Browsing by Subject "virulence"

Onion (Allium cepa L.) is an important vegetable, which provides major nutritional benefits. In Finland, basal rot caused by Fusarium fungal species (FBR) has become the most destructive disease of onion in recent years. It causes damping off and stunted growth on onion seedlings and root death and abscission and bulb rot on mature onions. The average onion crop loss caused by basal rot has been up to 10% in conventional farming and as high as 30% in organic farming. In this study, a seedling assay was conducted to test the virulence of different Fusarium isolates on a commercial onion cultivar. The study goals were, primarily, to find out which of the Fusarium isolates, originating from onion and crop rotation plants grown in Finland, are pathogenic, and secondly, to compare the virulence of different isolates. Altogether 115 Fusarium isolates were tested for virulence on onion seedlings in a greenhouse. Fifty-five of the tested isolates of F. oxysporum, F. proliferatum and F. redolens were more or less virulent. 19.4% of F. oxysporum isolates, 35.3% of F. proliferatum isolates and 18.2% of F. redolens isolates showed high virulence. Among the aggressive pathogens isolated from onion samples, F. oxysporum f.sp. cepae is still the dominant onion pathogen, F. proliferatum is a new pathogen on onion in Finland, and relatively more aggressive than F. oxysporum. The F. solani and F. tricinctum isolates tested did not have any detrimental effects on the onion seedling health or growth. In order to control FBR in Finland, avoiding planting onion in the infested soils, growing onions from local seedlings to avoid new contamination and storing onions at cold temperatures are recommended.

Fusarium proliferatum has recently become a major threat to onion, which is an important food crop for food security and has a significant role in the agricultural sector. This fungus is found causing rots and producing mycotoxin fumonisin that, if ingested, can cause carcinogenic effects in humans and fatal diseases in animals. F. proliferatum has been identified as a pathogen causing rots and wilts in many plants in several countries, and recently some isolates of this pathogen were also found in Finland causing basal rot in onion. Though F. proliferatum has wide adaptability and pathogenicity, there are research gaps on this newly emerging pathogen, which is mostly limited to some specific hosts such as maize. There is a lack of knowledge of its infection mechanisms and mycotoxin production dynamics in onion. This experimental work was conducted at the University of Helsinki, Finland, to study the nature of pathogenicity and toxin gene expression of F. proliferatum in laboratory conditions as a function of time. Spore suspensions of three isolates of F. proliferatum, Fpr047, Fpr049, and Fpr919, were prepared and used as inoculums that were injected into healthy organically grown onions, which were then stored in dark for five weeks. Control bulbs were inoculated with sterile water. Pathogen virulence, based on symptom development, and fungal colonization in the onion tissues were determined at five different time points and the toxin gene expression was determined at three time points. Colonization levels were determined by real-time PCR using primers binding to the intergenic spacer (IGS) region of F. proliferatum. A part of the IGS region of the three isolates was sequenced to study the diversity between the isolates. All the tested isolates were found to be virulent, and they colonized the onions after one week from inoculation. However, the isolate Fpr919 appeared different from the other two in terms of symptom severity. It was more aggressive than the other two, causing disease symptoms earlier and causing more severe rot symptoms in the infected bulbs. It also had nucleotide sequence variations in the IGS region in comparison with the other two isolates, suggesting genetic diversity. No significant differences were observed between the isolates in the fungal colonization levels. Expression of FUM1 gene and a putative virulence gene SIX2-1 was detected by RT-RT-PCR in most of the infected tissue samples. The results obtained signify that F. proliferatum is a pathogen with the potential of producing fumonisin toxin in onion, suggesting the need for further molecular study on this fungus to control the disease and prevent mycotoxin contamination in plant products.

Constantly increasing level of bacteria becoming resistant to clinically relevant antibiotics challenges the modern medical achievements made over the past century. In global scale, one of the most significant information gaps concerning the occurrence of resistant bacteria is located in West African countries. Klebsiella pneumoniae and Escherichia coli strains resistant to 3rd generation cephalosporins and carbapenems are a major risk to public health through infections with limited or no available treatment options. The resistance to these antibiotics among Enterobacteriaceae is mainly mediated by hydrolyzing enzymes such as extended-spectrum beta-lactamases (ESBL). The focus of this thesis is to study the genes encoding these enzymes and other resistance factors found in K. pneumoniae and E. coli isolated from human stool and waste water samples in Burkina Faso and Mali. Tree Enterobacteriaceae isolates were selected for whole genome sequence (WGS) analysis based on their phenotypic resistance profiles defined by disk diffusion method. Reads were assembled to draft genomes and the genomes were studied for their antibiotic resistance genes, virulence genes and their associations to mobile genetic elements found in these isolates’ genomes. Additionally a pan-genome was created to investigate species specific features of K. pneumoniae and their role in heavy load of antibiotic resistance genes among these isolates. Pan-genome consisted of two genomes sequenced in this study and 12 genomes from the publically available database. 16-month old Burkinabe child was a carrier of one ESBL-producing K. pneumoniae (isolate Burkina_1) and one ESBL-positive E. coli along with the resistance to multiple other antibiotics. With genome wide analysis the K. pneumoniae strain could be described as sequence type (ST) 45 representing, multidrug resistant and ESBL-gene CTX-M-15 carrying strain with highly similar virulence gene profile to strains previously described as pathogenic K. pneumoniae causing neonatal sepsis. K. pneumoniae isolated from the stool sample of an adult living in Burkina Faso was found to be multidrug resistant, though non-ESBL-producer strain (isolate Burkina_2). The isolate showed no similarity to any previously described sequence type. CTX-M-15 encoding E. coli of ST38 (isolate Mali_1) carried by Malian child showed resistance to five different classes of antibiotics in addition to the 3rd generation cephalosporins. At the same time the isolate showed hybrid virulence gene profile with virulence genes associated to many different E. coli pathotypes including neonatal meningitis causing E. coli (NMEC). The exceptional plasticity of K. pneumoniae genome could be recognized as one of the putative explanations for the high number of resistance genes found among the isolates studied in this work. Antibiotic resistance genes were found to be associated to mobile genetic elements (MGE) and as the genetic plasticity is caused by the acquisition of external genetic material via MGEs such as plasmids, this can lead to indirect accumulation of resistance genes in these genomes. The results in this thesis work show alarming examples of pathogens that potentially cause severe infections, have extremely narrow or no treatment options and are carried by infants. These findings are in line with the few data about the level of faecal carriage of ESBL-producing strains by people in Burkina Faso and Mali reported previously.