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APPLICATION FORM DETERMINATION Determine if an organism is a new organism under the Hazardous Substances and New Organisms Act 1996 Send by post to: Environmental Protection Authority, Private Bag 63002, Wellington 6140 OR email to: [email protected] Application number APP201895 Applicant Neil Pritchard Key contact NPN Ltd www.epa.govt.nz 2 Application to determine if an organism is a new organism Important This application form is used to determine if an organism is a new organism. If you need help to complete this form, please look at our website (www.epa.govt.nz) or email us at [email protected]. This application form will be made publicly available so any confidential information must be collated in a separate labelled appendix. The fee for this application can be found on our website at www.epa.govt.nz. This form was approved on 1 May 2012. May 2012 EPA0159 3 Application to determine if an organism is a new organism 1. Information about the new organism What is the name of the new organism? Briefly describe the biology of the organism. Is it a genetically modified organism? Pseudomonas monteilii Kingdom: Bacteria Phylum: Proteobacteria Class: Gamma Proteobacteria Order: Pseudomonadales Family: Pseudomonadaceae Genus: Pseudomonas Species: Pseudomonas monteilii Elomari et al., 1997 Binomial name: Pseudomonas monteilii Elomari et al., 1997. Pseudomonas monteilii is a Gram-negative, rod- shaped, motile bacterium isolated from human bronchial aspirate (Elomari et al 1997). They are incapable of liquefing gelatin. They grow at 10°C but not at 41°C, produce fluorescent pigments, catalase, and cytochrome oxidase, and possesse the arginine dihydrolase system. They are capable of respiratory but not fermentative metabolism. (Masua et al 2007) Rhodococcus pyridinivorans Kingdom: Bacteria Phylum: Actinobacteria Class: Actinobacteria Order: Actinomycetales Family: Nocardiaceae Genus: Rhodococcus Species: Rhodococcus pyridinivorans Yoon et al., 2000 The cells are non-spore-forming, non-motile and Gram-positive, but are Gram-variable in old cultures. Substrate mycelia that are on the surface and penetrate into the agar media are visible and fragment into short rod-to-coccus elements. The cells are rods and branched filaments during the early growth phase and then fragment into short rods or cocci. Colonies are light orange in colour, opaque and raised with slightly irregular edges on TSA. Colonies have irregularly round wrinkles. Grows well over a broad pH range (6.0-9.0); the optimal pH range is 7.5-8.5. Grows optimally at 30-37 °C; growth occurs at 10 and 45 °C but does not occur at 50 °C. Catalase- and urease- positive. Oxidase- and DNase-negative. Arbutin, aesculin, Tween 80, tyrosine and urea are hydrolysed. Casein, May 2012 EPA0159 4 Application to determine if an organism is a new organism hypoxanthine, starch and xanthine are not hydrolysed. Nitrate is reduced to nitrite. Voges- Proskauer and methyl red reactions are negative. H2S is produced. Acid is produced from d-fructose, glycerol, d-mannitol, d-mannose, d- ribose, salicin, d-sorbitol and starch. No acid is produced from larabinose, D-cellobiose, D-galactose, D-glucose, inulin, lactose, maltose, D-raffinose, L-rhamnose, trehalose or D-xylose. Acetate, citrate, fumarate and succinate are utilized. Benzoate, formate, hippurate and tartrate are not utilized. Degrades pyridine. The cell wall contains meso-diaminopimelic acid, arabinose and galactose (wall chemotype IV). The predominant menaquinone is MK- 8(H2). The major fatty acids are C16:0, C18:1 cis9, 10-methyl-C18:1 (TBSA). Mycolic acids with 36-46 carbon atoms are present. The GC content of the DNA is 66 mol% (as determined by HPLC). (Yoon et al. 2000) R. pyridinivorans has been characterized based on phylogenetic and chemotaxonomic descriptors such as G+C content, 16S rDNA sequences, and DNA-DNA relatedness. The 16S rDNA analysis revealed 99% nucleotide similarity to that of the type strain of R. rhodochrous, which is not known to be pathogenic. Paracoccus pantotrophus Kingdom: Bacteria Phylum: Proteobacteria Class: Alphaproteobacteria Order: Rhodobacterales Family: Rhodobacteraceae Genus: Paracoccus Species: Paracoccus pantotrophus (Robertson and Kuenen 1984) Rainey et al. 1999 Synonym: Thiosphaera pantotropha Some strains are capable of aerobic denitrification (simultaneous reduction of oxygen and nitrate) and heterotrophic nitrification (oxidation of ammonium to nitrite during heterotrophic growth); some strains are capable of aerobic growth on formate, aerobic chemolithoautotrophic growth with carbon disulfide or carbonyl sulfide as energy substrates, methylotrophic growth on methanol or methylated sulfides, and heterotrophic growth on diethyl sulfide, thioethanol, thioacetic acid or substituted thiophenes; some strains can grow anaerobically with denitrification on thiosulfate, carbon disulfide, methanol or formate as energy sources; some strains contain plasmids of 85-110 kb in size and megaplasmids greater than 450 kb in size; distinction from other species of Paracoccus can be confirmed by comparison of 16S rRNA gene sequence and DNA hybridization; the amino acid sequence of its cytochrome c-550 differs by about 16% from that of P. denitrificans; and the GC content of the DNA is 64-68 mol% (type strain GC content is 66 mol%). Kelly et al (2006) Nitrosomonas eutropha Kingdom: Bacteria Phylum: Proteobacteria May 2012 EPA0159 5 Application to determine if an organism is a new organism Class: Beta Proteobacteria Order: Nitrosomonadales Family: Nitrosomonadaceae Genus: Nitrosomonas Species: Nitrosomonas eutropha Koops et al., 1991 Cells tend to be pleomorphic, rod to pear-shaped (sometimes coccoid). One or both ends pointed. Cells 1.0-1.3 by 1.6-2.3 μm in size, occasionally in short chains. Cells are motile. Carboxysomes present. UtiIization of urea not observed. No salt requirement, high tolerance of increasing ammonia concentrations. The G+C content of the DNA is 47.9-48.5 mol % (Tm). The organism that is the subject of this application is also the subject of: a. an innovative medicine application as defined in section 23A of the Medicines Act 1981. Yes No b. an innovative agricultural compound application as defined in Part 6 of the Agricultural Compounds and Veterinary Medicines Act 1997. Yes No 2. What evidence do you have that the organism was present in New Zealand immediately prior to 29 July 1998? Pseudomonas monteilii The genus Pseudomonas encompasses the most diverse and ecologically significant group of bacteria on earth. Members of the genus are found in all of the major natural environments (terrestrial, freshwater and marine) and also form associations with plants and animals. This universal distribution suggests a remarkable degree of physiological and genetic adaptability (Spiers et al. 2000). Pseudomonas is a genus with widespread occurrence in water and in plant seeds such as dicots. Many members of the genus have been reported in New Zealand, including Pseudomonas putida and Pseudomonas oryzihabitans (http://nzfungi2.landcareresearch.co.nz), which are taxonomically closely related with Pseudomonas monteilii. Pseudomonas monteilii is a versatile bacterium found in various niches (Remold et al. 2011). Pseudomonas monteilii strains are associated with degradation of aromatic and heterocyclic compounds. P. monteiliiis is an aspirate found in many international hospitals, and where organisms are carried by people, it is easy to argue that they have been carried in to NZ even if they haven’t been recorded. Some Pseudomonas monteilii strains were isolated from clinical specimens, yet no P. monteilii infection has been reported. The P. monteilii strains are phenotypically and genotypically homogeneous and can be differentiated from related nonphytopathogenic fluorescent members of section I of the genus Pseudomonas. May 2012 EPA0159 6 Application to determine if an organism is a new organism The clinical significance of P. monteilii is not known. Future isolates of this new organism should be investigated to determine their role in nosocomial infections. At present, it is hypothesized that P. monteilii is a rare opportunistic pathogen or colonizer. Pseudomonas spp. are ubiquitous in the environment. As opportunistic pathogens, some Pseudomonas spp. could invade the host tissue and cause infection and bacteremia in immunocompromised hosts. P. aeruginosa is the most common agent associated with infection and inflammation during contact lens wear. Other Pseudomonas species are also opportunistic; however, cases of infection are rare. Rhodococcus pyridinivorans Members of Rhodococcus have been found to thrive in a broad range of environments, including soil, water, and eukaryotic cells. Several species of the genus have been found in New Zealand, such as Rhodococcus equi, Rhodococcus fascians, and Rhodococcus rhodochrous (http://nzfungi2.landcareresearch.co.nz) Members of the genus Rhodococcus are mostly soil saprophytes although they are common in many environments, and a large number of them are known as toxic-material decomposing strains. Rhodococcus pyridinivorans was originally isolated as an extremely efficient pyridine-degrading coryneform bacterium from industrial wastewater in Korea The species R. pyridinivorans comprises strains that are metabolically versatile and are able to degrade numerous aromatic compounds. The pathogenicity of the bacteria that belonged to Rhodococcus was investigated based on information from the literature. Previous reports revealed that R. equi and R. facians are
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