Search results for: root-knot nematode

Authors: Yuhei Kunihiro, Sorin V. Sabau, Kazuhiro Shibuya

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We study the molecular evolution of insulin from the metric geometry point of view. In mathematics, and particularly in geometry, distances and metrics between objects are of fundamental importance. Using a weaker notion than the classical distance, namely the weighted quasi-metrics, one can study the geometry of biological sequences (DNA, mRNA, or proteins) space. We analyze from the geometrical point of view a family of 60 insulin homologous sequences ranging on a large variety of living organisms from human to the nematode C. elegans. We show that the distances between sequences provide important information about the evolution and function of insulin.

Keywords: metric geometry, evolution, insulin, C. elegans

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Authors: Francesco La Torre, Angela Di Cesare, Donato Traversa

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The mosquito-transmitted nematode Dirofilaria repens is the causative agent of subcutaneous filariosis in dogs, other animals and humans. Adults and circulating microfilariae may cause different forms of skin conditions, and various allergic reactions. The infection is distributed in several countries and spreading in several areas of Europe. The control of D. repens is pivotal to reduce the transmission in dogs and to minimize the risk of infection in humans, but only few information is available for the chemoprevention of subcutaneous filariosis of dogs. A recent clinical field study showed the efficacy and safety of a monthly administration of an oral formulation containing milbemycin oxime (Milbemax®, Novartis Animal Health) in the chemoprevention of D. repens infection in dogs. Most recent and focused insights into epidemiology and control of zoonotic canine subcutaneous filariosis are here discussed.

Keywords: Dirofilaria repens, epidemiology, zoonosis, control

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Authors: Ekatereine Abashidze, Nino Nazarashvili, Dali Gaganidze, Oleg Gorgadze, Mariam Aznarashvili, Eter Gvritishvili

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Potato is one of the leading agricultural crops in Georgia. Georgia produces early and late potato varieties in almost all regions. Potato production is equal to 25,000 ha and its average yield is 20-25 t/ha. Among the plant pests that limit potato production and quality, the potato cyst nematodes (Globodera pallida (Stone) Behrens and Globodera rostochiensis (Wollenveber) Behrens) are harmful around the world. PCN is among the most difficult plant pests to control. Cysts protected by a durable wall can survive for over 30 years . Control of PCN (G. pallida and G. rostochiensis) is regulated by Council Directive 2007/33/EE C. There was no legislative regulation of these pests in Georgia before 2016. By Resolution #302 from July 1, 2016, developed within the action plan of the DCFTA (Deep and Comprehensive Free Trade Area) the Government of Georgia established control over potato cyst nematodes. The Agreement about the legal acts approximation to EU legislation concerns the approval of rules of PCN control and research of these pests. Taking into consideration the above mentioned, it is necessary to study PCN (G. pallida and G. rostochiensis) in the potato-growing areas of Georgia. The aim of this research is to conduct survey of potato cyst nematodes (Globodera rostochiensis and G. pallida) in two geographically distinct regions of Georgia - Samtskhe - Javakheti and Svanetii and to identify the species G. Rostochiensis and G. Pallida by the morphological - morphometric and molecular methods. Soil samples were taken in each village, in a zig-zag pattern on the potato fields of the private sector, using the Metlitsky method. Samples were taken also from infested potato plant roots. To extract nematode cysts from soil samples Fanwick can be used according to standard methods by EPPO. Cysts were measured under a stereoscopic microscope (Leica M50). Identification of the nematod species was carried out according to morphological and morphometric characteristics of the cysts and larvae using appropriate protocols EPPO. For molecular identification, a multiplex PCR test was performed by the universal ITS5 and cyst nematodes’ (G. pallida, G. rostochiensis) specific primers. To identify the species of potato cyst nematodes (PCN) in two regions (Samtskhe-Javakheti and Svaneti) were taken 200 samples, among them: 80 samples in Samtskhe-Javakheti region and 120 in Svaneti region. Cysts of Globiodera spp. were revealed in 50 samples obtained from Samtskhe-Javakheti and 80 samples from Svaneti regions. Morphological, morphometric and molecular analysis of two forms of PCN found in investigated regions of Georgia shows that one form of PCN belongs to G. rostoshiensi; the second form is the different species of Globodera sp.t is the subject of future research. Despite the different geographic locations, larvae and cysts of G. rostoshiensi were found in both regions. But cysts and larvae of G. pallida were not reported. Acknowledgement: The research has been supported by the Shota Rustaveli National Scientific Foundation of Georgia: Project # FR17_235.

Keywords: cyst nematode, globodera rostochiensis, globodera pallida, morphologic-morphometric measurement

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Authors: Vassilena Dakova, Mariana Panayotova-Pencheva

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The present work focused on the study of pulmonary helminth-fauna of the stone marten in Bulgaria in terms of which the data are little. For the purpose, four stone martens were helminthologically necropsied according to the common technique. In addition, some of the injured lung parts were investigated after their boiling in lactic acid and subsequent compression. Four nematode species from different families of order Strongylida and Trichocephalida were found in the lungs. These were Crenosoma petrowi Morosov, 1939; Eucoleus aerophilus Creplin, 1839; Filaroides martis Werner, 1782 and Sobolevingylus petrowi Romanov, 1952. Some of the parasite structures with taxonomic importance were measured and described. According to our best knowledge, the species F. martis and S. petrowi are recorded for the first time as a part of the helminth-fauna of Southeast Europe and Bulgaria in particular.

Keywords: Bulgaria, Crenosoma petrowi, Eucoleus aerophilus, Filaroides martis, lung parasites, Sobolevingylus petrowi, stone martens

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Authors: Zakheleni P. Dube, Phatu W. Mashela

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Minimum inhibition concentration (MIC) is the lowest concentration of a chemical that brings about significant inhibition of target organism. The conventional method for establishing the MIC for phytonematicides is tedious. The objective of this study was to use the Curve-fitting Allelochemical Response Data (CARD) to determine the MIC for pure cucurbitacin A on Meloidogyne incognita second-stage juveniles (J2) hatch, immobility and mortality. Meloidogyne incognita eggs and freshly hatched J2 were separately exposed to a series of pure cucurbitacin A concentrations of 0.00, 0.25, 0.50, 0.75, 1.00, 1.25, 1.50, 1.75, 2.00, 2.25 and 2.50 μg.mL⁻¹for 12, 24, 48 and 72 h in an incubator set at 25 ± 2°C. Meloidogyne incognita J2 hatch, immobility and mortality counts were determined using a stereomicroscope and the significant means were subjected to the CARD model. The model exhibited density-dependent growth (DDG) patterns of J2 hatch, immobility and mortality to increasing concentrations of cucurbitacin A. The average MIC for cucurbitacin A on M. incognita J2 hatch, immobility and mortality were 2.2, 0.58 and 0.63 µg.mL⁻¹, respectively. Meloidogyne incognita J2 hatch had the highest average MIC value followed by mortality and immobility had the least. In conclusion, the CARD model was able to generate MIC for cucurbitacin A, hence it could serve as a valuable tool in the chemical-nematode bioassay studies.

Keywords: inhibition concentration, phytonematicide, sensitivity index, threshold stimulation, triterpenoids.

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Authors: M. A. Ahmed, N. Basha, I. V. Nsahlai

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This study determined the effectiveness of selected anthementics (Ivermectin 1% (IVM), Closantel 7.5% (CST) and a combination Abamectin 0.08% and Praziquantel 1.5% (CAP) currently being used in SA. Gender, initial egg per gram (EPG) and initial live weight aided in blocking animals into groups, each group was randomly treated with one of four drug treatments comprising: the untreated control (D0), IVM, CST, and CAP. Animals grazed throughout on infested pasture. Rectal faeces were collected on days 0, 7, 14, and 21 for determining EPG. Faeces were mixed per group and incubated to identify and determine the abundance of larval forms of Haemonchus, Trichostrongylus, Strongyloides, Namatodirus, and Cooperia species. Differences between treatments changed over time. On day7 IVM, CST, and CAP depressed EPG to 0.66, 0.37 and 0.80 of their respective starting values whilst EPG increased 1.39 times for D0. Thereafter, EPG increased consistently for all drugs; CST recorded the lowest values. Haemonchus, Trichostrongylus, Strongyloides, Namatodirus and Coperia species contributed respectively 60%, 30%, 6%, 3%, and 1% of the larval forms on day 0; and 78%, 8%, 11%, 1%, 2% on day 21. Larval forms increased for Haemonchus species but decreased for Trichostrongylus species over time. Closantel was the most effective dewormer. Haemonchus Spp. were least affected whilst Trichostrongylus Spp. were the most affect by all drugs.

Keywords: anthementics, faecal egg count, L3 larvae, sheep

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Authors: Zakia Alioua, Amira Soumia, Zerouali-Khodja Fatiha

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In spite of a wide variety of contaminants such as heavy metals and organic compounds in addition to the importance of extended pollution, the deep-sea and its species are not in haven and being affected through contaminants exposure. This investigation is performed in order to provide data on the presence of pathological changes in the liver and gonads of the greater forkbeard. A total of 998 specimens of the teleost fish Phycis blennoides Brünnich, 1768 ranged from 5,7 to 62,7 cm in total length, were obtained from the commercial fisheries of Algerian ports. The sampling has been carried out monthly from December 2013 to June 2015 and from January to June 2016 caught by trawlers and longlines between 75 and 600 fathoms in the coast of Algeria. Individuals were sexed their gonads, and their livers were removed and processed for light microscopy and one case of atresia was identified. In whole, overall 0,002% of the specimens presented some degree of liver steatosis. For the gastric section, 442 selected stomachs contents were observed looking for parasitic infestation and enumerate 212 nematodes. A prospecting survey for metal contaminant was performed on the liver by atomic absorption spectrophotometry analysis.

Keywords: atresia, coast of Algeria, histopathology, nematode, Phycis blennoides, steatosis

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Authors: Seyed Hassan Moosa-kazemi, Hassan Vatandoost

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Malaria, Dirofilaria immitis (dog heart worm), and D. repens (dirofilariasis), which are transmitted by mosquitoes, have been reported in Iran. The Iranian mosquito fauna includes seven genera, 65 species, and three subspecies. Aedes albopictus has been reported since. West Nile, Sindbis, Dengue, Japanese encephalitis viruses, and the nematode Setaria (setariasis) has been reported in the country but there are no information about their vectors in Iran. Iran is malaria elimination phase. Insecticides residual spraying (IRS), distributed of insecticides long lasting treated nets (ITNs), fogging, release of larvivours fishes and Bacillus thuringiensis, chemical larviciding, as well as case finding and manipulation and modification of breeding places carried out thought the IVM program in the country. Prolonged exposure to insecticides over several generations of the vectors, develop resistance, a capacity to survive contact with insecticides. However, use of insecticides in agriculture has often been implicated as contributing to resistance in mosquito’s vectors. Resistance of mosquitoes to some insecticides has been documented just within a few years after the insecticides were introduced. Some enzymes such as monooxygenases, esterases and glutathione S-transferases have been considered as a reason for resistance to pyrethroid insecticides. In conclusion, regarding to documented resistance and tolerance of mosquitoes vectors to some insecticides, resistance management is suggested by using new insecticide with novel mode of action.

Keywords: control, Iran, resistance, vector

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Authors: Mariana Panayotova-Pencheva, Anetka Trifonova, Vassilena Dakova

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790 lungs from brown hares (Lepus europeus L.) from different regions of Bulgaria were investigated during the period 2009-2017. The parasitological status and pathomorphological features in the lungs were recorded. The following parasite species were established: one nematode - Protostrongylus tauricus (7.59% prevalence), one tapeworm – larva of Taenia pisiformis – Cysticercus pisiformis (3.04% prevalence) and one arthropod – larva of Linguatula serrata – Pentastomum dentatum (0.89% prevalence). Macroscopic lesions in the lungs were different depending on the causative agents. The infections with C. pisiformis and P. dentatum were attended with small, mainly superficial changes in the lungs. Protostrongylid infections were connected with different in appearance and burden macroscopic changes. In 77.7%, they were nodular, and in the rest of cases, they diffuse. The consistency of the lesions was compact. In most of the cases, alterations were grey in colour, rarely were dark-red or marble-like. In 91.7% of these cases, they were spread on the apical parts of large lung lobes. In 36.7% middle parts of the large lung lobes, and, in 26.7% small lung lobes, were also affected. The small lung lobes were never independently infected.

Keywords: Cysticercus pisiformis, Lepus europeus, lung lesions, Pentastomum dentatum, Protostrongylus tauricus

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Authors: Freitas Vania Moreira, Rodrigues B. B., Araujo M.B., Silva D. R., Sousa A. C., Araujo K. P., Pimentel R. R., Cares J. E., Junqueira N. T. V.

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The Pitahayas are cactus native from America and abundant in arid regions. The cultivation is based mainly on the following species: Hylocereus undatus, H. polyrhizus, H. setaceus and H. megalanthus, being H. undatus the most cultivated in Brazil. The pitahaya cultivation is recent in Brazil and is concentrated in São Paulo. Meloidogyne enterolobii is of unknown origin being distributed in several countries. This nematode has recently been detected in Brazil causing damage in several crops. Similarly, M. incognita is a widely distributed pathogen in Brazil. The objective of this study is to evaluate the following accesses of pitahaya to M. enterolobii: CPAC- Py H. hundatus 01, 02, 03, 04, 05, 06, 07 and 08; CPAC - Py H. costaricense 8A; CPAC - Py Selenicereus setaceus 17 and CPAC - Py S. megalantus 22. And the following accesses to M. incognita: CPAC- Py H. hundatus 05; CPAC - Py H. costaricense 8A; CPAC - Py S. setaceus 17 and CPAC - Py S. megalantus 22. According to the results, CPAC – Py H. hundatus 01, 02, 03, 04 and 07 were considered resistant. While CPAC - Py 05 and 08 was susceptible. CPAC-Py 06 also was considered susceptible, because there was the reaction of susceptibility in one of the trials. Given this wide diversity in H. hundatus and being this species the most cultivated in Brazil it is suggested to work more with this material in Embrapa Cerrados. CPAC - Py H. costaricense 8A behaved as susceptible in one of the trials. CPAC - Py S. setaceus 17 and CPAC – Py S. megalantus 22 were considered highly susceptible. The susceptibility of S. megalantus is widely described in the literature. In relation to M. incognita, there were differences between the results in both experiments, but all behaved as susceptible in at least one of the tests.

Keywords: pitaya, meloidogyne, fruit, resistance

Procedia PDF Downloads 182

Authors: M. Benothman, M. Stear, S. Mitchel, O. Abuargob, R. Vijayan, Sateesh Kumar

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Parasitic gastroenteritis caused by gastrointestinal nematodes (GIN) is a serious pathological complication in lambs. The dispersion pattern of GIN influences their transmission dynamics. There is no proper study on this aspect in Scottish Blackface lambs in Scotland. This study undertaken on 758 naturally infected, weaned, straight bred Scottish Blackface lambs in high land pasture in East Scotland extending over three months (August, September and October) in a year, and for three successive years demonstrated that the distribution of faecal egg counts (FEC) followed negative binomial distribution, with the exception of a few samples. The inverse index of dispersion (k) ranged between 0.19 ± 0.51 and 1.09 ± 0.08. Expression of low k values resulting from aggregation in a few individuals, suggested that a small proportion of animals with heavy parasitic influx significantly influenced the level of pasture contamination and parasite transmission. There was no discernible trend in the mean faecal egg count (FEC) and mean herbage larval population (HLP) in different months and in different years. Teladorsagia was the highest pasture contaminant (85.14±14.30 L3/kdh) followed by Nematodirus (53.00±13.96), Ostertagia (28.21±10.18) and Cooperia (11.43±5.55). The results of this study would be useful in instituting gastrointestinal nematode control strategies for sheep in cool temperate agro-ecological zones.

Keywords: blackface lamb, faecal egg count, Gastrointestinal nematodes, herbage larval population, Scotland

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Authors: Ravi Raghav Sonani, Niraj Kumar Singh, Jitendra Kumar, Datta Madamwar

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Phycobiliproteins (PBPs) are light harvesting proteins showing very strong absorbance and fluorescence in the visible range of the solar spectrum. Phycoerythrin (PE) and phycocyanin (PC) are majorly found PBPs in the cyanobacteria and red algae. In the present study, we have investigated the reactive oxygen species (ROS)-averting capacity of purified PE and PC of cyanobacterial origin. Furthermore, the possibility - whether the ROS-averting potential of PBPs can be explored in the therapeutics of oxidative stress associated physiological anomalies including aging and neurodegenerative diseases. The nematode Caenorhabditis elegans has been used as model organism in this study. PE and PC treatment moderated normal aging and associated physiological functionalities like pharyngeal pumping and locomotion of C. elegans. Moreover, PE-treatment enhanced the stress (oxidative and heat) tolerance upon PE and PC treatment. Specifically, PE treatment was also noted to moderate the progression of Alzheimer’s disease in transgenic C. elegans CL4176. However, PC-treatment curtailed the polyQ aggregation mediated proteotoxicity in C. elegans AM141 (Huntington disease model) under stressed (paraquat stress) as well as normal conditions. The effectiveness of PE and PC in expanding the lifespan of mutant C. elegans knockout for some up- (daf 16) and down- (daf-2 and age-1) stream regulators of insulin/IGF-1 signalling (IIS) shows the independency of their effects from DAF-2–AGE-1–DAF-16 signalling pathway. In conclusion, the present report demonstrates the anti-aging and neuro-protective potential of cyanobacterial PE and PC.

Keywords: phycobiliproteins, aging, alzheimer, huntington, C. elegans

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Authors: Saifullah K., Muhammad Naziruddin Saifullah, Muhammad N.

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The bio control potential and mechanism of P. chlamydosporia against Meloidogyne incognita was evaluated in the present study. Under invitro conditions, P. chlamydosporia was tested for parasitism of eggs and females of M. incognita. The results indicated that this fungus parasitized 87% eggs and 82% females. Culture filtrate (CF) of P. chlamydosporia was tested for its larvicide activity against M. incognita 2nd stage juvenile. The maximum mortality was 97.3% at 100% concentration of the culture filtrate while minimum mortality was 7.3% in 25% concentration after 24 hrs. The result of the pot experiment proved that P. chlamydosporia has reduced the incidence of RKN and improved all tested agronomic growth parameters. The treatment with inoculated M. incognita alone reduced plant height, fresh shoot, and fresh root weight by 44.7%, 29.8%, and 32.8% respectively over uninoculated healthy control. Histopathological studies on the interaction of Pochonia chlamydosporia and Meloidogyne incognita on tomato roots revealed anatomical changes among treatments. Less number of galls with small in size and scarcer abnormalities in the vascular cylinder was observed in plants inoculated with P. chlamydosporia and M. incognita than the plants treated with nematode only. The fungus was seen in in the intercellular spaces of cortical and epidermal cells while the vascular bundles of the plant remain intact, inoculated only with P. chlamydosporia. In the infected roots, many mature females were seen which feed on giant cells. The findings also revealed that control healthy plants were not affected and no histological changes were noted.

Keywords: histopathology, Pochonia chlamydosporia, Meloidogyne incognita, tomato

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Authors: Saqib Ali, Man-Qun Wang

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The Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae), is a major pest of pines and it is also the key vector of the exotic pinewood nematode in China. In the present study, we cloned, expressed, and purified a chemosensory protein (CSP) in M. alternatus. We surveyed its expression in various developmental stages of male and female adult tissues and determined its binding affinities for different pine volatiles using a competitive binding fluorescence assay. A CSP known as CSP5 in M. alternatus was obtained from an antennal cDNA library and expressed in Escherichia coli. Quantitative reverse transcription polymerase chain reaction results indicated that the CSP5 gene was mainly expressed in male and female antennae. Competitive binding assays were performed to test the binding affinity of recombinant CSP5 to 13 odour molecules of pine volatiles. The results showed that CSP5 showed very strong binding abilities to myrcene, (+)-β-pinene, and (−)-isolongifolene, whereas the volatiles 2-methoxy-4-vinylphenol, p-cymene, and (+)-limonene oxide have relatively weak binding affinity at pH 5.0. Three volatiles myrcene, (+)-β-pinene, and (−)-isolongifolene may play crucial roles in CSP5 binding with ligands, but this needs further study for confirmation. The sensitivity of insect to host plant volatiles can effectively be used to control and monitor the population through mass trapping as part of integrated pest management programs.

Keywords: olfactory-specific protein, volatiles, competitive binding assay, expression characteristics, qPCR

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Authors: Laurentius J. M. Rumokoy, Jimmy Posangi, Wisje Lusia Toar, Julio Lopez Aban

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The threat of pathogen infection agents to the neonates is a major health problem to the new born life livestock. Neonate losses became an important case in the world as well as in Indonesia. This condition can be triggered by an infection with nematode in conjunction with a failure of immunoglobulin passive transfer. The study was conducted to evaluate the role of the curcuma combined with IgG colostrum on the development of parasites in the gut of CD1 mice. Animal experiments were divided in four groups (G) based on the treatment: G1 (infection only); G2 (curcuma+infection), G3 (IgG + infection) and G4 (curcuma+IgG+infection). The parameters measured were EPG (eggs per gram) and female in the intestine. The results obtained showed that the treatment has no a significant influence on the number of eggs per gram of feces in the group infected compared to the control group without receiving IgG nor curcuma. However, the EGP response tended to decrease at day 6 in G3 and G4 with a minimum number at zero eggs. This performant showed that the immunoglobulin-G and curcuma substances could slightly decreased the number of eggs in animal infected with Strongyloides. The results obtained showed also that the treatment has no significant difference (P > 0.05) on female larva in the gut of MCD1 experimental. In other side, we found that the best performance to inhibit the female quantity in the gut was the treatment with IgG and infection of parasite in G3. In this treatment, the minimum number was five female only in the gut. The results described IgG response was better than the curcuma single use in reducing the female parasite in the gut. This positive response of IgG compared to other controls group was associated with the function of colostrum antibodies.

Keywords: parasites, livestock, curcuma, colostrums

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Authors: Carolina Nunes, Sónia Ribeiro, Hélio Faustinho, Hélia Sales, Rita Pontes, João Nunes

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Pine resin activity in Portugal was one of the most important and major non-wood forestry, representing a strategic natural resource for Portuguese Bioeconomy and an important social activity for rural regions. Pine forests representing a stock of atmospheric carbon, contributing to greenhouse effect mitigation and social and environmental important services returns. They are important sources of numerous useful products, including not only wood and cellulose but also nonwood products used by the chemical, food, and pharmaceutical industries, as well as for biorefineries. Portuguese pine forest area decreases from 1 million hectares to 400 mil hectares in the last 20 years. Portugal, in 80´s decade, was one of the world´s TOP 3 producers, with a middle annual production of 140 mil tones.year-1. With the pressure of the social desertification, forest fires, phytosanitary problems (e.g. nematode of the pine wood) and the decrease of economic value and competitivity of the Portuguese forest, the actual middle annual production is less than 10 mil tones.year-1 (lesser 92%). This significant decrease representing an annual economic loss of approximately 130-140 million Euros. year⁻¹ for forest primary sector in Portugal. The Biopinus project design new forestry approach and strategic biotechnologies knowledge to increase the economic value of Pine resin in Portugal, with an impact on the growth of the economic value of Pine resin from 1,1 to 1,5 Euros/kg.

Keywords: pine resin, bioeconomy, economic value, biotecnology

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Authors: D. S. Malatesh, K. J. Ananda, C. Ansar Kamran, K. Ganesh Udupa

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Dirofilaria repens is a mosquito-borne filarioid nematode of dogs and other carnivores and accidentally affects humans. D. repens is reported in many countries, including India. Subcutaneous dirofilariosis caused by D. repens is a zoonotic disease, widely distributed throughout Europe, Asia, and Africa, with higher prevalence reported in dogs from Sri Lanka (30-60%), Iran (61%) and Italy (21-25%). Dirofilariasis in dogs was diagnosed by detection of microfilariae in blood. Identification of different Dirofilaria species was done by using molecular methods like polymerase chain reaction (PCR). Even though many researchers reported molecular evidence of D. repens across India, to our best knowledge there is no data available on molecular diagnosis of D. repens in dogs and its zoonotic implication in Karnataka state a southern state in India. The aim of the present study was to identify the Dirofilaria species occurring in dogs from Karnataka, India. Out of 310 samples screened for the presence of microfilariae using traditional diagnostic methods, 99 (31.93%) were positive for the presence of microfilariae. Based on the morphometry, the microfilariae were identified as D. repens. For confirmation of species, the samples were subjected to PCR using pan filarial primers (DIDR-F1, DIDR-R1) for amplification of internal transcribed spacer region 2 (ITS2) of the ribosomal DNA. The PCR product of 484 base pairs on agarose gel was indicative of D. repens. Hence, a single PCR reaction using pan filarial primers can be used to differentiate filarial species found in dogs. The present study confirms that dirofilarial species occurring in dogs from Karnataka is D. repens and further sequencing studies are needed for genotypic characterization of D. repens.

Keywords: Dirofilaria repens, molecular characterization, polymerase chain reaction, Karnataka, India

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Authors: Dingiswayo Likhona, Arko-Cobbah Emmanuel, Carolina Pohl, Nthabiseng Z. Mokoena, Jolly Musoke

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A distinct strain of Klebsiella pneumoniae (K. pneumoniae), referred to as hypervirulent (hvKp), is associated with invasive infections such as an invasive pyogenic liver abscess in young and healthy individuals. In South Africa, limited information is known about the prevalence and virulence of this hvKp strain. Thus, this study aimed to determine the prevalence of hvKp and virulence-associated factors in K. pneumoniae isolates from one of the largest Tertiary hospitals in a South African province. A total of 74 K. pneumoniae isolates were received from Pelonomi National Health Laboratory Services (NHLS), Bloemfontein. Virulence-associated genes (rmpA, capsule serotype K1/K2, iroB, and irp2) were screened, and the virulence of hvKp vs. classical Klebsiella pneumoniae (cKp) was investigated using Caenorhabditis elegans nematode model. The iutA (aerobactin transporter) gene was used as a primary biomarker of hvKp. An average of 12% (9/74) of cases were defined as hvKp. Moreover, hvKp was found to be significantly more virulent in vivo Caenorhabditis elegans relative to cKp. The virulence-associated genes (rmpA, iroB, hmv phenotype, and capsule K1/K2) were significantly (p< 0.05) associated with hvKp. Findings from this study confirm the presence of hvKp in one large Tertiary hospital in South Africa. However, the low prevalence and mild to moderate clinical presentation suggest a marginal threat to public health. Further studies in different settings are required to establish the true potential impact of hvKp in developing countries.

Keywords: hypervirulent klebsiella pneumoniae, virulence, caenorhabditis elegans, aerobactin (iutA)

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Authors: Jose D. Zapata-Torres, Esteban Naranjo-Gutiérrez, Angela M. Martínez-Valencia, Jenny J. Chaparro-Gutiérrez, David Villar-Argaiz

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Reports of drug resistance have been made in every livestock host and to every anthelmintic class. In some regions of world, the extremely high prevalence of multi-drug resistance in nematodes of sheep and goats threatens the viability of small-ruminant industries. In the region of Antioquia, Colombia, no reports of nematode resistance have been documented due to a lack of veterinary diagnostic laboratories. The objective of this study was to evaluate the efficacy of albendazole, fenbendazole, and levamisole to control gastrointestinal nematodes in goat farms of Antioquia by doing fecal egg count reduction tests. A total of 139 crossbreed goats from four separate farms were sampled for feces prior to, and 14 days following anthelmintc treatments. Individual fecal egg counts were performed using the modified three chamber McMaster technique. The anthelmintics administered at day 0 were albendazole (farm 1, n=63), fenbendazole (farm 2, n=20), and levamisole (farm 3 and 4, n= 37, and 19). Larval cultures were used to identify the genus of nematodes using Baermann`s technique and the morphological keys for identification of L3 in small ruminants. There was no difference in fecal egg counts between 0 and 14, with means (±SD) of 1681,5 ± 2121,5 and 1715,12 ± 1895,4 epg (eggs per gram), respectively. The egg count reductions for each anthelmintic and farm were 25,86% for albendazole (farm 1), 0% for fenbendazole (farm 2), 0% (farm 3), and 5,5% (farm 4) for levamisole. The genus of nematodes identified was predominantly Haemonchus spp., with 70,27% and 82,81% for samples from day 0 and 14, respectively. These results provide evidence of a total state of resistance to 3 common anthelmintics. Further research is needed to design integrate management programs to control nematodes in small ruminants in Colombia.

Keywords: anthelmintics, goat, haemonchus, resistance

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Authors: Maryam Monazzah, Ronak Samadpour, Mehdi Nasr-esfahani, Fatemeh Qalavand, Marziye Motamedi

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Bipolaris sorokiniana, and Heterodera filipjevi, are important wheat diseases that lead to yield losses worldwide. Identifying novel resistant sources helps us combat these devastating diseases. In this study, we studied the role of Cre3 gene and antioxidant enzymes in the immune responses of wheat genotypes to H. filipjevi and B. sorokiniana. Therefore, real-time PCR analysis using Cre3 gene marker, a resistant gene to cereal cyst nematodes, was conducted on leaves and roots, along with changes ‎in the activity of antioxidant enzymes, peroxidase, and catalase. Enzyme activity assay was performed on roots attacked by nematode and in leaves infected with Bipolaris. Wheat accessions including “Bam” (resistant), “Parsi” (moderately-resistant), “Azar2”, “Ohadi”, “Homa” (highly-susceptible) were previously screened against both stresses under greenhouse and field conditions. Results showed that Cre3 expression against cyst nematodes was significantly higher in resistant cultivars compared to susceptible cultivars. Cre3 was used in marker-assisted selection programs to identify genotypes carrying resistant genes to cyst nematodes. Interestingly, Cre3 was also up-regulated in both tissues of resistant cultivars to B. sorokiniana. Therefore, Cre3 in wheat similarly modulates immunity against B. sorokiniana and might be one of the central components of the induced immune system in wheat. The activity of antioxidant enzymes also indicated the highest increase in resistant genotypes upon both stresses that subsequently neutralize oxidative stress in tissues and decrease damage. Further studies on these resistance components may help us gain insight into the molecular basis of resistance and shed new light on the interaction and overlap between different forms of stress.

Keywords: Bipolaris sorokiniana, Heterodera filipjevi, resistant gene expression, wheat

Procedia PDF Downloads 95

Authors: M. Manzoor, J. N. Ahmad, R. M. Giblin Davis, N. Javed, M. S. Haider

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The invasive Red Palm Weevil (RPW) (Rhynchophorus ferrugineus [Olivier] (Coleoptera: Curculionidae) is one of the most destructive palm pests in the world. Synthetic pesticides are environmentally hazardous pest control strategies being used in the past with emerging need of eco-friendly biological approaches including microbial entomopathogens for RPW management. The sublethal effects of a single entomopathogenic fungus (EPF) Beauveria bassiana (WG-11) (Ascomycota: Hypocreales) and two entomopathogenic nematode (EPN) species Heterorhabditis bacteriophora (Poinar) and Steinernema carpocapsae (Weiser) (Nematoda: Rhabditida) were evaluated in various combinations against laboratory-reared 3rd, 5th and 8th instar larvae of RPW in laboratory assays. Individual and combined effects of both entomopathogens (EP) were observed after the pre-application of B. bassiana fungus at 1-2-week intervals. A number of parameters were measured after the application of sub-lethal doses of EPF such as diet consumption, development, frass production, mortality, and weight gain. Combined treatments were tested for additive and synergistic effects. Synergism was more frequently observed in B. bassiana and S. carpocapsae combined treatments than in B. bassiana and H. bacteriophora combinations. Early instar larvae of RPW were more susceptible than older instars. Synergistic effects were observed in the 3rd and 5th instars exposed to B. bassiana and S. carpocapsae at 0, 7 and 14-day intervals. Whereas, in 8th instar larvae, the synergistic effect was observed only in B. bassiana and S. carpocapsae treatments after 0 and 7 days intervals. EPN treatments decreased pupation, egg hatching and emergence of adults. Lethal effects of nematodes were also observed in all growth stages of R. ferrugineus. Reduced larval weight, increased larval, pre-pupal and pupal duration, reduced adult weight and life span were observed. Sub-lethal concentrations of both entomopathogens induced variations in the different developmental stages and reduced food consumption, frass production, growth, and weight gain. So, on the basis of results, it is concluded that synthetic pesticides should be replaced with environmentally friendly sustainable biopesticides.

Keywords: H. bacteriophora, S. carpocapsae, B. bassiana, mortality

Procedia PDF Downloads 169

Authors: Valeria Dilcheva, Svetlozara Petkova, Ivelin Vladov

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Nemathodes of the genus Trichinella are zoonotic parasites with a cosmopolitan distribution. More than 100 species of mammals, birds and reptiles are involved in the natural cycle of this nematode. At present, T. spiralis, T. pseudospiralis, and T. britovi have been found in Bulgaria. The existence of natural wildlife and domestic reservoirs of Trichinella spp. can be a serious threat to human health. Three trichinella isolates caused human trichinella infection outbreaks from three regions of Sofia City Province were used for the research: sample No. 1 - Ratus norvegicus, sample No. 2 – domestic pig (Sus scrofa domestica), sample No. 3 - domestic pig (Sus scrofa domestica). Trichinella larvae of the studied species were isolated via digestive method (pepsin, hydrochloric acid, water) at 37ºC by standard procedure and were determined by gender (male and female) based on their morphological characteristics. As a reference trichinella species were used: T. spiralis, T. pseudospiralis, T. nativa and T. britovi. Single male and female larvae of the three isolates were crossed with single male and female larvae of the reference trichinella species as well as reciprocally. As a result of cross-breeding, offspring of muscular larvae with T. spiralis and T. britovi were obtained, while in experiments with T. pseudospiralis and T. nativa, trichinella larvae were not found in the laboratory mice. The results obtained in the control groups indicate that the trichinella larvae used from the isolates and the four trichinella species are infective. Also, the infective ability of the F1 offspring from the successful cross-breeding between isolates and reference species was investigated. Through the data obtained in the experiment was found that isolates No. 1 and No. 2 belong to the species T. spiralis, and isolate No. 3 belongs to the species T. britovi. The results were confirmed by PCR and real-time PCR analysis. Thus the presence and circulation of the species T. spiralis and T. britovi in Bulgaria was confirmed. Probably the rodents (rats) are involved in the distribution of T. spiralis in urban environment. The species T. britovi found in a domestic pig speaks of some contact with wild animals for which T. britovi is characteristic. The probable reason is that a large number of farmers in Bulgaria practice the free-range breeding of domestic pigs. Part of the farmers also used as food for domestic pigs waste products from the game (foxes, jackals, bears, wolves) and probably thus the infection was obtained. The distribution range of trichinella species in Bulgaria is not strictly outlined. It is believed that T. spiralis is most common in domestic animals and T. britovi and T. pseudospiralis are characteristic of wildlife. To answer the question whether wild and synanthropic animals are infected with the same or different trichinella species, which species predominate in nature and what their distribution among different hosts is, further research is required.

Keywords: cross-breeding, Sofia, trichinellosis, Trichinella britovi, Trichinella spiralis

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Authors: Dali Gaganidze, Ekaterine Abashidze

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Potato is one of the main agricultural crops in Georgia. Georgia produces early and late potato varieties in almost all regions. In traditional potato growing regions (Svaneti, Samckhet javaheti and Tsalka), the yield is higher than 30-35 t/ha. Among the plant pests that limit potato production and quality, the potato cyst nematodes (PCN) are harmful around the world. Yield losses caused by PCN are estimated up to 30%. Rout surveys conducted in two geographically distinct regions of Georgia producing potatoes - Samtskhe - Javakheti and Svaneti revealed potato cyst nematode Globodera rostochiensi. The aim of the study was the Phylogenetic analyses of Globodera rostochiensi revealed in Georgia by the amplification and sequencing of 28S gen in the D3 region and intergenic ITS1-15.8S-ITS2 region. Identification of all the samples from the two Globodera populations (Samtskhe - Javakheti and Svaneti), i.e., G. rostochiensis (20 isolates) were confirmed by conventional multiplex PCR with ITS 5 universal and PITSp4, PITSr3 specific primers of the cyst nematodes’ (G. pallida, G. rostochiensis). The size of PCR fragment 434 bp confirms that PCN samples from two populations, Samtskhe- Javakheti and Svaneti, belong to G. rostochiensi . The ITS1–5.8S-ITS2 regions were amplified using prime pairs: rDNA1 ( 5’ -TTGATTACGTCCCTGCCCTTT-3’ and rDNA2( 5’ TTTCACTCGCCGTTACTAAGG-3’), D3 expansion regions were amplified using primer pairs: D3A (5’ GACCCCTCTTGAAACACGGA-3’) and D3B (5’-TCGGAAGGAACCAGCTACTA-3’. PCR products of each region were cleaned up and sequenced using an ABI 3500xL Genetic Analyzer. Obtained sequencing results were analyzed by computer program BLASTN (https://blast.ncbi.nlm.nih.gov/Blast.cg). Phylogenetic analyses to resolve the relationships between the isolates were conducted in MEGA7 using both distance- and character-based methods. Based on analysis of G.rostochiensis isolate`s D3 expansion regions are grouped in three major clades (A, B and C) on the phylogenetic tree. Clade A is divided into three subclades; clade C is divided into two subclades. Isolates from the Samtckhet-javakheti population are in subclade 1 of clade A and isolates in subclade 1 of clade C. Isolates) from Svaneti populations are in subclade 2 of clade A and in clad B. In Clade C, subclade two is presented by three isolates from Svaneti and by one isolate (GL17) from Samckhet-Javakheti. . Based on analysis of G.rostochiensis isolate`s ITS1–5.8S-ITS2 regions are grouped in two main clades, the first contained 20 Georgian isolates of Globodera rostochiensis from Svaneti . The second clade contained 15 isolates of Globodera rostochiensis from Samckhet javakheti. Our investigation showed of high genetic variation of D3 and ITS1–5.8S-ITS2 region of rDNA of the isolates of G. rostochiensis from different geographic origins (Svameti, Samckhet-Javakheti) of Georgia. Acknowledgement: The research has been supported by the Shota Rustaveli National Scientific Foundation of Georgia : Project # FR17_235

Keywords: globodera rostochiensi, PCR, phylogenetic tree, sequencing

Procedia PDF Downloads 195

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

Procedia PDF Downloads 714

Authors: Getahun Lemma

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Introduction Guinea Worm Disease (GWD), also known as dracunculiasisis, is one of the oldest diseases in the history of mankind. Dracunculiasis is caused by a parasitic nematode, Dracunculus medinensis. Infection is acquired by drinking contaminated water with copepods containing infective Guinea Worm (GW) larvae). Almost one year after the infection, the worm usually emerges out through the skin on a lower, causing severe pain and disabilities. Although there is no effective drug or vaccine against the disease, the chain of transmission can be effectively prevented with simple and cost effective public health measures. Death due to dracunculiasis is very rare. However, it results in a wide range of physical, social and economic sequels. The disease is usually common in the rural, remote places of Sub-Saharan African countries among the marginalized societies. Currently, GWD is one of the neglected tropical diseases, which is on the verge of eradication. The global Guinea Worm Eradication Program (GWEP) was started in 1980. Since then, the program has achieved a tremendous success in reducing the global burden and number of GW case from 3.5 million to only 28 human cases at the end of 2018. However, it has recently been shown that not only humans can become infected, with a total of 1,105 animal infections have been reported at the end of 2018. Therefore, the objective of this study was to identify the existing challenges in the last mile of the GWEP in order To inform Policy makers and stakeholders on potential measures to finally achieve eradication. Method Systematic literature review on articles published from January 1, 2000 until May 30, 2019. Papers listed in Cochrane Library, Google Scholar, ProQuest PubMed and Web of Science databases were searched and reviewed. Results Twenty-five articles met inclusion criteria of the study and were selected for analysis. Hence, relevant data were extracted, grouped and descriptively analyzed. Results showed the main challenges complicating the last mile of global GWEP: 1. Unusual mode of transmission; 2. Rising animal Guinea Worm infection; 3. Suboptimal surveillance; 4. Insecurity; 5. Inaccessibility; 6. Inadequate safe water points; 7. Migration; 8. Poor case containment measures, 9. Ecological changes; and 10. New geographic foci of the disease. Conclusion This systematic review identified that most of the current challenges in the GWEP have been present since the start of the campaign. However, the recent change in epidemiological patterns and nature of GWD in the last remaining endemic countries illustrates a new twist in the global GWEP. Considering the complex nature of the current challenges, there seems to be a need for a more coordinated and multidisciplinary approach of GWD prevention and control measures in the last mile of the campaign. These new strategies would help to make history by eradicating dracunculiasis as the first ever parasitic disease.

Keywords: dracunculiasis, eradication program, guinea worm, last mile

Procedia PDF Downloads 132

Authors: Shiva Mardani, Mehdi Nasr-Esfahani, Majid Olia, Hamid Molahosseini, Hamed Hassanzadeh Khankahdani

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Plant-parasitic nematodes cause serious diseases on plants and effectively reduce food production in quality and quantity worldwide, with at least 17 nematode species in the three important and major genera, including Meloidogyne, Heterodera, and Pratylenchus. Root-knot nematodes (RKN), Meloidogyne spp. with the dominant species, Meloidogynejavanica, are considered as the important plant pathogens of agricultural products globally. The hosts range can be vegetables, bedding plants, grasses, shrubs, numerous weeds, and trees, including forests. In this study, chemical management was carried out on RKN, M. javanica, to investigate the efficacy of Iranian Abamectin insecticide product [acaricide Abamectin (Vermectin® 2% EC, Gyah Corp., Iran)] verses imported normal Abamectin available in the Iran markets [acaricide Abamectin (Vermectin® 1.8% EC, Cropstar Chemical Industry Co., Ltd.)] each of which at the rate of 8 L./ha, on Tomatoes, Solanumlycopersicum L., (No. 29-41, Dutch company Siemens) as a test plant, and the controls (infested to RKN and without any chemical pesticides treatments); and (sterile soil without any RKN and chemical pesticides treatments) at the greenhouse in Isfahan, Iran. The trails were repeated thrice. The results indicated a highly significant reduction in RKN population and an increase in biomass parameters at 1% level of significance, respectively. Relatively similar results were obtained in all the three experiments conducted on tomato root-knot nematodes. The treatments of Gyah-Abamectin (51.6%) and external Abamectin (40.4%) had the highest to least effect on reducing the number of larvae in the soil compared to the infected controls, respectively. Gyah-Abamectin by 44.1% and then external one by 31.9% had the highest effect on reducing the number of larvae and eggs in the root and 31.4% and 24.1% reduction in the number of galls compared to the infected controls, respectively. Based on priority, Gyah-Abamectin (47.4 % ) and external Abamectin (31.1 %) treatments had the highest effect on reducing the number of egg- masses in the root compared to the infected controls, with no significant difference between Gyah-Abamectin and external Abamectin. The highest reproduction of larvae and egg in the root was observed in the infected controls (75.5%) and the lowest in the healthy controls (0.0%). The highest reduction in the larval and egg reproduction in the roots compared to the infected controls was observed in Gyah-Abamectin and the lowest in the external one. Based on preference, Gyah-Abamectin (37.6%) and external Abamectin (26.9%) had the highest effect on the reduction of the larvae and egg reproduction in the root compared to the infected controls, respectively. Regarding growth parameters factors, the lowest stem length was observed in external Abamectin (51.9 cm), with nosignificantly different from Gyah-Abamectin and healthy controls. The highest root fresh weight was recorded in the infected controls (19.81 gr.) and the lowest in the healthy ones (9.81 gr.); the highest root length in the healthy controls (22.4 cm), and the lowest in the infected controls and external Abamectin (12.6 and 11.9 cm), respectively. Conclusively, the results of these three tests on tomato plants revealed that Gyah-Abamectin 2% compared to external Abamectin 1.8% is competitive in the chemical management of the root nematodes of these types of products and is a suitable alternative in this regard.

Keywords: solanum lycopersicum, vermectin, biomass, tomato

Procedia PDF Downloads 96

Authors: Catarina Ferreira Rebelo, Luis Madeira de Carvalho, Fernando González González

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The order Passeridae corresponds to the richest and most abundant group of birds, with approximately 6500 species, making it possible to assert that two out of every three bird species are passerines. They are globally distributed and exhibit remarkable morphological and ecological variability. While numerous species of parasites have been identified and described in wild birds, there has been little focus on passeriformes. Seventeen passerines admitted to GREFA, a Wildlife Rehabilitation Center, throughout the months of October, November and December 2022 were analyzed. The species included Aegithalos caudatus, Anthus pratensis, Carduelis chloris, Certhia brachydactyla, Erithacus rubecula, Fringilla coelebs, Parus ater, Passer domesticus, Sturnus unicolor, Sylvia atricapilla, Turdus merula and Turdus philomelos. Data regarding past history was collected, and necropsies were conducted to identify the cause of death and body condition and determine the presence of parasites. Additionally, samples of intestinal content were collected for direct/fecal smear, flotation and sedimentation techniques. Sixteen (94.1%) passerines were considered positive for the presence of parasitic forms in at least one of the techniques used, including parasites detected in necropsy. Adult specimens of both sexes and tritonymphs of Monojoubertia microhylla and ectoparasites of the genus Ornithonyssus were identified. Macroscopic adult endoparasitic forms were also found during necropsies, including Diplotriaena sp., Serratospiculum sp. and Porrocaecum sp.. Parasitism by coccidia was observed with no sporulation. Additionally, eggs of nematodes from various genera were detected, such as Diplotriaena sp., Capillaria sp., Porrocaecum sp., Syngamus sp. and Strongyloides sp., eggs of trematodes, specifically the genus Brachylecithum and cestode oncospheres, whose genera were not identified. To our knowledge, the respiratory nematode Serratospiculum sp. found in this study is being reported for the first time in passerines in the Iberian Peninsula, along with the application of common coprological techniques for the identification of eggs in the intestinal content. The majority of parasites identified utilize intermediary hosts present in the diet of the passerines sampled. Furthermore, the discovery of certain parasites with a direct life cycle could potentially exert greater influence, particularly in specific scenarios such as within nests or during the rehabilitation process in wildlife centers. These parasites may impact intraspecific competition, increase susceptibility to predators or lead to death. However, their cost to wild birds is often not clear, as individuals can endure various parasites without significant harm. Furthermore, wild birds serve as important sources of parasites across different animal groups, including humans and other mammals. This study provides invaluable insights into the parasitic fauna of these birds, not only serving as a cornerstone for future epidemiological investigations but also enhancing our comprehension of these avian species.

Keywords: birds, parasites, passerines, wild, spain

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