Search results for: entomopathogenic nematodes

Authors: Hamed A. El-Serehy, Khaled A. Al-Rasheid, Fahad A. Al-Misned

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The composition and distribution of the benthic meiofauna assemblages of the Egyptian coasts along the Red Sea are described in relation to abiotic variables. Sediment samples were collected seasonally from twelve stations chosen along the northern part of the Red Sea to observe the meiofaunal community structure, its temporal distribution and horizontal fluctuation in relation to environmental conditions of the Red Sea marine ecosystem. The meiofaunal assemblage in the area of study was well diversified including 140 taxa. The temperature, salinity, pH, dissolved oxygen, and redox potential were measured at the time of collection. The water content of the sediments, total organic matters and chlorophyll a values were determined, and sediment samples were subjected to granulometric analysis. A total of 10 meiofauna taxa were identified, with the meiofauna being primarily represented by nematodes (on annual average from 42% to 84%), harpacticoids, polycheates and ostracodes; and the meiofauna abundances ranging from 41- to 167 ind. / 10 cm2. The meiofaunal population density fluctuated seasonally with a peak of 192.52 ind. / 10 cm2 during summer at station II. The vertical zonation in the distribution of meiofaunal community was significantly correlated with interstitial water, chlorophyll a and total organic matter values. The present study indicates that the existing of well diversified meiofaunal group which can serve as food for higher trophic levels in the Red Sea interstitial environment.

Keywords: benthos, diversity, meiofauna, Red Sea

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Authors: A. Ridall, J. Ingels

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Microplastics (MPs; < 5 mm) have been cited as exceptionally detrimental to marine organisms and ocean health. They can carry other pollutants and abundant microbes that can serve as food for other organisms. Their small particle size and high abundance means that non-discriminatory feeders may ingest MPs involuntarily and microbial colonization of the particles (a niche coined ‘Plastisphere’) could facilitate particle ingestion. To assess how marine nematodes, the most abundant member of the meiofauna (32-500 um), are affected by microbe-MP associations, an experiment was conducted with three MP concentrations (low, medium, and expected high values of MPs in a local bay system), and two levels of microbe-MP associations (absence or presence). MPs were introduced into sediment microcosms and treatments were removed at three distinct time points (0, 3, and 7 days) to measure mean MP consumption/individual nematode. The quantitative results from this work should inform on microbial facilitation of MP ingestion and MP effects on seafloor ecology. As most MP feeding experiments use straight-from-package or sterile MPs, this work represents an important step in realizing the effects of MPs and their plastispheres in coastal sediments where they likely accumulate microbial biofilms prior to their ingestion by marine metazoans. Furthermore, the results here convey realistic effects of MPs on faunal behaviors, as the MP concentrations used are based on field measurements rather than artificially high levels.

Keywords: ecosystem function, microbeads, plastisphere, pollution, polyethylene

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Authors: Fumiko Kojima, Shuji Fujimoto

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Anisakiasis is a disease caused by infection with an anisakid larvae, mostly Anisakis simplex. The larvae commonly infect in marine fish and the disease is frequently reported in areas of the world where fish is consumed raw, lightly pickled or salted. In Japan, people have the habit of eating raw fish such as ‘sushi’ or ‘sashimi’, so they have more chance of infection with larvae of anisakid nematodes. There are three sibling species in A. simplex larvae, namely, A. simplex sensu stricto (Asss), A. pegreffii (Ap) and A. simplex C. It was revealed that Ap is dominant among the larvae from fish (Scomber japonics) in the Japan Sea side and Asss is dominant among those of the Pacific Ocean side conversely. Although anisakiasis has happened in Japan among both the Japan Sea side area and the Pacific Ocean side area. The aim of this study was to investigate genetic variations between the siblings (Asss and Ap) and within the same sibling species by random amplified polymorphic DNA (RAPD) technique. In order to investigate the genetic difference among the each A. simplex larvae, we used RAPD technique to differentiate individuals of A. simplex obtained from Scomber japonics fish those were caught in the Japan sea (Goto Islands in Nagasaki Prefecture) and the cost of Pacific Ocean (Kanagawa Prefecture). The RAPD patterns of the control DNA (Genus Raphidascaris) were markedly different from those of the A. simplex. There were differences in amplification patterns between Asss and Ap. The RAPD patterns for larvae obtained from fish of the same sea were somewhat different and variations were detected even among larvae from the same fish. These results suggest the considerable high genetic variability between Asss and Ap and the possible existence of genetic variation within the sibling species.

Keywords: Anisakiasis in Japan, Anisakis simplex, genomic identification, PCR-RAPD

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Authors: Anil F. Felek, Mehmet M. Ozcan, Faruk Akyazi

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Volatile oils of medicinal and aromatic plants are important for managing nematological problems in agriculture. In present study, volatile oils extracted from five medicinal and aromatic plants including Origanum onites (flower+steam+leaf), Salvia officinalis (leaf), Lippia citriodora (leaf+seed), Mentha spicata (leaf) and Mentha longifolia (leaf) were tested for egg hatching inhibition activity against root-knot nematode Meloidogyne hapla under laboratory conditions. The essential oils were extracted using water distillation method with a Clevenger system. For the homogenisation process of the oils, 2% gum arabic solution was used and 4 µl oils was added into 1ml filtered gum arabic solution to prepare the last stock solution. 5 ml of stock solution and 1 ml of M. hapla egg suspension (about 100 eggs) were added into petri dishes. Gum arabic solution was used as control. Seven days after exposure to oils at room temperature (26±2 °C), the cumulative hatched and unhatched eggs were counted under 40X inverted light microscope and Abbott’s formula was used to calculate egg hatching inhibition rates. As a result, the highest inhibition rate was found as 54% for O. onites. In addition, the other inhibition rates varied as 31.4%, 21.6%, 23.8%, 25.67% for the other plants, S. officinalis, M. longifolia, M. spicata and L. citriodora, respectively. Carvacrol was found as the main component (68.8%) of O. onites followed by Thujone 27.77% for S. officinalis, I-Menthone 76.92% for M. longifolia, Carvone 27.05% for M. spicata and Citral 19.32% for L. citriodora.

Keywords: egg hatching, Meloidogyne hapla, medicinal and aromatic plants, root-knot nematodes, volatile oils

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Authors: Abera Hailu Degaga, Emana Getu Degaga

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Spodoptera frugiperda is an invasive insect pest that infests and feeds various crops, particularly affecting maize yields. However, nature has its own way of maintaining balance, and in this case, natural enemies play a crucial role in regulating the population of S. frugiperda. Locally available and easily prepared botanical sources, bio-pesticides, are also important. The objectives of the study were to investigate the natural enemies of S. frugiperda in the Gurage zone and to compare Neem aqueous extracts against its larvae in central Ethiopia. S. frugiperda larvae and egg masses were collected randomly from smallholder maize farms infested with pests between June and August 2023. Our findings revealed the existence of diverse types of parasitoids, predators, and entomopathogenic fungi associated with S. frugiperda. Notably, we documented three species of parasitoids, namely Exorista xanthaspis and Tachina spp. (Diptera: Tachinidae) and Charops annulipes (Hymenoptera: Ichneumonidae). All three species of parasitoids were recorded from Ethiopia for the first time. The overall parasitism rate was 5.3%, with individual rates ranging from 1.3 to 4%. Additionally, we identified ten species of predator insects from four different orders, including Hemiptera, Dermaptera, Coleoptera, and Mantodea, in the maize farms infested with S. frugiperda. Aqueous extract of Neem seed and leaf powder and green leaf exhibited similar mortality rates of S. frugiperda larvae at 72 hours even though there was a significant difference at 24 and 48 hours of the test. For effective management of S. frugiperda further research is necessary to fully exploit the potential of these natural enemies and additionally to use botanical source pesticides like Azadirachta indica.

Keywords: bio-pesticide, natural enemy, parasitoids, predators, Tachinid flies

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Authors: G. Quiroga-Cubides, M. Cruz, E. Grijalba, J. Sanabria, A. Ceballos, L. García, M. Gómez

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Beauveria sp. has been used as an entomopathogenic microorganism for biological control of various plant pests such as whitefly, thrips, aphids and chrysomelidaes (including Cerotoma tingomariana species), which affect soybean crops in Colombia´s Altillanura region. Therefore, a biopesticide prototype based on B. bassiana strain Bv060 was developed at Corpoica laboratories. For the production of B. bassiana conidia, a baseline fermentation was performed at laboratory in a solid medium using broken rice as a substrate, a temperature of 25±2 °C and a relative humidity of 60±10%. The experimental design was completely randomized, with a three-time repetition. These culture conditions resulted in an average conidial concentration of 1.48x10^10 conidia/g, a yield of 13.07 g/kg dry substrate and a productivity of 8.83x10^7 conidia/g*h were achieved. Consequently, the objective of this study was to evaluate the influence of the particle size reduction of rice (<1 mm) and the addition of a complex nitrogen source over conidia production and efficiency parameters in a solid-state fermentation, in a completely randomized experiment with a three-time repetition. For this aim, baseline fermentation conditions of temperature and humidity were employed in a semisolid culture medium with powdered rice (10%) and a complex nitrogen source (8%). As a result, it was possible to increase conidial concentration until 9.87x10^10 conidia/g, yield to 87.07 g/g dry substrate and productivity to 3.43x10^8 conidia/g*h. This suggested that conidial concentration and yield in semisolid fermentation increased almost 7 times compared with baseline while the productivity increased 4 times. Finally, the designed system for semisolid-state fermentation allowed to achieve an easy conidia recovery, which means reduction in time and costs of the production process.

Keywords: Beauveria bassiana, biopesticide, solid state fermentation, semisolid medium culture

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Authors: Fundime Miri, Emanuela Sulaj

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Benthic ecosystems constitute important ecological habitats, providing fundamental services for spawning, foraging, and sheltering aquatic organisms. Benthic faunal communities are characterized by a large biological diversity, supported by a great physical variety of benthic habitats. Until late, the study of meiobenthic communities in Albania has been neglectedthus excluding an important component of benthos. The present study aims to bring characteristics of distribution pattern of meiofaunal communities with further focus on nematode genus-based diversity from different intertidal habitats along Albanian Adriatic Sea Coast. The investigation area is extended from Shkodra to Vlora District, including six sandy sampling sites in beaches and areas near river estuaries. Sediment samples were collected manually in low intertidal zone by using a cylindrical corer, with an internal diameter of 5 cm. The richness onmeiofaunalmajor taxon level did not show any significant change between different sampling sites compare to significant changes in nematode diversity at genus level, with distinct nematode assemblages per sampling sites and presence of exclusive genera. All meiofaunal communities under study were dominated by nematodes. Further assessment of functional diversity on nematode assemblages exhibited changes as well on trophic groups and life strategies due to diverse feeding behaviors and c-p values of nematode genera. This study emphasize the need for lower level taxonomic identification of meiofaunal organisms and extending of ecological assessments on trophic diversity and life strategies to understanding functional consequences.

Keywords: benthos, meiofauna, nematode genus-based diversity, functional diversity, intertidal, albanian adriatic coast

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Authors: Phatu William Mashela, Pontsho Edmund Tseke, Kgabo Martha Pofu

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‘Inconsistent results’ in nematode suppression post-application of botanical-based products created credibility concerns. Relative to untreated control, sampling for nematodes post-application of botanical-based products suggested significant increases in nematode population densities. ‘Inconsistent results’ were confirmed in Tylenchulus semipenetrans on Citrus jambhiri seedlings when sampling was carried out at 120 days post-application of a granular Nemarioc-AG phytonematicide. The objective of this study was to determine post-application effects of untreated control, Nemarioc-AG phytonematicide and aldicarb to T. semipenetrans population densities on C. jambhiri seedlings. Two hundred and ten seedlings were each inoculated with 10000 T. semipenetrans eggs and second-stage juveniles (J2) in plastic pots containing 2700 ml growing mixture. A week after inoculation, seedlings were equally split and subjected to once-off treatment of 2 g aldicarb, 2 g Nemarioc-AG phytonematicide and untreated control. Five seedlings from each group were randomly placed on greenhouse benches to serve as a sampling block, with a total of 14 blocks. The entire block was sampled weekly and assessed for final nematode population density (Pf). After the final assessment, post-regression of untreated Pf to increasing sampling intervals exhibited positive quadratic relations, with the model explaining 90% associations, with optimum Pf of 13804 eggs and J2 at six weeks post-application. In contrast, treated Pf and increasing sampling interval exhibited negative quadratic relations, with the model explaining 95% and 92% associations in phytonematicide and aldicarb, respectively. In the phytonematicide, Pf was 974 eggs and J2, whereas that in aldicarb was 2205 eggs and J2 at six weeks. In conclusion, temporal cyclic nematode population growth provided an empirically-based explanation of ‘inconsistent results’ in nematode suppression post-application of the two nematode management strategies.

Keywords: nematode management, residual effect, slow decline of citrus, the citrus nematode

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Authors: Iyabode A. Kehinde, Joshua O. Oyekanmi, Jumoke T. Abimbola, Olajumoke E. Ayanda

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African yam bean (AYB), (Sphenostylis stenocarpa) is a leguminous crop that provides nutritionally rich seeds, tubers and leaves for human consumption. AYB potentials as an important food security crop is yet to be realized and thus classified as underutilized crop. Underutilization of the crop has been partly associated with scarce information on the incidence and characterization of fungal endophytes infecting vascular parts of AYB. Accurate and robust detection of these endophytic fungi is essential for diagnosis, modeling, surveillance and protection of germplasm (seed) health. This work aimed at isolating and identifying fungal endophytes associated with leaves, stems and roots of AYB in Ogun State, Nigeria. This study investigated both cultural and molecular properties of endophytic fungi in AYB for its characterization and diversity. Fungal endophytes were isolated and culturally identified. DNA extraction, PCR amplification using ITS primers and analyses of nucleotide sequences of ribosomal DNA fragments were conducted on selected isolates. BLAST analysis was conducted on consensus nucleotide sequences of 28 out of 30 isolates and results showed similar homology with genera of Rhizopus, Cunninghamella, Fusarium, Aspergillus, Penicillium, Alternaria, Diaporthe, Nigrospora, Purpureocillium, Corynespora, Magnaporthe, Macrophomina, Curvularia, Acrocalymma, Talaromyces and Simplicillium. Slight similarity was found with endophytes associated with soybean. Phylogenetic analysis by maximum likelihood method showed high diversity among the general. These organisms have high economic importance in crop improvement. For an instance, Purpureocillium lilacinum showed high potential in control of root rot caused by nematodes in tomatoes. Though some can be pathogens, but many of the fungal endophytes have beneficial attributes to plant in host health, uptake of nutrients, disease suppression, and host immunity.

Keywords: molecular characterization, African Yam Bean, fungal endophyte, plant parts

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Authors: M. Younus, Muhammad Shafique, M. Athar Khan, Tanveer Akhtar , M. Moeen Athar

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A total of 48 apparently healthy weaned sheep lambs (Ovis aries) of 8-10 weeks old weighing 7-10 Kg were purchased from the contractors, maintained in the experimental station of University of the Punjab, Quaid-e-Azam Campus at Lahore, Pakistan. They were dewormed against nematodes with levamisole (ICI) at recommended dose rates. The feces were tested against the parasitic eggs, no helminths ova were seen. All the 48 sheep lambs were divided into two groups i.e. group A & group B. Group 'A' comprising of 40 sheep, kept as infected groups whereas group 'B' comprising of 08 sheep & kept as a new infected control group. Each sheep lamb of group A was given 3-4 fresh gravid segments contains 2-3 thousand eggs of Echinococcus granulosus. These were collected from experimentally infected dogs by feeding fresh hydrated cysts collected from liver & lungs of sheep after slaughtering. Each lamb was fed with fresh gravid segments for a total period of 5 days or each alternate day. Coagulated blood was collected before the start of infected diet and after every month by jugular phlebotomy of each sheep lamb from the infected & new infected control group. One lamb each from group A & group B was slaughtered at the end of each month for the presence of macroscopic hydatid cyst in viscera & abdominal cavity. After 180 days of the experiment, hydatid cysts were confirmed in the abdominal cavity. Hematological parameters of zero days & then at the end of every month revealed that there was a gradual increase (PL 0.05) in the White Blood Cell (WBC), Mean Corpuscular Volume (MCV), Mean Corpuscular Hemoglobin Concentration (MCHC) and Erythrocyte Sedimentation Rates (ESR). The increasing trend was probably due to inflammatory response and lytic effect of the newly developing E. granulosus hydatid cysts. The red blood cell (RBC), Hemoglobin (HB), Packed Cell Volume (PCV) and Mean Corpuscular Hemoglobin (MCH) infected groups were decreased significantly as compared to the control group (PL 0.05). The experiment was terminated at the end of the 7th month. It can be concluded that Echinococcus granulosus can damage livestock and other intermediate hosts such as horses, the development of hydatid cysts affect the organs due to the growing cysts pressuring the organ tissues. Parts of the tissue die, which impairs the functioning of the affected organ. The clinical signs depend on the affected organ. The major damage for livestock is organ condemnation at slaughter.

Keywords: echinococcus granulosus, hydatidosis, sheep, hematology

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Authors: Valeria T. Dilcheva, Svetlozara L. Petkova, Ivelin Vladov

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Trichinellosis is a food-borne parasitic disease caused by nematodes of the genus Trichinella which are zoonotic parasites with cosmopolitan distribution and major socio-economic importance. Human infection is acquired through consumption of undercooked meat from domestic or wild animal. Penetration of Trichinella larvae into striated skeletal muscle cells results in ultrastructural and metabolic changes. Migration of larvae causes the typical symptoms and signs of the disease. The severity of the symptoms depends on the number of ingested Trichinella larvae and the immune response of the host. Eosinophilia is present, with few exceptions, in most cases of human trichinellosis, inasmuch as it is the earliest and most important host response. Even in human asymptomatic cases, increases in eosinophilia of up to 15% have been observed. Eosinophilia appears at an early stage of infection between the second and fifth weeks of infection. By 2005 it was considered that only two species of Trichinella genus were found in the country. After routine trichinelloscopy procedure disseminated single muscle larvae in samples of wild boars and badger were PCR-identified as T. pseudospiralis. The study aimed to observed hematological changes occurring during experimentally induced infection with Trichinella spiralis, T. britovi and T. pseudospiralis in mice. We performed hematological blood profile, tracking 15 blood indicators. In statistical analysis made by Two-way ANOVA, there were significant differences of HGB, MCHC, PLT, Lymph%, Gran% in all three types of trichinellosis compared to control animals. Capsule-forming T. spiralis showed statistically significant differences in HGB, MCHC, Lymph% and PLT compared to the other two species. Non capsule-forming T. pseudospiralis showed statistically significant differences in Lymph%, Gran% relative to the control and in Gran% relative to T. spiralis. It appears rather substantial the process of capsule formation for prolonged immune response and retention of high content of percentage of lymphocytes(Lymph%) and low of granulocyte(Gran%) in T. pseudospiralis, which is contrary to studies for T. spiralis and eosinophilia. Studies and analyzes of some specific blood profile parameters can provide additional data in favor of early diagnosis and adequate treatment as well as provide a better understanding of acute and chronic trichinosis.

Keywords: hematological test, T. britovi, T. spiralis, T. pseudospiralis

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Authors: Gunjan Srivastava, Shamila Kalia

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Dalbergia sissoo Roxb., (Family- Leguminosae; Subfamily- Papilionoideae), is an economically and ecologically important tree species having medicinal value. Of the rich complex of insect fauna, ten have been recognized as potential pests of nurseries and plantations. Present study was conducted to explore an effective ecofriendly control of Dichomeris eridantis Meyrick, an important defoliator pest of D. sissoo. Health and environmental concerns demanded devising a bio-intensive pest management strategy and employing ecofriendly measures. In the present laboratory bioassay two entomopathogenic fungi Acremonium perscinum and Beauveria bassiana were tested and compared for evaluating the efficacy of their seven different concentrations (besides control) against the 3rd, 4th and 5th instar larvae of D. eridantis, on the basis of mean percent mortality data recorded and tabulated for seven days after treatment application. Analysis showed that both treatments vary significantly among themselves. Also, variations amongst instars and duration with respect to their mortality were highly significant (p < .001). All their interactions were found to vary significantly. B. bassiana at 0.25x107 spores / ml spore concentration caused maximum mean percent mortality (62.38%) followed by mean percent mortality at its 0.25x106 spores / ml concentration (56.67%). Mean percent mortality at maximum spore concentration (0.054x107 spores / ml) and next highest spore concentration (0.054 x106 spores / ml) due to A. perscinum treatment were far less effective (mean percent mortality of 45.40% and 31.29%, respectively). At 168 hours mean percent mortality of larval instars due to both fungal treatment applications reached its maximum (52.99%) whereas, at 24 hours mean percent mortality remained least (5.70%). In both cases, treatments were most effective against 3rd instar larvae and least effective against 5th instar larvae. A comparative acccount of efficacy of B. bassiana and A. perscinum on the 3rd, 4th and 5th instar larvae of D. eridantis on 5th, 6th and 7th post treatment observation days after their application, on the basis of their median lethal concentrations (LC50) proved B. bassiana to be more potential microbial pathogen of the two fungal microbes, for all the three instars (3rd, 4th and 5th) of D. eridantis, on all the three days (5th, 6th and 7th post observation days after application of both treatments). Percent mortality of D. eridantis increased in a dose dependent manner. Koch’s Postulates tested positive, thus confirming the pathogenicity of B. bassiana against the larval instars of D. eridantis. LC90 values of 0.280x1011 spores/ml, 0.301x108 spores/ml and 0.262x108 spores/ml concentrations of B. bassiana were standardized which can effectively cause mortality of all the larval instars of D. eridantis in the field after 5th, 6th and 7th day of their application, respectively. Therefore, these concentrations can be safely used in nurseries as well as plantations of D. sissoo for effective control of D. eridantis larvae.

Keywords: Acremonium perscinum, Beauveria bassiana, Dalbergia sissoo, Dichomeris eridantis

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Authors: P. Serrano-Pérez, M. C. Rodríguez-Molina, C. Palo, E. Palo, A. Lacasa

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Phytophthora nicotianae is the principal causal agent of root and crown rot disease of red pepper plants in Extremadura (Western Spain). There is a need to develop a biologically-based method of soil disinfestation that facilitates profitable and sustainable production without the use of chemical fumigants. Anaerobic Soil Disinfestation (ASD), as well know as biodisinfestation, has been shown to control a wide range of soil-borne pathogens and nematodes in numerous crop production systems. This method implies soil wetting, incorporation of a easily decomposable carbon-rich organic amendment and covering with plastic film for several weeks. ASD with rapeseed cake (var. Tocatta, a glucosinolates-free variety) used as C-source was assayed in spring 2014, before the pepper crop establishment. The field experiment was conducted at the Agricultural Research Centre Finca La Orden (Southwestern Spain) and the treatments were: rapeseed cake (RCP); rapeseed cake without plastic cover (RC); control non-amendment (CP) and control non-amendment without plastic cover (C). The experimental design was a randomized complete block design with four replicates and a plot size of 5 x 5 m. On 26 March, rapeseed cake (1 kg·m-2) was incorporated into the soil with a rotovator. Biological probes with the inoculum were buried at 15 and 30-cm depth (biological probes were previously prepared with 100 g of disinfected soil inoculated with chlamydospores (chlam) of P. nicotianae P13 isolate [100 chlam·g-1 of soil] and wrapped in agryl cloth). Sprinkler irrigation was run until field capacity and the corresponding plots were covered with transparent plastic (PE 0.05 mm). On 6 May plastics were removed, the biological probes were dug out and a bioassay was established. One pepper seedling at the 2 to 4 true-leaves stage was transplanted in the soil from each biological probe. Plants were grown in a climatic chamber and disease symptoms were recorded every week during 2 months. Fragments of roots and crown of symptomatic plants were analyzed on NARPH media and soil from rizospheres was analyzed using carnation petals as baits. Results of “survival” were expressed as the percentage of soil samples where P. nicotianae was detected and results of “infectivity” were expressed as the percentage of diseased plants. No differences were detected in deep effect. Infectivity of P. nicotianae chlamydospores was successfully reduced in RCP treatment (4.2% of infectivity) compared with the controls (41.7% of infectivity). The pattern of survival was similar to infectivity observed by the bioassay: 21% of survival in RCP; 79% in CP; 83% in C and 87% in RC. Although ASD may be an effective alternative to chemical fumigants to pest management, more research is necessary to show their impact on the microbial community and chemistry of the soil.

Keywords: biodisinfestation, BSD, soil fumigant alternatives, organic amendments

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Authors: S. A. Bhat, Ahmad Arif, Muneeb U. Rehman, Manzoor R Mir, S. Bilal, Ishraq Hussain, H. M Khan, S. Shanaz, M. I Mir, Sabhiya Majid

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Background: Geologically Climatic conditions of the state range from sub-tropical (Jammu), temperate (Kashmir) to cold artic (Ladakh) zones, which exerts significant influence on its agro-climatic conditions. Gastrointestinal parasitism is a major problem in sheep production worldwide. Materials and Methods: The present study was to evaluate the resistance status of sheep breeds reared in Kashmir Valley for natural resistance against Haemonchus contortus by natural pasture challenge infection. Ten microsatellite markers were used in the study for evaluation of association of Ovar-MHC with parasitic resistance in association with biochemical and parasitological parameters. Following deworming, 500 animals were subjected to selected contaminated pastures in a vicinity of the livestock farms of SKUAST-K and Sheep Husbandry Kashmir. For each animal about 10-15 ml blood was collected aseptically for molecular and biochemical analysis. Weekly fecal samples (3g) were taken, directly from the rectum of all experimental animals and examined for Fecal egg count (FEC) with modified McMaster technique. Packed cell volume (PCV) was determined within 2-5 h of blood collection, all the biochemical parameters were determined in serum by semi automated analyzer. DNA was extracted from all the blood samples with phenol-chloroform method. Microsatellite analysis was done by denaturing sequencing gel electrophoresis Results: Overall sheep from Bakerwal breed followed by Corriediale breed performed relatively better in the trial; however difference between breeds remained low. Both significant (P<0.05) and non-significant differences with respect to resistance against haemonchosis were noted at different intervals in all the parameters.. All the animals were typed for the microsatellites INRA132, OarCP73, DRB1 (U0022), OLA-DQA2, BM1818, TFAP2A, HH56, BM1815, IL-3 and BM-1258. An association study including the effect of FEC, PCV, TSP, SA, LW, and the number of alleles within each marker was done. All microsatellite markers showed degree of heterozygosity of 0.72, 0.72, 0.75, 0.62, 0.84, 0.69, 0.66, 0.65, 0.73 and 0.68 respectively. Significant association between alleles and the parameters measured were only found for the OarCP73, OLA-DQA2 and BM1815 microsatellite marker. Standard alleles of the above markers showed significant effect on the TP, SA and body weight. The three sheep breeds included in the study responded differently to the nematode infection, which may be attributed to their differences in their natural resistance against nematodes. Conclusion: Our data confirms that some markers (OarCP73, OLA-DQA2 and BM1815) within Ovar-MHC are associated with phenotypic parameters of resistance and suggest superiority of Bakerwal sheep breed in natural resistance against Haemonchus contortus.

Keywords: Ovar-Mhc, ovine leukocyte antigen (OLA), sheep, parasitic resistance, Haemonchus contortus, phenotypic & genotypic markers

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