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Item Temporal reconstruction of a Salmonella Enteritidis ST11 outbreak in New Zealand(Microbiology Society, 2025-10-30) Strydom H; Wright J; Bromhead C; Welch D; Williams E; Mulqueen K; de Ligt J; Biggs PJ; Paine S; Jefferies S; French NOutbreaks caused by Salmonella Enteritidis are commonly linked to eggs and poultry meat internationally, but this serovar had never been detected in Aotearoa New Zealand (NZ) poultry prior to 2021. Locally designated genomic cluster Salmonella Enteritidis_2019_C_01, was implicated in a 2019 outbreak associated with a restaurant in Auckland. Four Enteritidis_2019_C_01 sub-clusters have since been identified, two retrospectively, in the Auckland region. Authorities initiated a formal outbreak investigation after genomically indistinguishable S. Enteritidis was isolated from the NZ poultry production environment. This study analysed 231 S. Enteritidis genomes obtained from the outbreak using Bayesian phylodynamic tools to gain insight into the outbreak's dynamics and origin. We used Bayesian integrated coalescent epoch plots to estimate the change of the Enteritidis ST11 population size over time and marginal structured coalescent approximation to estimate transmission between poultry producers. We investigated human and poultry isolates to elucidate the time and location of the most recent common ancestor of the outbreak and transmission pathways. The median most recent common ancestor was estimated to be February 2019. We found evidence of amplification and spread of strain Enteritidis_2019_C_01 within the poultry industry, as well as transmission events throughout the production chain. The intervention by the public health and food safety authorities coincided with a drop in the effective population size of the S. Enteritidis ST11 as well as notified human cases. This information is crucial for understanding and preventing the transmission of S. Enteritidis in NZ poultry to ensure poultry meat and eggs are safe for consumption.Item Longitudinal survey investigating vectors and reservoirs for Campylobacter colonization of chickens on a New Zealand broiler poultry farm(American Society for Microbiology, 2025-09-17) Kingsbury JM; French N; Midwinter A; Lucas R; Callander M; Hird CP; Smith S; Mulqueen K; Biggs R; Biggs PJ; Ercolini DThis longitudinal survey followed the life cycle of a New Zealand broiler flock to investigate sources of flock colonization by Campylobacter. Samples were collected at frequent intervals from potential Campylobacter reservoirs and sources, transmission routes for Campylobacter ingress into the broiler shed, and to monitor flock colonization. Of the 738 samples, 200 (27%) tested positive for Campylobacter. Campylobacter species from sample isolates included 316 Campylobacter jejuni, 39 Campylobacter coli, and 8 Campylobacter lari isolates; only C. jejuni was isolated from chickens. C. jejuni isolates (n = 199) were sequenced and consisted of seven sequence types (STs); the most abundant was ST6964 (105 isolates). Most flock isolates were ST6964 (44 isolates) or ST50 (27 isolates). ST6964 isolates closely matched those from the previous flock and another age-matched flock on the same farm, supporting a role for an on-farm reservoir contaminating flocks. There were six STs from catching crew and equipment isolates; the most prevalent were ST6964 (19 isolates) and ST50 (21 isolates). The close genetic match, high Campylobacter prevalence in catching samples (59/130, 45%), and the timing of flock colonization occurring closely following catcher presence in the shed support that catchers and equipment might also contaminate the shed and flock from prior flocks that they visited. There was no evidence for wildlife, feed, drinking water, breeder flock, or shed litter as sources of the Campylobacter genotypes colonizing the flock. Taken together, this study identified key areas where the poultry industry might focus on-farm risk management practices to reduce colonization of broiler flocks by Campylobacter.IMPORTANCECampylobacteriosis is the most frequently notified enteric disease in New Zealand, and New Zealand has one of the highest rates of campylobacteriosis among industrialized countries. Reducing Campylobacter colonization of poultry at the farm level would reduce reliance on processing interventions for reducing Campylobacter contamination of broiler meat. This study aimed to identify on-farm sources of Campylobacter contamination in New Zealand broiler chicken flocks. No evidence was found that wildlife, chicken feed, drinking water, or parent breeder flocks were contaminating sources. Instead, carryover of Campylobacter from the previous flock or other farm flocks, and/or contamination from chicken catching crews and their equipment, may have contributed Campylobacter strains that colonized the study flock. These are key areas where the poultry industry might focus on-farm risk management practices to reduce colonization of broiler flocks by Campylobacter.Item Genomic analysis of the 2017 Aotearoa New Zealand outbreak of Mycoplasma bovis and its position within the global population structure(Frontiers Media S.A., 2025-07-23) Binney BM; Gias E; Foxwell J; Little A; Biggs PJ; French N; Lambert CL; Ha HJ; Carter GP; Gyuranecz M; Pardon B; De Vliegher S; Boyen F; Bokma J; Krömker V; Wente N; Mahony TJ; Gibson JS; Barnes TS; Wawegama N; Legione AR; Heller M; Schnee C; Pelkonen S; Autio T; Higuchi H; Gondaira S; McCulley M; Cloeckaert AIn 2017 an outbreak of Mycoplasma bovis (M. bovis), an infectious agent of cattle, was identified in Aotearoa New Zealand. This study characterizes the genomic population structure of the outbreak in New Zealand and compares it with the known global population structure using multilocus sequence typing (MLST) and genomic analysis. The New Zealand outbreak strain was MLST genotyped as ST21. A comprehensive collection of 840 genomes from the New Zealand outbreak showed a pattern of clonal expansion when characterized by MLST, core genome MLST (cgMLST) and whole genome MLST (wgMLST). A lineage of genomes was found with no in silico identifiable pta2 locus, a housekeeping gene used in the MLST scheme. We compared a sample set of 40 New Zealand genomes to 47 genomes from other countries. This group had 79 ST21 genomes and eight genomes that were single nucleotide polymorphism (SNP) variants within the MLST loci of ST21. Two of the 47 international genomes showed signs of extensive unique recombination. Unique alleles in six genes were identified as present only in the New Zealand genomes. These novel variants were in the genes; haeIIIM encoding for cytosine-specific methyltransferase, cysC encoding for cysteinyl tRNA synthetase, era encoding for GTPase Era, metK encoding for S-adenosylmethionine synthase, parE encoding for DNA topoisomerase, and hisS encoding for histidine-tRNA ligase. This finding could be due to a population bottleneck, genetic drift, or positive selection. The same sample set of 40 New Zealand genomes were compared using MLST to 404 genomes from 15 other countries and 11 genomes without a known country. A FastBAPS analysis of 455 genomes showed a global population structure with 11 clusters. Some countries, such as Canada, Denmark and Australia contained both internally closely related genomes and some genomes that were more closely related to genomes found in other countries. Our results support the need for Whole Genome Sequencing (WGS) as well as MLST genotyping in M. bovis outbreaks. They also support the importance of understanding the national and international movement patterns of cattle and their genetic material, as possible routes of transmission, when managing the spread of M. bovis.Item Heritability estimates and genetic and phenotypic correlations of skin thickness and skin temperature with key production traits in FocusPrime, Texel, Romney and Highlander sheep(Oxford University Press on behalf of the American Society of Animal Science., 2025-06-16) Graña-Baumgartner A; Dukkipati VSR; Biggs PJ; Kenyon PR; Blair HT; Pickering NK; Van der Linden DS; López-Villalobos NSkin thickness was found to be moderately heritable and genetically associated with lamb survival in a previous study on Romney sheep. The aims of this study were to estimate the heritabilities of skin thickness and skin temperature at around five and 11 months of age, and determine genetic and phenotypic correlations between them and with production traits such as fat depth, loin-eye muscle depth and width, live weights at weaning, scanning, and 12 months, and 12-month fleece weight, in FocusPrime (n=2,088), Texel (n=732), Romney (n=825) and Highlander (n=1,801) sheep breeds. Heritability estimates of skin thickness at 5-month old were moderate in FocusPrime (0.39 ± 0.12) and low in Texel and Highlander (0.11 ± 0.15 and 0.13 ± 0.09, respectively). Heritability estimates of skin thickness at 11-month old were moderate in all breeds (ranged from 0.19 ± 0.07 to 0.29 ± 0.15). Heritability estimates of skin temperature were high in FocusPrime (0.39 ± 0.11), low in Texel (0.17 ± 0.11) and Highlander (0.12 ± 0.06) and almost zero in Romney (0.04 ± 0.03). A tendency in all breeds for negative and favourable correlations was found between skin thickness and skin temperature at 11-month old sheep. Skin thickness at 11-months tended to have a positive genetic correlation with fat depth in all breeds except in Texel where the correlation tended to be negative (-0.10 ± 0.34). Genetic correlations of skin thickness at 11-month old with the weight traits were variate. There tended to be a positive correlation with weaning weight in Texel (0.14 ± 0.34) and Highlander (0.29 ± 0.22). However, there tended to be negative correlations with live weight at scanning and at 12-month of age in FocusPrime (-0.03 ± 0.18 and -0.13 ± 0.22 , respectively) and tended to be positive in Romney (0.09 ± 0.25 and 0.10 ± 0.24, respectively) and Highlander (0.26 ± 0.22 and 0.39 ± 0.21, respectively). Moreover, genetic correlations of skin thickness at 11-month of age with FW12 tended to be positive in both Romney (0.20 ± 0.22) and Highlander (0.55 ± 0.19). Further studies on the genetic correlations of skin thickness and skin temperature with lamb survival in these breeds are warranted.Item Evidence for a Role of Extraintestinal Pathogenic Escherichia coli, Enterococcus faecalis and Streptococcus gallolyticus in the Aetiology of Exudative Cloacitis in the Critically Endangered Kākāpō (Strigops habroptilus)(John Wiley and Sons Ltd, 2025-04-19) French RK; Waller SJ; Wierenga JR; Grimwood RM; Hodgkinson-Bean J; Digby A; Uddstrom L; Eason D; Kākāpō Recovery Team; Argilla LS; Biggs PJ; Cookson A; French NP; Geoghegan JLThe kākāpō is a critically endangered flightless parrot which suffers from exudative cloacitis, a debilitating disease resulting in inflammation of the vent margin or cloaca. Despite this disease emerging over 20 years ago, the cause of exudative cloacitis remains elusive. We used total RNA sequencing and metatranscriptomic analysis to characterise the infectome of lesions and cloacal swabs from nine kākāpō affected with exudative cloacitis, and compared this to cloacal swabs from 45 non-diseased kākāpō. We identified three bacterial species—Streptococcus gallolyticus, Enterococcus faecalis and Escherichia coli—as significantly more abundant in diseased kākāpō compared to healthy individuals. The genetic diversity observed in both S. gallolyticus and E. faecalis among diseased kākāpō suggests that these bacteria originate from exogenous sources rather than from kākāpō-to-kākāpō transmission. The presence of extraintestinal pathogenic E. coli (ExPEC)-associated virulence factors in the diseased kākāpō population suggests that E. coli may play a critical role in disease progression by facilitating iron acquisition and causing DNA damage in host cells, possibly in association with E. faecalis. No avian viral, fungal nor other parasitic species were identified. These results, combined with the consistent presence of one E. coli gnd sequence type across multiple diseased birds, suggest that this species may be the primary cause of exudative cloacitis. These findings shed light on possible causative agents of exudative cloacitis, and offer insights into the interplay of microbial factors influencing the disease.Item The Host Adaptation of Staphylococcus aureus to Farmed Ruminants in New Zealand, With Special Reference to Clonal Complex 1(John Wiley and Sons Ltd, 2025-06) Nesaraj J; Grinberg A; Laven R; Chanyi R; Altermann E; Bandi C; Biggs PJGenetic features of host adaptation of S. aureus to ruminants have been extensively studied, but the extent to which this adaptation occurs in nature remains unknown. In New Zealand, clonal complex 1 (CC1) is among the most common lineages in humans and the dominant lineage in cattle, enabling between-, and within-CC genomic comparisons of epidemiologically cohesive samples of isolates. We assessed the following genomic benchmarks of host adaptation to ruminants in 277 S. aureus from cattle, small ruminants, humans, and pets: 1, phylogenetic clustering of ruminant strains; 2, abundance of homo-specific ruminant-adaptive factors, and 3, scarcity of heterospecific factors. The genomic comparisons were complemented by comparative analyses of the metabolism of carbon sources that abound in ruminant milk. We identified features fulfilling the three benchmarks in virtually all ruminant isolates, including CC1. Data suggest the virulomes adapt to the ruminant niche sensu lato accross CCs. CC1 forms a ruminant-adapted clade that appears better equipped to utilise milk carbon sources than human CC1. Strain flow across the human–ruminant interface appears to only occur occasionally. Taken together, the results suggest a specialisation, rather than mere adaptation, clarifying why zoonotic and zoo-anthroponotic S. aureus transmission between ruminants and humans has hardly ever been reported.Item Genomic epidemiology of ESBL-producing Escherichia coli from humans and an Aotearoa New Zealand river(2024-05-05) Gray HA; Biggs PJ; Midwinter AC; Rogers LE; Fayaz A; Akhter RN; Burgess SAItem Antibiotic and Heavy Metal Resistance in Bacteria from Contaminated Agricultural Soil: Insights from a New Zealand Airstrip(MDPI (Basel, Switzerland), 2025-02) Heydari A; Kim ND; Biggs PJ; Horswell J; Gielen GJHP; Siggins A; Bromhead C; Meza-Alvarado JC; Palmer BR; Abia ALKBACKGROUND/OBJECTIVES: Agricultural soils accumulate inorganic contaminants from the application of phosphate fertilisers. An airstrip located at Belmont Regional Park (BRP), near Wellington, New Zealand, has been found to have a gradient of cadmium contamination due to spillage of superphosphate fertiliser. METHODS: Soil samples from the BRP airstrip with a gradient of cadmium contamination, were used as a novel source to explore bacterial communities' resistance to heavy metals (HMs) and any co-selected antibiotic (Ab) resistance. RESULTS: Differences between BRP soil samples with higher levels of HMs compared to those with lower HM concentrations showed significantly more bacterial isolates resistant to both HMs (40.6% versus 63.1% resistant to 0.01 mM CdCl2, p < 0.05) and Abs (23.4% versus 37.8% resistant to 20 μg/mL tetracycline, p < 0.05) in soils with higher initial levels of HMs (1.14 versus 7.20 mg kg-1 Cd). Terminal restriction fragment length polymorphism (TRFLP) and 16S rDNA next-generation sequencing profiling investigated changes in HM-induced bacterial communities. Significant differences were observed among the bacterial community structures in the selected BRP soil samples. Conjugative transfer of cadmium resistance from 23-38% of cadmium-resistant isolates to a characterised recipient bacterial strain in vitro suggested many of these genes were carried by mobile genetic elements. Transconjugants were also resistant to zinc, mercury, and Abs. Higher levels of HMs in soil correlated with increased resistance to HMs, Abs, and elevated levels of HMs thus disturbed the bacterial community structure in BRP soil significantly. CONCLUSIONS: These findings suggest that HM contamination of agricultural soil can select for Ab resistance in soil bacteria with potential risks to human and animal health.Item Metabarcoding captures genetic diversity and links cases in outbreaks of cryptosporidiosis in New Zealand.(Elsevier Ltd on behalf of The British Infection Association, 2025-01-30) Ogbuigwe P; Biggs PJ; Garcia-Ramirez JC; Knox MA; Pita A; Velathanthir N; French NP; Hayman DTSCryptosporidiosis is a disease caused by the parasite Cryptosporidium. Globally, it is a leading cause of diarrhoea and a notifiable disease in New Zealand. Molecular analyses of Cryptosporidium isolated from notified cases do not always provide support for epidemiological links between individuals. We hypothesised this could be due to undetected diversity and the use of consensus Sanger sequence analyses. Here, we analysed 105 Cryptosporidium samples from outbreaks and sporadic cases occurring between 2010 and 2018 in New Zealand using both Next-Generation Sequencing (NGS) and Sanger sequencing of the glycoprotein 60 (gp60) locus. NGS metabarcoding at the gp60 locus uncovered significant intra- and inter-sample genotypic diversity in outbreaks and identified subtypes shared by epidemiologically linked cases, along with rare subtypes, suggesting it may be a useful tool for epidemiological investigations.Item Genomic epidemiology of extended-spectrum beta-lactamase-producing Escherichia coli from humans and a river in Aotearoa New Zealand.(Microbiology Society, 2025-01-10) Gray HA; Biggs PJ; Midwinter AC; Rogers LE; Fayaz A; Akhter RN; Burgess SAIn Aotearoa New Zealand, urinary tract infections in humans are commonly caused by extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli. This group of antimicrobial-resistant bacteria are often multidrug resistant. However, there is limited information on ESBL-producing E. coli found in the environment and their link with human clinical isolates. In this study, we examined the genetic relationship between environmental and human clinical ESBL-producing E. coli and isolates collected in parallel within the same area over 14 months. Environmental samples were collected from treated effluent, stormwater and multiple locations along an Aotearoa New Zealand river. Treated effluent, stormwater and river water sourced downstream of the treated effluent outlet were the main samples that were positive for ESBL-producing E. coli (7/14 samples, 50.0%; 3/6 samples, 50%; and 15/28 samples, 54%, respectively). Whole-genome sequence comparison was carried out on 307 human clinical and 45 environmental ESBL-producing E. coli isolates. Sequence type 131 was dominant for both clinical (147/307, 47.9%) and environmental isolates (11/45, 24.4%). Only one ESBL gene was detected in each isolate. Among the clinical isolates, the most prevalent ESBL genes were bla CTX-M-27 (134/307, 43.6%) and bla CTX-M-15 (134/307, 43.6%). Among the environmental isolates, bla CTX-M-15 (28/45, 62.2%) was the most prevalent gene. A core SNP analysis of these isolates suggested that some strains were shared between humans and the local river. These results highlight the importance of understanding different transmission pathways for the spread of ESBL-producing E. coli.
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