Zoonotic parasitic organisms on vegetables: Impact of production system characteristics on presence, prevalence on vegetables in northwestern Iran and washing methods for removal.

Fresh vegetables are essential components of a healthy and nutritious diet, but if consumed raw without proper washing and/or disinfection, can be important agents of transmission of enteric pathogens. This study aimed to determine the prevalence of zoonotic parasites on vegetables freshly harvested and "ready to eat" vegetables from greengrocers and markets in northwestern Iran. In addition, the effect of cropping system and season on contamination levels were assessed as well as the efficacy of washing procedures to remove parasites from the vegetables. A total of 2757 samples composed of field (n = 1, 600) and "ready to eat" (n = 1157) vegetables were analyzed. Vegetables included leek, parsley, basil, coriander, savory, mint, lettuce, cabbage, radish, dill, spinach, mushroom, carrot, tomato, cucumber and pumpkin. Normal physiological saline washings from 200 g samples were processed using standard parasitological techniques and examined microscopically. A total of 53.14% of vegetable samples obtained from different fields and 18.23% of "ready to eat" vegetables purchased from greengrocers and markets were contaminated with different parasitic organisms including; Entamoeba coli cysts, Giardia intestinalis cysts, Cryptosporidium parvum oocysts, Fasciola hepatica eggs, Dicrocoelium dendriticum eggs, Taenia spp. eggs, Hymenolepis nana eggs, Ancylostoma spp. eggs, Toxocara cati eggs, Toxocara canis eggs, Strongyloides stercoralis larvae, and Ascaris lumbricoides eggs. In both field and "ready to eat" vegetables, the highest parasitic contamination was observed in lettuce with a rate of 91.1% and 55.44%, respectively. The most common parasitic organism was Fasciola hepatica. A seasonal difference in contamination with parasitic organisms was found for field and "ready to eat" vegetables (P < 0.05). There was a significant difference in the recovery of parasitic organisms depending on the washing method with water and dishwashing liquid being the least effective. Proper washing of vegetables is imperative for a healthy diet as the results of this study showed the presence of zoonotic parasites from field and ready to eat vegetables in Iran.

Genome-wide analysis of a putative lipid transfer protein LTP_2 gene family reveals CsLTP_2 genes involved in response of cucumber against root-knot nematode (Meloidogyne incognita).

Plant lipid transfer proteins (LTPs) are small basic proteins that play important roles in the regulation of various plant biological processes as well as the response to biotic and abiotic stresses. However, knowledge is limited on how this family of proteins is regulated in response to nematode infection in cucumber. In the present study, a total of 39 CsLTP_2 genes were identified by querying databases for cucumber-specific LTP_2 using a Hidden Markov Model approach and manual curation. The family has a five-cysteine motif (5CM) with the basic form CC-Xn-CXC-Xn-C, which differentiates it from typical nsLTPs. The members of CsLTP_2 were grouped into six families according to their structure and their phylogenetic relationships. Expression data of CsLTP_2 genes in 10 cucumber tissues indicated that they were tissue-specific genes. Two genes showed significant expression change in roots of resistant and susceptible lines during nematode infection, indicating their involvement in response to Meloidogyne incognita. This systematic analysis provides a foundation of knowledge for future studies of the biological roles of CsLTP_2 genes in cucumber in response to nematode infection and may help in the efforts to improve M. incognita-resistance breeding in cucumber.

A Phenotyping Method of Giant Cells from Root-Knot Nematode Feeding Sites by Confocal Microscopy Highlights a Role for CHITINASE-LIKE 1 in Arabidopsis.

Most effective nematicides for the control of root-knot nematodes are banned, which demands a better understanding of the plant-nematode interaction. Understanding how gene expression in the nematode-feeding sites relates to morphological features may assist a better characterization of the interaction. However, nematode-induced galls resulting from cell-proliferation and hypertrophy hinders such observation, which would require tissue sectioning or clearing. We demonstrate that a method based on the green auto-fluorescence produced by glutaraldehyde and the tissue-clearing properties of benzyl-alcohol/benzyl-benzoate preserves the structure of the nematode-feeding sites and the plant-nematode interface with unprecedented resolution quality. This allowed us to obtain detailed measurements of the giant cells' area in an Arabidopsis line overexpressing CHITINASE-LIKE-1 (CTL1) from optical sections by confocal microscopy, assigning a role for CTL1 and adding essential data to the scarce information of the role of gene repression in giant cells. Furthermore, subcellular structures and features of the nematodes body and tissues from thick organs formed after different biotic interactions, i.e., galls, syncytia, and nodules, were clearly distinguished without embedding or sectioning in different plant species (Arabidopsis, cucumber or Medicago). The combination of this method with molecular studies will be valuable for a better understanding of the plant-biotic interactions.

High tunnels: protection for rather than from insect pests?

BACKGROUND: High tunnels are a season extension tool creating a hybrid of field and greenhouse growing conditions. High tunnels have recently increased in the USA and thus research on their management is lacking. One purported advantage of these structures is protection from common field pests, but evidence to support this claim is lacking. We compared insect pest populations in high tunnels with field production over two years for three crops: tomato, broccoli and cucumber. RESULTS: Greenhouse pests (e.g. aphids, whiteflies) were more prevalent in high tunnels, compared to field plots. Hornworms (tobacco (Manduca sexta L.) and tomato (M. quinquemaculata Haworth)), a common field pest on tomato, were also more abundant in high tunnels, requiring chemical control while field populations were low. The crucifer caterpillar complex (imported cabbageworm (Pieris rapae L.), diamondback moth (Plutella xylostella L.) and cabbage looper (Trichoplusia ni Hübner)) was also more abundant in high tunnels in 2010. Cucumber beetle (striped (Acalymma vittatum F.) and spotted (Diabrotica undecimpunctata Mannerheim)) densities were higher in high tunnels in 2010 and field plots in 2011. CONCLUSION: The common assumption that high tunnels offer protection from field pests was not supported. Instead, high tunnel growing conditions may facilitate higher pest populations. © 2017 Society of Chemical Industry.

Identification of Aphis gossypii Glover (Hemiptera: Aphididae) Biotypes from Different Host Plants in North China.

BACKGROUND: The cotton-melon aphid, Aphis gossypii Glover (Hemiptera: Aphididae), is a polyphagous species with a worldwide distribution and a variety of biotypes. North China is a traditional agricultural area with abundant winter and summer hosts of A. gossypii. While the life cycles of A. gossypii on different plants have been well studied, those of the biotypes of North China are still unclear. RESULTS: Host transfer experiments showed that A. gossypii from North China has two host-specialized biotypes: cotton and cucumber. Based on complete mitochondrial sequences, we identified a molecular marker with five single-nucleotide polymorphisms to distinguish the biotypes. Using this marker, a large-scale study of biotypes on primary winter and summer hosts was conducted. All A. gossypii collected from three primary hosts-hibiscus, pomegranate, and Chinese prickly ash-were cotton biotypes, with more cotton-melon aphids found on hibiscus than the other two species. In May, alate cotton and cucumber biotypes coexisted on cotton and cucumber seedlings, but each preferred its natal host. Both biotypes existed on zucchini, although the cucumber biotype was more numerous. Aphids on muskmelon were all cucumber biotypes, whereas most aphids on kidney bean were cotton biotypes. Aphids on seedlings of potato and cowpea belong to other species. In August, aphids on cotton and cucumber were the respective biotypes, with zucchini still hosting both biotypes as before. Thus, the biotypes had different fitnesses on different host plants. CONCLUSIONS: Two host-specialized biotypes (cotton and cucumber) are present in North China. Hibiscus, pomegranate, and Chinese prickly ash can serve as winter hosts for the cotton biotype but not the cucumber biotype in North China. The fitnesses of the two host-specialized biotypes differ on various summer hosts. When alate aphids migrate to summer hosts, they cannot accurately land on the corresponding plant.

Prevalence of Parasitic Contamination in Salad Vegetables Collected from Supermarkets and Street Vendors in Amman and Baqa'a - Jordan.

One of the main ways in transmitting parasites to humans is through consuming contaminated raw vegetables. The aim of this study was to evaluate the prevalence of parasitological contamination (helminthes eggs, Giardia and Entamoeba histolytica cysts) of salad vegetables sold at supermarkets and street vendors in Amman and Baqa'a ­ Jordan. A total of 133 samples of salad vegetables were collected and examined for the prevalence of parasites. It was found that 29% of the samples were contaminated with different parasites. Of the 30 lettuce, 33 tomato, 42 parsley and 28 cucumber samples examined the prevalence of Ascaris spp. eggs was 43%, 15%, 21% and 4%; Toxocara spp. eggs was 30%, 0%, 0% and 4%; Giardia spp. cysts was 23%, 6%, 0% and 0%; Taenia/Echinococcus eggs was 20%, 0%, 5% and 0%; Fasciola hepatica eggs was 13%, 3%, 2% and 0%; and E. histolytica cysts was 10%, 6%, 0% and 0%, respectively. There was no significant difference in the prevalence of parasite in salad vegetables either between supermarkets and street vendors, or between Amman and Baqa'a, Ascaris spp. was found to be the highest prevalent parasite in salad vegetables from supermarkets and street vendors and from Amman and Baqa'a. Our results pointed out that, the parasitic contamination of salad vegetables found in our study might be caused by irrigating crops with faecal contaminated water. We concluded that salad vegetables sold in Amman and Baqa'a may cause a health risk to consumers.

Cucumis sativus L-type lectin receptor kinase (CsLecRK) gene family response to Phytophthora melonis, Phytophthora capsici and water immersion in disease resistant and susceptible cucumber cultivars.

L-type lectin receptor kinase (LecRK) proteins are an important family involved in diverse biological processes such as pollen development, senescence, wounding, salinity and especially in innate immunity in model plants such as Arabidopsis and tobacco. Till date, LecRK proteins or genes of cucumber have not been reported. In this study, a total of 25 LecRK genes were identified in the cucumber genome, unequally distributed across its seven chromosomes. According to similarity comparison of their encoded proteins, the Cucumis sativus LecRK (CsLecRK) genes were classified into six major clades (from Clade I to CladeVI). Expression of CsLecRK genes were tested using QRT-PCR method and the results showed that 25 CsLecRK genes exhibited different responses to abiotic (water immersion) and biotic (Phytophthora melonis and Phytophthora capsici inoculation) stresses, as well as that between disease resistant cultivar (JSH) and disease susceptible cultivar (B80). Among the 25 CsLecRK genes, we found CsLecRK6.1 was especially induced by P. melonis and P. capsici in JSH plants. All these results suggested that CsLecRK genes may play important roles in biotic and abiotic stresses.

Multiplex PCR for the simultaneous identification and detection of Meloidogyne incognita, M. enterolobii, and M. javanica using DNA extracted directly from individual galls.

Meloidogyne incognita, M. enterolobii, and M. javanica are the most widespread species of root-knot nematodes in South China, affecting many economically important crops, ornamental plants, and fruit trees. In this study, one pair of Meloidogyne universal primers was designed and three pairs of species-specific primers were employed successfully to rapidly detect and identify M. incognita, M. enterolobii, and M. javanica by multiplex polymerase chain reaction (PCR) using DNA extracted from individual galls. Multiplex PCR from all M. incognita, M. enterolobii, and M. javanica isolates generated two fragments of ≈500 and 1,000, 500 and 200, and 500 and 700 bp, respectively. The 500-bp fragment is the internal positive control fragment of rDNA 28S D2/D3 resulting from the use of the universal primers. Other Meloidogyne spp. included in this study generated only one fragment of ≈500 bp in size. Using this approach, M. incognita, M. enterolobii, and M. javanica were identified and detected using DNA extracted directly from individual galls containing the Meloidogyne spp. at various stages of their life cycle. Moreover, the percentage of positive PCR amplification increased with nematode development and detection was usually easy after the late stage of the second-stage juvenile. The protocol was applied to galls from naturally infested roots and the results were found to be fast, sensitive, robust, and accurate. This present study is the first to provide a definitive diagnostic tool for M. incognita, M. enterolobii, and M. javanica using DNA extracted directly from individual galls using a one-step multiplex PCR technique.

[Effects of root-knot nematodes on cucumber leaf N and P contents, soil pH, and soil enzyme activities].

A pot experiment was conducted to study the effects of inoculation with root-knot nematodes on the cucumber leaf N and P contents, and the rhizospheric and non-rhizospheric soil pH and enzyme activities. The rhizospheric soil pH didn't have a significant decrease until the inoculation rate reached 6000 eggs per plant. With the increase of inoculation rate, the leaf N and P contents, rhizospheric soil peroxidase activity, and rhizospheric and non-rhizospheric soil polyphenol oxidase activity all decreased gradually, rhizospheric soil catalase activity was in adverse, non-rhizospheric soil pH decreased after an initial increase, and non-rhizospheric soil catalase activity had no regular change. After inoculation, rhizospheric soil urease activity decreased significantly, but rhizospheric and non-rhizospheric soil phosphatase activity and non-rhizospheric soil peroxidase activity only had a significant decrease under high inoculation rate. In most cases, there existed significant correlations between rhizospheric soil pH, enzyme activities, and leaf N and P contents; and in some cases, there existed significant correlations between non-rhizospheric soil pH, enzyme activities, and leaf N and P contents.

Efeito de extrato bruto e óleo essencial de Achillea millefolium em desenvolvimento in vitro de Corynespora cassiicola e proteção de pepino à mancha de corinespora / Effect of crude extract and essential oil of Achillea millefolium L. on the in vitro development of Corynespora cassiicola and protection of cucumber against corinespora leaf spot

Medidas de controle de doenças por meio do uso de extratos e óleos essenciais vêm sendo investigadas como alternativa aos fungicidas convencionais. Este trabalho objetivou estudar o efeito antimicrobiano de extratos brutos aquosos e óleo essencial de Achillea millefolium no crescimento micelial (CM), na esporulação e na germinação de conídios de Corynespora cassiicola bem como a atividade de peroxidase em plantas de pepino. Para isto, o extrato bruto aquoso (EBA) (1; 5; 10; 20 e 25%) foi incorporado ao meio de cultura BDA (batata-dextrose-ágar) e o óleo essencial (OE) (20, 40, 60, 100 e 200 mL) foi distribuído sobre a superfície do meio de cultura. Disco de micélio foi repicado para os diferentes meios e o crescimento micelial avaliado, diariamente, por 7 dias. Para os ensaios in vivo, plantas de pepino foram pulverizadas com EBAs nas concentrações 1, 10 e 25%, Saccharomyces cereviseae 20% e água, aos quatro e dois dias antes e concomitantemente à inoculação com C. cassiicola. Foi avaliada a severidade da doença e a atividade da enzima peroxidase. Para EBA, não houve inibição do crescimento micelial, da germinação e da esporulação em todas as concentrações testadas. Entretanto, nos tratamentos com OE, houve inibição de até 63% do crescimento micelial. Na esporulação e germinação, a inibição foi de 100% e 98%, respectivamente, na alíquota de 200 ?L para OE. No controle da doença, o tratamento mais efetivo foi EBA a 25% quando realizado concomitante à inoculação. No intervalo de tempo avaliado, não se constatou atividade da enzima peroxidase.

The control of diseases by using crude extracts (CE) and essential oils (EO) are being investigated as an alternative to conventional fungicides. The objective of this work was to evaluate the antimicrobial effect of aqueous extracts and essential oil of Achillea millefolium on mycelial growth, sporolution and spores germination of Corynespora cassiicola, as well as peroxidase activity in cucumber plants. For this, crude extract (1; 5; 10; 20 and 25%) was incorporated into potato dextrose agar medium (PDA), and essential oil (20, 40, 60, 100 and 200 mL) was distributed over the surface of the PDA. The mycelial growth was measured daily for 7 days. In the in vivo test, cucumber plants were sprayed with crude extracts at concentrations 1; 10 and 25%, and Saccharomyces cereviseae 20% (control), 4 and 2 days before and the same time as inoculation with C. cassiicola. The severity of disease and peroxidase activity were evaluated. For crude extract, neither mycelial growth nor spore germination and sporulation was inhibited at any of the tested concentrations. In the treatments with essential oil there was inhibition up to 63% of the mycelial growth. In the sporulation and spores germination the inhibition was 100% and 98%, respectively, for 200uL of EO. In the control of the disease the mosteffective treatment was the crude extract 25% when carried out at the same time as the inoculation. In the evaluated period, no peroxidase activity was found.

Sampling plan for Thrips (Thysanoptera: Thripidae) on cucumber / Plano de amostragem para Tripes (Thysanoptera: Thripidae) em pepino

Este trabalho determina a melhor técnica, unidade amostral e o número de amostras para compor um plano de amostragem para os tripes Frankliniella schultzei (Trybom) e Thrips palmi (Karny) em pepino. A eficácia de três técnicas de amostragem: batida de folhas em bandeja plástica branca, contagem direta de insetos na face inferior das folhas e coleta de folhas em sacola plástica foi comparada em nove cultivos comerciais de pepino, empregando três unidades amostrais (uma folha de ramo localizado nos terços apical, mediano e basal do dossel da planta). Baseado na variância relativa e precisão econômica dos melhores sistemas amostrais (melhores técnicas e unidades amostrais), calculou-se o número de amostras para a composição do plano de amostragem convencional. O sistema mais adequado para a amostragem de F. schultzei em pepino foi a contagem direta de insetos em folha do terço apical do espaldeiramento. Para tanto, deve-se amostrar uma folha por planta em 38 plantas/lavoura. Para a amostragem de T. palmi, o melhor sistema foi a batida de folha do terço apical do espaldeiramento em bandeja plástica. Na amostragem conjunta das duas espécies, o sistema mais adequado foi a contagem direta dos insetos em folha do terço apical do espaldeiramento. Nesse caso, amostra-se uma folha por planta, em 35 plantas/lavoura. Estes resultados facilitam a tomada de decisão no manejo de tripes em pepino, agregando os benefícios da decisão correta para adoção ou não de práticas de redução populacional.

This work determines the best technique, sampling unit and the number of samples to compose a sampling plan for Frankliniella schultzei (Trybom) and Thrips palmi (Karny) (Thysanoptera: Thripidae) on cucumber. The efficacy of three sampling techniques: leaf beating on a plastic tray, direct counting of insects on the lower leaf surface, and whole leaf collection in bags were compared in nine commercial cucumber crops using three sampling units (a leaf from a branch located in the apical, median or basal third of the canopy). The number of samples was determined based on the relative variance and the economic precision for the best technique and sampling unit. The direct counting of insects on the apical third of the plant canopy was the best sampling technique for F. schultzei based on one leaf surveyed per plant using 38 plants per field. The best sampling technique for T. palmi was the leaf beating on a tray using one leaf of the apical third per plant and 35 plants per field. When joining both species, the best sampling system was the direct counting on the apical third, and it requires sampling one leaf per plant using at least 35 plants per field. These results facilitate the decision-making for the management of thrips on cucumber and aggregate the benefits of the correct decision for the adoption of strategies for population reduction.

Influência do substrato alimentar sobre o desenvolvimento de Diaphania hyalinata L. (Lepidoptera: Crambidae) / Influence of nutritional substrates on the development of Diaphania hyalinata L. (Lepidoptera: Crambidae)

Aspectos biológicos de Diaphania hyalinata L. foram estudados quando as lagartas foram criadas em dietas natural e artificial em condições de laboratório, a 25 ± 1ºC, umidade relativa de 70 ± 10 por cento e fotofase de 14h. Como dieta natural foi utilizada abóbora jacaré, pepino e abobrinha caserda e uma dieta artificial utilizada em criações de Diatraea saccharalis (Fabr.), desenvolvida por Hensley & Hammond. As lagartas de D. hyalinata foram colocadas nos diferentes substratos e criadas por todo o ciclo. Observaram-se diferenças na duração do período larval, sobrevivência das larvas e pupas, longevidade do adulto e número de ovos entre os substratos. A dieta artificial a que mostrou de modo geral, mais adequada para a criação de D. hyalinata. Entre as dietas naturais, o pepino foi o que se apresentou mais adequado para criação desta praga.

The aim of this research was to evaluate the biological behaviour of melonworm at different natural and artificial diets. Squash cultivar jacaré, cucumber, summer squash and an artificial diets developed by Hensley & Hammond for the sugarcane borer were tried. The research was carried out in the Laboratório de Entomologia do Centro de Ciências Agrárias of the Universidade Federal do Espírito Santo, in climatized room at 25 ± 1ºC, relative humidity of 70 ± 10 percent and 14h photofase. The caterpillars fed on each substrate during the whole life cycle. Differences in duration of the larval period, larvae and pupae survival, adult longevity and number of eggs were registered between the substrates. The results show that the best diet for rearing is the artificial one. Between the natural diets the cucumber showed the best results.