School of Science - Te Kura Pūtaiao
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Research at AUT's School of Science - Te Kura Pūtaiao is focused on key scientific issues with regional and global significance. The common theme connecting all research areas is sustainability – in the broadest sense as it relates to environmental and human health. Our research is closely allied to teaching and learning opportunities at undergraduate and postgraduate level within the school.
Research is organised in five main areas:
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Browsing School of Science - Te Kura Pūtaiao by Author "Alfaro, Andrea C"
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- ItemBehavioural and Physiological Responses of Juvenile Geoduck (Panopea zelandica) Following Acute Thermal Stress(Elsevier, 2023-08-11) Sharma, Shaneel S; Venter, Leonie; Frost, Emily J; Alfaro, Andrea C; Ragg, Norman LC; Zamora, Leonardo NClimate extremes, such as heatwaves, are expected to become more intense and of longer duration in the near future. These climatic conditions may have a significant impact on the prospects of establishing a new aquaculture industry for the endemic New Zealand geoduck, Panopea zelandica. This study focused on characterising animal behaviour, haemocyte characteristics, and heat shock protein (HSP70 & HSP90) mRNA expression following exposure to elevated temperatures, such as those encountered during marine heatwaves around 20 °C and an extreme scenario of 25 °C, contrasted to an ambient temperature of 17 °C. After 24 h of heat challenge, P. zelandica were found to be significantly influenced by the thermal changes, as there were differences recorded in all the responses examined. With increasing temperatures, juvenile geoducks were observed to fully emerge from the sediment a behaviour that has not previously been quantified nor associated with stress in this species. The ability of P. zelandica juveniles to re-bury still warrants further investigation, as adults are unable to do so. Haemocyte analyses revealed an increase in the abundance of granulocytes, cellular aggregations, and size of these aggregations at the highest temperature exposure. Increased expression of the hsp70 gene in the haemolymph after exposure at 25 °C for 24 h was detected and attributed to attempts to mitigate protein denaturation caused by thermal stress. The inter-individual variability in the response of heat shock proteins recorded could aid in future selective breeding programs if it is reflected in net thermotolerance. P. zelandica shows great potential for growing in subtidal habitats around New Zealand, and this study highlights the importance of temperature considerations when selecting potential farm and reseeding locations.
- ItemEffect of Dietary Insect Meal and Grape Marc Inclusion on Flavor Volatile Compounds and Shell Color of Juvenile Abalone Haliotis iris(Hindawi Limited, 2023-07-18) Bullon, Natalia; Alfaro, Andrea C; Hamid, Nazimah; Masoomi Dezfooli, Sara; Seyfoddin, Ali; Dawood, MahmoudAlmost 60% of the fish meal produced globally is used in aquaculture feeds. Fish meal production relies on finite wild-marine resources and is considered as an unsustainable ingredient. Insect meal (IM) is considered a sustainable source with high levels of protein suitable for growth promotion. Grape marc (GM) is a waste byproduct of the winery industry rich in pigments with antioxidant capacity. However, the inclusion of both ingredients can affect the flavor of the meat of abalone and the color of the shell due to different nutritional profiles. The aim of this study was to evaluate the effect of the dietary inclusion of IM and GM on the flavor volatile compounds and shell color of the juvenile Haliotis iris in a 165-days feeding trial. Abalone were offered four experimental diets with different levels of IM and GM inclusion and a commercial diet (no IM or GM). Soft bodies of abalone were used to characterize volatile compounds using solid-phase microextraction gas chromatography–mass spectrometry, and color changes were analyzed in ground powder of abalone shells using color spectrophotometry 400–700 nm (visible). The results showed 18 volatile compounds significantly different among the dietary treatments. The inclusion of IM did not significantly affect the flavor volatile compounds detected, whereas the inclusion of GM reduced volatile compounds associated with lipid-peroxidation in abalone meat. The inclusion of IM and GM did not significantly affect the lightness nor the yellowness, blueness, redness, and greenness of the ground shells. The supplementation of abalone feeds with GM can help to reduce off-flavour compounds which may extend shelf-life of raw abalone meat.
- ItemGut Microbiome Resilience of Green-Lipped Mussels, Perna canaliculus, to Starvation(Viguera Editores, S.L., 2023-07-31) Li, Siming; Young, Tim; Archer, Stephen; Lee, Kevin; Alfaro, Andrea CHost gut microbiomes play an important role in animal health and resilience to conditions, such as malnutrition and starvation. These host-microbiome relationships are poorly understood in the marine mussel Perna canaliculus, which experiences significant variations in food quantity and quality in coastal areas. Prolonged starvation may be a contributory factor towards incidences of mass mortalities in farmed mussel populations, resulting in highly variable production costs and unreliable market supplies. Here, we examine the gut microbiota of P. canaliculus in response to starvation and subsequent re-feeding using high-throughput amplicon sequencing of the 16S rRNA gene. Mussels showed no change in bacterial species richness when subjected to a 14-day starvation, followed by re-feeding/recovery. However, beta bacteria diversity revealed significant shifts (PERMANOVA p-value < 0.001) in community structure in the starvation group and no differences in the subsequent recovery group (compared to the control group) once they were re-fed, highlighting their recovery capability and resilience. Phylum-level community profiles revealed an elevation in dominance of Proteobacteria (ANCOM-BC p-value <0.001) and Bacteroidota (ANCOM-BC p-value = 0.04) and lower relative abundance of Cyanobacteria (ANCOM-BC p-value = 0.01) in the starvation group compared to control and recovery groups. The most abundant genus-level shifts revealed relative increases of the heterotroph Halioglobus (p-value < 0.05) and lowered abundances of the autotroph Synechococcus CC9902 in the starvation group. Furthermore, a SparCC correlation network identified co-occurrence of a cluster of genera with elevated relative abundance in the starved mussels that were positively correlated with Synechococcus CC9902. The findings from this work provide the first insights into the effect of starvation on the resilience capacity of Perna canaliculus gut microbiota, which is of central importance to understanding the effect of food variation and limitation in farmed mussels.
- ItemHistological Examination of Perna canaliculus Mussels During a Summer Mortality Event in New Zealand(Hindawi Limited, 2024-04-12) Muznebin, Farhana; Van Nguyen, Thao; Webb, Stephen C; Alfaro, Andrea C; Abdelrahman, HishamThe New Zealand Greenshell™ mussel (Perna canaliculus) is endemic to New Zealand and contributes to the success of the country’s aquaculture industry. However, summer mortality and potential disease outbreak events are having an increasing effect on the growth of this industry. The cause of these mortalities remains unknown, and histopathological studies of the pathogen and parasites in mussels are still incomplete. In the present study, a histological approach was used to identify pathogens and parasites, as well as immunological tissue responses in unhealthy- and healthy-looking P. canaliculus during a summer mortality event in 2018. A highly significant association between health conditions and the presence of Perkinsus olseni in mussels was observed. A higher prevalence of P. olseni, Apicomplexan-X (APX), and bacterial (rods and cocci) infections were noted in the unhealthy-looking mussels than in the healthy-looking mussels. In an assessment of stains, hematoxylin and eosin (H&E) staining appeared to be the best method for general pathological and anatomical characterization, while Giemsa provided the clearest visual definition of bacteria. In this aspect, it was comparable to Ziehl–Neelsen (ZN) in apparent sensitivity. Although Gram and ZN staining revealed bacterial cells marginally better than with H&E, their differential staining could not be assessed as no Gram-positive or acid-fast bacteria were seen, and no mussel-positive controls were available for comparison. This study also provides an illustrated guide to some significant mussel health indicators.
- ItemHistopathological Changes in the Greenshell Mussel, Perna canaliculus, in Response to Chronic Thermal Stress(Elsevier, 2023-09-01) Copedo, Joanna S; Webb, Stephen C; Ragg, Norman LC; Ericson, Jessica A; Venter, Leonie; Schmidt, Alfonso J; Delorme, Natalí J; Alfaro, Andrea CClimate change associated temperature challenges pose a serious threat to the marine environment. Elevations in average sea surface temperatures are occurring and increasing frequency of marine heatwaves resulting in mortalities of organisms are being reported. In recent years, marine farmers have reported summer mass mortality events of the New Zealand Greenshell mussel, Perna canaliculus, during the summer months; however, the etiological agents have yet to be determined. To elucidate the role of thermal stress, adult P. canaliculus were exposed to three chronic temperature treatments: a benign control of 17 °C and stressful elevations of 21 °C and 24 °C. Eight mussels per treatment were collected each month throughout a 14-month challenge period to identify and investigate histopathological differences among P. canaliculus populations exposed to the three temperatures. Histopathology revealed several significant deleterious alterations to tissues associated with temperature and exposure time. Increasing temperature and progression of time resulted in 1) an increase in the number of focal lipofuscin-ceroid aggregations, 2) an increase in focal hemocytosis, 3) an increase in the thickness of the sub-epithelial layer of the intestinal tract and 4) a decreased energy reserve cell (glycogen) coverage in the mantle. Prolonged exposure, irrespective of temperature, impacted gametogenesis, which was effectively arrested. Furthermore, increased levels of the heat shock protein 70 kDa (HSP 70) were seen in gill and gonad from thermally challenged mussels. The occurrence of the parasite Perkinsus olseni at month 5 in the 24 °C treatment, and month 7 at 21 °C was unexpected and may have exacerbated the fore-mentioned tissue conditions. Prolonged exposure to stable thermal conditions therefore appears to impact P. canaliculus, tissues with implications for broodstock captivity. Mussels experiencing elevated, temperatures of 21 and 24 °C demonstrated more rapid pathological signs. This research provides further insight into the complex host-pathogen-environment interactions for P. canaliculus in response to prolonged elevated temperature.
- ItemInteractive Effects of Elevated Temperature and Photobacterium swingsii Infection on the Survival and Immune Response of Marine Mussels (Perna canaliculus): A Summer Mortality Scenario(Elsevier BV, 2024-02-03) Azizan, Awanis; Venter, Leonie; Zhang, Jingjing; Young, Tim; Ericson, Jessica A; Delorme, Natalí J; Ragg, Norman LC; Alfaro, Andrea CThe New Zealand Greenshell™ mussel (Perna canaliculus) is an economically important aquaculture species. Prolonged increases in seawater temperature above mussel thermotolerance ranges pose a significant threat to mussel survival and health, potentially increasing susceptibility to bacterial infections. Using challenge experiments, this study examined the combined effects of increased seawater temperature and bacterial (Photobacterium swingsii) infection on animal survival, haemocyte and biochemical responses of adult mussels. Mussels maintained at three temperatures (16, 20 and 24 °C) for seven days were either not injected (control), injected with sterile marine broth (injection control) or P. swingsii (challenged with medium and high doses) and monitored daily for five days. Haemolymph and tissue samples were collected at 24, 48, 72, 96, 120 h post-challenge and analysed to quantify bacterial colonies, haemocyte responses and biochemical responses. Mussels infected with P. swingsii exhibited mortalities at 20 and 24 °C, likely due to a compromised immune system, but no mortalities were observed when temperature was the only stressor. Bacterial colony counts in haemolymph decreased over time, suggesting bacterial clearance followed by the activation of immune signalling pathways. Total haemocyte counts and viability data supports haemocyte defence functions being stimulated in the presence of high pathogen loads at 24 °C. In the gill tissue, oxidative stress responses, measured as total antioxidant capacity and malondialdehyde (MDA) levels, were higher in infected mussels (compared to the controls) after 24h and 120h post-challenge at the lowest (16 °C) and highest temperatures (24 °C), indicating the presence of oxidative stress due to temperature and pathogen stressors. Overall, this work confirms that Photobacterium swingsii is pathogenic to P. canaliculus and indicates that mussels may be more vulnerable to bacterial pathogens under conditions of elevated temperature, such as those predicted under future climate change scenarios.
- ItemInvestigating the Effect of Bacterial Coinfections on Juvenile and Adult Green-Lipped Mussels (Perna canaliculus)(Wiley, 2023-10-03) Azizan, Awanis; Carter, Jack; Venter, Leonie; Young, Tim; Sharma, Shaneel S; Chen, Tony; Alfaro, Andrea CThe New Zealand's Greenshell™ mussel (Perna canaliculus) aquaculture industry is being affected by summer mortality events associated with increasing seawater temperatures and pathogens. In this study, challenge experiments were conducted to investigate, for the first time, the effects of pathogen coinfection on the survivability and haemolymph immune responses of juvenile and adult mussels. Animals were injected with marine broth (control), Vibrio mediterranei, Photobacterium swingsii, or a mixture of V. mediterranei and P. swingsii. Then, mussel survival was monitored for 72 h, and haemolymph was sampled for bacterial quantification and metabolomics analyses at 24- and 48-h post challenge. Coinfected adults and juveniles showed 100% mortality. Bacterial colony counts in haemolymph decreased as infection time continued, especially in juveniles. The haemolymph metabolome of mussels exposed to single bacterial species and coinfection showed response changes largely within energy metabolism. Mussels infected with V. mediterranei exhibited increased metabolites linked to the glutathione pathway, branched-chain amino acids, and others over time, supporting structural functions. Conversely, mussels infected with P. swingsii showed no metabolic differences over time. The coinfection group exhibited large decreases in important metabolites, such as fatty acids as an alternative energy source and amino acids to support immune functions and protein synthesis.
- ItemMetabolic Regulation of Copper Toxicity during Marine Mussel Embryogenesis(MDPI AG, 2023-07-11) Young, Tim; Gale, Samantha L; Ragg, Norman LC; Sander, Sylvia G; Burritt, David J; Benedict, Billy; Le, Dung V; Villas-Bôas, Silas G; Alfaro, Andrea CThe development of new tools for assessing the health of cultured shellfish larvae is crucial for aquaculture industries to develop and refine hatchery methodologies. We established a large-volume ecotoxicology/health stressor trial, exposing mussel (Perna canaliculus) embryos to copper in the presence of ethylenediaminetetraacetic acid (EDTA). GC/MS-based metabolomics was applied to identify potential biomarkers for monitoring embryonic/larval health and to characterise mechanisms of metal toxicity. Cellular viability, developmental abnormalities, larval behaviour, mortality, and a targeted analysis of proteins involved in the regulation of reactive oxygen species were simultaneously evaluated to provide a complementary framework for interpretative purposes and authenticate the metabolomics data. Trace metal analysis and speciation modelling verified EDTA as an effective copper chelator. Toxicity thresholds for P. canaliculus were low, with 10% developmental abnormalities in D-stage larvae being recorded upon exposure to 1.10 μg·L-1 bioavailable copper for 66 h. Sublethal levels of bioavailable copper (0.04 and 1.10 μg·L-1) caused coordinated fluctuations in metabolite profiles, which were dependent on development stage, treatment level, and exposure duration. Larvae appeared to successfully employ various mechanisms involving the biosynthesis of antioxidants and a restructuring of energy-related metabolism to alleviate the toxic effects of copper on cells and developing tissues. These results suggest that regulation of trace metal-induced toxicity is tightly linked with metabolism during the early ontogenic development of marine mussels. Lethal-level bioavailable copper (50.3 μg·L-1) caused severe metabolic dysregulation after 3 h of exposure, which worsened with time, substantially delayed embryonic development, induced critical oxidative damage, initiated the apoptotic pathway, and resulted in cell/organism death shortly after 18 h of exposure. Metabolite profiling is a useful approach to (1) assess the health status of marine invertebrate embryos and larvae, (2) detect early warning biomarkers for trace metal contamination, and (3) identify novel regulatory mechanisms of copper-induced toxicity.
- ItemMetabolite Changes of Perna canaliculus Following a Laboratory Marine Heatwave Exposure: Insights from Metabolomic Analyses(MDPI AG, 2023-07-03) Azizan, Awanis; Venter, Leonie; Jansen van Rensburg, Peet J; Ericson, Jessica A; Ragg, Norman LC; Alfaro, Andrea CTemperature is considered to be a major abiotic factor influencing aquatic life. Marine heatwaves are emerging as threats to sustainable shellfish aquaculture, affecting the farming of New Zealand's green-lipped mussel [Perna canaliculus (Gmelin, 1791)]. In this study, P. canaliculus were gradually exposed to high-temperature stress, mimicking a five-day marine heatwave event, to better understand the effects of heat stress on the metabolome of mussels. Following liquid chromatography-tandem mass spectrometry analyses of haemolymph samples, key sugar-based metabolites supported energy production via the glycolysis pathway and TCA cycle by 24 h and 48 h of heat stress. Anaerobic metabolism also fulfilled the role of energy production. Antioxidant molecules acted within thermally stressed mussels to mitigate oxidative stress. Purine metabolism supported tissue protection and energy replenishment. Pyrimidine metabolism supported the protection of nucleic acids and protein synthesis. Amino acids ensured balanced intracellular osmolality at 24 h and ammonia detoxification at 48 h. Altogether, this work provides evidence that P. canaliculus has the potential to adapt to heat stress up to 24 °C by regulating its energy metabolism, balancing nucleotide production, and implementing oxidative stress mechanisms over time. The data reported herein can also be used to evaluate the risks of heatwaves and improve mitigation strategies for aquaculture.
- ItemQuantification of Photobacterium swingsii and Characterisation of Disease Progression in the New Zealand Greenshell™ Mussel, Perna canaliculus(Elsevier BV, 2024-01-19) Azizan, Awanis; Alfaro, Andrea C; Venter, Leonie; Jaramillo, Diana; Bestbier, Mark; Bennett, Peter; Foxwell, Jonathan; Young, TimGreenshell™ mussels (Perna canaliculus) are endemic to New Zealand and support the largest aquaculture industry in the country. Photobacterium swingsii was isolated and identified from moribund P. canaliculus mussels following a mass mortality event. In this study, a challenge experiment was used to characterise, detect, and quantify P. swingsii in adult P. canaliculus following pathogen exposure via injection into the adductor muscle. A positive control (heat-killed P. swingsii injection) was included to account for the effects of injection and inactive bacterial exposure. Survival of control and infected mussels remained 100% during 72-hour monitoring period. Haemolymph was sampled for bacterial colony counts and haemocyte flow cytometry analyses; histology sections were obtained and processed for histopathological assessments; and adductor muscle, gill, digestive gland were sampled for quantitative polymerase chain reaction (PCR) analyses, all conducted at 12, 24, 48 h post-challenge (hpc). The most profound effects of bacterial injection on mussels were seen at 48 hpc, where mussel mortality, haemocyte counts and haemolymph bacterial colony forming were the highest. The quantification of P. swingsii via qPCR showed highest levels of bacterial DNA at 12 hpc in the adductor muscle, gill, and digestive gland. Histopathological observations suggested a non-specific inflammatory response in all mussels associated with a general stress response. This study highlights the physiological effects of P. swingsii infection in P. canaliculus mussels and provides histopathological insight into the tissue injury caused by the action of injection into the adductor muscle. The multi-technique methods used in this study can be applied for use in early surveillance programs of bacterial infection on mussel farms.
- ItemSustainable Aquafeed Formulations Containing Insect Larval Meal and Grape Marc for the New Zealand Farmed Abalone(Hindawi Limited, 2023) Bullon, Natalia; Alfaro, Andrea C; Manivannan, Moganakumaar; Dezfooli, Seyedehsara Masoomi; Seyfoddin, Ali; Dawood, MahmoudThe aquaculture industry has been criticised for the excessive use of fish meal (FM) in feeds due to the utilisation of wild fish in the formulation and the exacerbation of overfishing marine resources. Land-based abalone aquaculture mainly uses commercial feeds (CFs) to promote faster growth, which include FM as a primary protein component. Alternative ingredients, such as insect meal (IM) and grape marc (GM) are potential candidates for FM replacement due to their suitable nutritional profile and sustainable production. This paper reports on a novel nutritional approach for the New Zealand farmed abalone, which replaces FM with IM by 10% and includes a waste by-product (GM) by 30% as a potential prebiotic source. The study was performed in two stages: (a) physico-chemical determination of diets delivered in an alginate matrix (experimental diets) and their stability in seawater compared to CF and (b) evaluation of growth and feed intake for the New Zealand black-foot abalone. There were significant differences between experimental diets and CF in terms of sinking rate, particle weight, and microscopic observations. Water stability of the experimental diets was increased by 50% in 24 and 48 hr compared to CF, producing less solid waste, and potentially reducing cleaning efforts in the farm. The inclusion of IM and GM did not compromise overall animal growth or their feed conversion ratio, however, further evaluation need to be explored in the future research. The findings revealed that the developed encapsulated feeds are a more stable food delivery method for Haliotis iris compared to the CF. Furthermore, both IM and GM can be included in feed formulations as a more sustainable strategy without compromising weight and shell gains in the abalone farming.