and 18.95% for the ascarid infection, and percent accuracies (mean count/number of eggs spiked) of 97.53, 88.39 and 74.18% for the strongylid infection and 65.53, 83.18 and 90.28% for the ascarid infection, respectively. Accuracy depended greatly on the type of nematode, but precision did not. The Mini-FLOTAC technique was more precise than the Simple and Concentration McMaster techniques regardless of nematode type. Simple McMaster was the most accurate technique for detecting strongylid eggs, and Mini-FLOTAC was the most accurate technique for detecting ascarid eggs. Our results indicated that none of the current techniques were universally and sufficiently reliable for the simultaneous quantification of both of these common equine nematodes.

In vitro and in vivo Areca catechu crude aqueous extract as an anthelmintic against Ascaridia galli infection in chickens.

Aim:Intestinal parasites cause severe diseases at poultry farms, especially in developing countries, including Indonesia. Infections cause significant decreases in productivity at poultry farms. Ascaridia galli is a common nematode found in chickens with a prevalence of up to 92%. This study evaluates the in vitro and in vivo effects of Areca catechu crude aqueous extract (AAE) as an anthelmintic against A. galli.Materials and Methods:In the in vitro study, already-infected slaughtered chickens were collected from local slaughterhouses in Yogyakarta. The chicken intestines were carefully examined and transported to the Parasitological Laboratory of Gadjah Mada University, Yogyakarta. A. galli was collected from the intestines and put into a Petri dish containing 0.62% saline solution. Sixty A. galli individuals were soaked in different concentrations of the AAE for 3-6 h. The number of deceased A. galli was recorded by ascertaining whether they had ceased movement using a stereo microscope. The morphological characteristics of the parasites were studied. The in vivo study used 40 female chickens that were 1.5 months old and infected with the eggs of A. galli. Approximately 3000 infective eggs were found in each chicken. There were four treatment groups; Group A served as the control and was treated with Aquadestilata, Group B was treated with 26 mg/ml of AAE, Group C was treated with 79 mg/ml of AAE, and Group D was treated with 50 mg/ml of pyrantel pamoate. The AAE was applied using a pipette for 14 days. The observed parameters include the effect of the treatments on hematology, body weight (BW), and the number of A. galli eggs within each gram of fecal matter.Results:The AAE damaged the morphology, including the anterior end, posterior end, and vulva, of A. galli in vitro. The tegument of the anterior and posterior ends was slightly rougher, wrinkled, and damaged. The vulva and spicule ducts were irregular and indistinct. The average eggs per gram (EPG) in the untreated control increased in vivo from 1450±355.90 to 1975±325.96. The average EPG decreased from 1485±386.62 to 0±0.00 during 14 days of treatment of 79 mg/ml of AAE. The mean BW gain in the treated chickens was significantly (p<0.5) higher than the control. The treatment with 79 mg/ml of AAE increased the total erythrocyte count, total leukocyte count, hemoglobin, and packed cell volume values in the chickens.Conclusion:AAE can cause morphological changes and subsequent death in adult A. galli in vitro and effectively decrease ascariasis severity in chickens, therefore increasing overall BW in vivo.