The Veterinarian Magazine September 2023

TheVeterinarian ■ September 2023

■ www.theveterinarian.com.au

Cruel Sea’s kind hearts benefit Byron Bay Wildlife Hospital One of Australia’s best-known bands will play their first show in 10 years as part of a fundraiser for the Byron Bay Wildlife Hospital, after the hospital lost a $6 million NSW funding grant.

The Cruel Sea, led by wildlife ambassador Tex Perkins, will headline the fundraiser, Wild Aid 2023 on Saturday 21 October at the Green Room at Byron Events Farm, with the event aiming to raise awareness and funds for the hospital, which was opened in 2020 after the Black Summer bushfires and has treated over 4,000 animals since it was launched.

The hospital’s services, which are under threat since the loss of the funding grant, are provided by licensed wildlife carers, NSW National Parks and Wildlife Service and members of the community, seven days a week, free of charge.

Byron Bay Wildlife Hospital CEO Stephen Van Mil said his entire team were deeply grateful for the support of such a brilliant group of musicians.

“We were left reeling after the NSW government told us they wouldn’t approve the funding they announced in February. Having The Cruel Sea play their first show in ten years to help raise money for us is mind-blowing.”

Licenced by the Veterinary Practitioners Board of NSW, the hospital is the only all species wildlife hospital in NSW outside of Taronga Wildlife Hospitals in Sydney and Dubbo.

Developed to mee the need for a large-scale mobile clinic to respond to natural disasters, the hospital was custom built inside a semi-trailer and is capable of being deployed anywhere in NSW or interstate to provide emergency triage, treatment and care for native animals.

Perkins said he was shocked when he heard the hospital had lost the funding and contacted the hospital’s management team offering his support.

The Cruel Sea

“I’ve seen what these incredibly caring humans do to help wildlife,” Perkins said. “Losing that vital funding was a brutal shock, and I really wanted to help,” he said. “The Cruel Sea had started to jam again, it feels and sounds really good, the guys were keen, so playing a benefit gig made sense to kick start getting back on the road to play a bunch of shows.”

Bluesfest Executive Chairman Peter Noble

Byron Bay Wildlife Hospital, welcomed the review’s recommendations, after learning in June that a $6 million, four-year funding grant for the hospital, announced by the NSW Government in February, was not approved, citing inadequate value for money.

“This review is very timely because Australian wildlife is in crisis,” Van Mil said.

Australia has the world's highest mammal extinction rate and the review notes that only 50 per cent of NSW threatened species are expected to survive in 100 years.

Environmental impacts on wildlife, such as wildfires, flooding and storm events, add to the workload of volunteers and vets, as well as to the out-of-pocket expenses they

■ JULIA GARDINER

Australians support a legislated end date for the live sheep export trade

Review of funding comes as Australian wildlife is in crisis The NSW Government needs to consider changing finding models to support vets, volunteers, wildlife hospitals, and accredited specialist rehabilitation hospitals, according to an independent review of the Biodiversity Conservation Act. The review, led by Ken Henry AC, was tabled in NSW Parliament in September and made 58 recommendations to conserve biodiversity more effectively at a state scale.

Legally, native animals are the property of the State, yet the State bears little of the cost for rehabilitation of sick, injured and orphaned animals, which the review found come at a great financial and emotional cost to professional service providers and volunteers.

Stephen Van Mil, CEO of

said he was delighted to provide the venue for the fundraiser for an important cause.

“Byron Bay Wildlife Hospital is unique in Australia, and we’re lucky to have it in the richly biodiverse Northern Rivers. We simply cannot let more wildlife suffer and die needlessly in our backyards,” Noble said.

Stephen Van Mil

are left to cover, often without government assistance.

“Volunteer carers and veterinarians with the skills and knowledge of the physiology and anatomy of wildlife are demonstrably effective in rescuing, treating, caring for and releasing native animals into the wild. These people are doing essential work to prevent further loss of biodiversity and species extinction. They need support,” Van Mil said.

A Parliamentary e-petition that called for a legislated end date to live sheep export, and that was initiated by the RSPCA, has received almost 44,000 signatures, making it the fifth largest e-petition in the 47th Parliament.

Richard Mussell, RSPCA Australia’s CEO said the petition showed there was overwhelming community support for the phase out of live sheep export particularly since the petition was only open for a four-week period in August.

“It builds on recent surveys showing 78 per cent of Australians would support a phase out if farmers were supported through the transition.

Given there are clear signs of strong political will from this government, and its continued commitment to improve animal welfare, the next step is to legislate the end date in the current parliamentary term so there can be certainty for farmers, and the community’s expectations are met,” he said.

In its May submission to the independent panel the RSPCA made 17 recommendations.

This included legislation being passed in the current parliamentary term that To page 30

PIGEON POST 10 I MANAGEMENT 12 I ABSTRACTS 14 I CLINICAL REVIEW 20 I BUSINESS 29

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News

Lumpy skin disease found in more cattle exported from Australia Concern around lumpy skin disease (LSD) continues to threaten cattle export, one of the northern Australia’s primary industries, after the Indonesian government moved to block the live export of cattle from the region in September, after Indonesian officials reported eight cases of the highly infectious virus in imported cattle.

The move came only two months after Australia’s chief veterinarian, Mark Schipp, released a statement confirming the country is still free of lumpy skin disease (LSD), despite Indonesia and Malaysia ceasing importation of cattle from four Australian export yards, after a small number of the cattle tested positive for the disease after reaching their destination in June.

LSD occurs in cattle and water buffalo and is mainly spread by biting insects including flies, mosquitos, midges, and ticks. If infected, the animals develop hard nodules under the skin, which develop into open wounds highly prone to infection. Infected stock quickly lose condition as the infection spreads, as they become reluctant to move or eat.

Australia's Acting Chief Veterinary Officer, Beth Cookson said the investigation into the

July results had tested more than 1,000 cattle across Western Australia, Queensland and the Northern Territory, which found no trace of the disease, and Australia remained free of LSD. “Our disease status has not changed. The negative test results further highlight Australia's robust systems for the ongoing monitoring of animal diseases, including LSD,” Cookson said.

Indonesia bought 337,000 cattle worth $550 million in 2022 alone, making it the largest importer of the Australian livestock. LSD was first detective in Indonesia in March 2022, with cases since confirmed in 14 Indonesian provinces, which Western Australia Farmers’ livestock president, Geoff Pearson believes indicates the source of the infection. “[The Australian cattle] clearly … contacted it from inside Indonesia”, Pearson said, noting the Indonesian government’s response was an understandable precautionary measure.

With producers in northern Australia still struggling with the aftermath of January's floods, freight interruptions and the COVID pandemic, there is concern that further bans on export yards, as well as stricter protocols needed to use the yards, will become cost prohibitive.

Outfoxing predators In experimenting with non-lethal control methods for introduced animals, researchers from the Australian National University and the University of South Australia found that if baits deliberately laid for red foxes (Vulpes vulpes) are spiked with capsules containing a chemical that causes vomiting and nausea, the reaction is similar to that of humans who consume foods that make them feel unwell. Those foods are avoided in future.

The world first experiment was conducted at 30 stations across open landscape in southeastern Australia where foxes are responsible for countless wildlife deaths. The scientific team buried baits of fried deboned chicken, some of which contained capsules of levamisole, a chemical that induces nausea and vomiting when consumed by predators.

Tim Andrewartha, lead author of the paper published recently in the journal Conservation Science and Practice, and a PhD student at ANU’s Fenner School of Environment and Society, said using non-lethal tactics for keeping predators at bay could potentially be more effective than shooting, trapping or poison baiting in certain contexts as these could lead to adaptations in the predator population that counteract the control method used.

“Some techniques become less effective over time, poison baits for example can lead to bait resistance or shyness, so the potential for non-lethal methods is something we hope to explore in the future,” he said.

A range of encapsulated nausea-inducing

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agents can be injected into food sources such as eggs or animal carcasses so the predator associates the symptoms with food odour and not the chemical.

Co-author Anton Blencowe, an Associate Professor, senior lecturer and Applied Chemistry and Translational Biomaterials Group Leader at UniSA’s School of Pharmacy and Medical Sciences, said invasive predators were responsible for almost 60 per cent of all bird, mammal, and reptile extinctions globally, so finding ways to mitigate their impacts on native wildlife was critical.

The NSW Department of Primary Industries recommends producers implement an insect management plan for tick control, and to target all stages of the stable fly and mosquito life cycles to break the breeding cycle, including environment management to reduce the number of standing water sites where breeding can occur.

“Existing lethal control methods can be counterproductive in some situations, with predators capable of adapting their behaviour and becoming harder to control, so the conditioned taste aversion tactic employed in this experiment has shown promise as a tool to reduce the predation of vulnerable wildlife,” Blencowe said.

Over three sequential periods researchers laid untreated baits followed by encapsulated levamisole baits, and then untreated baits again two weeks later. During the final period baits taken fell by at least 30 per cent, suggesting foxes had consumed the treated baits, fallen ill, and were reluctant to go back for seconds, despite the absence of levamisole. The reduction in baits taken continued for at least 68 days after treatment.

“This evidence shows it’s possible to condition foxes to avoid food sources based on their unique odour, and ultimately this strategy may be applicable to protecting vulnerable wildlife living in that habitat, although more research is needed to determine the best chemical and dosage used,” Andrewartha said.

Future research will involve determining if the conditioning aversion tactic is more effective in foxes through using live prey or a carcass, as well as the time between consumption and onset of symptoms.

■ ANNE LAYTON-BENNETT ‘Landscapes of nausea: successful conditioned taste aversion in a wild red fox population’ is available at doi.org/10.1111/csp2.12984

SEPTEMBER 2023 TheVeterinarian 3

News

RSPCA provides safe beds Recognising companion animals can often be the overlooked victims of homelessness, as well as family violence, the Tasmanian government has extended the Flexible Support Program it established in 2021. The program was designed to provide support for people with pets who are affected by family violence but following the release of the National Partnership on Family, Domestic and Sexual Violence Responses 2021-23, RSPCA Tasmania has been provided with $100,000 to pilot the Safe Beds Program to include people experiencing homelessness.

Seed funding for the program was also received from the Petbarn Foundation, an organisation with an existing partnership with the RSPCA nationally, and that will continue to provide a portion of funding for the Safe Beds Program on an annual basis.

Several studies have been conducted that show there is a link between animal abuse and domestic violence with several jurisdictions recognising legislation needs to be updated to incorporate this.

Andrea Dawkins, RSPCA Tasmania’s Community Engagement Manager and Deputy Chief Executive Officer said the organisation is currently

lobbying the Tasmanian government so that provision for animals in family violence laws is included in revised legislation.

“Although the program initially focused on those fleeing domestic violence situations, we’re also now working with case workers and individuals to ensure that the companion animals of people experiencing homelessness are given the time and space to regroup and get back on their feet. There were people staying in unsafe circumstances trying to shelter their animals, and we had people going into hospital.

We knew we had to broaden the program because since the pandemic people were becoming homeless more rapidly,” Dawkins said.

Jayla Sainty joined the Safe Beds Program in April as the RSPCA’s inaugural Community Awareness Officer. After completing an animal studies course at TAFE several years ago and working in a range of jobs for several years, Sainty is now combining her work at the RSPCA with studying social work at Deakin University. Her interest in animal welfare issues led to research showing most jurisdictions rarely include animal welfare in their codes of practice for social workers.

This omission is one that Sainty would like Tasmania to rectify, with models already in place in both Victoria and New Zealand that could be adapted.

“Animal welfare isn’t only a personal problem, it’s a systemic problem and highlights the challenges we have in our housing crisis. It’s not just domestic violence issues but it’s being unable to afford housing or rentals anymore.

Surrendering their pets is a very tough decision for people to make,” she said.

Although those who join the program understand they will not be allowed to access their pets, Sainty said every effort was made to provide photos of the animals in their foster homes, and to reassure owners their pets were content and being well cared for. This was especially important if children were involved since separation from a pet can add to a child’s trauma.

“The main reason people aren’t allowed to visit their pets is the safety aspect, particularly if domestic violence is involved. It can be confusing and stressful for the animal to see their owner - who then leaves again - but if domestic violence is involved then pets are usually moved to another area of the state. The emphasis is always on the safety element,” Sainty said.

Andrea Dawkins

Although the program’s arbitrary timeframe is two weeks, an animal’s stay with a foster carer can be up to a year, which can provide an ongoing challenge. Sainty said care was taken to ensure foster carers know they are helping someone experiencing a difficult period in their life and the pets they take in are from loving and caring owners.

“It’s important carers have that understanding and knowledge they’re helping someone, and the decision to surrender their pets was made because they wanted the best care for their animals,” she said.

■ ANNE LAYTON-BENNETT

Koalas under threat from Artificial habitat structures for wildlife conservation: cars, disease and dogs Research has shown the southeast Queensland koala population is under threat, with hopes that the data will pave the way for proactive measures to protect the native species.

KoalaBASE, a database developed by the University of Queensland, has been collecting data on koala hospital admissions since 1997 across 15 local government areas from the Fraser Coast to the Scenic Rim and Darling Downs and the South Burnett.

14 per cent were because of a dog attack,” Henning said, noting that the numbers of un-reported deaths would likely make these numbers significantly higher.

Mark Stevenson of the University of Melbourne helped supervise the study, which will now focus on minimising the impact of the known threats.

“This analysis is a useful starting point for more pro-active approaches to managing preventable wild

Close to 3,000 of the reported koala deaths were analysed by Joerg Henning and his colleagues at The University of Queensland School of Veterinary Science, and the finding showed a large threat to koala populations from car strikes, dog attacks and chlamydiainduced illnesses.

“In the five years between 2009 and 2014, 52 per cent of reported wild koala deaths were caused by car strike, 34 per cent were from a chlamydia-related disease and

koala mortality risk,” Stevenson said.

The research group hopes to work with policymakers to identify specific areas where intervention efforts would be best suited, including improvements to road signage in areas with known koala habitat and activity areas, the construction of over and underpasses for their safe movement, and education campaigns for dog owners about the threat dogs pose to koalas.

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■ JULIA GARDINER

use with caution

An increase in the use of artificial habitat structures for wildlife species under threat from habitat loss or disturbance is not without risk according to the results of a study published recently in the journal Austral Ecology.

Co-author Tenaya Duncan, a Conservation and Wildlife Biology PhD student at Western Australia’s Murdoch University, said artificial structures could be a double-edged sword unless their use was backed by sound ecological science that considered the design, construction, and positioning of the structures.

“Many of the world’s natural environments have been irreparably modified and damaged, so species have lost critical habitat components they need to survive. Increasingly conservationists and developers seek to supplement these destroyed components with artificial habitat structures that are now used in marine, freshwater and terrestrial environments,” she said.

Duncan and her fellow PhD student researchers from Deakin University and the University of Western Australia, found that unless appropriate controls were implemented artificial structures could lead to an increase in predation, as well as exposure to fatal environmental conditions, disease, the spread of invasive species, and even a decline in genetic diversity. They can also be embraced as a ‘green-

washing mechanism’ so unsustainable practices appear to be more environmentally responsible.

“While these structures can certainly have positive conservation outcomes and help the recovery of populations, they need to be carefully planned with appropriate knowledge of the critical features animals use to bask, hibernate, reproduce and take refuge from environmental stressors and predators,” she said.

In addition to the appropriate design, construction, and positioning of artificial habitat structures in the landscape the study also found it was essential that wildlife population conservation was properly supported and monitored over the long-term.

“If done badly, artificial structures may actually facilitate further impingement on natural areas and habitat destruction. Importantly we also mustn’t lose sight of the underlying threats that are causing species decline in the first place.

Artificial habitats generally don’t address these underlying threats, and once destroyed, natural habitats take a long time to recover - if they’re able to recover at all,” she said.

■ ANNE LAYTON-BENNETT ‘The risks and rewards of using artificial habitat structures for wildlife conservation’ is available at doi.org/10.1111/aec.13376 SEPTEMBER 2023 TheVeterinarian 5

T

F O E F I L E H T N I Y A D A : R E T S O F E C I D CAN ILL

RSPC

H N E K O BR A VET,

his month our Day in the Life feature heads to the far western NSW town of Broken Hill, which is home to Candice Forster, a veterinarian with the Royal Society for Prevention of Cruelty to Animals (RSPCA) NSW. While much of what Forster does would be familiar to many vets, living and working in a regional area means her veterinary practice appears typical – but with several twists.

Originally hailing from Adelaide, Forster worked as a laboratory technician at Flinders University prior to studying a Bachelor of Veterinary Science at the University of Sydney, from which she graduated in 2016. “I decided to trade in laboratory animals for companion animals, but I still love seeing rodents and rabbits in general practice,” she said.

Forster is currently employed as the Managing Veterinarian at RSPCA NSW Broken Hill, which is a small animal practice servicing the Broken Hill community. “I am the senior clinician at the practice, working primarily in clinical veterinary work, but I am also involved with clinic management and mentoring junior veterinarians,” she explained. “At the clinic, we try to foster a supportive working and learning environment where we discuss cases and learn from each other. Even I still have a lot to learn, and I am constantly working on my own development as a veterinary clinician and manager.”

RSPCA NSW’s Broken Hill Clinic is classed as a small animal general practice, and Forster describes the work as being similar most of the other clinics she has worked at. “There’s never a dull moment in general practice, so my day always begins with a coffee!” Forster said.

Morning consultations and routine procedures such as desexing, lumpectomies and dental surgeries tend to follow, with staff then rotating through lunch and continuing with surgical procedures and afternoon consultations. A typical day for Forster then concludes with inpatient checks and client updates.

Forster enjoys the variety of general practice and has not yet undertaken any formal postgraduate study – largely because she is yet to choose one thing upon which to focus her attention. “The last short course I completed was on guinea pigs after I brough home some strays from work,” she said. “I realised I would have to learn how to treat them if any problems came up, as there aren’t any vets who work with small animal exotics for hundreds of kilometres.”

Being hundreds of kilometres from specialist veterinary support in a town of nearly 18,000 people is the first clue Forster provides to how her work is different from regular small animal practice.

“With no emergency clinics in town, we need to be prepared to deal with all kinds of emergency presentations,” she said. “We see trauma cases, snake bites, GDVs, toxicities – the whole gamut of emergency medicine and surgery.”

At times, the work can be difficult, not least because there are no 24-hour veterinary facilities in the region for Forster and her team to fall back on either. “The most challenging aspect of my job is managing hospital cases after hours,” she said. “We will often drop back into the clinic after hours to check on our patients.” Despite the crazy hours, Forster is quick to point out how satisfying it is to see a case through from initial presentation to completion. “I find the afterhours cases very rewarding,” she said. “It’s generally more peaceful in the clinic, and I find I can focus all my attention on the task in front of me.”

The other important distinction between Forster’s work at RSPCA NSW Broken Hill and the veterinary clinics she has experienced previously is the level of community involvement. “In Broken Hill, we often find ourselves being part of important community work,” she said.

■ www.theveterinarian.com.au

“For example, back in 2021 the RSPCA NSW Broken Hill branch provided the funds for us to set up a free parvovirus vaccination drive, resulting in over 250 vaccinated dogs and a significant drop in parvovirus cases in the Broken Hill area.” The clinic has also collaborated with Broken Hill City Council Rangers, assisting them with dogs and cats in their care. “We also see kangaroos, birds and reptiles, and many other wildlife cases for Rescue and Rehabilitation of Australian Native Animals (RRANA), our local wildlife rescue group,” Forster said.

For what is supposedly a small animal general practice, RSPCA NSW Broken Hill Clinic is clearly much more, attending to emergency, exotic and wildlife cases in addition regular domestic

animals, as well as creating and nurturing important community connections. And for Forster, getting to know members of the Broken Hill community is what she loves most about the job.

“Even after just 18 months out here, I’ve become familiar with many of the locals and their animals,” she said. “The people I’ve met here are amazing, and they’re happy to lend a hand if we need any help tending to their animals.” It sounds like RSPCA NSW Broken Hill is very lucky to have her.

And now, as always, we conclude our Day in the Life of Candice Forster with our Creature Questions: Do you currently have any pets?

Oh, yes. I have six cats, a cheeky little mixed terrier dog, a guinea pig family of four, some tropical fish and a goldfish pond, and, last but not least, a very old green tree frog that has travelled with me from Adelaide to Sydney and now out to Broken Hill.

What is your favourite animal?

I’m definitely more in tune with felines but I’m enjoying learning how to live with a dog.

It’s been a bit of an adjustment after being surrounded by just cats for the past fifteen years or so.

If you could transform into any animal, what would you be and why?

A cat, for sure. I love to nap!

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A role for drones in saving wildlife Spring – not summer

Diane Colombelli-Négrel Picture Flinders University

The jury is still out on the benefits of adopting drones to assess and manage wildlife populations in the aftermath of floods or bushfires, or to assess how species are adapting to a changing climate, but behavioural responses are known to vary according to individual species, type of drone and flight distances employed.

While drones – or Unmanned Aerial Vehicles – can be a cost-effective tool that ‘eliminates human bias and is able to produce data over high spatial and temporal resolution, even for species that inhabit locations difficult to access’, researchers from Flinders University found drone-based surveying can be a vital conservation tool in monitoring the distribution and abundance of koalas. As a sedentary cryptic species, koala populations are generally spatially fragmented but the species response to drones has not previously been measured.

8 TheVeterinarian SEPTEMBER 2023

In a drone study published recently in the journal Applied Animal Behaviour Science, scientists studied the heart rate and responses of captive koalas to test the effects of drone activity near their trees.

The koala study coincided with another update from the Flinders University’s Bird Lab research group on a population of the threatened little penguin (Eudyptula minor) at Granite Island, a popular South Australian tourist location near Victor Harbour.

Lead author and team leader Diane Colombelli-Négrel, a lecturer in animal behaviour and penguin ecologist from FU’s College of Science and Engineering, said technologies such as audio and visual recordings were emerging as key methods in helping to understand factors that may influence wildlife survival, as populations, particularly near large urban areas, become threatened or endangered.

“Drones have become a popular conservation tool, especially when monitoring cryptic species or animals in remote or inaccessible locations.

We developed a non-invasive way to measure heart rate in koalas using a Fitbit, and to record their behaviour and breathing rate with mobile GoPro cameras when we flew drones about 15m above their heads in South Australia’s Cleland Wildlife Park enclosure,” she said.

The koalas responded to the drone flight with a short-term in vigilance but there was no change in heart or breathing rates. This suggests drones may not have a long-term detrimental effect on the animals’ fitness or energy demands, which due to the species’ specific diet of eucalyptus leaves means koalas require up to 20 hours of rest per day to conserve energy, so any disturbance can affect their fitness.

To page 30

– is peak tick time

As we approach spring, new research from the University of Queensland has shown that the season is the peak time for tick paralysis in dogs and cats, rather than the hotter months of summer.

A team led by Stephen Barker at UQ’s School of Chemistry and Molecular Biosciences analysed over 20,000 cases across 20 years of veterinary records in four regions along Australia’s east coast.

The researchers found spring appears to be when eggs laid by ticks the previous summer hatch, posing the biggest threat; however, ticks are a year round issue for pet owners.

“We found that there were tick cases in pets at veterinary hospitals all year in all of the regions we studied, even in the coldest winter months,” Barker said.

“The number of cases actually drops off through the hottest summer months of January and February in all regions.”

The research also showed that studying the climate could be a good indicator for the severity of the next year’s outbreak.

“We lined up the vet data to weather records and saw that variations in temperature and moisture in summer determines how bad the tick season is the following spring and summer,” Barker said.

“If the weather is quite dry and hot in January and February, many eggs will die but a mild and wet summer means a high hatching rate and a bad tick season in the following spring.”

The team has used their finding to develop computer model to make predictions for future tick seasons, allowing for warnings to pet owners and to help vets be prepared.

■ JULIA GARDINER

■ www.theveterinarian.com.au

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Pigeon post from the UK

Ian Neville BA(Hons) BVSc MRCVS reports from Britain.

The British summer is drawing to a close, but unlike last year’s exceptionally hot and dry conditions (an unprecedented maximum of 40.3°C recorded), 2023 will be remembered as a mostly cool and very wet summer. Measured rainfall in my home region (Yorkshire) during July was 218 per cent of the long-term average. The conditions seemed reminiscent of my boyhood and the national news was duly plastered with images of families sheltering from wind and rain on camping grounds or beachfronts as they endured a ‘traditional’ British summer holiday. The vagaries of our maritime climate are never far from the national consciousness or daily chit-chat, and even more so since climate change has made accurate forecasting even more problematic.

Likewise, the activities of corporate veterinary groups have remained headline news across all veterinary media outlets and a hot topic in staff room conversation over recent months as their growing influence on the profession continues to evolve. In May the Competition and Markets Authority (CMA) announced its initial findings into acquisitions by the Medivet group concluding that a ‘realistic prospect’ existed of a loss of competition and out-of-hours provision in some areas of England and Northern Ireland if proposed buyouts went ahead. This is the fourth investigation by the CMA into veterinary groups over the last two years which have previously

involved: CVS, IVC Evidensia, Vet Partners and the Eville and Jones group. The CMA is concerned that the progressive acquisition of independent practices is reducing competition and raising prices or lowering the quality of service offered at a time when household budgets are already squeezed by the ‘cost of living crisis’. The CMA has authority to investigate mergers when any acquired business generates over £70 million (A$137 million) of its turnover in the UK or where the combined entity has at least a 25 per cent market share nationally or in a region. In June the devolved Welsh government (the Senedd) began considering a petition, signed by vets and members of the public calling for an investigation into the ‘corporate takeover’ of the veterinary sector in Wales. Comments by petitioners included claims that the profession had ‘lost its principles’ and was colluding with corporate business at the expense of animal welfare.

An inquiry is to be held in the autumn when the Royal College of Veterinary Surgeons and veterinary representatives will have an opportunity to give evidence to the Senedd. Corporate takeovers have also attracted opposition in the Republic of Ireland since 2021 where rationalisation has left some rural clients without local veterinary services or a more expensive provision.

Around the same time the IVC Evidensia group was at the centre of a furore over restructuring

proposals that could potentially see hundreds of redundancies in support and financial roles.

The announcement caused widespread outrage among threatened employees and criticism from the Veterinary Management Group and the British Veterinary Union. There were even calls for clinical staff to consider strike action in support of their company colleagues and to protect their own future working conditions. Subsequently part of the CVS group, which was itself previously subject to a competition enquiry, also announced restructuring plans that may result in redundancies of support staff.

It has been reported that some clinical staff in corporate practices now worry that the proposed redundancies mark a sea change away from prioritising clinical outcomes to chasing financial goals, and that the job losses are a direct result of companies overextending themselves in the race to expand. In July CVS group released its results for the year ending 30 June which revealed that it had spent £55million (A$108 million) acquiring a further 11 UK practices. It also announced its first acquisitions of independent small animal practices in Australia: three in Brisbane and one in Sydney for an estimated combined value of £16.8 million (A$40 million). The stated CVS future strategy for Australia is to acquire more practices in the major urban conurbations.

The imperative to control costs in the veterinary sector is not

restricted to the handful of very large, UK corporate practices but is a result of generalised and continuing inflationary pressure on household disposable incomes. The rate of inflation has continued to fall slowly over the summer, but most prices have not come down, and goods and services generally remain far more expensive than they were just a couple of years ago. Speakers at the Veterinary Management Group congress in June reported deep-seated problems in practice as more clients become unable to afford treatment and more vets decided to leave the profession. The remaining practitioners are faced with the challenging task of managing the workload while pacifying an increasingly worried and belligerent clientele. Reducing their range of services, not accepting new clients and not appointing staff to fill vacancies are strategies practices have adopted to try and cope.

There were also calls for practices to offer a more pragmatic, affordable approach to treatment to maximise pet welfare outcomes from more restricted spending.

Long established pet charities like the PDSA and RSPCA are also struggling to provide their services as they have staff shortages and their income depends largely upon donations and legacies which are falling as the demand for their help rises. The consequence is that a tranche of the pet owning public are increasingly unable to access or afford adequate veterinary care.

DO YOU HAVE SOMETHING TO SAY? Is there a topic you’ve wanted to tell everyone about?

Have you ever thought you would like to write a clinical review but don’t know where to start? Do you have a hobby or history that you think vets everywhere would like to learn more about? Then don’t delay – email The Veterinarian at editor@vetmag.com.au and let us know – because your words could be in the pages of next month’s issue!

10 TheVeterinarian SEPTEMBER 2023

■ www.theveterinarian.com.au

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Management

Frequently asked questions: Insurance for practice purchases There are many misconceptions about the role that insurance plays in your practice purchase.

Often, buyers will be too busy with business plans, arranging finance, renovation or reinvestment to spend much time thinking about insurance…

When insurance is thought about during the purchase process, it is often dismissed as discretionary, optional and overly pessimistic … that is, until the buyer gets close to the finish line and realises that their finance provider and/or landlord may only approve your loan or tenancy if insurance cover has been arranged.

We asked the experts at Credabl finance and Experien Insurance Services (Experien), to give us clarity on what insurances a buyer may need to have in place to complete a practice purchase.

Q1. What insurances would you typically need to have to get a bank loan to purchase a vet practice?

In many situations, a bank will want life insurance, income protection and/or business insurance in place as a condition of the loan. The insurances required can vary on a

12 TheVeterinarian SEPTEMBER 2023

deal-by-deal basis: a high loan-tovalue ratio, for example, may trigger this. Unfortunately, the risk of illness, injury and other perils may not be insignificant over a long period of time, and so the bank will want to know that insurance is there to protect the ability for the loan to be repaid, should this happen. This would protect the borrower and the lender and, most importantly, your family and your investment.

If applicable, the bank will most likely: ■ want the insurance in place prior to the loan being settled and/or in the process of being established ■ require ongoing confirmation that the insurance is in place, and, in some situations, they may own the policy (with the borrower paying the premiums) to ensure any claims paid go to the intended purpose ■ require to be named on the policy as an interested party to give them adequate protection.

Q2. What insurance would you typically need to have in a commercial lease of a vet practice premises?

Landlords will usually request

$20m of public liability and glass insurance as a minimum requirement, in order to approve a lease of their premises. Some may want comfort that there is also insurance for the contents (including the fit out). And, in single-tenanted buildings (a stand-alone house converted into a practice, for example) the landlord may require the tenant to pay the cost of the building’s own insurance.

Since most practice sales are subject to the purchaser either purchasing or leasing the premises, and the premises lease is dependent upon insurance, it is a good idea to be looking into insurance for the premises with plenty of time up your sleeve, so as not to hold up settlement of practice purchase.

Q3. If you are planning on buying the real estate premises with the practice would you need additional insurance?

For lending purposes, you will need building insurance to cover the replacement value of the property.

This is a figure typically stated in a valuation report. In other words,

Kevin Koton

cover and purchase price are not necessarily correlated.

An exception to this will be if it is a body corporate, then it will automatically be included in the body corporate fee.

Q4 What information would you need to provide in order to obtain these insurances and how long will it take?

Depending on the type of insurance required, getting insurance can take as little as a couple of days to as much as a few weeks to set up.

Gathering the necessary information to apply for insurance can take time, for example: ■ life insurance policies may need a medical assessment ■ Premises lease: an insurer might need construction information for the property.

■ www.theveterinarian.com.au

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■ EDITED BY JENI HOOD BSc (Hons) BVMS PhD

Abstracts

Companion animal veterinarians discuss aspects of one health with pet owners during most veterinary appointments

Sustainability policies and practices at veterinary centres in the UK and Republic of Ireland

Objective: To examine the prevalence and context of one-health conversations between veterinarians and clients in companion animal practice.

Background: Veterinary professionals operate at the human-animal-environment interface and are concerned about sustainability issues. This study examined the extent to which sustainability is represented in policy and enacted in veterinary practice settings, as reported by practice representatives.

Sample: A random selection of 60 companion animal veterinarians; a convenience sample of 917 interactions from Southern Ontario, Canada. Of these, 100 audio-video-recorded interactions including 47 of 60 veterinarians were randomly selected for inclusion in this study.

Methods: An online survey was completed by 392 veterinary centre representatives in the UK and Republic of Ireland to identify existing policies and practices around the environmental impacts of veterinary services and animal husbandry, responsible medicine use, animal welfare and social wellbeing.

Methods: Audio-video recordings were made of veterinarianclient-patient interactions between November 2017 and January 2019. A researcher-generated coding framework was developed and used to assess the prevalence and content of one-health topics communicated during veterinary appointments.

Results: A minority of respondents were aware of an environmental policy at their practice (17 per cent, 68/392). Many others were undertaking waste reduction initiatives, but wider environmental interventions were infrequently reported. The majority were aware of medicine stewardship and animal welfare policies or guidelines, but a minority reported social wellbeing policies (40 per cent, 117/289) and the provision of advice to clients on the environmental impacts of animal husbandry (31 per cent, 92/300).

Results: Of the interactions assessed, 60 were preventive care and 40 were health problem appointments. Further, 78 per cent (78/100) included at least 1 discussion related to one health. One-health topics included zoonoses (28 per cent [28/100]), animal behavior (25 per cent [25/100]), illness/disease (20 per cent [20/100]), activity level/ exercise (16 per cent [16/100]), nutrition (16 per cent [16/100]), dentistry (6 per cent [6/100]), body weight (3 per cent [3/100]), animal welfare (3 per cent [3/100]), dog/cat bites (2 per cent [2/100]), cannabis (2 per cent [2/100]), and aging (1 per cent [1/100]). Zoonotic diseases were mentioned in 65 appointments, 28 of which evolved into a one-health discussion. Antibiotics were discussed in 27 appointments, none of which were discussed in relation to one health (eg, antimicrobial resistance).

Clinical relevance: Findings suggest that one-health topics are raised within most veterinary appointments. Opportunities exist for more comprehensive one-health conversations between veterinarians and their clients, particularly in relation to zoonotic diseases and antimicrobials.

Natasha Janke1, Elizabeth A Stone2, Jason B Coe1, Cate E Dewey1 J Am Vet Med Assoc. 2023 Aug 22; 1-9.doi: 10.2460/javma.23.05.0287.

1Department of Population Medicine, Ontario Veterinary College,

University of Guelph, Guelph, ON, Canada.

2Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada.

The Gordon Memorial Lecture: Laying hen welfare Preference tests remain a useful tool in the assessment of laying hen welfare and have been used to establish what types of resources and enrichments are most likely to meet the birds' needs.

Evidence on the underlying structure of bird preference suggests that hens make stable and reliable choices across time and context.

This means that their preferences can also be used as a benchmark in the validation of other welfare indicators. Hens have sophisticated cognitive abilities. They are quick to form associations between events and they are flexible in how they apply their knowledge in different contexts. However, they may not form expectations about the world in the same way as some mammalian species. Limited research in this area to date seems to show that hens judge situations in absolute terms rather than evaluating how a situation may be improving or deteriorating. The proportion of hens housed in cage-free systems is increasing globally, providing birds with greater behavioural freedom. Many of the problems associated with cage-free systems, such as keel bone fractures, mortality and injurious pecking, are slowly reducing due to improved experience and appropriate changes in rearing practices, diet, To page 30

14 TheVeterinarian SEPTEMBER 2023

Limitations: The bias arising from the small convenience sample of practice representatives and potential discrepancies between the claims of survey respondents and their practices' policies and activities are acknowledged.

Conclusion: Results depict a value-action gap between the concern of veterinary professionals towards sustainability and the policies and practices at their workplaces. Building on progress in the sector, wider adoption of comprehensive policies and practices, with guidance, could enhance veterinary contributions to the sustainability agenda, in particular to mitigate the environmental externalities of veterinary services and animal care and ensure safe, fair and inclusive workplaces.

Laura E Higham1 2, Zoe J Halfacree1, Jo Stonehewer3, David H Black1 3, Gudrun Ravetz1 3, Dominic Moran2, Lisa Boden2, Catherine Oxtoby3 Vet Rec. 2023 Aug 5;193(3): e2998.doi: 10.1002/vetr.2998.

1Vet Sustain, Carlisle, UK. 2Global Academy of Agriculture and Food Systems, Royal (Dick)

School of Veterinary Studies, Easter Bush Campus, Midlothian, UK.

3The Veterinary Defence Society, Knutsford, UK.

Insect Bite Hypersensitivity in horses: Causes, diagnosis, scoring and new therapies Insect Bite Hypersensitivity (IBH, Queensland itch, sweet itch, equine summer eczema) is the most common pruritic disease of horses. It is most often caused by sensitivity to the saliva of Culicoides spp. of biting midges; however, it can also be caused by hypersensitivity to other insect species. The prevalence of IBH in horses is reported to be as high as 60 per cent in some parts of the world. Due to the severe pruritus and effects of secondary self-trauma, IBH has animal welfare concerns, and there is currently no cure. Management of this condition is life-long, time consuming and costly. New grading systems to document disease severity are being validated, which will allow the comparison of clinical trial results of new and existing therapies. Management involves the minimisation of insect bites by use of stabling, fans, rugs and repellents. Symptomatic therapy involves the administration of systemic or topical corticosteroids, systemic antihistamines, and creams and sprays to promote skin healing and decrease inflammation. New immune-mediated To page 30

■ www.theveterinarian.com.au

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Dr. Aine and Dr. Mark are always available for information or support via contact numbers as shown or via our website contact page Contact details For more information about incorporating this advanced therapy into your practice: Visit: Vets1Laser.com Email:info@vets1laser.com Ph: Australia: 02 8213 2581 Europe/Ireland: + 353, 15395086 Or Come see us next month at Booth D62 at The Melbourne 2023 Veterinary Expo Laser/Photobiomodulation Therapy’s exceptional healing properties have garnered global recognition and acceptance, and now Australian & European veterinarians can access this innovative technology through the affordable

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This key clinical tool expands veterinary therapeutic options using a safe, proven, precise noninvasive, ‘drug-equivalent’ treatment modality applicable across a wide range of medical conditions i.e.: acute and chronic pain, wounds, otitis, most inflammatory presentations, rehab, exercise pre-conditioning etc.

Having seen first-hand the benefits of this therapy herself, both as a patient and as a veterinary surgeon in practice on her own veterinary patients, Dr Seavers is excited to share this true game-changer in the consult room with her veterinary colleagues.

“We are delighted to introduce Vets1Laser to Australia and Europe, where it will have a meaningful positive impact on frontline veterinary care; that in turn reduces the emotional and financial health stress burden on our veterinary practitioner colleagues, said Dr Aine Seavers.

Our affordable technology provides a non-invasive, drug-free approach

that enables vets to suppress pain, get ahead of neuropathic pain, decrease healing times, inflammation, tissue oedema, muscle spasms etc.

As a practising Veterinarian, I personally chose the Vets1Laser machine because initially I was attracted by the lack of spin and hype around the purchase; but then convinced by the practicality, versatility and ease-of-use of the machines themselves. The Vets1Laser machines arrive with a full library of pre-loaded treatment protocols, which combined with a user-friendly interface and fast treatment times, allows Vets to almost immediately and effortlessly start delivering exceptional adjunct care to their patients.

In addition to very little training being needed, the affordable price sees these machines outperform just about any other piece of capital equipment in regard to return on investment. Typically, we would expect a full ROI in 12 months, but some Practices can see this ROI even at six months.”

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TheVeterinarian 15

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Feature

Demystifying deworming: a parasitologist’s perspective An interview with Professor Rebecca Traub, BSc BVMS (Hons) PhD. Professor of Veterinary Parasitology, Melbourne Veterinary School, University of Melbourne.

Professor Rebecca Traub BSc BVMS (Hons) PhD Professor of Veterinary Parasitology, Melbourne Veterinary School, University of Melbourne Rebecca graduated as a veterinarian from Murdoch University and after spending time in small animal practice, completed a PhD in canine parasitic zoonoses. Rebecca was subsequently awarded a Fellowship to continue her research in this field by the Australian Research Council. In 2007, she gained employment as a Lecturer in Veterinary Public Health at the University of Queensland and in 2014 moved to the Melbourne Veterinary School, where she became a Professor of Veterinary Parasitology and Australian Research Council Future Fellow.

Rebecca has published over 170 international peer-reviewed papers and book chapters that have been cited over 8800 times, covering the diagnosis, zoonotic potential, epidemiology and control of canine endoparasites and vectorborne diseases, with much of her research based in the Asia Pacific. Rebecca’s research expertise has been formally recognised through consultations for the WHO, FAO, OIE, The Gates Foundation, the veterinary pharmaceutical industry, and not-forprofit organisations. In 2015, Prof. Traub founded the Tropical Council for Companion Animal Parasites. In 2019, she was awarded the Bancroft Mackerras Medal of Excellence by the Australian Society for Parasitology and is currently serving as the President of that organisation.

■ www.theveterinarian.com.au

Are you confused about what you should be recommending to dog owners when it comes to deworming? In this interview, Professor Rebecca Traub shares her expert insights on best practice deworming recommendations for dogs in Australia.

This article was supplied by Boehringer Ingelheim.

What should we be thinking about when it comes to deworming recommendations for dogs?

When considering deworming recommendations for dogs, there are a few things we need to factor in.

For example, we need to consider our patient’s age, their environment, diet, and their lifestyle. This will provide an idea of the worms they are at risk of being exposed to. Here is where remembering those parasite lifecycles becomes important! Worms like hookworm, roundworm and whipworm are canine soil-transmitted helminths. These worms have a primarily direct lifecycle. Infected dogs shed eggs in their faeces, which require an essential period in the soil, usually between 1-3 weeks, depending on conditions, to mature, before becoming infectious to a new host. So, the primary risk factor for infection with these worms is access to the outdoors. Here, contact with, sniffing or ingesting grass or soil causes dogs to ingest the mature eggs or larvae of these worms. In the case of hookworm, larvae can also infect the dog by penetrating the skin, most commonly through the paws; so, unless dogs evolve the ability to levitate, they are all essentially at risk.

In addition, microscopic larvae of roundworm encysted in raw free-range chicken, beef, lamb, and kangaroo meat can also result in dogs acquiring roundworm infection. Contrast this with hydatid tapeworm, which follows a strictly indirect lifecycle involving grazing animals as an intermediate host. The only way a dog can be infected with hydatid tapeworm is through ingesting raw meat or offal of an infected intermediate host, such as a sheep or kangaroo which has not been subjected to abattoir inspection for hydatid cysts. This might be a

risk factor for some dogs, particularly in rural areas, however for a typical suburban pet dog, the risk of infection with hydatid tapeworm is zero, and therefore this should be factored into a preventative health plan.

Similarly, dogs can only be infected with the fleatapeworm Dipylidium caninum by ingesting infected fleas, so in this case, the primary risk factor for dogs is flea infestation.

Age is another important consideration. Puppies are at greater risk of being infected with intestinal worms, as these may be transmitted in utero via the placenta for roundworms or via milk for both roundworms and hookworms. Because of their lack of immunity, low physiological reserves, and potentially high worm burdens, puppies are also more prone to severe consequences such as intestinal blockage or bloody diarrhoea, which in some cases may be fatal. This is why recommended fortnightly deworming regimes for puppies are more intensive than those for older dogs. We should not be complacent with our recommendations for older dogs however – while there is an age-related decrease in worm burdens and prevalence for some worm species, older dogs do not become resistant to infection, and in fact, for whipworm, prevalence is actually higher in older dogs.

The final consideration is the one which is perhaps the least obvious, but nevertheless important.

Historically, the default answer for many in the animal health industry as to the goal of deworming, is that it is about maximising the health and well-being of our patients. That is a very good reason for regular deworming; there is no doubt worms can have a negative impact on the health of dogs of all ages. Apart from the animal health benefits however, the other key aim of regular deworming is to keep the pet’s owners, their family and the public safe, as many worms infesting dogs pose a zoonotic risk. Regular deworming of dogs is an important aspect of minimising this risk, and in my opinion, should be the driving factor behind deworming protocols. This is particularly the case when a dog is living with individuals that are at greater risk of infection or likely to suffer severe consequences of zoonotic infections, such as young children, the elderly, or those that are immunocompromised.

SEPTEMBER 2023

TheVeterinarian 17

Feature Many clinics would say they don’t see worminfested dogs frequently in their practice.

What is the prevalence of intestinal worm infestations in Australian dogs?

This is a common question, and an important one to think about as this information informs the risk that these parasites pose. If we were considering the prevalence of say diabetes in dogs, we can get a good ballpark estimate of prevalence based on our clinical experience – dogs with diabetes present with overt clinical signs and diagnosis is relatively simple. On the other hand, gauging the prevalence of canine worms in a similar manner provides a gross underestimate of the prevalence as a healthy adult dog on a good plane of nutrition can tolerate low to moderate worm burdens without overt clinical signs. Additionally, the diagnosis of infestations can be tricky. Although performing an annual standard faecal flotation is strongly recommended, the test is relatively low in sensitivity when light infection burdens are present and may also miss tapeworm infections altogether. So, our intuitive clinical impression of worm prevalence is not a good guide with which to make recommendations around deworming.

Instead, we need to rely on prevalence studies, and in this, we are fortunate to have good data from a study published last year which looked at the prevalence of canine soil-transmitted helminths in faecal samples from dog parks across Australia. This study provides a useful real-world indicator of the risk of infestation in urban areas. This study showed that over 40% of parks across Australia were contaminated with faecal samples positive for canine soiltransmitted helminths, with hookworm being the most prevalent, followed by whipworm, threadworm, and roundworm. Apart from whipworm, these are all potential zoonoses, so there is definitely a public health issue there.

It is clear that worm infestations remain common in dogs. What about zoonotic infections in people – how common are they?

This is an area where there is limited data; these aren’t notifiable conditions. If we consider roundworm as an example though, based on seroprevalence data, it is estimated that 7% of people in Australia have been exposed to roundworm or Toxocara spp., which with our current population equates to almost 2 million people exposed. Clinical manifestations are most common in children owing to their increased likelihood of ingesting dirt-laden roundworm eggs in soil; these may be severe and include vision loss due to ocular larva migrans in children and even in adults, and encephalitis due to neural larva migrans. Fortunately, these severe manifestations represent a very small proportion of people infected, with the vast majority apparently asymptomatic or with non-specific signs including headache, fever, wheezing and abdominal pain; a classic low incidence, high consequence condition. There is, however, more recent research demonstrating a link between Toxocara seropositivity and asthma, epilepsy and potentially reduced cognitive performance in children. Similarly, infection with the dog hookworm Ancylostoma caninum may be more common than reported. In northern Australia, 165 cases of A. caninum attributed eosinophilic enteritis were reported over an eight-year period in the mid-1990s, with severe intermittent abdominal pain, vomiting, diarrhoea, and high blood-eosinophil counts being the primary presenting signs. Since dog worms infecting humans don’t usually produce eggs, diagnosis is often challenging; therefore, gastroenterologists rule out hookworms as the cause of this syndrome by observing the patient’s response to treatment with a dewormer. Given the frequency with which humans are infected, the overall public health impact of parasitic zoonoses may be considerably greater than suggested by the more overt manifestations of these diseases.

18 TheVeterinarian SEPTEMBER 2023

Cutaneous larva migranscaused by migration of hookworm larvae through human skin

Bringing this all together, and considering the goal of minimising the risk of zoonoses, what is an appropriate deworming protocol?

Before talking about specific protocols, it is important to reflect on the critical role veterinarians play in helping reduce the risk of zoonotic disease in our clients. The AVA policy on the role of vets in the management of zoonotic disease is really clear on this.

When it comes to sourcing knowledge on locally endemic zoonoses and strategies to minimise the risk of infection, the Australian Companion Animal Zoonoses Guidelines are an excellent starting point. The deworming recommendations in these guidelines are those that I would recommend. Puppies should be dewormed fortnightly starting at 2 weeks until 8 weeks of age (or two weeks post-weaning), and following this, all dogs should be dewormed monthly with a product effective against hookworms

and roundworm.2 The increased frequency of deworming from quarterly as historically practiced, to monthly, is based upon our aim of minimising the shedding of worm eggs that can contaminate the environment and pose a zoonotic risk. The benefits of monthly deworming extend to whipworm also, which although not zoonotic, has a prepatent period shorter than the traditionally recommended quarterly deworming interval. Given whipworm eggs are highly environmentally resilient, we really want to minimise environmental contamination with this parasite. In the most recent prevalence study, whipworm was the second most common worm identified, with one in 10 parks in temperate regions containing contaminated faecal samples, demonstrating there is clearly work to be done regarding controlling this parasite.

For that reason, I recommend monthly deworming with a product effective against all three of the so called “unholy trinity” – hookworm, roundworm, and whipworm.

Regular deworming with praziquantel is not required unless a dog is infested with fleas, fed raw offal from on-farm slaughtering procedures, or scavenging on carcasses. In those cases, quarterly deworming is still not appropriate. Due to the pre-patent period of flea- and hydatid tapeworms, these dogs should be dewormed every 4-6 weeks with a praziquantel- containing product.

Is monthly deworming all that we should be recommending?

No. Monthly deworming is a key part of the solution, but it is not everything. Owners should be reminded of the importance To page 30

■ www.theveterinarian.com.au

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Clinical Review

Characterising circulating nucleosomes in the plasma of dogs with hemangiosarcoma Heather Wilson-Robles1, Tasha Miller1, Jill Jarvis1, Jason Terrell2, Theresa Kathleen Kelly1,2, Thomas Bygott3 and Mhammed Bougoussa4

Abstract

biobanks. All control samples were recruited from the TAMU-SAC.

Background: Nucleosomes consist of DNA wrapped around a histone octamer core like thread on a spool to condense DNA as chromatin into chromosomes. Diseases such as cancer or inflammation lead to cell death, chromatin fragmentation and release of nucleosomes into the blood. The Nu.Q™ platform measures circulating nucleosomes in the blood of humans that result from disease and has been used to detect and identify cancer even at early stages. The objectives of this study are to quantify and better characterize nucleosomes in dogs with various stages of hemangiosarcoma (HSA) using this ELISA-based assay.

Samples from 77 dogs with a confirmed diagnosis of hemangiosarcoma and 134 healthy controls were utilized for this study. The HSA samples were recruited from the Texas A&M University Small Animal Clinic (TAMU-SAC) or purchased from

Introduction Liquid biopsy (LB) techniques refer to the minimally invasive approach of sampling tumour specific biomarkers in the blood of diseased patients [1]. LB has been most studied in the field of oncology; however, literature also exists on the use of LB for the prognostication of inflammation, sepsis and trauma in humans and dogs [2–5]. In general, LB techniques are used to identify intact circulating tumour cells, extracellular nucleic acids such as cell free DNA (cfDNA), messenger RNA (mRNA), microRNA (miRNA), extracellular vesicles (exosomes), nucleosomes and glycoproteins or antigens in the circulation (e.g. PSA, CEA, CA125) [1].

The detection of cfDNA in the blood of cancer patients is a rapidly emerging field in cancer diagnostics [6]. These techniques allow for the minimally invasive detection of cancer DNA as an adjunct or even as an alternative to standard biopsy approaches. cfDNA can be found in the cell free component of the blood and is typically in fragments that are 120–200 bp long [6]. This cfDNA is often released into the bloodstream through apoptosis or necrosis, therefore, healthy individuals typically have low levels of cfDNA in their plasma [4, 7]. Plasma concentrations of cfDNA are also very dynamic as these fragments are often cleared from the plasma within an hour [4, 7]. Many conditions can influence plasma concentrations of cfDNA including cancer, heavy exercise, inflammation, surgery or tissue injury [8].

Nucleosomes are a particular form of cfDNA released during apoptosis or necrosis in a variety of diseases. They are small fragments of chromosomes that are composed of a 147 bp segment of DNA wrapped around a histone core made of four histones present in duplicate forming an octamer [9]. Nucleosomes have been used as epigenetic biomarkers for the detection or monitoring of a variety of cancers in humans including pancreatic, lung, and colorectal cancer [10–13]. Letendre et al. published some of the first work describing the use of nucleosomes in dogs with trauma and sepsis in 2018 [14, 15]. Recently, our group published data demonstrating elevated nucleosome concentrations in a small cohort of dogs with lymphoma [16]. However, the nucleosome compartment is a relatively uninvestigated area of circulating tumour biomarkers in dogs.

The purpose of this study was to determine if dogs with hemangiosarcoma (HSA), an aggressive mesenchymal tumour derived from malignant endothelial cell precursors, 20 TheVeterinarian SEPTEMBER 2023

Results: Dogs with hemangiosarcoma had a 6.6-fold increase in their median plasma nucleosome concentrations compared to controls (AUC 92.9 %). Elevated nucleosome concentrations were seen at all stages of disease and nucleosome concentrations increased with the stage of the disease.

Conclusions: Plasma nucleosome concentrations are a reliable way to differentiate dogs with hemangiosarcoma from healthy dogs.

Further testing is underway to better characterize cancer associated HSA circulating nucleosomes and optimize future diagnostics for canine HSA detection.

Keywords: Hemangiosarcoma, Dog, Canine, Nucleosome, H3.1, Cell free DNA, Plasma, H3

demonstrated elevated concentrations of plasma nucleosomes when compared to healthy dogs [17]. HSA is a devastating disease of dogs and represents 12–21 per cent of the mesenchymal tumours in this species [18, 19]. This cancer is characterised by rapid progression and early metastasis with a one-year survival rate of < 10 per cent [20–23]. The most common primary tumour site is the spleen with HSA representing 45–51 per cent of all splenic malignancies in dogs [20, 21]. The most common presenting complaint for this disease is acute hypovolemic shock due to a spontaneous bleeding episode, however, splenic masses may be found incidentally in many older dogs. For these reasons, HSA is a prime candidate for the development of non-invasive liquid biopsy diagnostics as it would enable a non-invasive cancer screening method that would prevent unnecessary exploratory surgeries which have associated morbidities.

Results Canine patient characteristics A total of 134 healthy dogs were recruited for this study ranging in age from 10 months to 14 years (median 6 years). Forty-eight of the 134 dogs were aged 7 years or older (35.8 per cent). The healthy control cohort had a male to female ratio of 1.05 and a sex distribution including 4 intact females, spayed females (n = 61), intact males (n = 3), and castrated males (n = 65). The mean body weight for all healthy dogs was 23kg (range 2.5 to 55.8 kg). The majority of these dogs were mixed breed dogs (29/134, 21.6 per cent). Other common breeds included Labrador retriever (n = 15), Australian cattle dog (n = 10), pit bull terrier (n = 7), border collie (n = 6), golden retriever (n = 5), dachshund (n = 4), and German shepherd (n = 3). There was a total of 77 dogs in the HSA cohort. These dogs ranged in age from 4 to 15 years (median age 10 years). There were 33 spayed females (42.9 per cent), 2 intact females (2.6 per cent), 33 neutered males (42.9 per cent) and 9 intact males (11.7 per cent) in this cohort. The majority of these dogs were also mixed breed dogs (23/77, 29.9 per cent). Other common breeds in this cohort included golden retrievers (10/77, 13 per cent), German Shepherd Dogs (5/77, 6.5 per cent), Labrador retrievers (4/77, 5.2 per cent) and 3 Boxers (3.9 per cent). The remaining dog breeds contained two or fewer patients or were undefined. As expected, the majority of the dogs in this cohort presented with primary splenic HSA (53/77, 68.8 per cent). Other common sites included

the right atrium (7/77, 9.1 per cent) and bone (5/77, 6.5 per cent), primary renal (5/77, 6.5 per cent), subcutaneous/cutaneous (5/77, 6.5 per cent) and 1 each primary hepatic and skin tumours (1/77, 1.3 per cent). HSA was confirmed via histopathology in all 77 cases. Tumour stage was available for 64 of the dogs in this cohort. There were 9 dogs with stage I disease (13.8 per cent), 25 of the cases were considered stage II (38.5 per cent) and 30 were considered stage III (40.0 per cent) at diagnosis.

Nucleosome concentrations are elevated in dogs with HSA Total H3.1 nucleosome concentrations were measured in duplicate for all cases. The median nucleosome concentration for the healthy dogs was 31.1 ng/mL (mean 32.1 ng/ mL, SEM 1.1).

The dogs with confirmed HSA had a 6.6-fold increase in the median nucleosome concentration at 204.6 ng/mL (mean 420.0 ng/mL, SEM 53.6, p < 0.0001) when compared to the healthy dogs (Fig. 1). A receiver operator characteristic (ROC) curve was generated using these data and the area under the curve (AUC) was calculated to be 92.9 per cent. For a specificity of 100 per cent the cut off for the healthy range was set at 67.5 ng/mL (nucleosome range for all healthy dogs was 6.3–67.4 ng/mL). At this concentration, the sensitivity of this assay was 80.5 per cent and the specificity was 100 per cent (Fig. 2).

Elevated nucleosome concentrations vary by location in dogs with HSA Plasma nucleosome concentrations were evaluated for those locations with at least five cases each (Fig. 3). The median nucleosome concentration for dogs with primary splenic HSA regardless of stage was 214.8 ng/mL (n = 53, mean 461.6 ng/mL, SEM 69.4, p < 0.0001). For dogs with primary cardiac HSA the median nucleosome concentration was 129.9 ng/mL (n = 7, mean 300.9 ng/mL, SEM 172.9, p = 0.0001. For those with primary osseous HSA the median nucleosome concentration was 592.8 ng/mL (n = 5, mean 549.8 ng/mL, SEM 191.5, p = 0.00015.

Though the median nucleosome concentration for dogs with subcutaneous HSA was high, this group was not significantly different from the healthy cohort (n = 5, median 331.0 ng/mL, mean 331.0 ng/mL, SEM 184.8, p = 0.904). The group with the lowest median nucleosome concentration were those with primary renal HSA (n-=5, median 55.5 ng/mL, mean 115 ng/mL, SEM 72.2, p = 0.068. With the designated cut off of 67.5 ng/ mL the test was able to positively ■ www.theveterinarian.com.au

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Clinical Review predict HSA in 47/53 (88.7 per cent) cases of splenic HSA, 5/7 (71.4 per cent) primary cardiac HSA cases, 5/5 (100 per cent) primary bone HSA cases, 3/ 5 (60 per cent) primary SQ HSA cases and 1/5 (20 per cent) primary renal HSA cases.

Elevated nucleosome concentrations are seen in all three stages of HSA The 64 dogs with HSA for whom staging information was available were assessed to determine if nucleosome concentrations could be used to identify those with early stage disease. Using the previously determined healthy range cut off of 67.5 ng/mL, the assay was able to detect 6 of 9 cases with stage I disease (66.7 per cent), 19/25 cases with stage II disease (76 per cent) and 27 of 30 cases with stage III disease (90 per cent). Interestingly, the 2 stage III dogs not detected by the assay both had primary renal HSA. Median nucleosome concentrations increased with the stage of disease. The median nucleosome concentration for dogs with stage I disease was 124.2 ng/mL (mean 276.3 ng/mL, SEM 122.3, p = 0.04, AUC 78.7 per cent). The median nucleosome concentration for dogs with stage II disease was 144 ng/mL (mean 361.6 ng/mL, SEM 100.8, p < 0.0001, AUC 92.9 per cent) and the median plasma concentration for dogs with stage III disease was 413.1 ng/mL (mean 521.9 ng/mL, SEM 89.4, p < 0.0001, AUC 95.7 per cent) (Fig. 4).

Nucleosome concentrations in dogs with HSA are not affected by age, sex or weight All 77 dogs had a reported age and gender.

Sixty-nine of the dogs had a reported body weight (kg). Dogs with HSA were separated into 3 groups based on age (4–7 years n = 18, 8–11 years n = 39 and 12–15 years n = 20). All three age groups had significantly higher mean and median nucleosome concentrations than healthy dogs (Table 1). Dogs with HSA were separated into 4 groups based on gender (female spayed n = 33, female intact n = 2, neutered male n = 33, and male intact n = 9).

There were too few cases in the female intact group for a meaningful comparison, however there were enough cases in the other groups for comparisons to be made (Table 2). Finally, dogs with HSA were separated into 4 groups based on weight. The 4 weight groups were < 15 kg n = 5, 15–30 kg n = 25, 30–45 kg n = 34, and > 45 kg n = 5. All groups had significantly higher median nucleosome concentrations except for the dogs > 45 kg (p = 0.057) than the control dogs, however, there were very few dogs in the < 15 kg and > 45 kg groups (Table 3). When compared to each other, there was no significant difference between the groups of HSA cases in any category (p > 0.999 for all comparisons).

Nucleosome concentrations are altered with response to therapy Serial plasma samples from one dog with recurrent stage III HSA were evaluated throughout the course of treatment (Fig. 5). This patient was a 12-year-old MN Catahoula hog dog that had previously been diagnosed and treated for subcutaneous HSA in 2017. Through routine monitoring (whole body CT scan every 2–3 months) he was found to have progressive disease in April of 2019 with multiple pulmonary and intramuscular lesions as well as an intraseptal intracardiac lesion. Cytology on an intramuscular metastatic lesion was consistent with HSA and this was later confirmed at necropsy. This patient underwent several therapeutic interventions including stereotactic radio-body therapy, whole lung radiation, chemotherapy (doxorubicin, dacarbazine, chlorambucil and vinblastine), alpha- and beta- adrenergic blockade and immunomodulatory therapy over the course of the next year. He was in clinical remission in January of 2020 until April of 2020 where progressive disease was noted once more on imaging. Interestingly, his plasma nucleosome 22 TheVeterinarian SEPTEMBER 2023

Fig. 1 Mean nucleosome concentrations in the plasma of healthy dogs and dogs with HSA

Fig. 2 ROC curve demonstrating the ability of the assay to discriminate between healthy dogs and those with HSA. The AUC for this assay was determined to be 92.9 %

concentrations began to increase in February, 2 months before progression was noted on imaging and remained high even though additional therapies were attempted. Unfortunately, the patient failed to respond to the additional therapies and was euthanised secondary to complications of central nervous system metastases at the end of May 2020. Necropsy confirmed widespread metastatic disease.

Plasma samples were collected before progression was noted in October of 2018 and throughout treatment. These samples were processed according to predetermined optimal criteria and frozen at -80 C [16]. The samples were batched and all run together after the patient’s death in conjunction with C reactive protein (CRP) and thymidine kinase (TK) assays.

Interestingly, plasma nucleosome concentrations reflect the clinical course of disease more accurately than CRP or TK, at least in this one patient. There is a spike in CRP noted with a small increase in nucleosome concentrations in May of 2020 for this patient which coincided with an episode of severe gastroenteritis that required ICU hospitalisation for 2 days.

Discussion HSA is a common mesenchymal malignancy often affecting larger breed dogs. This disease is characterized by a primary cavitated lesion in the viscera (spleen, heart, liver), muscular or subcutaneous tissue with a high propensity to spread early and often to other locations in the body such as the lungs, lymph nodes and other viscera [20, 21]. Because this is a tumour of blood

Fig. 3 Median nucleosome concentrations in dogs with HSA based on primary location.

Dogs with primary splenic, cardiac and bone HSA had significantly higher median nucleosome concentrations than healthy dogs

vessels, a sudden onset of hypovolemic shock due to acute spontaneous bleeding maybe the first clinical sign observed by the pet owner [24].

Non-invasive diagnostics may be highly suggestive of HSA; however, it is very difficult to collect cytologic or biopsy samples of these tumours without a surgical approach in many cases due to the fragility of the tumour and the extraordinary amount of blood flow the tumours possess [21, 25]. For this reason, new tests that can help direct client decision making without increasing the risk of acute hemorrhage are needed.

Though nucleosomes have DNA wrapped around them, they stand apart from other forms of circulating DNA through their pathophysiology.

Nucleosomes play an active role in sepsis as part of neutrophil extracellular traps (NETs) which serve to remove extracellular pathogens from the blood [26]. In cancer, they are suspected to play a role in metastasis and injections of nucleosomes into mice with xenografts led to increased metastatic lesions and new tumour manifestations [27]. They have also been shown to enhance endothelial cell expression of interleukin 8 leading to neoangiogenesis in tumour tissue and ultimately tumour progression [28].

Finally, they have also been shown to play a role in tumour cell immune evasion by inhibiting natural killer cellmediated tumour lysis [29].

Though not specific for HSA, elevated nucleosome levels in the plasma of patients with HSA may provide additional information that can be useful to the client and veterinarian alike. In this cohort of 77 dogs, 80.5 per cent of them (62 ■ www.theveterinarian.com.au

Clinical Review of 77) had mean plasma nucleosome concentrations above the 67.5 ng/mL designated cut off which is higher than the nucleosome levels of all 134 healthy dogs included in this study.

A previously published manuscript evaluating the nucleosome compartment of healthy dogs determined that factors such as fasting, processing time, sample type and storage conditions can alter nucleosome concentrations in the plasma samples of healthy dogs [16]. For this reason, special care was taken in the selection and handling of these samples to ensure that all samples were handled the same way. When properly prepared, the plasma samples collected from normal dogs showed very little variability, similarly to what is seen in humans7.

The cohorts of both healthy dogs and those diagnosed with HSA were fairly diverse representing a variety of ages, breeds, body sizes and genders. However, nucleosome levels were unaffected by these factors. Only a diagnosis of HSA was able to differentiate between the two groups.

Furthermore, the mean and median plasma nucleosome concentrations in the healthy dogs were surprisingly consistent across a variety of ages, body sizes and genders (supplement data) with SEMs < 5 ng/mL in all categories evaluated except body weight where dogs weighing more than 45 kg had an SEM of 14.7 ng/mL. These data demonstrate that the nucleosome compartment is consistently low in healthy dogs across the board.

In this study nucleosome concentrations varied by location; however, these results should be viewed with caution for all locations except splenic HSA due to the low numbers of cases in the other groups. Interestingly, 100 per cent (5/5) of the primary bone HSA cases were elevated. A study evaluating 48 cases of osteosarcoma (OSA) by this group demonstrated elevated nucleosome concentrations in only 17/48 cases (35 per cent) with the highest levels in those cases with metastasis (data not shown). More cases of primary osseous HSA are needed; however, this assay may be able to differentiate HSA from OSA when it is localised to the bone.

Nucleosome concentrations are predictive of stage in humans in gastrointestinal cancer, while there is no correlation to stage for other cancer types [30]. In humans with lung and breast cancer, even the early stages of disease were found to have elevated serum nucleosome concentrations, similar to what was found here in dogs with HSA [31]. In the present study, elevated nucleosome concentrations were detected in 67 per cent of dogs with stage I disease, 76 per cent of dogs with stage II disease and 90 per cent of dogs with stage III disease.

This ability to detect early stage disease can be particularly useful as a screening test for older dogs by possibly detecting HSA before there is an acute bleeding episode.

Like most types of cfDNA, nucleosomes are cleared from the plasma quickly, usually within an hour [3, 4]. In healthy individuals’ nucleosomes are often cleared within minutes through endonuclease degradation, immunocomplexing with anti-nucleosome antibodies, phagocytosis and lysosomal degradation, hepatic metabolism or direct excretion through the kidneys [4].

However, in diseased patients, nucleosome elimination may be delayed when they bind to acute phase proteins [4]. Though cancer is not the only cause of elevated nucleosome concentrations in humans or dogs, the short half-life of this type of cfDNA can provide a real time window into the amount of apoptosis or cell turn over in that patient at any given time.

Elevations in plasma nucleosomes have been used in humans for monitoring response to therapy or recovery in a variety of diseases such as autoimmune disease, cancer, sepsis, viral infections and trauma [3, 30, 32–34]. For this reason, plasma nucleosome concentrations may ■ www.theveterinarian.com.au

Fig. 4 Mean Nucleosome concentrations from dogs of all stages with HSA compared to healthy controls. The assay was able to detect 67 % of dogs with stage I, 76 % of dogs with stage II and 90 % of dogs with stage III disease

be a reliable and effective way to screen otherwise healthy canine patients at risk for HSA. Caution should be used; however, when evaluating plasma nucleosome concentrations of clinically ill dogs as nucleosomes are not specific for HSA or other common cancers.

Nucleosome concentrations have been used to monitor treatment response in rodents with HeLa xenografts as well as humans with cervical cancer and were found to reliably predict responders in both groups [35]. Circulating nucleosome concentrations went down significantly after one cycle of platinum-based chemotherapy in the women with cervical cancer who had measurable responses. In patients with acute myeloid leukemia undergoing induction chemotherapy, nucleosome concentrations were an early predictor of treatment responders [36]. Plasma nucleosome concentrations also correlated with response to therapy in patients with non-small cell lung cancer [37, 38]. Here, we present a single case as a proof of concept that plasma nucleosome concentrations may serve as a surrogate for therapeutic response in patients with HSA. Though many more data points are needed before the utility of this test can be applied as a treatment monitoring tool for dogs with HSA, it is promising that nucleosome concentrations in dogs may mirror that which is seen in humans with a variety of cancers.

Other proteins, such as C-reactive protein (CRP) and Thymidine kinase have also been used to diagnose and monitor disease status in dogs with HSA. CRP is an acute phase protein that has been previously reported as a useful tool for evaluating remission status in canines with lymphoma and HSA [39–41]. Thymidine kinase is an enzyme involved in pyrimidine synthesis and increases in extracellular TK activity could indicate the overall degree of DNA synthesis and dying cells. TK has also been reported as a useful marker of remission status in canine lymphoma patients [39]. We evaluated both CRP and TK along with nucleosome concentrations in this patient as a proof of concept and found that CRP and TK values did not significantly change during the course of therapy and were not correlated with the clinical response in this one patient. Additional cases are needed to determine how effective nucleosomes concentrations are for disease monitoring when compared to CRP and TK. Conclusions Plasma nucleosome concentrations are an effective way to differentiate dogs with HSA from their healthy counterparts. Plasma nucleosome

concentrations are fairly consistent in healthy dogs regardless of age, gender or body size and elevated nucleosome concentrations were seen even in early-stage disease in over 80 per cent of the patients evaluated. Common signalment characteristics did not affect nucleosome concentrations in dogs with HSA when compared to healthy dogs. Measuring circulating nucleosome levels provides significant promise as a non-invasive tool for detecting HSA while limiting the likelihood for complications that are common with vascular tumours.

Materials and methods Animal samples All animal studies were approved by the Texas A&M University Institutional Animal Care and Use Committee (AUP# 2019 ? 0211 and AUP# 2017 - 0350). Additional samples were purchased from the National Cancer Institute Division of Cancer Treatment and Diagnosis (NCI-DCTD) biospecimen repository. A total of 134 healthy dog volunteers were recruited from TAMU-SAC staff and client pets. A total of 77 samples from dogs with confirmed HSA were also recruited or purchased for this study. Informed consent was obtained from all pet owners before participation in the study.

Sample collection and processing All blood samples were collected from a peripheral or jugular vein using standard blood collection techniques. All dogs were fasted for a minimum of 6 h before samples were collected.

A total of 3–5 mL (depending on the size of the dogs) were placed in EDTA primed (lavender top) tubes (Becton, Dickinson and Company, Franklin Lakes, NJ). Samples were centrifuged within one hour of collection at room temperature at 3000xg for 10 min [16]. Plasma was then immediately removed without disrupting the buffy coat layer, placed in prelabeled cryovials and frozen at -80°C to run in batches. All samples were run in duplicate.

All samples were tested using the Nu.Q H3.1 assay (Belgian Volition, SRL, Isnes, Belgium).

This enzyme-linked immunosorbent assay (ELISA) contains a capture antibody directed at histone 3.1 and a nucleosome specific detection antibody [42]. Frozen samples were thawed and allowed to come to room temperature for at least 30 min prior to analysis. Assays were performed according to the manufacturer’s protocol.

Briefly, a standard curve was generated using the positive control stock (recombinant H3.1 nucleosomes) provided. Samples were vortexed and centrifuged at 11,000 x g for 3 min at 4°C. HSA SEPTEMBER 2023

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Clinical Review samples were diluted 3-fold to ensure that they could be measured within the detection limits of the assay. The nucleosomes were bound to the capture antibody and the plates were washed 3 times using 1x wash buffer. Twenty microliters of each diluted sample were pipetted in duplicate into wells on the 96 well plates. Next, 90µL of the assay buffer was added to each well.

The plate was covered with sealing film and incubated on an orbital shaker for 2.5 h at 700 rpm. Plates were then emptied and washed 3 times using 1x washing buffer. Next, 100 uL of the detection antibody was added to each well, the plate was resealed and incubated for 1.5 h on the orbital shaker. The plates were then washed as described above. Streptavidin HRP conjugate was incubated for 30 min in each well and washed before applying the colorimetric substrate solution and incubating the plates in the dark for 20 min. A stop solution was added to the wells and the plates were read on a plate reader at 450 nm (BioTek Synergy H1 plate reader, BioTek Instruments, Winooski, VT). The standard curve was linearized and fitted to a 4-parameter logistic curve using statistical software (Graphpad Software, version 8, San Diego, CA).

One patient was followed longitudinally through his treatment protocol with serial sampling. C-reactive protein (CRP) and thymidine kinase (TK) were also measured on the samples collected at these appointments. The CRP assays were performed by the Texas A&M University Gastrointestinal Laboratory and the TK assay was performed using the Canine Thymidine Kinase 1 soluble ELISA assay (My Biosource Inc, San Diego, CA) as long as sufficient sample was available to do so.

CRP Assays For the longitudinally followed patient, samples were submitted to the Texas A&M University Gastrointestinal Laboratory for c-reactive protein (CRP) assays if sufficient sample quantity was present. If sample quantity was not sufficient for both nucleosome and CRP analysis, nucleosome assays were given priority.

Thymidine kinase assays The Canine Thymidine Kinase 1 soluble ELISA assay (MyBiosource Inc, San Diego, CA) was used to evaluate TK1 levels in the dog followed longitudinally. The assay was performed according to the manufacturer’s protocol.

Briefly, 40 µl of sample was added to wells followed by 10 µl anti-TK1 antibody. Then 50 µl streptavidin-HRP was added to each well except the blank control well. The plate was mixed well, covered with sealer, and incubated for 60 min at 37°C. The plate was then washed 5 times with wash buffer and the wells were soaked with at least 0.35 ml wash buffer for 30 s to 1 min for each wash. Next 50 µl of substrate solution A was added to each well followed by 50 µl of substrate solution B to each well. The plate was covered with a fresh sealer for 10 min at 37°C in the dark. Finally, 50 µl of Stop Solution was added to each well.

Plates were read at an absorbance of 450nm (BioTek Synergy H1 plate reader, BioTek Instruments, Winooski, VT) within 10 min of stop solution being added. The standard curve was linearised and fitted to a 5-parameter logistic curve using statistical software (Graphpad Software, version 8, San Diego, CA).

Intra and intergroup comparisons HSA samples were separated out by stage, age, weight and gender and compared used the statistical methods below. Stage I HSA is characterised by a solitary lesion that is < 5 cm in diameter, no evidence of hemorrhage and no evidence of metastasis (locoregional or distant) [43]. Stage II HSA is characterised by a lesion ■ www.theveterinarian.com.au

that is either 5 cm or greater or one that has ruptured with or without locoregional metastasis and no distant metastasis [43]. These tumours may also be invading into the subcutaneous tissues. Stage III HSA is characterised by highly invasive tumours with or without distant metastasis or tumours of any size with distant metastasis [43]. Age was divided into 4 groups: <3 years, 4–7 years, 8–11 years and > 12 years. Gender was also divided into 4 groups: spayed females, intact females, neutered males and intact males. Finally weight was also divided into 4 groups: <15 kg, 15–30 kg, 30–45 kg and > 45 kg.

Statistical analysis Descriptive statistics for the patient populations were performed using Microsoft Excel for Mac (v. 16.16.27, 2016). For data sets containing only two cohorts, such as the healthy controls versus all HSA cases, a Wilcoxon rank sum test was used to compare the medians of the data sets. For data sets where multiple conditions were compared, such as disease stage, a two-way ANOVA for repeat measures with a Tukey’s multiple comparisons test was performed. This part of the analysis was performed using GraphPad Prism version 8.0.0 for Macintosh, GraphPad Software, San Diego, California USA, www.graphpad.com.

Wilcoxon rank sum tests, ROC curves and specificity/sensitivity calculations were performed using R version 3.4.3 and the pROC package [44, 45].

Abbreviations AUC: Area under the curve; CA125: Cancer antigen 125; CEA: Carcinoembryonic antigen; cfDNA: Cell free deoxyribose nucleic acid; CRP: C-reactive protein; EDTA: Ethylenediaminetetraacetic acid; ELISA: Enzyme linked immunosorbent assay; HSA: Hemangiosarcoma; ICU: Intensive care unit; LB: Liquid biopsy; NCI-DCTD: National Cancer Institute Division of Cancer Treatment and Diagnosis; NETs: Neutrophil extracellular traps; PSA: Prostate

specific antigen; ROC: Receiver operator characteristic; SEM: Standard error of mean; TK: Thymidine kinase Author details

1College of Veterinary Medicine, Small Animal

Clinical Sciences Department, Texas A&M University, College Station, TX 77843, USA. 2Volition America & Volition Veterinary Diagnostic Development, 13215 Bee Cave Parkway, Galleria Oaks B, Suite 125, Austin, Texas 78738, USA. 3Volition Diagnostics UK Ltd, 93-95 Gloucester Place, London W1U 6JQ, UK. 4Belgian Volition SRL, 22 Rue Phocas Lejeune, Parc Scientifique Crealys, 5032 Isnes, Belgium.

This article originally appeared in BMC Veterinary Research and was supplied to The Veterinarian by Heska.

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14. Letendre JA, Goggs R. Determining prognosis in canine sepsis by bedside measurement of cell-free DNA and nucleosomes. J Vet Emerg Crit Care (San Antonio). 2018;28(6):503–11.

15. Letendre JA, Goggs R. Concentrations of Plasma Nucleosomes but Not CellFree DNA Are Prognostic in Dogs Following Trauma. Front Vet Sci. 2018;5: 180.

16. Wilson-Robles H, Miller T, Jarvis J, Terrell J, Dewsbury N, Kelly T, et al. Evaluation of nucleosome concentrations in healthy dogs and dogs with cancer. PLoS One. 2020;15(8):e0236228.

17. Lamerato-Kozicki AR, Helm KM, Jubala CM, Cutter GC, Modiano JF. Canine hemangiosarcoma originates from hematopoietic precursors with potential for endothelial differentiation.

Exp Hematol. 2006;34(7):870–8.

18. Bastianello SS. A survey of neoplasia in domestic species over a 40-year period from 1935 to 1974 in the Republic of South Africa. III. Tumours occurring in pigs and goats.

Onderstepoort J Vet Res. 1983;50(1):25–8.

19. MacVean DW, Monlux AW, Anderson PS, Jr., Silberg SL, Roszel JF. Frequency of canine and feline tumours in a defined population. Vet Pathol. 1978;15(6): 700–15.

20. Spangler WL, Culbertson MR. Prevalence, type, and importance of splenic diseases in dogs: 1,480 cases (1985–1989). J Am Vet Med Assoc.

1992;200(6): 829–34.

21. Spangler WL, Kass PH. Pathologic factors affecting postsplenectomy survival in dogs. J Vet Intern Med. 1997;11(3):166–71.

22. Brown NO, Patnaik AK, MacEwen EG. Canine hemangiosarcoma: retrospective analysis of 104 cases. J Am Vet Med Assoc.

1985;186(1):56–8.

23. Prymak C, McKee LJ, Goldschmidt MH, Glickman LT. Epidemiologic, clinical, pathologic, and prognostic characteristics of splenic hemangiosarcoma and splenic hematoma in dogs: 217 cases (1985). J Am Vet Med Assoc.

1988; 193(6):706–12.

24. Aronsohn MG, Dubiel B, Roberts B, Powers BE. Prognosis for Acute Nontraumatic Hemoperitoneum in the Dog: A Retrospective Analysis of 60 Cases (2003–2006). J Am Anim Hosp Assoc. 2009;45(2):72–7.

25. Wendelburg KM, Price LL, Burgess KE, Lyons JA, Lew FH, Berg J. Survival time of dogs with splenic hemangiosarcoma treated by splenectomy with or without adjuvant chemotherapy: 208 cases (2001–2012). J Am Vet Med Assoc. 2015;247(4):393–403.

26. Krogh AKH, Lyngby JG, Bjørnvad CR, Nielsen LN. Presence of nucleosomes in plasma and increased thrombin generation in dogs ■ www.theveterinarian.com.au

Fig. 5 Longitudinal assessment of nucleosome concentrations in a patient with stage III HSA throughout the course of treatment. The green circles represent clinical remission, red circles represent progressive disease and the orange circle represent a partial clinical response. All assessments were performed by a board-certified veterinary medical oncologist. All imaging was interpreted by a board-certified veterinary radiologist

with acute and chronic gastroenteropathies.

Res Vet Sci. 2021;135:504–10 https://doi.org/ 10.1016/j.rvsc.2020.11.012.

27. García-Olmo D, García-Olmo DC, Ontañón J, Martinez E, Vallejo M. Tumour DNA circulating in the plasma might play a role in metastasis.

The hypothesis of the genometastasis. Histol Histopathol. 1999;14(4):1159–64.

28. Tanner JE. Nucleosomes activate NF-kappaB in endothelial cells for induction of the proangiogenic cytokine IL-8. Int J Cancer.

2004;112(1):155–60.

29. Le Lann-Terrisse AD, Fournié GJ, Benoist H. Nucleosome-dependent escape of tumour cells from natural-killer-mediated lysis: nucleosomes are taken up by target cells and act at a postconjugation level. Cancer Immunol Immunother.

1997;43(6):337–44.

30. Holdenrieder S, Stieber P, Bodenmüller H, Busch M, Fertig G, Fürst H, et al. Nucleosomes in serum of patients with benign and malignant diseases. Int J Cancer. 2001;95(2):114–20.

31. Holdenrieder S, Stieber P, Bodenmüller H, Busch M, Von Pawel J, Schalhorn A, et al.

Circulating nucleosomes in serum. Ann N Y Acad Sci. 2001;945:93–102.

32. Holdenrieder S, Stieber P. Therapy control in oncology by circulating nucleosomes. Ann N Y Acad Sci. 2004;1022:211–6.

33. Holdenrieder S, von Pawel J, Dankelmann E, Duell T, Faderl B, Markus A, et al. Nucleosomes, ProGRP, NSE, CYFRA 21 – 1, and CEA in monitoring firstline chemotherapy of small cell lung cancer. Clin Cancer Res. 2008;14(23): 7813–21.

34. Holdenrieder S, von Pawel J, Dankelmann E, Duell T, Faderl B, Markus A, et al. Nucleosomes and CYFRA 21 – 1 indicate tumour response after one cycle of chemotherapy in recurrent non-small cell lung cancer. Lung Cancer.

2009;63(1):128–35.

35. Trejo-Becerril C, Perez-Cardenas E, TrevinoCuevas H, Taja-Chayeb L, GarciaLopez P, SeguraPacheco B, et al. Circulating nucleosomes and response to chemotherapy: an in vitro, in vivo and clinical study on cervical cancer patients.

Int J Cancer. 2003;104(6):663–8.

36. Mueller S, Holdenrieder S, Stieber P,

Haferlach T, Schalhorn A, Braess J, et al. Early prediction of therapy response in patients with acute myeloid leukemia by nucleosomal DNA fragments. BMC Cancer. 2006;6:143.

37. Holdenrieder S, Stieber P, J VONP, Raith H, Nagel D, Feldmann K, et al. Early and specific prediction of the therapeutic efficacy in nonsmall cell lung cancer patients by nucleosomal DNA and cytokeratin-19 fragments. Ann N Y Acad Sci. 2006;1075:244–57.

38. Holdenrieder S, Stieber P, von Pawel J, Raith H, Nagel D, Feldmann K, et al. Circulating nucleosomes predict the response to chemotherapy in patients with advanced non-small cell lung cancer. Clin Cancer Res. 2004;10(18 Pt 1): 5981–7.

39. Selting KA, Ringold R, Husbands B, Pithua PO. Thymidine Kinase Type 1 and C-Reactive Protein Concentrations in Dogs with Spontaneously Occurring Cancer. J Vet Intern Med. 2016;30(4):1159–66.

40. Nakamura M, Takahashi M, Ohno K, Koshino A, Nakashima K, Setoguchi A, et al. Creactive protein concentration in dogs with various diseases. J Vet Med Sci. 2008;70(2):127–31.

41. Thamm DH, Kamstock DA, Sharp CR, Johnson SI, Mazzaferro E, Herold LV, et al.

Elevated serum thymidine kinase activity in canine splenic hemangiosarcoma*. Vet Comp Oncol. 2012;10(4):292–302.

42. Arimura Y, Ikura M, Fujita R, Noda M, Kobayashi W, Horikoshi N, et al. Cancer-associated mutations of histones H2B, H3.1 and H2A.Z.1 affect the structure and stability of the nucleosome. Nucleic Acids Res. 2018;46(19): 10007–18.

43. Batschinski K, Nobre A, Vargas-Mendez E, Tedardi MV, Cirillo J, Cestari G, et al. Canine visceral hemangiosarcoma treated with surgery alone or surgery and doxorubicin: 37 cases (2005–2014). Can Vet J. 2018;59(9):967– 72.

44. Team RC. R: A language and environment for statistical computing. 2017.

45. Xavier Robin NT, Alexandre Hainard, et al. pROC: an open-source package for R and S + to analyze and compare ROC curves. BMC Bioinformatics. 2011;7(77).

SEPTEMBER 2023

TheVeterinarian 27

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Introducing NexGard SPECTRA Cats are the masters of multiparasitism, with their natural behaviours and instincts placing them at high risk of ecto- and endoparasites.

For example, roaming exposes cats to fleas, ticks and intestinal parasite eggs and larvae shed by other pets and wildlife; grooming behaviour can result in ingestion of fleas containing flea tapeworm larvae; and hunting exposes cats to roundworm, lungworm and Taenia tapeworms carried by intermediate or paratenic hosts.

More than 80% per cent of owned cats in Australia have some access to the outdoors, increasing their risk of exposure to parasites and infectious diseases.1 However, parasites aren’t only a risk for outdoor cats. Indoor cats may be exposed due to rodents or other intermediate/paratenic hosts entering the home, environmentally resilient worm eggs being carried in on shoes, heartworm-infected mosquitoes entering the home, or other pets bringing ecto- or endoparasites into the home environment. In fact, in one study, 20 per cent of cats diagnosed with intestinal worms were described by their owners as “indoor only”.2 Broad-spectrum control of the important endoparasites and ectoparasites of cats has traditionally required the use of multiple products – potentially with different dosing intervals and methods of administration.

This can be confusing for pet owners and may reduce compliance. NexGard SPECTRA Spot-On for Cats helps address these

challenges – it is Australia’s first and only all-in-one parasiticide for cats, providing protection against fleas, ticks, mites, heartworm, lungworm and intestinal worms (including tapeworms).

NexGard SPECTRA Spot-On for Cats’ unique combination of three active ingredients (esafoxolaner, eprinomectin and praziquantel) provides the most complete protection available against both external and internal parasites, all in one easy, monthly application.

The product is applied topically with an innovative, easy-to-use applicator to reduce stress and mess for cats and their owners. It can be used on all cats from eight weeks of age and weighing at least 0.8 kg, including breeding, pregnant and lactating queens.

Parasite protection is not just about keeping our pets healthy. Cat fleas can transmit zoonotic pathogens and parasites, and roundworms and hookworms can infect and cause disease in people. The monthly dosing regimen of NexGard SPECTRA® aligns perfectly with the recommendations from the Australian Companion Animal Zoonoses Guidelines which recommend monthly deworming and year-round flea control for all cats to help reduce this risk.3 For more product information and other technical resources, please visit Animal Health Academy at www.animalhealthacademy.com.au/ products/nexgardspectraforcats. When registering for the first time, please use the access code: myAcademy

CVS enters Australian market UK-based multinational CVS has entered the Australian veterinary market with the purchase of several practices in Queensland and New South Wales. The company, which already has around 500 practices across the UK, the Netherlands and Ireland, intends to purchase more Australian practices in the future. “Last November we outlined that we wished to grow and offer independent vets the opportunity to join our group in new countries,” Deputy CEO of CVS Ben Jacklin said. “Since then, we’ve seen that the veterinary sector in Australia gives us an opportunity to stand out by providing a high level of clinical care, great facilities and equipment, and for being a great place to work and have a career.”

Founded in 1999, CVS has gradually expanded its global presence. In addition to the more than 500 practices it has acquired worldwide, the group operates ten specialist referral hospitals, 35 dedicated out of hours sites providing 24/7 emergency care, two laboratories and MiLab offering fast diagnostic services, seven pet crematoria, a Healthy Pet Club scheme aimed at providing preventative care and spreading the cost of regular health checks, MiPet pet insurance, an online retail business and a VetDirect veterinary buying group. “Our purpose is to provide the best clinical care every time, and we believe this is what drives the success of our practices,” Jacklin said. “Alongside great facilities and equipment, we aim to deliver the highest standard of healthcare and empower our colleagues to make the right clinical decisions for our patients.”

Australia has proved to be a natural fit for CVS’s recent expansion, with Jacklin citing strong similarities in practice operations and an emphasis on veterinarians’ wellbeing, together with shared language and culture, as being important factors underpinning the

■ www.theveterinarian.com.au

acquisition of practices in this country. “All the practices we have acquired to date in Australia are of a very high quality, so the UK could learn as much from them as we can bring to the Australian market here,” Jacklin said.

“For us, it's not about making Australia an extension of the UK. It’s about focussing on Australia itself and supporting our Australian colleagues with operational enhancements from Europe.”

CVS is primarily aiming to acquire small animal practices in Australia but has not ruled out the possibility of adding practices with farm and equine clients as well. The main feature Jacklin is looking for in practices interested in joining CVS is cultural similarity and a strong alignment to the company’s values. “Our culture is veterinary led and we are people focussed,” he explained. “We’re particularly interested in practices who believe in our purpose, which is to give the best possible care to animals, and our vision, which is to be the veterinary company people most want to work for.” That said, Jacklin is quick to point out that practices who choose to join the group will maintain their own identity and autonomy. “We won’t change their practice, practice name, clinical care or culture, and we’ll also maintain and protect entitlements, wages and the conditions of their team,” he said. “CVS will supply expert teams in Recruitment, HR, Accounts and Procurement – which leaves practices free to focus on patients. We’ll look to invest in great facilities and equipment, and we’ll support and develop colleagues to be their best by offering industry leading CPD, including our renowned Knowledge Hub learning platform.”

CVS is initially focusing on acquiring veterinary practices in major centres, including Sydney, Melbourne, Brisbane, Perth, Adelaide, Canberra and Newcastle, and Jacklin expects

Two applicator sizes are available: For cats over 7.5 kg, the appropriate combination of applicators can be used.

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*Retail single dose packs in vet only.

References 1. Johnston, L., et al (2017) Demographics, lifestyle and veterinary care of cats in Australia and New Zealand. J Feline Med Surg, 1-7.

2. Coati, N. et al (2003) Recent investigation on the prevalence of gastrointestinal nema todes in cats from France and Germany.

Parasitol Res. 90, S146-S147.

3. Australian Companion Animal Zoonoses Advisory Panel (2021) Companion Animal Zoonoses Guidelines.

See product label for full claim details and directions for use.

CVS Australia to be run by Australian clinicians rather than UK based personnel in the short to medium term. The company also plans to have dedicated Australian-based recruitment support and operational support teams. “In addition, our front-line general practice vets will be supported by specialist CVS veterinary teams and subject matter experts – from our Regional Clinical Hubs, Advanced Clinical Network, and Vet Oracle ‘virtual animal hospital’ – who will offer unique expertise, experience and advice to practices, so they are never alone when dealing with a case,” Jacklin said. “Added to this, we have a culture of sharing best practice company-wide, whilst giving our colleagues freedom to make the best clinical decisions.”

While it is difficult to ascertain precisely when CVS will become a national presence in Australia, Jacklin said the company is “looking at all opportunities in the Australian veterinary market” and, despite current veterinary staff shortages, believes that Australia represents an attractive place to work for veterinarians. “We have dramatically reduced attrition in CVS UK over the last five years and will want to have the same impact in Australia,” he said. “We also have a focus on helping the next generation of vets. In the UK we operate outstanding student Intra Mural Studies and Extra Mural Studies, and we recruit up to 250 graduations annually, who all pass through our industry-leading two-year graduate induction programme.”

Jacklin is excited about the prospect of the CVS group making a positive contribution to the Australian veterinary marketplace. “We are people focussed, not bottom line focussed, and we think this will be appreciated in Australia,” he said. “We are not necessarily in it to be the biggest. We are in this for the long term in Australia; it is a long-term investment.”

■ JAI HUMEL

SEPTEMBER 2023 TheVeterinarian 29

News

Australians support a legislated end A role for drones date for the live sheep export trade in saving wildlife TheVeterinarian From page 1

would see an end to Australia’s live sheep export trade. The submission also recommended a short-term phase out period of no longer than three years from the passage of legislation to avoid adverse animal

welfare outcomes from a longer-term phase out.

The independent panel is due to lodge its final report and recommendations to the federal government by 30 September.

■ ANNE LAYTON-BENNETT

Demystifying deworming: a parasitologist’s perspective From page 19

of personal and environmental hygiene. The role of personal hygiene is, or at least should be, pretty obvious. Environmental hygiene on the other hand may be underappreciated. Pet owners should promptly remove and dispose of dog faeces, at least daily.

If we consider that hookworms require several days at a minimum in the environment before becoming infectious to a new host, canine or human, the benefit of daily removal, and appropriate disposal of faeces becomes apparent.

Even with monthly deworming, we should also be recommending annual faecal testing for all dogs to ensure owner compliance. While standard faecal flotations will detect moderate and heavy worm burdens, antigen-based techniques are more sensitive and preferred for this.

Is there a risk that more frequent deworming of pet dogs will promote anthelmintic resistance?

There is no doubt that if you look at production animal medicine, intensive deworming regi-mens have resulted in the development of anthelmintic resistance. Will the same thing happen in companion animals? The short answer, based upon our knowledge of the drivers of anthelmintic resistance, is that this is considered unlikely. The key to understanding why this is the case is ‘refugia’. Think of refugia as the proportion of the worm population not exposed to anthelmintics, and which are therefore susceptible or killed by their action. Refugia comes from eggs or larvae in the environment passed by untreated animals (domestic or wild), or parasite stages in the host not exposed to treatment (e.g., encysted tissue stages). In livestock, animals are typically treated simultaneously, on-mass, in a closed population, resulting in very little refugia. Under this scenario, animals become reinfested with worms which survived the initial anthelmintic treatment, so that the 30 TheVeterinarian SEPTEMBER 2023

worms that are acquired are already partially or wholly resistant. As this process repeats, it selects for aparasite population with ever greater resistance.

Compare this to deworming in dogs. Pet-owned dogs are treated on an individual basis, in an open environment, with refugia provided by untreated pet-owned dogs and wildlife (foxes, feral dogs) in that environment. Proper environmental hygiene further reduces the risk, with potentially resistant worms ending up in landfill rather than a new host, thereby eliminating those resistant genes from the gene pool.

Monthly deworming in the context of the typical husbandry practices of pet-owned dogs in Australia is considered likely to have a negligible impact on refugia and therefore as a driver for anthelmintic resistance.

The importance of appropriate husbandry practices and the role of refugia in minimising the risk of anthelmintic resistance is highlighted by the fact that the rare, reported cases of anthelmintic resistance in dogs almost invariably involve animals managed more like livestock than pet dogs – that is in racing greyhounds, the husbandry of which typically involves regular mass treatment of a closed population in circumstances of lessthan-ideal environmental hygiene.

If we avoid treating dogs like livestock, we may avoid the unintended negative consequences of anthelmintic treatment seen in production animals.

References 1. Australian Veterinary Association (2020) The role of veterinarians in the management of zoonotic disease. www.ava.com.au/policyadvocacy/policies/professionalpractices-for-veterinarians/the-roleof-veterinarians-in-the-management-of-zoonotic-disease/, accessed 7 May 20232. Australian Companion Animal Zoonoses Advisory Panel (2021) Companion Animal Zoonoses Guidelines.

From page 8

Colombelli-Négrel, who is also the principal investigator at FU’s Bird Lab, said the research formed part of an ongoing development of tools such as drones and software loaded on UAVs, able to compile unbiased data over large geographic areas with high spatial and temporal resolution. The use of acoustic software recording, and night-time videoing of Granite Island’s little penguin population was also being trialled by her team of researchers and community volunteers.

“We found the videos were also critical in identifying other threats, such as land predators of little penguins. These results add to a growing body of literature that suggests human activity and the consequent disturbances may affect the numbers and behaviours of wildlife, and appropriate measures need to be developed to limit such impacts, particularly when populations are declining or in peril,” she said.

‘Koalas showed limited behavioural response and no physiological response to drones’ is available at doi.org/10.1016/j.applanm.2023.

105963 ■ ANNE LAYTON-BENNETT

Abstracts - laying hen welfare From page 14

housing design and alignment of breeding goals. However, much remains to be done. The design and performance of veranda-based systems which provide hens with fresh air and natural light is a promising avenue for future research aimed at optimising hen welfare and improving sustainability.

Christine Nicol1 Br Poult Sci. 2023 Aug;64(4): 441447.doi: 10.1080/00071668.2023.

2211891.

Sydney Magazine Publishers Pty Ltd ACN 102752787 ISSN 1447-9768 PO Box 305 Millthorpe NSW 2798 Tel: 02 9487 6627 Director - Clem Martin Email: clem.martin@vetmag.com.au Editorial Advisory Board Thomas Donnelly BVSc DipVP DipACLAM Nicholas Kannegieter BVSc DipVetClinStud PhD FACVSc David Lidbetter BVSc MVS FACVSc DipECVS Dip ACVS Angus Martin BVSc Mary Porter BVSc (Hons) MACVSc Jeffrey Smith BVSc FACVSc Dip ACVO David Vella BSc BVSc (Hons) DABVP Editor - Luke Martin Tel: 0418 698 228 Email: luke.martin@vetmag.com.au National Sales Manager James Martin Tel: 0432 575 877 Email: james.martin@vetmag.com.au Designer - Anne Norrell Email: annenorrelldesign@gmail.com Printed by Newstyle Printing 41 Manchester St, Mile End SA 5031 Post Print Approved PP255003/06488 All material in The Veterinarian is subject to copyright and must not be reproduced wholly or in part without the written permission of the Editor.

Views expressed in The Veterinarian are not necessarily those of The Veterinarian or Sydney Magazine Publishers Pty Ltd. Subscriptions - Clem Martin Tel: 02 9487 6627 Email: clem.martin@vetmag.com.au

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1Department of Pathobiology and

Population Sciences, Royal Veterinary College, London, UK. The Gordon Memorial Lecture was given at the Spring Meeting of the WPSA UK Branch meeting held at the University of Nottingham, 13 April 2022.

Abstracts - insect bite

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From page 14

therapeutics including vaccines, in addition to desensitisation procedures, show promise at controlling hypersensitivity reactions. This article will review aetiologic agents, pathophysiology, scoring systems and current and new therapies.

Abbey Cox1, Allison J Stewart1 Animals (Basel). 2023 Aug 4;13(15): 2514.doi: 10.3390/ani 13152514.

1School of Veterinary Science, University of Queensland Gatton, 5391 Warrego Highway, Gatton, QLD 4343, Australia.

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Free PMC article ■ www.theveterinarian.com.au

New Zealand sea lion (Phocarctos hookeri) Once known as Hooker’s sea lion, this species is endemic to New Zealand and primarily breeds on the subantarctic Auckland and Campbell islands.

With a population of only around 12,000 individuals, it is one of the world’s rarest sea lion species. Picture Hase

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Best clinical practice is to identify causative organism by cytology or culture and sensitivity before, or as soon as possible after treatment is commenced. For single application treatment of otitis externa in dogs associated with susceptible strains of yeast (Malassezia pachydermatis) and bacteria (S. pseudintermedius, S. canis, E. coli and Proteus spp). © 2021 Elanco or its affiliates. Elanco Australasia Pty Ltd (ABN 64 076 745 198), Level 3, 7 Eden Park Drive, Macquarie Park NSW 2113. All trade marks are the property of their respective owners. For further information contact: 1800 678 368 from anywhere in Australia Monday to Friday or email enquiries_au@elancoah.com. ELAN0146 11/21. PM-AU-21-0854.