Sequence Analysis Of Region Research Articles

Evaluation the pathogenicity and control of Penicillium citrinum isolates affecting the white fungus Agaricus bisporus

Five isolates of Penicillium citrinumwere isolated from casing soils, copmpost and fruiting bodies of Agaricusbisporus from several culture stations in Iraq. According to the pathogenicity test, the highest percentage of inhibition was recorded for isolate P.citrinum(B1). The causal agent was molecularly diagnosed to the level of P. citrinumaccording to the nucleotide sequence analysis of the ITS genome region and it was recorded globally on the NCBI data base under the accission number ON738710.1.The effect of the hot aqueous extract of cloves and conocarpus, and the efficacy of the biocide VEROX on the production and phenotypic characteristics of the white edable fungus Agaricusbisporus was studied with the presence of the pathogenic fungus. The results showed that the treatment of the combination of plant extracts + biocide + pathogenic fungus led to the highest rate of growth inhibition of the pathogenic fungus, increasing number of fruiting bodies up to 435 compared to the control treatment of pathogenic fungus 140. The total yield of the same treatment was 16.398 kg compared to 3.343 kg in the control treatment, and the biological efficiency was recorded by 37.2% compared to 10.2% in the control treatment. In general, this combination led to an increase in production and an improvement in the qualitative characteristics of the fruiting bodies, as the average diameter of the cap was 40.6 mm and the stalk length was 37.3 mm compared to 27.8 mm and 23.5 mm in the control treatment, respectively.

Prevalence of fluoroquinolone resistance-associated mutations in Mycoplasma genitalium among clients of two sexual health centres in the Netherlands: a cross-sectional study

ObjectiveThis study aimed to determine the prevalence of fluoroquinolone resistance-associated mutations (QRAMs) in Mycoplasma genitalium (MG) among clients of two sexual health centres (SHCs) in the Netherlands.DesignA cross-sectional study.Setting and...

Utilisation of a mitochondrial intergenic region for species differentiation of fruit flies (Diptera: Tephritidae) in South Africa

BackgroundFruit flies (Diptera: Tephritidae) comprise species of agricultural and economic importance. Five such fruit fly species are known to affect commercial fruit production and export in South Africa: Ceratitis capitata, Ceratitis cosyra, Ceratitis rosa, Ceratitis quilicii, and Bactrocera dorsalis. Management practices for these pests include monitoring, application of pest control products, post-harvest disinfestation measures and inspection of consignments both prior to shipment and at ports of entry. In activities relating to monitoring and inspection, accurate identification of these pests to species level is required. While morphological keys for adult stages of these fruit fly species have been well developed, morphological keys for earlier life stages remain problematic. In instances where closely related species cannot be reliably distinguished morphologically, there is a need for molecular tools to assist in identifying these five fruit fly species during surveillance practices, where sequencing-based approaches would be beneficial.ResultsTwo complete mitochondrial genomes were assembled for each fruit fly species investigated using high throughput sequencing data generated in this study. A single primer set was designed to amplify a region between tRNAile and tRNAmet. The amplicon consists of a partial segment of tRNAile, intergenic region I (tRNAile - tRNAgln), the complete sequence of tRNAgln, intergenic region II (tRNAgln - tRNAmet), and a partial segment of tRNAmet. PCR amplicons were generated for 20 specimens of each species, five of which were colony adult males, five colony larvae, and 10 wild, trap-collected specimens. Upon analysis of the amplicon, intergenic region I was identified as the most informative region, allowing for unambiguous identification of the five fruit fly species. The similarity in intergenic region II was too high between C. rosa and C. quilicii for accurate differentiation of these species.ConclusionThe identity of all five fruit flies investigated in this study can be determined through sequence analysis of the mitochondrial intergenic regions. Within the target amplicon, intergenic region I (tRNAile - tRNAgln) shows interspecific variation sufficient for species differentiation based on multiple sequence alignment. The variation in the length of intergenic region I is proposed as a potential tool for accurately identifying these five fruit flies in South Africa.

STOX1 promotor region -922 T > C polymorphism is associated with Early-Onset preeclampsia

Preeclampsia (PE), affecting 5–8% of pregnancies, is a common pregnancy disease that has harmful effects on mother and foetus. It has been found that the STOX1 (Storkhead Box 1), which is a transcription factor, carries variants associated with PE. Previous studies showed that, there was a strong relationship between PE and STOX1 variants. Therefore, we hypothesised that variants in the promoter region of the gene may be related to the onset of PE. The aim of this study is to investigate the contribution of STOX1 gene promoter region variants to PE. The blood samples taken from 118 PE patients and 96 healthy pregnant women were analysed by Sanger sequencing method. Sequence analysis results showed that, there is a-922 T > C polymorphism (rs884181) in the promoter region of the STOX1 gene. This polymorphism was found to be statistically significant in individuals with early onset PE (p = 0.02) and in PE (p = 0.014) compared to the control group. IMPACT STATEMENT What is already known on this subject? As a result of whole-exon studies on the STOX1 gene, polymorphisms were found to disrupt the structure/expression/function of the gene and strengthen its relationship with PE and HELLP syndrome. A previous study by our team found an association between Y153H, the most common polymorphism of STOX1, and early onset PE. What do the results of this study add? In our study, it was aimed to investigate the effect of genetic modifications in STOX1 gene promoter region on PE through the maternal genotype. Because any change in the promoter region affects the expression level of the gene. Also, for the first time, sequence analysis of the promoter region of STOX1 is investigated in PE. The variations in STOX1 appear to be important in PE especially in Early Onset PE. What are the implications of these findings for clinical practice and/or further research? Although PE is a disease that occurs with pregnancy and shows its effects most during this period, women and children with a history of PE are more prone to various disorders, especially cardiovascular diseases in the following years. Therefore, understanding the pathogenesis of the disease is important for both prevention and treatment process. Variations on STOX1 appear to be important in terms of disease risk.

New insight into morphological and genetic diversity of Phlyctema vagabunda and Neofabraea kienholzii causing bull's eye rot on apple and pear

AbstractFungi of genera Phlyctema and Neofabraea are the causal agents of bull's eye rot, a major postharvest disease of pome fruits. To investigate their morphological and genetic diversity, isolates obtained in Italy and Chile from decayed fruit and rainwater between 2014 and 2019 were grown on two agar media, inoculated onto four fruit cultivars and compared using four marker genes. Consistent intra‐ and interspecies phenotypic differences were recorded among isolates identified as P. vagabunda (two main morphotypes, PvM‐I and PvM‐II, were distinguished) and N. kienholzii. In particular, the Chilean isolates belonging to PvM‐I showed low sporulation in vitro, while isolates belonging to PvM‐II showed the most abundant sporulation and also formed conidiomata deep within fruit tissue. Host cultivar influenced the disease incidence in unwounded, inoculated fruit. Cripps Pink and Golden Delicious apples favoured the formation of P. vagabunda conidiomata and macroconidia, while Granny Smith apples and/or Kaiser pears restricted sporulation of some isolates of PvM‐I. Mycelial cords of P. vagabunda and N. kienholzii were consistently recorded in inoculated fruit, suggesting their possible involvement as a source of inoculum. Propagules of P. vagabunda were present in rainwater collected from apple plants from September to October in Italy. According to sequence analysis of ITS, EF‐1α, TUB2 and ACT1 regions of the fungi, 12 distinct sequence types were identified, three of which were characteristic of isolates from the Southern Hemisphere. The condensed maximum‐likelihood phylogenetic tree separated the 50 P. vagabunda isolates into six phylogroups, suggesting a correlation with their geographical distribution.

Prevalence, hematological, serum biochemical, histopathology, and molecular characterization of Sarcoptes scabiei in naturally infected rabbits from Minoufiya Governorate, Egypt.

Sarcoptic mange is a highly contagious ectoparasitic disease that causes significant economic losses in the rabbit industry. The current study intended to reveal the infection rate, histopathology, and genetic characterization of Sarcoptes scabiei (S. scabiei) in naturally infected rabbits in Minoufiya governorate, Egypt. A total of 1120 rabbits were physically inspected for sarcoptic mange lesions and infections were confirmed microscopically. In addition, the various hematologic and serum biochemical parameters in naturally infected and non-infected rabbits were evaluated. A histopathological examination was performed. Genomic DNA was isolated from skin scraping samples and amplified using PCR primers targeting the ITS-2 region and Cox1 and Actin genes, which were then sequenced and phylogenetically analyzed. The overall prevalence of S. scabiei was 5.98%. Although the infection was higher in females than males, the analysis showed no statistically significant difference. White blood cells, lymphocytes, liver enzymes (GOT and GPT), urea, total antioxidant capacity, glutathione peroxidase, and malondialdehyde dramatically increased whereas RBCs, Hb, and MCV significantly decreased. There were epidermal thickening and hyperkeratosis, inflammation, and homogenous faint pink edematous lesions, and the S. scabiei was attached to the stratum corneum and/or burrowing through it, causing tunnels. PCR and sequence analysis of the ITS-2 region and Cox1 and Actin genes showed that the sequences in the present study were highly identical to the homologous sequences from several countries and confirmed that the mite was S. scabiei. This study presented the first molecular characterization of S. scabiei in rabbits from Minoufiya Governorate, Egypt.

Characterization and biocontrol potential of Trichoderma longibrachiatum TL-RD-01 against plant pathogens

A study was conducted to isolate efficient biocontrol agent against agriculturally important plant pathogens. Isolated fungus was identified using morphological and molecular characters and evaluated against plant pathogens. Isolated culture was identified as Trichoderma longibrachiatum based on morphological and cultural characteristics. The isolate (TL-RD-01) was able to solubilize the zinc and able to produce the indole 3 acetic acid of about 32.01 μg/ml. To optimize the growth media different carbon and nitrogen sources were studied. Maltose, Corn flour, Peptone and Yeast Extract used media attained maximum growth of T. longibrachiatum. The pH (5 to 6) and temperature (25 °C to 30 °C) was optimized for the growth of T. longibrachiatum TL-RD-01. Molecular identification based on internal transcribed spacer 1 and 2 region (ITS1 and ITS2) sequence analysis have been further confirmed as T. longibrachiatum. The phylogenetic analysis of above ITS gene sequences were carried out. T. longibrachiatum isolate was clearly separated with ITS sequence’s analysis with other species of Trichoderma. Mycoparasitic effect of T. longibrachiatum was carried out by dual plate technique against agriculturally important plant pathogens viz., Macrophomina phaseolina, Phytopthora infestans, Collectrotrichum gleosporides (Mango) and Colletotrichum falcatum. Pathogen inhibition effect of T. longibrachiatum isolate was in between 50% to 81.94%. As per the investigation, Trichoderma longibrachiatum (TL-RD-01) could be deployed as a biocontrol agent and a biostimulant.

Selection and characterization of a yeast strain for the suppression of brown spot on Tru Long pummelo (Citrus maxima)

Tru Long pummelo is an endemic plant of Quang Nam province with high economic value. However, many different diseases, especially brown spot disease caused by Alternaria fungus on pummelos, have affected the quality of pummelos. Pesticides are overused and misused for the environment and human health. Therefore, there is an urgent need to reduce the use of synthetic chemicals. Biological control offers an alternative to the use of pesticides. Eight yeast strains have been isolated from healthy leaves, pummelo fruits, and healthy mulberries. They were screened for biological activity against Alternaria sp. by the dual culture method. The inhibitory potency ranged from 10.46% to 59.86%. The strain with the highest percentage (59.86%) was identified as Candida tropicalis using sequence analysis of the ITS region.

Graphiola phoenicis associated with false smut on Canary Island date palm (Phoenix canariensis) in Korea

AbstractLeaf spots and smut‐like symptoms have been observed on the Canary Island date palm Phoenix canariensis in Korea since 2021. The fungus associated with the false smut was identified as Graphiola phoenicis based on the morphological characteristics and molecular sequence analysis of the large subunit region of the rDNA. This study reports the novel occurrence of G. phoenicis on P. canariensis in Korea and provides mycological information of the Korean isolates.

Evolution of surrogate light chain in tetrapods and the relationship between lengths of CDR H3 and VpreB tails.

In the mammalian immune system, the surrogate light chain (SLC) shapes the antibody repertoire during B cell development by serving as a checkpoint for production of functional heavy chains (HC). Structural studies indicate that tail regions of VpreB contact and cover the third complementarity-determining region of the HC (CDR H3). However, some species, particularly bovines, have CDR H3 regions that may not be compatible with this HC-SLC interaction model. With immense structural and genetic diversity in antibody repertoires across species, we evaluated the genetic origins and sequence features of surrogate light chain components. We examined tetrapod genomes for evidence of conserved gene synteny to determine the evolutionary origin of VpreB1, VpreB2, and IGLL1, as well as VpreB3 and pre-T cell receptor alpha (PTCRA) genes. We found the genes for the SLC components (VpreB1, VpreB2, and IGLL1) only in eutherian mammals. However, genes for PTCRA occurred in all amniote groups and genes for VpreB3 occurred in all tetrapod groups, and these genes were highly conserved. Additionally, we found evidence of a new VpreB gene in non-mammalian tetrapods that is similar to the VpreB2 gene of eutherian mammals, suggesting VpreB2 may have appeared earlier in tetrapod evolution and may be a precursor to traditional VpreB2 genes in higher vertebrates. Among eutherian mammals, sequence conservation between VpreB1 and VpreB2 was low for all groups except rabbits and rodents, where VpreB2 was nearly identical to VpreB1 and did not share conserved synteny with VpreB2 of other species. VpreB2 of rabbits and rodents likely represents a duplicated variant of VpreB1 and is distinct from the VpreB2 of other mammals. Thus, rabbits and rodents have two variants of VpreB1 (VpreB1-1 and VpreB1-2) but no VpreB2. Sequence analysis of VpreB tail regions indicated differences in sequence content, charge, and length; where repertoire data was available, we observed a significant relationship between VpreB2 tail length and maximum DH length. We posit that SLC components co-evolved with immunoglobulin HC to accommodate the repertoire – particularly CDR H3 length and structure, and perhaps highly unusual HC (like ultralong HC of cattle) may bypass this developmental checkpoint altogether.

Keratinic amyloid deposition in canine hair follicle tumors.

Keratinic primary localized cutaneous amyloidosis is a disease in humans; however, no similar condition has been reported in animals. This study aimed to investigate cutaneous keratinic amyloid deposition in dogs and elucidate its etiology. Canine hair follicle tumor tissues were histopathologically analyzed. Immunohistochemistry and mass spectrometry-based proteomic analyses were performed to identify precursor protein candidates. Structural prediction and in vitro fibrillization analyses were conducted to determine the amyloidogenic region and gene sequencing analysis was performed to assess mutations. Of the 266 samples, 16 had amyloid deposition. Amyloid deposits were found in the stroma of tumors and in the margins of keratin debris and around normal hair follicles. Cytokeratin 5 (CK5) was identified as a precursor protein candidate. C-terminal truncation of CK5 was observed in amyloid deposits, and the truncation sites varied depending on the deposition pattern. There was a significantly higher incidence of amyloid deposition in Shiba dogs, and CK5 amino acid polymorphisms were identified in these dogs. A part of the C-terminal region of both canine and human CK5 exhibited highly amyloidogenic properties in vitro. This study revealed the existence of cutaneous keratinic amyloid deposition in animals and identified CK5 as an amyloid precursor protein, providing novel insights into understanding the etiology of cutaneous amyloidosis.

Bartonella spp. detection in laelapid (Mesostigmata: Laelapidae) mites collected from small rodents in Lithuania.

The genus Bartonella contains facultative Gram-negative intracellular bacteria from the family Bartonellaceae that can cause diseases in humans and animals. Various Bartonella species have been detected in rodents' ectoparasites, such as fleas, ticks, mites, and lice. However, the role of laelapid mites (Mesostigmata: Laelapidae) as carriers of Bartonella spp. needs to be confirmed. We aimed to investigate the presence of Bartonella spp. in laelapid mites collected from small rodents in Lithuania using real-time PCR targeting the transfer-messenger RNA/tmRNA (ssrA) gene and to characterize Bartonella strains using nested PCR and sequence analysis of the 16S-23S rRNA intergenic transcribed spacer region (ITS). A total of 271 laelapid mites of five species (Laelaps agilis, Haemogamasus nidi, Eulaelaps stabularis, Myonyssus gigas, and Hyperlaelaps microti) were collected from five rodent species (Apodemus flavicollis, Apodemus agrarius, Clethrionomys glareolus, Micromys minutus, and Microtus oeconomus) during 2015-2016. Bartonella DNA was detected in three mite species L. agilis, M. gigas, and Hg. nidi with an overall prevalence of 11.4%. Sequence analysis of the 16S-23S rRNA ITS region revealed the presence of Bartonella taylorii in L. agilis, Hg. nidi, and M. gigas, and Bartonella grahamii in L. agilis. Our results suggest that laelapid mites are involved in the maintenance of rodent-associated Bartonella spp. in nature. To the best of the authors' knowledge, this is the first study to demonstrate the presence of Bartonella spp. DNA in laelapid mites from small rodents.

First Report of Sclerotinia minor on lettuce in Serbia.

Lettuce (Lactuca sativa L.) is the world's most popular leafy salad vegetable. One of the major challenges facing lettuce producers are fungal diseases that could, under favorable conditions, devastate the harvest (Raid, 2004). During February 2021, poor growth, plant stunning and blanching of leaves of lettuce plants, cultivated in unheated plastic tunel in Potočanje (Zlatibor region), Serbia, were observed. The crowns were softened with spreading decaying lesions covered with white mycelium, particulary on the leaves near the soil surface. Approximately 2 to 3 weeks before harvest, diseased plants began to wilt and collapse. Estimated disease incidence was 50-55%. In order to identify the causal agent, symptomatic tissues from diseased plants were cut into small pieces, surface sterilized with 70% ethanol for 1 min, rinsed three times in sterile distilled water and placed on potato dextrose agar (PDA). Five isolates with uniform morphology were derived from infected tissue. The colonies had fast-growing, white, cottony aerial mycelium, producing profuse numbers (184 sclerotia/ Petri plate in average) of small, black, irregularly shaped sclerotia, less than 2 mm in diameter. Based on morfological features, the isolates were identified as Sclerotinia minor Jagger (Kohn, 1979). To confirm the species identity, the internal transcribed spacer region of nuclear rDNA of a representative isolate 15-2 was amplified using the primer pair ITS1/ITS4 (White et al. 1990). Sequence analysis of ITS region revealed 100% nucleotide identity between the isolate 15-2 (GenBank Accession No. OL423632.1) and 14 isolates of S. minor from different parts of the world (e.g., accession Nos. MK356551.1, KY707828.1, JF2798801.1). Pathogenicity tests were conducted by artificial inoculation of 55-day-old lettuce plants cv. 'Majska kraljica', grown on commercial growth substrate in l L pots. The obtained isolates were grown on PDA for 7 days and mycelial plugs, 5 mm in diameter, were cut from the margin of the colony and placed mycelium-side down on undamaged ground-level leaves of lettuce plants. Two plugs per isolate were placed onto five plants each for a total of ten replicates per isolate. Negative control plants (5) were inoculated similarly with sterile PDA plugs. Inoculated plants were covered with transparent plastic bags, sprayed with water (under the plastic) twice a day for 3 to 5 days to maintain high humidity, and kept in a growth chamber at 22°C (13 h light). After 7 to 10 days, all pathogen-inoculated plants developed lettuce drop disease symptoms, whereas the control plants remained symptomless. The pathogen was reisolated from symptomatic leaves and Koch's postulates were completed by confirming the identity of the isolates. To our knowledge, this is the first report of S. minor on lettuce in Serbia. More research is needed to better understand this disease, establish control strategies and minimize the spread of the pathogen to other lettuce-producing areas of the country.

Detection and characterization of bopiviruses in domestic and wild ruminants.

Highly divergent picornaviruses (PVs) classified in the genus Bopivirus have been recently discovered on faecal samples from sheep and goats in Hungary and from fallow and red deer in Australia. In this study, we investigated the epidemiology of these novel viruses in domestic and wild ruminants from Northwestern Italian Alps by testing archival faecal samples collected from 128 sheep, 167 goats, 61 red deer (Cervus elaphus), 77 roe deer (Capreolus capreolus), 43 chamois (Rupicapra rupicapra) and 32 Alpine ibex (Capra ibex). Bopivirus RNA was detected in a total of 19 animals, including 14 sheep (10.9%), 2 red deer (3.3%), 1 roe deer (1.3%), 1 chamois (2.3 %) and 1 Alpine ibex (3.3 %), but not in goats. Upon sequence analysis of the 3DRdRp region, the sequences generated from chamois, roe deer, Alpine ibex and ovine faecal samples showed the highest nucleotide identity (96.8-100%) to bopiviruses detected in goats and sheep from Hungarian farms, whereas strains found in red deer displayed the closest relatedness (90.8%-91.2%) to bopiviruses identified in fallow and red deer in Australia. The nearly complete genome sequence of strains 12/2020/ITA (ON497046) and 14-73/2020/ITA (ON497047) detected in an Alpine ibex and in a sheep, respectively, was determined by combining a modified 3'-RACE protocol with Oxford Nanopore Technologies sequencing platform. On phylogenetic analysis based on the complete polyprotein, both strains segregated into the candidate species Bopivirus B along with ovine and caprine strains detected in Hungary (90.0-94.6% nucleotide and 94.6-98.0% amino acid identities). The findings of this study expand the host range of these novel viruses and hint to a possible virus circulation between domestic ruminants and wild animals.

Molecular Detection and Genotyping of Enterocytozoon bieneusi in Pigs in Shanxi Province, North China.

Enterocytozoon bieneusi is a common opportunistic intestinal pathogen that can cause acute diarrhea in immunosuppressed humans and animals. Though E. bieneusi has been widely detected in pigs around the world, little is known of its prevalence and genotype distribution in pigs in Shanxi province, north China. In this study, a total of 362 fecal samples were collected from pigs in three representative counties in north, south, and central Shanxi province, China. The prevalence and genotypes of E. bieneusi were investigated by nested PCR amplification of the ribosomal internal transcribed spacer (ITS) region of the ribosomal RNA (rRNA) gene. Overall, the prevalence of E. bieneusi in pigs in Shanxi province was 54.70% (198/362). Statistical analysis showed the difference in prevalence was statistically significant between regions (χ2 = 41.94, df = 2, P < 0.001) and ages (χ2 = 80.37, df = 1, P < 0.001). In addition, 16 genotypes of E. bieneusi were identified in this study by sequence analysis of the ITS region, including 15 known genotypes (EbpC, EbpA, EbpB, pigEb4, PigEBITS5, I, Henan-I, G, WildBoar 7, SH10, EbpD, CHC5, PigSpEb1, PigSpEb2, and CHG19) and one novel genotype (designated as PigSX-1). Phylogenetic analysis revealed that 14 known genotypes and the novel genotype were clustered into Group 1, whereas genotype I belonged to Group 2. To the best of our knowledge, this is the first report on the prevalence and genotypes of E. bieneusi in pigs in Shanxi province. These findings enrich the genetic diversity of E. bieneusi and provide the baseline data for the prevention and control of E. bieneusi in pigs in the study regions.

Cambisol Mycobiome in a Long-Term Field Experiment with Korean Pine as a Sole Edificator: A Case Study

A culture-independent mycobiome survey in Haplic Cambisol under Korean pine in a long-term field experiment in the Russian Far East was conducted using sequence analysis of the ITS region amplified with ITS3/ITS4 primers using the metagenomic DNA as a matrix. Overall 758 fungal OTUs were identified, representing 15 phyla, 47 classes, 104 orders, 183 families, and 258 genera. More OTUs represented the Ascomycota phylum (513) than Basidiomycota (113), with both phyla together comprising 95% of the relative abundance. The Leotiomycetes class was ultimately prevailing; apparently contributing significantly to the organic matter decomposition and microbial biomass in soil, as shown by a PCA. Only two dominant OTUs (Pseudogymnoascus sp. and Hyaloscyphaceae, both Ascomycota) were common in the studied samples. The presented high mycobiome diversity in soil under the monospecies artificial forest, where Korean pine had been the sole edificator for forty years, allows concluding that plant chemistry diversity is the main factor shaping the soil mycobiome in such an environment. The obtained data provide a reference for further studies of soil mycobiota, especially under Korean pine with its aesthetic, as well as nut-producing, potential. The results can be helpful in the targeted creating of a soil mycobiome beneficial for pines in afforestation and remediation contexts.

First report of Globisporangium attrantheridium causing root rot of Gynostemma pentaphyllum

Gynostemma pentaphyllum, also known as Jiao-Gu-Lan, is a perennial liana belonging to the family Cucurbitaceae. This plant has been used as a traditional folk medicine for the treatment of various human diseases. In May 2020, G. pentaphyllum cultivated at the Research Station of Medicinal Plants at Tam Dao, Vinh Phuc, Vietnam was affected severely by root rot disease, resulting in leaf yellowing, wilting, and death. Based on morphology and sequence analysis of the ITS region, the pathogen was identified as Globisporangium attrantheridium. Koch’s postulates were fulfilled. This is the first report of Gl. attrantheridium as a pathogen causing root rot on G. pentaphyllum. This finding will be of importance for effective disease management.

Characterization of some fungal pathogens causing anthracnose disease on yam in Cross River State, Nigeria.

Yam anthracnose is one of the most serious fungal diseases affecting white and water yam production. Screening of available landraces for new sources of durable resistance to the pathogen is a continuous process. In the present study, the pathogens causing anthracnose in Dioscorea alata and Dioscorea rotundata farms in Cross River State yam belt region were characterized. Diseased yam leaves with anthracnose symptoms collected from the farms were used in the isolation, purification and, identification of C. alatae strains using morphological, cultural, and molecular methods. Leaf chlorosis, leaf edge necrosis, blights, dark brown to black leaf spots, shot holes, necrotic vein banding and vein browning were the predominantly observed symptoms. Seven isolates of C. alatae, Ca5, Ca14, Ca16, Ca22, Ca24, Ca32 and Ca34, and one isolate of Lasidioplodia theobromae, Lt1 were found to be associated with yam infection in Cross River State, with Lt1 as the most prevalent, occurring in all the locations. These isolates were classified into four forms which included the slow-growing grey (SGG), the fast-growing grey (FGG), the fast-growing salmon (FGS), and the fast-growing olive (FGO). Sequence analysis of the ITS region revealed <80% nucleotide identity between the isolates and the reference C. gloeosporioides. Pathogenicity test showed that all the isolates displayed typical symptoms of anthracnose disease as were observed in the field, but Lt1 was the most virulent. Inoculation of 20 D. alata and 13 D. rotundata landraces with isolate Lt1, showed that 63.64% of the landraces were susceptible while 36.36%were resistant. D. alata landraces were the most susceptible. This study revealed that anthracnose is prevalent and may assume an epidemic dimension in the yam growing communities of the state. There is need for increased effort in the breeding of yam for anthracnose resistance.

Functional Variants Associated With CMPK2 and in ASB16 Influence Bovine Digital Dermatitis.

Bovine digital dermatitis (BDD) is an infectious disease of the hoof in cattle with multifactorial etiology and a polygenic influence on susceptibility. With our study, we identified genomic regions with the impact on occurrence and development of BDD. We used 5,040 genotyped animals with phenotype information based on the M-stage system for genome-wide association. Significant associations for single-nucleotide polymorphisms were found near genes CMPK2 (chromosome 11) and ASB16 (chromosome 19) both being implicated in immunological processes. A sequence analysis of the chromosomal regions revealed rs208894039 and rs109521151 polymorphisms as having significant influence on susceptibility to the disease. Specific genotypes were significantly more likely to be affected by BDD and developed chronic lesions. Our study provides an insight into the genomic background for a genetic predisposition related to the pathogenesis of BDD. Results might be implemented in cattle-breeding programs and could pave the way for the establishment of a BDD prescreening test.

Characterization of infectious laryngotracheitis virus isolates from laying hens during 2019-2020 outbreaks in Tamil Nadu, India.

Infectious laryngotracheitis (ILT) is an acute respiratory disease in chickens that is a serious threat to poultry-producing countries worldwide. In the present study, we isolated and characterized infectious laryngotracheitis (ILTV) virus isolates by sequencing and restriction fragment length polymorphism analysis of PCR-amplified products (PCR-RFLP). A total of 26 ILTV outbreaks were investigated that occurred between 2019 and 2020 in flocks that had not been vaccinated against ILTV. ILTV was isolated by cultivating tracheal samples in embryonated chicken eggs, which showed multiple opaque pock lesions and thickening of the chorioallantoic membrane after 120 hours of infection. The ILTV isolates were identified and characterized by PCR and sequencing a portion of the ICP4 and TK genes. Phylogenetic analysis based on the ICP4 region showed that the sequences clustered with chicken-embryo-origin vaccine-like strains. Sequence analysis of the ICP4 region differentiated chicken-embryo-origin (CEO), tissue-culture-origin (TCO), and field ILTV strains, with significant differences in nucleotide and amino acid sequences. Furthermore, PCR-RFLP analysis of the TK gene showed that the patterns were identical to those obtained with low-virulence and vaccine strains. In conclusion, sequencing of a portion of the ICP4 region of ILTV allowed differentiation of ILTV field, CEO, and TCO vaccine strains. In this study, CEO-vaccine-like strains were found to be the cause of ILTV outbreaks between 2019 and 2020 in Tamil Nadu in southern India.