Search results for: subspecies
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 28

Search results for: subspecies

28 Morphometrics Study of Apis florea and Apis mellifera from Different Locations in Sudan

Authors: Mohammed M. Ibrahim, A. A. Yusuf, Manuel Du, Fiona Mumoki

Abstract:

The traditional honey bee species of Sudan is Apis mellifera, but in 1985, the dwarf bee Apis florea was introduced to the country, so now there are two species present. However, there are conflicting assessments regarding the subspecies of Apis mellifera colonies in Sudan. Likewise, it is unclear if, in the 40 years since its introduction, Apis florea has already developed regional differences or ecotypes. To shed light on these questions, we performed a morphology study on Sudanese honeybees. Samples of 10 to 20 honeybee workers per colony of the two species were collected from 16 locations, spanning different climatic zones in Sudan during 2021. Measurements were taken from 16 morphometric characteristics using a stereo-microscope equipped with an Image Analysis System (Moticam Image Plus 5.0 Digital Microscope Camera) to study their variability. The results indicate that in both species, the general means of various characters showed significant differences (p < 0.05) within a species between different locations, indicating that there might indeed be regional differences. However, more taxonomic investigation and, ideally also, molecular studies are needed in order to confirm the proper identification of subspecies and their ecotypes.

Keywords: Apis, subspecies, morphology, Sudan

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27 An Atlantic Canadian Case of Disseminated Streptococcus equi Subspecies zooepidemicus Infection

Authors: Albert Chang, Duncan Webster

Abstract:

Streptococcus equi subspecies zooepidemicus infections in humans can be contracted through contact with domestic animals or unpasteurized dairy products. Although infection in humans is rare, the course can be fulminant. We describe the case of a 75-year-old, immunocompetent male, who developed disseminated disease with bacteremia, native aortic valve endocarditis, suppurative pericarditis with cardiac tamponade, meningitis and bilateral endopthalmitis. Despite treatment with pericardial drain placement, intravenous ceftriaxone and rifampin the patient, unfortunately, did not survive. To date, reported cases of disseminated infection by S. zooepidemicus are few. Furthermore, with the review of the literature, this case demonstrates the broadest organ system involvement reported. Of interest, previous studies have suggested an affinity of this organism for certain organ systems and this case corroborates an emerging association of S. zooepidemicus with endopthalmitis. In addition, this is the second Canadian case of documented human infection with both cases being similar in clinical features, presentation, and geographical location. A discussion regarding previous S. zooepidemicus outbreaks and the potential for zoonotic outbreaks to occur is included. In short, this case report should serve to warn clinicians regarding complications and sites of haematogenous seeding in the setting of disseminated S. zooepidemicus infections.

Keywords: endopthalmitis, endocarditis, meningitis, Streptococcus equi subspecies zooepidemicus

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26 Molecular Evidence for Three Species of Giraffa

Authors: Alice Petzold, Alexandre Hassanin

Abstract:

The number of giraffe species has been in focus of interest since the exploration of sub-Saharan Africa by European naturalists during the 18th and 19th centuries, as previous taxonomists, like Geoffroy Saint-Hilaire, Richard Owen or William Edward de Winton, recognized two or three species of Giraffa. For the last decades, giraffes were commonly considered as a single species subdivided into nine subspecies. In this study, we have re-examined available nuclear and mitochondrial data. Our genetic admixture analyses of seven introns support three species: G. camelopardalis (i.e., northern giraffes including reticulated giraffes), G. giraffa (southern giraffe) and G. tippelskirchi (Masai giraffe). However, the nuclear alignments show small variation and our phylogenetic analyses provide high support only for the monophyly of G. camelopardalis. Comparisons with the mitochondrial tree revealed a robust conflict for the position and monophyly of G. giraffa and G. tippelskirchi, which is explained firstly by a mitochondrial introgression from Masai giraffe to southeastern giraffe, and secondly, by gene flow mediated by male dispersal between southern populations (subspecies angolensis and giraffa). We conclude that current data gives only moderate support for three giraffe species and point out that additional nuclear data need to be studied to revise giraffe taxonomy.

Keywords: autosomal markers, Giraffidae, mitochondrial introgression, taxonomy

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25 Re-Introduction of the Red-Necked Ostrich (Struthio camelus camelus) in Fenced Protected Area of Central Semi-Arid Area in Saudi Arabia

Authors: M. Zafar-ul Islam

Abstract:

The Arabian Ostrich Struthio camelus syriacus is a distinct subspecies that became extinct in the wild during the mid-20th century, due to over-hunting and commercial exploitation. The extant of S. c. camelus, a red-necked form that occurs in Northeastern Africa and is considered the most closely related, and possibly the same subspecies as the extinct Arabian form has been chosen for the reintroduction in 1988-89 by obtaining red-necked ostrich from Sudan from a private collection. Few birds were translocated to Mahazat as-Sayd protected area in 25 ha fenced enclosure in 1994. Until now a total of 96 red-necked ostrich have been released in fenced Mahazat as-Sayd, and the estimated population is between 125 to 150 individuals. Since captive flock of ostriches were translocated to Mahazat, their survival rate increased (>41%) by the end of 2000. On an average 22-30 chicks are hatched annually. A total of 137 ostriches recorded dead over the period of 13 years during the drought period. One of the key questions is what proportion of birds makes use of the supplementary food and water provisions, and what happens to those birds that do not use it? Captive-bred and wild-born adult and young ostriches died of starvation and thirst, despite being provisioned with alfalfa and water in several years. The present population of ostrich in Mahazat is more than 300.

Keywords: red-necked ostrich, Struthio camelus camelus, reintroduction, Saudi Arabia, drought

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24 Complete Chloroplast DNA Sequences of Georgian Endemic Polyploid Wheats

Authors: M. Gogniashvili, I. Maisaia, A. Kotorashvili, N. Kotaria, T. Beridze

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Three types of plasmon (A, B and G) is typical for genus Triticum. In polyploid species - Triticum turgidum L. and Triticum aestivum L. plasmon B is detected. In the forthcoming paper, complete nucleotide sequence of chloroplast DNA of 11 representatives of Georgian wheat polyploid species, carrying plasmon B was determined. Sequencing of chloroplast DNA was performed on an Illumina MiSeq platform. Chloroplast DNA molecules were assembled using the SOAPdenovo computer program. All contigs were aligned to the reference chloroplast genome sequence using BLASTN. For detection of SNPs and Indels and phylogeny tree construction computer programs Mafft and Blast were used. Using Triticum aestivum L. subsp. macha (Dekapr. & Menabde) Mackey var. paleocolchicum Dekapr. et Menabde as a reference, 5 SNPs can be identified in chloroplast DNA of Georgian endemic polyploid wheat. The number of noncoding substitutions is 2, coding substitutions - 3. In comparison with reference DNA two - 38 bp and 56 bp inversions were observed in paleocolchicum subspecies. There were six 1 bp indels detected in Georgian polyploid wheats, all of them at microsatellite stretches. The phylogeny tree shows that subspecies macha, carthlicum and paleocolchicum occupy different positions. According to the simplified scheme based on SNP and indel data, the ancestral, female parent of the all studied polyploid wheat is unknown X predecesor, from which four lines were formed. 1 SNP and two inversions (38 bp and 56 bp) caused the formation of subsp. paleocolchicum. Three other lines are macha, durum and carthlicum lines. Macha line is further divided into two sublines (M_1 and M_4). Carthlicum line includes subsp.carthlicum and T.aestivum - C_1 - C_2 - A_1. One of the central question of wheat domestication is which people(s) participated in wheat domestication? It is proposed that the predecessors of Georgian peoples (Proto-Kartvelians) must be placed, on the evidence of archaic lexical and toponymic data, in the mountainous regions of the western and central part of the Little Caucasus (the Transcaucasian foothills) at least 4,000 years ago. One of the possibility to explain the ‘wheat puzzle’ is that Kartvelian speakers brought domesticated wheat species and subspecis from Fertile Crescent further north to South Caucasus.

Keywords: chloroplast DNA, sequencing, SNP, triticum

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23 Effects of a Dwarfing Gene sd1-d (Dee-Geo-Woo-Gen Dwarf) on Yield and Related Traits in Rice: Preliminary Report

Authors: M. Bhattarai, B. B. Rana, M. Kamimukai, I. Takamure, T. Kawano, M. Murai

Abstract:

The sd1-d allele at the sd1 locus on chromosome 1, originating from Taiwanese variety Dee-geo-woo-gen, has been playing important role for developing short-culm and lodging-resistant indica varieties such as IR36 in rice. The dominant allele SD1 for long culm at the locus is differentiated into SD1-in and SD1-ja which are harbored in indica and japonica subspecies’s, respectively. The sd1-d of an indica variety IR36 was substituted with SD1-in or SD1-ja by recurrent backcrosses of 17 times with IR36, and two isogenic tall lines regarding the respective dominant alleles were developed by using an indica variety IR5867 and a japonica one ‘Koshihikari’ as donors, which were denoted by '5867-36' and 'Koshi-36', respectively. The present study was conducted to examine the effect of sd1-d on yield and related traits as compared with SD1-in and SD1-ja, by using the two isogenic tall lines. Seedlings of IR36 and the two isogenic lines were transplanted on an experimental field of Kochi University, by the planting distance of 30 cm × 15 cm with two seedlings per hill, on May 3, 2017. Chemical fertilizers were supplied by basal application and top-dressing at a rate of 8.00, 6.57 and 7.52 g/m², respectively, for N, P₂O₅ and K₂O in total. Yield, yield components, and other traits were measured. Culm length (cm) was in the order of 5867-36 (101.9) > Koshi-36 (80.1) > IR36 (60.0), where '>' indicates statistically significant difference at the 5% level. Accordingly, sd1-d reduced culm by 41.9 and 20.1 cm, compared with SD1-in and SD1-ja, respectively, and the effect of elongating culm was higher in the former allele than in the latter one. Total brown rice yield (g/m²), including unripened grains, was in the order of IR36 (611) ≧ 5867-36 (586) ≧ Koshi-36 (572), indicating non-significant differences among them. Yield-1.5mm sieve (g/m²) was in the order of IR36 (596) ≧ 5867-36 (575) ≧ Koshi-36 (558). Spikelet number per panicle was in the order of 5867-36 (89.2) ≧ IR36 (84.7) ≧ Koshi-36 (79.8), and 5867-36 > Koshi-36. Panicle number per m² was in the order of IR36 (428) ≧ Koshi-36 (403) ≧ 5867-36 (353), and IR36 > 5867-36, suggesting that sd1-d increased number of panicles compared with SD1-in. Ripened-grain percentage-1.5mm sieve was in the order of Koshi-36 (86.0) ≧ 5867-36 (85.0) ≧ IR36 (82.7), and Koshi-36 > IR36. Thousand brown-rice-grain weight-1.5mm sieve (g) was in the order of 5867-36 (21.5) > Koshi-36 (20.2) ≧ IR36 (19.9). Total dry weight at maturity (g/m²) was in the order of 5867-36 (1404 ) ≧ IR36 (1310) ≧ Kosihi-36 (1290). Harvest index of total brown rice (%) was in the order of IR36 (39.6) > Koshi-36 (37.7) > 5867-36 (35.5). Hence, sd1-d did not exert significant effect on yield in indica genetic background. However, lodging was observed from the late stage of maturity in 5867-36 and Koshi-36, particularly in the former, which was principally due to their long culms. Consequently, sd1-d enables higher yield with higher fertilizer application, by enhancing lodging resistance, particularly in indica subspecies.

Keywords: rice, dwarfing gene, sd1-d, SD1-in, SD1-ja, yield

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22 Investigation on Single Nucleotide Polymorphism in Candidate Genes and Their Association with Occurrence of Mycobacterium avium Subspecies Paratuberculosis Infection in Cattle

Authors: Ran Vir Singh, Anuj Chauhan, Subhodh Kumar, Rajesh Rathore, Satish Kumar, B Gopi, Sushil Kumar, Tarun Kumar, Ramji Yadav, Donna Phangchopi, Shoor Vir Singh

Abstract:

Paratuberculosis caused by Mycobacterium avium subspecies paratuberculosis (MAP) is a chronic granulomatous enteritis affecting ruminants. It is responsible for significant economic losses in livestock industry worldwide. This organism is also of public health concern due to an unconfirmed link to Crohn’s disease. Susceptibility to paratuberculosis has been suggested to have genetic component with low to moderate heritability. Number of SNPs in various candidates genes have been observed to be affecting the susceptibility toward paratuberculosis. The objective of this study was to explore the association of various SNPs in the candidate genes and QTL region with MAP. A total of 117 SNPs from SLC11A1, IFNG, CARD15, TLR2, TLR4, CLEC7A, CD209, SP110, ANKARA2, PGLYRP1 and one QTL were selected for study. A total of 1222 cattle from various organized herds, gauhsalas and farmer herds were screened for MAP infection by Johnin intradermal skin test, AGID, serum ELISA, fecal microscopy, fecal culture and IS900 blood PCR. Based on the results of these tests, a case and control population of 200 and 183 respectively was established for study. A total of 117 SNPs from 10 candidate genes and one QTL were selected and validated/tested in our case and control population by PCR-RFLP technique. Data was analyzed using SAS 9.3 software. Statistical analysis revealed that, 107 out of 117 SNPs were not significantly associated with occurrence of MAP. Only SNP rs55617172 of TLR2, rs8193046 and rs8193060 of TLR4, rs110353594 and rs41654445 of CLEC7A, rs208814257of CD209, rs41933863 of ANKRA2, two loci {SLC11A1(53C/G)} and {IFNG (185 G/r) } and SNP rs41945014 in QTL region was significantly associated with MAP. Six SNP from 10 significant SNPs viz., rs110353594 and rs41654445 from CLEC7A, rs8193046 and rs8193060 from TLR4, rs109453173 from SLC11A1 rs208814257 from CD209 were validated in new case and control population. Out of these only one SNP rs8193046 of TLR4 gene was found significantly associated with occurrence of MAP in cattle. ODD ratio indicates that animals with AG genotype were more susceptible to MAP and this finding is in accordance with the earlier report. Hence it reaffirms that AG genotype can serve as a reliable genetic marker for indentifying more susceptible cattle in future selection against MAP infection in cattle.

Keywords: SNP, candidate genes, paratuberculosis, cattle

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21 Both Floristic Studies and Molecular Markers Are Necessary to Study of the Flora of a Region

Authors: Somayeh Akrami, Vali-Allah Mozaffarian, Habib Onsori

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The studied region in this research, watershed Kuhkamar river, is about 112.66 square kilometers, it is located between 45º 48' 9" to 45º 2' 20" N and 38º 34' 15" to 38º 40' 28" E. The gained results of the studies on flora combinations, proved 287 plant species in 190 genera and 51 families. Asteracea with 49 and Lamiaceae with 27 plant species are the major plant families. Among collected species one interesting plant was found and determined as a new record Anemone narcissiflora L. for flora of Iran. This plant is known as a complex species that shows intraspecific speciation and is classified into about 12 subspecies and 10 varieties in world. To identify the infraspecies taxons of this species, in addition to morphological characteristics, the use of appropriate molecular markers for the better isolation of the individuals were needed.

Keywords: Anemone narcissiflora, floristic Study, kuhkamar, molecular marker

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20 Influence of Laser Treatment on the Growth of Sprouts of Different Wheat Varieties

Authors: N. Bakradze, T. Dumbadze, N. Gagelidze, L. Amiranashvili, A. D. L. Batako

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Cereals are considered as a strategic product in human life and it demand is increasing with the growth of world population. There is always shortage of cereals in various areas of the globe. For example, Georgia own production meets only 15-20% of the demand for grain, despite the fact that the country is considered one of the main centers of wheat origin. In Georgia, there are 14 types of wheat and more than 150 subspecies, and 40 subspecies of common wheat. Increasing wheat production is important for the country. One of the ways to solve the problem is to develop and implement new, environmentally and economically acceptable technologies. Such technologies include pre-sowing treatment of seed with a laser and associative nitrogen-fixing of the Azospirillum brasilensse bacteria. In the region there are Dika and Lomtagora which are among the most common in Georgia. Dika is a frost-resistant wheat, with a high ability to adapt to the environment, resistant to falling and it is sown in highlands. Dicka excellent properties are due to its strong immunity to fungal diseases; Dicka grains are rich in protein and lysine. Lomtagora 126 differs with its winter and drought resistance, and, it has a great ability to germinate. Lomtagora is characterized by a strong root system and a high budding capacity. It is an early variety, fall-resistant, easy to thresh and suitable for mechanized harvesting with large and red grains. The plant is moderately resistant to fungal diseases. This paper presents some preliminary experimental results where, a continuous CO2 laser at a power of 25-40 W/cm2 was used to radiate grains at a flow rate of 10-15 cm/sec. The treatment was carried out on grains of the Triticum aestivum L. var. of Lutescens (local variety name - Lomtagora 126), and Triticum carthlicum Nevski (local variety name - Dika). Here the grains were treated with Azospirillum brasilensse isolate (108-109 CFU / ml), which was isolated from the rhizosphere of wheat. It was observed that the germination of the wheat was not significantly influenced by either laser or bacteria treatment. In the case of the variety Lomtagora 126, when irradiated at an angle of 90°, it slightly improved the growth within 38 days of sawing, and in the case of irradiation at an angle of 90°+1, by 23%. The treatment of seeds with Azospirillum brazilense in both irradiated and non-irradiated variants led to an improvement in the growth of ssprouts. However, in the case of treatment with azospiril alone - by 22%, and with joint treatment of seeds with azospiril and irradiation - by 29%. In the case of the Dika wheat, the irradiation only led to an increase in growth by 8-9%, and the combine treatment of seeds with azospiril and irradiation - by 10-15%, in comparison with the control. Thus, the combine treatment of wheat of different varieties provided the best effect on the growth. Acknowledgment: This work was supported by Shota Rustaveli National Science Foundation of Georgia (SRNSFG) (Grant number CARYS 19-573)

Keywords: laser treatment, Azospirillum brasilensse, seeds, wheat varieties, Lomtagora, Dika

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19 Identifying Apis millefera Strains in Akkar District (North Lebanon) Using Mitochondrial DNA: A Step in Preserving the Local Strain A. m. Syriaca

Authors: Zeina Nasr, Bashar Merheb

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The honey bee is a social insect that had driven the human interest much more than any other organism. Beekeeping practices dated the appearance of Man on earth and now it provides a hobby or a secondary work that contributes to the family revenue and requires a little time engagement and money investment. Honey production is not the only contribution of honey bees to the economy, since honey bees play an important role in the pollination. Bee keeping in Lebanon is an important part of the agricultural economy. However, a growing concern about bees is spreading globally, due to an accelerated decline of bees colonies. This raises the alert to preserve and protect local bee strains against uncontrolled introduction of foreign strains and invasive parasitic species. Mitochondrial DNA (mtDNA) markers are commonly used in studying genetic variation in the Apis mellifera species. The DraI-COI-COII test is based on the analysis of the intergenic region between the two genes COI and COII. The different honey bee strains differ in the presence or absence of the p sequence and the number of Q sequences present. A. m. syriaca belonging to the lineage Z, is the native honey bee subspecies in Lebanon, Syria, Jordan, and Palestine. A. m. syriaca is known for its high defensiveness, even though it has many important advantages. However, commercial breeder strains, such as the Italian (A. m. ligustica), and Carniolan (A. m. carnica) strains, have been introduced by beekeepers and regularly used for honey production. This raises worries about the disappearance of the local subspecies. It is obvious that identifying A. m. syriaca colonies and protecting them against uncontrolled mating with other bee strains is a crucial step to protect and improve the original local strain. This study aims to reveal the existing sub-species of honey bee in Akkar – Lebanon and to assess the influence of introgression on the hybridization of the local strain. This will help to identify areas of pure A.m. syriaca population over this district to be considered in choosing syriaca reserves. We collected samples of bees from different regions of Akkar district in order to perform mtDNA analysis. We determined the restriction fragments length of the intergenic region COI-COII, using the restriction enzyme DraI. The results showed both the C and the Z lineages. Four restriction patterns were identified among the restriction maps of the studied samples. The most abundant mitochondrial lineage is the Z lineage constituting about 60% of the identified samples. Al-Dreib region reported the lowest introgression with foreign mtDNA of 21% making it the most suitable area for a genetic reserve of syriaca in Akkar based on its lowest introgression and suitable environment in addition to the attitude of local beekeepers to conserve the local strain. Finally, this study is the first step in constructing conservation programs for the preservation of the local strain and should be generalized to the whole Lebanese population, consistent with the effort done in neighboring countries.

Keywords: Akkar Lebanon, Apis millefera syriaca, DraI-COI-COII test, mitochondrial DNA

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18 Study on Butterfly Visitation Patterns of Stachytarpheta jamaicensis as a Beneficial Plant for Butterfly Conservation

Authors: P. U. S. Peiris

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The butterflies are ecologically very important insects. The adults generally feed on nectar and are important as pollinators of flowering plants. However, these pollinators are under threat with their habitat loss. One reason for habitat loss is spread of invasive plants. However, there are even beneficial exotic plants which can directly support for Butterfly Conservation Action Plan of Sri Lanka by attracting butterflies for nectar. Stachytarpheta jamaicensis (L.) is an important nectar plant which attracts a diverse set of butterflies in higher number. It comprises a violet color inflorescence which last for about 37 hours where it attracted a peak of butterflies around 9.00am having around average of 15 butterflies. There were no butterflies in early and late hours where the number goes to very low values as 2 at 1.00pm. it was found that a diverse group of butterflies were attracted from around 15 species including 01 endemic species, 02 endemic subspecies and 02 vulnerable species. Therefore, this is a beneficial exotic plant that could be used in butterfly attraction and conservation however with adequate monitoring of the plant population.

Keywords: butterflies, exotic plants, pollinators, Stachytarpheta jamaicensis (L.)

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17 Biodiversity And Ecosystem Services In Morocco: Current State And Human Development

Authors: Mohammed Taleb

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Morocco is characterized by an important genetic diversity represented by a rich and varied flora with 5211 species and subspecies and many natural ecosystems. Biodiversity and natural ecosystems provide the local population with highly diversified services represented by aromatic and medicinal plants, forage plants, melliferous plants, firewood, lumber, mushrooms, etc. Ecosystem services are currently subject to many pressures: overgrazing and deforestation, climate change, including increased drought, urbanization and forest fire. Conscious of the risks that weigh on biodiversity and ecosystem services, Morocco had made an important effort to reverse the tendencies by developing a consistent biodiversity conservation strategy focused on in-situ and ex-situ conservation. This presentation will be focused on the current state of biodiversity and ecosystem services and their role for the human development and their decline under the action of different pressures (grazing, timber harvest, harvesting of medicinal and aromatic plants, charcoal making...) while emphasizing efforts constructed by Morocco to conserve and sustainably manage biodiversity and ecosystem services.

Keywords: morocco, biodiversity, ecosystem services, local population

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16 Complete Genome Sequence Analysis of Pasteurella multocida Subspecies multocida Serotype A Strain PMTB2.1

Authors: Shagufta Jabeen, Faez J. Firdaus Abdullah, Zunita Zakaria, Nurulfiza M. Isa, Yung C. Tan, Wai Y. Yee, Abdul R. Omar

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Pasteurella multocida (PM) is an important veterinary opportunistic pathogen particularly associated with septicemic pasteurellosis, pneumonic pasteurellosis and hemorrhagic septicemia in cattle and buffaloes. P. multocida serotype A has been reported to cause fatal pneumonia and septicemia. Pasteurella multocida subspecies multocida of serotype A Malaysian isolate PMTB2.1 was first isolated from buffaloes died of septicemia. In this study, the genome of P. multocida strain PMTB2.1 was sequenced using third-generation sequencing technology, PacBio RS2 system and analyzed bioinformatically via de novo analysis followed by in-depth analysis based on comparative genomics. Bioinformatics analysis based on de novo assembly of PacBio raw reads generated 3 contigs followed by gap filling of aligned contigs with PCR sequencing, generated a single contiguous circular chromosome with a genomic size of 2,315,138 bp and a GC content of approximately 40.32% (Accession number CP007205). The PMTB2.1 genome comprised of 2,176 protein-coding sequences, 6 rRNA operons and 56 tRNA and 4 ncRNAs sequences. The comparative genome sequence analysis of PMTB2.1 with nine complete genomes which include Actinobacillus pleuropneumoniae, Haemophilus parasuis, Escherichia coli and five P. multocida complete genome sequences including, PM70, PM36950, PMHN06, PM3480, PMHB01 and PMTB2.1 was carried out based on OrthoMCL analysis and Venn diagram. The analysis showed that 282 CDs (13%) are unique to PMTB2.1and 1,125 CDs with orthologs in all. This reflects overall close relationship of these bacteria and supports the classification in the Gamma subdivision of the Proteobacteria. In addition, genomic distance analysis among all nine genomes indicated that PMTB2.1 is closely related with other five Pasteurella species with genomic distance less than 0.13. Synteny analysis shows subtle differences in genetic structures among different P.multocida indicating the dynamics of frequent gene transfer events among different P. multocida strains. However, PM3480 and PM70 exhibited exceptionally large structural variation since they were swine and chicken isolates. Furthermore, genomic structure of PMTB2.1 is more resembling that of PM36950 with a genomic size difference of approximately 34,380 kb (smaller than PM36950) and strain-specific Integrative and Conjugative Elements (ICE) which was found only in PM36950 is absent in PMTB2.1. Meanwhile, two intact prophages sequences of approximately 62 kb were found to be present only in PMTB2.1. One of phage is similar to transposable phage SfMu. The phylogenomic tree was constructed and rooted with E. coli, A. pleuropneumoniae and H. parasuis based on OrthoMCL analysis. The genomes of P. multocida strain PMTB2.1 were clustered with bovine isolates of P. multocida strain PM36950 and PMHB01 and were separated from avian isolate PM70 and swine isolates PM3480 and PMHN06 and are distant from Actinobacillus and Haemophilus. Previous studies based on Single Nucleotide Polymorphism (SNPs) and Multilocus Sequence Typing (MLST) unable to show a clear phylogenetic relatedness between Pasteurella multocida and the different host. In conclusion, this study has provided insight on the genomic structure of PMTB2.1 in terms of potential genes that can function as virulence factors for future study in elucidating the mechanisms behind the ability of the bacteria in causing diseases in susceptible animals.

Keywords: comparative genomics, DNA sequencing, phage, phylogenomics

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15 Ecobiological Study of Olivier in the Northern Slopes of the Mountains of Tlemcen, Western Algeria

Authors: Hachemi Nouria

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The olive tree is a Mediterranean tree, which belongs to the family Oleaceae. The Olea genus contains various species and subspecies, and the only species bearing edible fruit is Olea europaea. The desired issue in this study is to provide the current status of plant cover and especially the training in Olea europaea currently existing in the major centers of the region of Tlemcen. While based on the flora and biometric aspect of this plant germplasm. In order to make an assessment of the phytomass, we made measurements of the four parameters of the aerial part of the taxon: height, diameter, and canopy density to ten feet of the olive tree per station. The floristic analysis shows a certain floristic difference between the different stations. The vegetal formations reflect the biotic and abiotic conditions including climate affecting the ecosystem. Biometric study on the feet of Olea in the six study sites, has led us to conclude that the four measured parameters provides insight on the development or degradation of Olea feet depending on the layout of the stations and the factors environmental. We find that the terrains are havens for these assets. Also the local microclimate (Oued Thalweg) promotes the healthy development of this species.

Keywords: olivier, ecology, biometrics, Tlemcen, Algeria

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14 Current Status of Mosquitoes Vector Research and Control in Iran

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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13 Modelling and Management of Vegetal Pest Based On Case of Xylella Fastidiosa in Alicante

Authors: Maria Teresa Signes Pont, Jose Juan Cortes Plana

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Our proposal provides suitable modelling to the spread of plant pest and particularly to the propagation of Xylella fastidiosa in the almond trees. We compared the impact of temperature and humidity on the propagation of Xylella fastidiosa in various subspecies. Comparison between Balearic Islands and Alicante (Spain). Most sharpshooter and spittlebug species showed peaks in population density during the month of higher mean temperature and relative humidity (April-October), except for the splittlebug Clastoptera sp.1, whose adult population peaked from September-October (late summer and early autumn). The critical season is from when they hatch from the eggs until they are in the pre-reproductive season (January -April) to expand. We focused on winters in the egg state, which normally hatches in early March. The nymphs secrete a foam (mucilage) in which they live and that protects them from natural enemies of temperature changes and prevents dry as long as the humidity is above 75%. The interaction between the life cycles of vectors and vegetation influences the food preferences of vectors and is responsible for the general seasonal shift of the population from vegetation to trees and vice versa, In addition to the temperature maps, we have observed humidity as it affects the spread of the pest Xylella fastidiosa (Xf).

Keywords: xylella fastidiosa, almod tree, temperature, humidity, environmental model

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12 Clinicopathological Findings of Partuberclosis in Camels: Possible Steps for Control Strategy

Authors: A. M. Almujalli, G. M. Al-Ghamdi

Abstract:

Mycobacterium avium subspecies paratuberculosis causes paratuberculosis, a chronic debilitating granulomatous enteritis, in camels as well as domestic and wild ruminants. The clinical manifestation of the disease in camel is not well characterized, therefore this study was aimed to investigate the clinical and pathological pictures of camels that are suffering from partuberculosis. Twelve young camels that were presented to the Veterinary Teaching Hospital, King Faisal University were investigated. Clinical and pathological examination were performed. The results revealed highly significant increase in creatinine, blood urea nitrogen, magnesium, AST and ALT in diseased camels, while glucose, total protein and albumin were highly significantly decreased in diseased camels when compared to healthy ones. Post-mortem testing indicated thickening, corrugation of the intestinal wall, folded mucosa, enlarged and oedemated ileocaecal and mesenteric lymph nodes. The microscopic findings detected short, blunt and distorted intestinal villi with hyperactive goblet cells of the villi and the crypts of lieberkuhn contained mucin droplets. The lamina propria was heavily infiltrated with mononuclear cells mostly macrophages. This clinical picture of paratuberculosis may be used to initiate control strategy to limit the spread of the disease in camel herds.

Keywords: camel, partuberclosis, control, Saudi Arabia

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11 Epidemiological Survey of Feline Leukemia Virus in Domestic Cats on Tsushima Island, Japan: Tsushima Leopard Cats Are at Risk

Authors: Isaac Makundi, Kazuo Nishigaki

Abstract:

The Tsushima leopard cat (TLC) Prionailurus bengalensis euptilurus, designated a National Natural Monument of Japan, inhabits Tsushima Island, Nagasaki Prefecture, Japan. TLC is considered a subspecies of P. bengalensis, and lives only on Tsushima Island. TLCs are threatened by various infectious diseases. Feline leukemia virus (FeLV) causes a serious infectious disease with a poor prognosis in cats. Therefore, the transmission of FeLV from Tsushima domestic cats (TDCs) to TLCs may threaten the TLC population. We investigated the FeLV infection status of both TDCs and TLCs on Tsushima Island by screening blood samples for FeLV p27 antigen and using PCR to amplify the full-length FeLV env gene. The prevalence of FeLV was 6.4% in TDCs and 0% in TLCs. We also demonstrated that the virus can replicate in the cells of TLCs, suggesting its potential cross-species transmission. The viruses in TDCs were classified as genotype I/clade 3, which is prevalent on a nearby island, based on previous studies of FeLV genotypes and FeLV epidemiology. The FeLV viruses identified on Tsushima Island can be further divided into two lineages within genotype I/clade 3, which are geographically separated in Kamijima and Shimojima, indicating that FeLV may have been transmitted to Tsushima Island at least twice. Monitoring FeLV infection in the TDC and TLC populations is highly recommended as part of the TLC surveillance and management strategy.

Keywords: epidemiology, Feline leukemia virus, Tsushima Island, wildlife management

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10 Nucleotide Diversity and Bacterial Endosymbionts of the Black Cherry Aphid Myzus cerasi (Fabricus, 1775) (Hemiptera: Aphididae) from Turkey

Authors: Burcu Inal, Irfan Kandemir

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Sequences of mitochondrial cytochrome oxidase I (COI) gene of twenty-five Turkish and one Greek Myzus cerasi (Fabricus) (Hemiptera: Aphididae) in populations were collected from Prunus avium and Prunus cerasus. The partial coding region of COI studied is 605 bp for all the populations, from which 565 nucleotides were conserved, 40 were variable, 37 were singleton, and 3 sites were parsimony-informative. Four haplotypes were identified based on nucleotide substitutions, and the mean of intraspecific divergence was calculated to be 0.3%. Phylogenetic trees were constructed using Maximum Likelihood, Minimum Evolution, Neighbor-joining, and Unweighed Pair Group Method of Arithmetic Averages (UPGMA) and Myzus persicae (Sulzer) and Myzus borealis Ossiannilson were included as outgroups. The population of M. cerasi from Isparta diverged from the rest of the groups and formed a clade (Haplotype B) with Myzus borealis. The rest of the haplotype diversity includes Haplotype A and Haplotype C with individuals characterized as Myzus cerasi pruniavium and Haplotype D with Myzus cerasi cerasi. M. cerasi diverge into two subspecies and it must be reevaluated whether this pest is monophagous or oligophagous in terms of plant type dependence. The obligated endosymbiont Buchnera aphidicola was also found during this research, but no facultative symbionts could be found. It is expected further studies will be required for a complete barcoding and diversity of bacterial endosymbionts present.

Keywords: bacterial endosymbionts, barcoding, black cherry aphid, nucleotide diversity

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9 Investigation of Genetic Variation among Anemone narcissiflora L. Population Using PCR-RAPD Molecular Marker

Authors: Somayeh Akrami, Habib Onsori, Elham Tahmassebian

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Species of Anemone narcissiflora is belonged to Anemone genus of Ranunculaceae family. This species has two subspecies named narcissiflora and willdenowii which the latest is recorded in Iran in 2010. Some samples of A. narcissiflora is gathered from kuhkamar-zonouz region of East -Azerbaijan province, Iran to study the genetic diversity of the species by using RAPD molecular markers, and estimation of genetic diversity were evaluated with the using 10mer RAPD primers by PCR-RAPD method. 39 polymorphic bands were produced from the six primers used in this technique that the maximum band is related to the RP1 primer, the lowest band is related to the RP7 and the average band for all primers were 6.5 polymorphic bands. Cluster analysis of samples in done by UPGMA method in NTSYSpc 2.02 software. Dendrogram resulting from migrating bands showed that the studied samples can be divided into two groups. The first group includes samples with 1-2 flowers and the second group consists of two sub-groups which the first subgroup consists of samples with 3-5 flowers, and the second subgroup consists of samples with 6-7 flowers. The results of the comparison and analysis of the data obtained from RAPD technique and similarity matrix represents the genetic variation between collected samples. This study shows that RAPD markers can determine the polymorphisms between different genotypes of A. narcissiflora and their hybrids. So RAPD technique can serve as a suitable molecular method to determine the genetic diversity of samples.

Keywords: Anemone narcissiflora, genetic diversity, RAPD-PCR

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8 Molecular Diagnosis of Influenza Strains Was Carried Out on Patients of the Social Security Clinic in Karaj Using the RT-PCR Technique

Authors: A. Ferasat, S. Rostampour Yasouri

Abstract:

Seasonal flu is a highly contagious infection caused by influenza viruses. These viruses undergo genetic changes that result in new epidemics across the globe. Medical attention is crucial in severe cases, particularly for the elderly, frail, and those with chronic illnesses, as their immune systems are often weaker. The purpose of this study was to detect new subtypes of the influenza A virus rapidly using a specific RT-PCR method based on the HA gene (hemagglutinin). In the winter and spring of 2022_2023, 120 embryonated egg samples were cultured, suspected of seasonal influenza. RNA synthesis, followed by cDNA synthesis, was performed. Finally, the PCR technique was applied using a pair of specific primers designed based on the HA gene. The PCR product was identified after purification, and the nucleotide sequence of purified PCR products was compared with the sequences in the gene bank. The results showed a high similarity between the sequence of the positive samples isolated from the patients and the sequence of the new strains isolated in recent years. This RT-PCR technique is entirely specific in this study, enabling the detection and multiplication of influenza and its subspecies from clinical samples. The RT-PCR technique based on the HA gene, along with sequencing, is a fast, specific, and sensitive diagnostic method for those infected with influenza viruses and its new subtypes. Rapid molecular diagnosis of influenza is essential for suspected people to control and prevent the spread of the disease to others. It also prevents the occurrence of secondary (sometimes fatal) pneumonia that results from influenza and pathogenic bacteria. The critical role of rapid diagnosis of new strains of influenza is to prepare a drug vaccine against the latest viruses that did not exist in the community last year and are entirely new viruses.

Keywords: influenza, molecular diagnosis, patients, RT-PCR technique

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7 Analysis of Non-Coding Genome in Streptococcus pneumoniae for Molecular Epidemiology Typing

Authors: Martynova Alina, Lyubov Buzoleva

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Streptococcus pneumoniae is the causative agent of pneumonias and meningitids throught all the world. Having high genetic diversity, this microorganism can cause different clinical forms of pneumococcal infections and microbiologically it is really difficult diagnosed by routine methods. Also, epidemiological surveillance requires more developed methods of molecular typing because the recent method of serotyping doesn't allow to distinguish invasive and non-invasive isolates properly. Non-coding genome of bacteria seems to be the interesting source for seeking of highly distinguishable markers to discriminate the subspecies of such a variable bacteria as Streptococcus pneumoniae. Technically, we proposed scheme of discrimination of S.pneumoniae strains with amplification of non-coding region (SP_1932) with the following restriction with 2 types of enzymes of Alu1 and Mn1. Aim: This research aimed to compare different methods of typing and their application for molecular epidemiology purposes. Methods: we analyzed population of 100 strains of S.pneumoniae isolated from different patients by different molecular epidemiology methods such as pulse-field gel electophoresis (PFGE), restriction polymorphism analysis (RFLP) and multilolocus sequence typing (MLST), and all of them were compared with classic typing method as serotyping. The discriminative power was estimated with Simpson Index (SI). Results: We revealed that the most discriminative typing method is RFLP (SI=0,97, there were distinguished 42 genotypes).PFGE was slightly less discriminative (SI=0,95, we identified 35 genotypes). MLST is still the best reference method (SI=1.0). Classic method of serotyping showed quite weak discriminative power (SI=0,93, 24 genotypes). In addition, sensivity of RFLP was 100%, specificity was 97,09%. Conclusion: the most appropriate method for routine epidemiology surveillance is RFLP with non-coding region of Streptococcsu pneumoniae, then PFGE, though in some cases these results should be obligatory confirmed by MLST.

Keywords: molecular epidemiology typing, non-coding genome, Streptococcus pneumoniae, MLST

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6 A Study on Genus Carolia Cantraine, 1838: A Case Study in Egypt with Special Emphasis on Paleobiogeographic, and Biometric Context

Authors: Soheir El-Shazly, Gouda Abdel-Gawad, Yasser Salama, Dina Sayed

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Twelve species belonging to genus Carolia Cantraine, 1838 were recorded from nine localities in the Tertiary rocks of the Tethys, Atlantic and Eastern Pacific Provinces. During The Eocene two species were collected from Indian-Pakistani region, two from North Africa (Libya, Tunis and Algeria), one from Jamaica and two from Peru. The Oligocene shows its appearance in North America (Florida) and Argentina. The genus showed its last occurrence in the Miocene rocks of North America (Florida) before its extinction. In Egypt, the genus was diversified in the Eocene rocks and was represented by four species and two subspecies. The paleobiogeographic distribution of Genus Carolia Cantraine, 1838 indicates that it appeared in the Lower Eocene of West Indian Ocean and migrated westward flowing circumtropical Tethys Current to the central Tethyan province, where it appeared in North Africa and continued its dispersal westward to the Atlantic Ocean and arrived Jamaica in the Middle Eocene. It persisted in the Caribbean Sea and appeared later in the Oligocene and Miocene rocks of North America (Florida). Crossing Panama corridor, the genus migrated to the south Eastern Pacific Ocean and was collected from the Middle Eocene of Peru. The appearance of the genus in the Oligocene of the South Atlantic Coast of Argentina may be via South America Seaway or its southward migration from Central America to Austral Basin. The thickening of the upper valve of the genus, after the loss of its byssus to withstand the current action, caused inability of the animal to carry on its vital activity and caused its extinction. The biometric study of Carolia placunoides Cantraine, 1938 from thhe Eocene of Egypt, indicates that the distance between the muscle scars in the upper valve increases with the closure of the byssal notch.

Keywords: Atlantic, carolia, paleobiogeography, tethys

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5 Anatomical and Histological Analysis of Salpinx and Ovary in Anatolian Wild Goat (Capra aegagrus aegagrus)

Authors: Gulseren Kirbas, Mushap Kuru, Buket Bakir, Ebru Karadag Sari

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Capra (mountain goat) is a genus comprising nine species. The domestic goat (C. aegagrus hircus) is a subspecies of the wild goat that is domesticated. This study aimed to determine the anatomical structure of the salpinx and ovary of the Anatolian wild goat (C. aegagrus aegagrus). Animals that were taken to the Kafkas University Wildlife Rescue and Rehabilitation Center, Kars, Turkey, because of various reasons, such as traffic accidents and firearm injuries, were used in this study. The salpinges and ovaries of four wild goats of similar ages, which could not be rescued by the Center despite all interventions, were dissected. Measurements were taken from the right-left salpinx and ovary using digital calipers. The weights of each ovary and salpinx were measured using a precision scale (min: 0.0001 g − max: 220 g, code: XB220A; Precisa, Swiss). The histological structure of the tissues was examined after weighing the organs. The tissue samples were fixed in 10% formaldehyde for 24 h. Then a routine procedure was applied, and the tissues were embedded in paraffin. Mallory’s modified triple staining was used to demonstrate the general structure of the salpinx. The salpinx was found to consist of three different regions (infundibulum, ampulla, and isthmus). These regions consisted of tunica mucosa, tunica muscularis, and tunica serosa. The prismatic epithelial cells were observed in the lamina epithelialis of tunica mucosa in every region, but the prismatic fimbrae cells occurred most in the infundibulum. The ampulla was distinguished by its many mucosal folds. It was the longest region of the salpinx and was joined to the isthmus via the ampullary–isthmus junction. Isthmus was the caudal end of the salpinx joined to the uterus and had the thickest tunica muscularis compared with the other regions. The mean length of the ovary was 13.22 ± 1.27 mm, width was 8.46 ± 0.88 mm, the thickness was 5.67 ± 0.79 mm, and weight was 0.59 ± 0.17 g. The average length of the salpinx was 58.11 ± 14.02 mm, width was 0.80 ± 0.22 mm, the thickness was 0.41 ± 0.01 mm, and weight was 0.30 ± 0.08 g. In conclusion, the Anatolian wild goat, which is included in wildlife diversity in Turkey, has been disappearing due to illegal and uncontrolled hunting as well as traffic accidents in recent years. These findings are believed to contribute to the literature.

Keywords: Anatolian wild goat, anatomy, ovary, salpinx

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4 A Preliminary Survey on Butterfly Fauna at Rajagala Archaeological Site, Ampara, Sri Lanka

Authors: D. Eranda N. Mandawala, P. A. D. Mokshi V. Perera

Abstract:

The RajagalaArchaeological site (RAS) is located 26 km from Ampara town (7º29'25.22" N, 81º36'59.05" E) accessible through the Ampara-Uhana-MahaOya highway of the Eastern province of Sri Lanka. This site has recently been added to the tentative list of UNESCO world heritage site and is also a forest reserve. This dry zone forest consists of tropical mixed evergreen vegetation and scrublands on a rocky outcrop of elevation of about 350 meters above mean sea level. It is also scattered with several ponds of differing sizes on rocky outcrops, rocky cliffs, and about 50 cave dwellings. No comprehensive biodiversity survey of any sorts has been conducted at the RAS so far. Therefore, a preliminary survey was conducted to determine its butterfly fauna diversity. An opportunistic Visual Encounter Survey method was used to observe various butterfly species during the morning between 8:00am-12:00noon and in the evening between 2:00-6:00pm on 3 site visits in October 2017, February 2018, and November 2019. All encountered species were photographed using a Nikon D750 camera with Sigma 105mm f/2.8 EX DG OS HSM macro lens, and field guide books were used to identify them. Sri Lanka is home to 248 species of butterflies, of which are 26 are endemic. At RAS, we observed a total of 39 species (15%) of butterflies belonging to 5 Lepidoptera families. Out of these, one endemic species(4%) and 9 endemic subspecieswere also identified. The former was Troidesdarsius, also known as the Sri Lanka birdwing which is the national butterfly and the largest butterfly in Sri Lanka, and the latter were Plains cupid (Chiladespandavalanka), Yamfly (Loxuraatymnus arcuate), Common Cerulean (Jamidescelenotissama), Tawny Rajah(Charaxespsaphonpsaphon), Tamil Yeoman(Cirrochroathaislanka), Angled Castor(Ariadne ariadneminorata), GladeyeBushbrown(Mycalesispatnia patina), Common Crow (Euploea core asela)and Blue Mormon (Papiliopolymnestorparinda). The endemic subspecies belonged to 3 Lepidoptera families (3from Lycaenidae, 5 from Nymphalidae, and 1 from Papilionidae family). Anthropogenic activities such as unauthorized cattle farming, forest clearance, and man-made forest fires currently threaten this site. If such trends continue, it may lead to the reduction of butterfly fauna diversity within this area in the future.

Keywords: lepidoptera, rajagala, Sri Lanka birdwing, endemic

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3 Determination of Phenolic Contents and Antioxidant Activities of Chenopodium quinoa Willd. Seed Extracts

Authors: Nilgün Öztürk, Hakan Sabahtin Ali, Hülya Tuba Kıyan

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The genus Chenopodium belongs to Amaranthaceae, is represented by approximately 250 species in the world and 15 species and three subspecies in Turkey. Chenopodium species are traditionally used to treat chest and abdominal pain, shortness of breath, cough and neurological disorders. Chenopodium quinoa Willd. (Quinoa) is native to Andes region of South America (especially Peru and Bolivia) and cultivated in many countries include also Turkey in the world nowadays. The seeds of quinoa are rich in protein, and the phytochemical composition consists of antioxidant substances such as polyphenolic compounds, flavonoids, vitamins, and minerals; anticancer and neuroprotective compounds such as tocotrienols; anti-inflammatory compounds such as carotenoids and anthocyanins and also saponins and starch. Food products of quinoa such as quinoa cereal bar, pasta and cornflakes are used in the diet made during many disorders like obesity, cardiovascular disorder, hypertension and Celiac disease. Also quinoa seems to have antimicrobial, anti-inflammatory and cholesterol-lowering properties because of its bioactive compounds. In this present study, the aqueous ethanolic extracts of the seeds of three different coloured genotypes of quinoa were investigated for their antioxidant activities using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, ferrous ion-chelating effect, ferric-reducing antioxidant power, ABTS radical cation decolorization assays and total phenolic contents using Folin-Ciocalteu assay. Among the three genotypes of quinoa; the aqueous ethanolic extract of the red genotype had the highest total phenolic content (83.54 ± 2.12 mg gallic acid/100 g extract) whereas the extract of the white genotype had the lowest total phenolic content (70.66 ± 0.25 mg gallic acid/100 g). According to the antioxidant activity results; the extracts showed moderate reducing power effect whereas weak ABTS radical cation decolorization and ferrous ion-chelating effect and also too weak DPPH radical scavenging activity when compared to the positive standards.

Keywords: amaranthaceae, antioxidant activity, Chenopodium quinoa willd., total phenolic content

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2 Feeding Value Improvement of Rice Straw Fermented by Spent Mushroom Substrate on Growth and Lactating Performance of Dairy Goat

Authors: G. J. Fan, T. T. Lee, M. H. Chen, T. F. Shiao, B. Yu, C. F. Lee

Abstract:

Rice straw with poor feed quality and spent mushroom substrate are both the most abundant agricultural residues in Taiwan. Edible mushrooms from white rot fungi possess lignocellulase activity. It was expected to improve the feeding value of rice straw for ruminant by solid-state fermentation pretreatment using spent mushroom substrate. Six varieties or subspecies of spent edible mushrooms (Pleurotus ostreatus (blue or white color), P. sajor-caju, P. citrinopileatus, P. eryngii and Ganoderma lucidium) substrate were evaluated in solid-state fermentation process with rice straw for 8 wks. Quality improvement of fermented rice straw was determined by its in vitro digestibility, lignocellulose degradability, and cell wall breakdown checked by scanning electron microscope. Results turned out that Pleurotus ostreatus (white color) and P. sajor-caju had the better lignocellulose degradation effect than the others and was chosen for advance in vivo study. Rice straw fermented with spent Pleurotus ostreatus or Pleurotus sajor-caju mushroom substrate 8 wks was prepared for growing and lactating feeding trials of dairy goat, respectively. Pangolagrass hay at 15% diet dry matter was the control diet. Fermented or original rice straw was added to substitute pangolagrass hay in both feeding trials. A total of 30 head of Alpine castrated ram were assigned into three groups for 11 weeks, 5 pens (2 head/pen) each group. A total of 21 head of Saanen and Alpine goats were assigned into three treatments and individually fed in two repeat lactating trials with 28-d each. In castrated ram study, results showed that fermented rice straw by spent Pleurotus ostreatus mushroom substrate attributed the higher daily dry matter intakes (DMI, 1.53 vs. 1.20 kg) and body weight gain (138 vs. 101 g) than goats fed original rice straw. DMI (2.25 vs. 1.81 kg) and milk yield (3.31 vs. 3.02 kg) of lactating goats fed control pangolagrass diet and fermented rice straw by spent Pleurotus sajor-caju mushroom substrate were also higher than those fed original rice straw diet (P < 0.05). Milk compositions, milk fat, protein, total solid and lactose, were similar among treatments. In conclusion, solid-state fermentation by spent Pleurotus ostreatus or Pleurotus sajor-caju mushroom substrate could effectively improve the feeding value of rice straw. Fermented rice straw is a good alternative fiber feed resource for growing and lactating dairy goats and 15% in diet dry matter is recommended.

Keywords: feeding value, fermented rice straw, growing and lactating dairy goat, spent Pleurotus ostreatus and Pleurotus sajor-caju mushroom substrate

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1 Multilocus Phylogenetic Approach Reveals Informative DNA Barcodes for Studying Evolution and Taxonomy of Fusarium Fungi

Authors: Alexander A. Stakheev, Larisa V. Samokhvalova, Sergey K. Zavriev

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Fusarium fungi are among the most devastating plant pathogens distributed all over the world. Significant reduction of grain yield and quality caused by Fusarium leads to multi-billion dollar annual losses to the world agricultural production. These organisms can also cause infections in immunocompromised persons and produce the wide range of mycotoxins, such as trichothecenes, fumonisins, and zearalenone, which are hazardous to human and animal health. Identification of Fusarium fungi based on the morphology of spores and spore-forming structures, colony color and appearance on specific culture media is often very complicated due to the high similarity of these features for closely related species. Modern Fusarium taxonomy increasingly uses data of crossing experiments (biological species concept) and genetic polymorphism analysis (phylogenetic species concept). A number of novel Fusarium sibling species has been established using DNA barcoding techniques. Species recognition is best made with the combined phylogeny of intron-rich protein coding genes and ribosomal DNA sequences. However, the internal transcribed spacer of (ITS), which is considered to be universal DNA barcode for Fungi, is not suitable for genus Fusarium, because of its insufficient variability between closely related species and the presence of non-orthologous copies in the genome. Nowadays, the translation elongation factor 1 alpha (TEF1α) gene is the “gold standard” of Fusarium taxonomy, but the search for novel informative markers is still needed. In this study, we used two novel DNA markers, frataxin (FXN) and heat shock protein 90 (HSP90) to discover phylogenetic relationships between Fusarium species. Multilocus phylogenetic analysis based on partial sequences of TEF1α, FXN, HSP90, as well as intergenic spacer of ribosomal DNA (IGS), beta-tubulin (β-TUB) and phosphate permease (PHO) genes has been conducted for 120 isolates of 19 Fusarium species from different climatic zones of Russia and neighboring countries using maximum likelihood (ML) and maximum parsimony (MP) algorithms. Our analyses revealed that FXN and HSP90 genes could be considered as informative phylogenetic markers, suitable for evolutionary and taxonomic studies of Fusarium genus. It has been shown that PHO gene possesses more variable (22 %) and parsimony informative (19 %) characters than other markers, including TEF1α (12 % and 9 %, correspondingly) when used for elucidating phylogenetic relationships between F. avenaceum and its closest relatives – F. tricinctum, F. acuminatum, F. torulosum. Application of novel DNA barcodes confirmed the fact that F. arthrosporioides do not represent a separate species but only a subspecies of F. avenaceum. Phylogeny based on partial PHO and FXN sequences revealed the presence of separate cluster of four F. avenaceum strains which were closer to F. torulosum than to major F. avenaceum clade. The strain F-846 from Moldova, morphologically identified as F. poae, formed a separate lineage in all the constructed dendrograms, and could potentially be considered as a separate species, but more information is needed to confirm this conclusion. Variable sites in PHO sequences were used for the first-time development of specific qPCR-based diagnostic assays for F. acuminatum and F. torulosum. This work was supported by Russian Foundation for Basic Research (grant № 15-29-02527).

Keywords: DNA barcode, fusarium, identification, phylogenetics, taxonomy

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