Reports of Dirofilaria immitis infection vary by location in the USA and Europe, with an occurrence gradient increasing towards the equator and warmer climates. In Australia, heartworm preventative guidelines are not climate specific, implying homogenous risk of infection across the continent. We systematically reviewed the published literature to assess if the distribution of D. immitis infection varies in Australia. We sourced 41 relevant publications by searching multiple bibliographic databases and the citations of key articles. From those, we extracted the data from a total of 106 individual surveys of variable size and quality. The reported prevalences ranged from 0.0 % to 100.0 % and we meta-analysed the observed heterogeneity using survey location’s latitude, climate zone or temperature/humidity zone. Crude heterogeneity between surveys was partly explained (approximately 30 %) by temperature/humidity zone, and the quality of the surveys did not affect the proportional explanation of heterogeneity. There was significantly higher pooled prevalence in humid summer zones (18.7 %, 95 %CI: 10.9–30.1 %) compared to cool winter zones (2.1 %, 95 %CI: 1.0–4.3 %). Historical reports of D. immitis infection in Australia concur with the strong association to warmer climates observed in other continents. The current blanket recommendation for dirofilariosis preventative usage does not reflect the important variability of infection risk to canids across locations of Australia.

Frontline drug treatments for malaria are at risk of failing due to emerging resistance, meanwhile drugs used to treat toxoplasmosis have suboptimal efficacy and safety. As demonstrated by the success of clinically used antiparasitic drugs, the diverse structural complexity and biological activity of natural products holds great potential for drug discovery and development, to address the need for new compounds with novel mechanisms. Here we screened the BioAustralis Discovery Plates Series I library, a collection of 812 microbial natural product compounds including rare microbial metabolites, against Plasmodium falciparum erythrocytic stage and Toxoplasma gondii tachyzoite parasites. We identified 219 compounds that inhibited P. falciparum growth by at least 80 % at a concentration of 2 μg/mL (1-10 μM for >90 % of compounds), whilst 149 compounds demonstrated equivalent activity against T. gondii. The active compounds were assigned based on chemical structure to more than 50 compound classes. After triaging active compounds for those with low mammalian cytotoxicity, we defined the in vitro half maximal inhibitory concentration (IC50) of a selection of compounds against the parasites, identifying four compound groups with activity in the low nanomolar range. The macrocyclic lactone pladienolide B and cryptopleurine were found to be very potent against the parasites but also mammalian cells, warranting further structure-activity relationship investigation. Two groups, the monocyclic thiazole peptides, including micrococcin P1 and the thiocillins, and the pleuromutilins, exhibited both low antiparasitic IC50 and low cytotoxicity, highlighting their potential for further analysis. This study defines the activity of the BioAustralis Discovery Plates Series I against two apicomplexan parasites of significant global importance, providing potential new tools to study parasite biology and possible starting points for novel antiparasitic development.

Cryptosporidium spp. sporadically infect a range of Australian native mammals including koalas, red kangaroos, eastern grey kangaroos, bilbies and brush tailed possums and can range from asymptomatic to fatal infections. Traditionally considered a disease of the young or immuno-compromised, and resulting in profuse diarrhoea in other species, here we report an atypical clinical syndrome associated with Cryptosporidium in a captive population of koalas. All affected animals were in-contact adults, and demonstrated anorexia, dehydration and abdominal pain in the absence of diarrhoea. Following euthanasia on welfare grounds, Cryptosporidium infection was confirmed postmortem in three of four symptomatic animals via faecal floatation and/or intestinal histopathology, with enteritis also diagnosed in the fourth koala. Further screening of the captive colony found the outbreak had been contained. Based on sequencing the cause of the infection was C. fayeri, but the source was undetermined. In conclusion, Cryptosporidium should be considered as a possible cause of generalised illness in koalas.

Toxoplasma gondii (T. gondii) is a protozoan parasite of substantial impact to small ruminants, with reproductive failure a possible outcome of exposure. This observational study assessed T. gondii prevalence within the South Australian sheep population and investigated on-farm risk factors to T. gondii exposure via a cross-sectional survey. 1433 individual animals, consisting of 1282 mixed-age ewes and 151 mixed-age rams were blood sampled and serologically screened for T. gondii-specific antibodies. A risk-analysis questionnaire was conducted for each participating property. Of the 1433 animals sampled, 530 tested positive, with seroprevalence observed to be 39 % (95 % CI 28.7-49.3 %) after accounting for clustering within properties. All properties returned at least one positive result, indicating a flock level seroprevalence of 100 %. T. gondii seroprevalence was found to be higher in sheep on Kangaroo Island (46.6 %; 95 % CI 32.1-61.1 %) compared to the South Australian mainland (31.3 %; 95 % CI 18.4-44.2 %), however this difference was not statistically significant (P=0.125). A significant association was observed between T. gondii seroprevalence and age, with seroprevalence increasing from 30.2 % (95 % CI 17.7-42.6 %) in one year old sheep, to 69.7 % (95 % CI 47.0-92.5 %) in sheep older than six years (P=0.001). T. gondii seroprevalence was significantly higher in animals drinking water from surface water sources (55.9 %; 95 % CI 35.2-76.6 %), compared to those exclusively sourcing reticulated mains water (19.1 %; 95 % CI 0 %-39.0 %) (P=0.028). An individual animal exposed to a surface water source was found to be more than ten times as likely to be exposed to T. gondii, than an animal sourcing only reticulated mains water (odds ratio:10.68; 95 % CI 1.30-87.88). Water source is important in the transmission of T. gondii to South Australian sheep. Mitigation strategies should be developed and targeted at reducing contact between oocysts and water sources and reducing interaction between livestock and contaminated water.

The causative agent of canine heartworm disease, Dirofilaria immitis, requires specific temperature conditions to mature within its mosquito vector, and therefore (re-)infect a canid host. Suitable temperature conditions are not continuously met for locations where most (>97%) Australians and their pet dogs live. The length of the disruption in the transmissibility of D. immitis varies greatly across Australia, and to some degree, between years. We developed an online dashboard 'Transmission Tracker - Dirofilaria' that processes near real-time temperature records across Australia and allows users to enquire about historical and current weather suitability for canine heartworm transmission at any Australian postcode of their interest. This information allows veterinarians to access when, and for how long, heartworm may be transmitted at a specific location, assess the associated risk of infection and advise on a patient-dependent dirofilariosis prevention plan for their canine patients and guardians. Our dashboard is publicly accessible at: https://heartworm-mapping.adelaide.edu.au/shiny/.

Dirofilaria immitis is the causative agent of canine heartworm disease. We used the established heartworm development unit (HDU) principle to map the extrinsic incubation period (EIP) of D. immitis in Australia using historical weather data from 2013-2022. We found weather conditions suitable for EIP completion showed substantial seasonality and geographical variability. Whilst a considerable percentage of the Australian territory showed suitable weather conditions to always support EIP completion (17%), only 2.7% of the 2021 Australian human population lived in this region. Therefore, 97% of the population lived in an area that changed its EIP suitability within the study period. EIP completion is required prior to D. immitis transmission, meaning that infection risk of D. immitis is seasonal and location-dependent, being disrupted each year for most of the human population's dogs. We developed an online, open access tool allowing us to visualise EIP completion across Australia historically and in near real-time. We aim to support veterinarians to make risk-based recommendations for dirofilariosis prevention by using the tool, available at https://heartworm-mapping.adelaide.edu.au/shiny/.

The single-celled parasite Toxoplasma gondii uses mice as a vector to reach its definitive host, the cat, where it can accomplish its sexual reproduction and produce oocysts, which will contaminate the environment. In this study, we have captured 103 feral house mice (Mus musculus) on Kangaroo Island, Australia. We have measured the level of exposure to T.gondii serologically with the Modified Agglutination Test and conjointly with a T.gondii B1 gene PCR. We have included stringent quality control steps in the molecular analysis to reduce the risk of false positivity and false negativity. Our results indicated a low seroprevalence of 0.97%, 95%CI [-0.36; 0.58] associated with the detection of T.gondii genetic material in 51.46%, 95%CI [41.93, 60.88] of mice brains. Neither sex nor mice body weight had an effect on the PCR outcome. We postulate that both the transmission route, horizontal or vertical, and natural selection processes could lead to this discordance which has been observed elsewhere in wild mice. The question of the biological mechanisms allowing the chronic infection of wild mice in the absence of a measurable humoral immune response remains. Our findings indicate that serological studies should not be used to measure the level of exposure to T.gondii in feral house mice.

Parasitism of mice by Toxoplasma gondii reduces the host's aversion to cat odours, likely increasing predation and transmission of the parasite to its definitive host. This behavioural change suggests a parasitic manipulation where host behaviour becomes an extended phenotype of the parasite. Independently, epigenetic changes within an organism are now known to create behavioural change. The results described here provide an experimental connection between these disparate strands of extended phenotypes and the role of epigenetics in behavioural diversity. Using mice captured on Kangaroo Island in Australia, we demonstrate that Toxoplasma gondii infection leads to specific DNA hypomethylation events in the host brain. Previous laboratory studies have shown that these epigenetic changes underlie the central processing of cat odours. We posit that the concept of extended phenotype can be expanded to extended epiphenotype, thus linking parasite genes to host behaviour through epigenesis. This phenomenon has broad implications for inter-species relationships. Read the free Plain Language Summary for this article on the Journal blog.

Canine heartworm, Dirofilaria immitis, can cause severe disease and sometimes death of the host. Associated clinical signs, lack of preventative usage and regional endemicity are unlikely sufficient by themselves to reach a definitive diagnosis. Several point-of-care (POC) diagnostic tests are commercially available to aid in-clinic diagnosis, however, there is variable diagnostic accuracy reported and no synthesis of published evidence. This systematic review aims at meta-analysing the likelihood ratio of a positive result (LR+) to inform the selection and interpretation of POC tests in practice to rule-in heartworm infection when there is clinical suspicion. Three literature index interfaces (Web of Science, PubMed, Scopus) were searched on November 11th, 2022, for diagnostic test evaluation (DTE) articles assessing at least one currently commercialised POC test. Risk of bias was assessed adapting the QUADAS-2 protocol and articles with no evidence of high risk of bias were meta-analysed if deemed applicable to our review objective. Substantial between DTE heterogeneity was investigated including potential threshold or covariate effects. A total of 324 primary articles were sourced and 18 were retained for full text review of which only three had low risk of bias in all four QUADAS-2 domains. Of the nine heartworm POC tests evaluated, only three, IDEXX SNAP (n DTEs = 6), Zoetis WITNESS (n DTEs = 3) and Zoetis VETSCAN (n DTEs = 5) could be analysed. Both WITNESS and VETSCAN DTEs showed substantial heterogeneity due to a putative threshold effect and no summary point estimates could be reported. SNAP DTEs showed acceptable heterogeneity, and a summary LR+ was estimated at 559.0 (95%CI: 24.3-12,847.4). The quality and heterogeneity of heartworm POC test DTEs was highly variable which restricted our summary of the diagnostic accuracy to only the SNAP test. A positive result from the SNAP test provides strong evidence of the presence of an infection with adult heartworm(s) in a dog patient and this test is warranted to rule-in clinical suspicion(s) in clinics. However, our review did not appraise the literature to assess the fitness of SNAP test, or any other POC tests, to rule-out heartworm infection in dogs without clinical suspicion or following heartworm therapy.

This case-control study investigated associations between Campylobacter fetus or Campylobacter jejuni titre and reproductive outcomes in 22 flocks of Merino and non-Merino maiden ewes aged 1-2 years old. Campylobacter titres were also determined for multiparous ewes aged 3 years or older on the same farms. C. fetus 'positivity' (titre ≥1:80) was detected for 12% (57/462; 95% confidence interval [95% CI] 9.6 to 15.6) of maiden ewes and 31% (65/210; 95% CI 25.0 to 37.4) of mature ewes. The odds for failing to rear a lamb in C. fetus-'exposed' maiden ewes (titre ≥1:10) was 2.01 times that of seronegative ewes (95% CI 1.09 to 3.77; P = 0.027), but there was no association between C. fetus-'positivity' (titre ≥1:80) and failure to rise (OR 1.69; 95% CI 0.77 to 3.76; P = 0.191). C. fetus abortions were confirmed with microbial culture in one maiden ewe flock. In this flock, C. fetus titres fluctuated and often waned by lamb marking, highlighting the value of necropsies during abortion investigations. C. jejuni-'positivity' (titre ≥1:80) was detected for 44% (204/462; 95% CI 39.7 to 48.7) maiden ewes, but odds of failing to rear were decreased for C. jejuni-'positive' ewes (OR 0.52; 95% CI 0.32 to 0.83; P = 0.007). The association between Campylobacter serology and the reproductive outcome was inconsistent in these flocks. Serology should be considered in the context of other risk factors and used in conjunction with other strategies to investigate the impact of Campylobacter exposure on ewe reproductive performance such as monitoring for abortions and lamb necropsies to determine aetiological diagnosis, and vaccination trials.

Toxoplasma gondii is a ubiquitous parasite increasingly detected in marine mammals and suspected to contribute to limited recovery of endangered populations. This study reports on the exposure of the Australian sea lion ( Neophoca cinerea) to this protozoon using archived adult and pup sera from three island colonies in South Australia. Modified agglutination testing (MAT) detected a seroprevalence of 30.4% (95% CI 13.2-52.9: n = 23) and high antibody titers (512 to > 2048 IU/ml) in adult females (median age 9.5 y, range 5.5-14.5 y) at Dangerous Reef, a felid-free island. Antibodies weren’t detected in any surveyed pup (97.5% CI 0.0-2.0%, n = 184) at two felid-free islands (Dangerous Reef: n = 21; Olive Island: n = 65), nor at Kangaroo Island (n = 98), which has a high-density feral cat population. Kangaroo Island pups of known age were 7-104 d old, while standard length and bodyweight comparison suggested younger and older pup cohorts at Dangerous Reef and Olive Island, respectively. This study provides the first quantification of disease risk in this endangered species from T. gondii exposure. The absence of detectable pup seroconversion supports the lack of detectable congenital transmission, maternal antibody persistence or early post-natal infection in the sampled cohort yet to commence foraging. An extended serosurvey of N. cinerea colonies is recommended to confirm the hypothesis of predominantly forage-associated exposure to T. gondii in this species. Findings have implications for parasitic disease risk in wildlife inhabiting Australia’s islands and for the feral cat control program on Kangaroo Island.

Background Toxoplasma gondii causes reproductive losses in sheep worldwide, including Australia. The reproductive performance of primiparous ewes is typically lower than for mature, multiparous ewes, and younger ewes are more likely to be immunologically naïve and therefore more susceptible to reproductive disease if T. gondii infection occurs during pregnancy. The aim of this study was to assess the impact of infection with T. gondii on the reproductive performance of primiparous ewes in southern Australia using a prospective cohort study. This will inform the need for targeted control strategies for T. gondii in Australian sheep. Results Toxoplasma gondii seropositivity using indirect ELISA was detected at 16/28 farms located across southern Australia. Apparent seropositivity to T. gondii was lower in primiparous ewes (1.1, 95% confidence interval (CI) 0.6, 1.8) compared to mature, multiparous ewes (8.1, 95% CI 6.0, 10.5; P < 0.001). Toxoplasma gondii seroconversion during the gestation and lambing period was confirmed for 11/1097 (1.0, 95% CI 0.5, 1.7) of pregnant primiparous ewes that failed to raise a lamb, and 1/161 (0.6, 95% CI 0.1, 2.9) primiparous ewes with confirmed mid-pregnancy abortion. Conclusions Low frequency of detection of T. gondii seroconversion during gestation and low frequency of seropositivity to T. gondii suggests that toxoplasmosis was not an important contributor to reproductive losses in primiparous ewes on farms located over a wide geographical area in southern Australia.

The Kangaroo Island dunnart (Sminthopsis aitkeni) is a critically endangered marsupial species with an estimated population of ~ 500 individuals found only on the western end of Australia's third largest island. Severe bushfires recently burnt more than 98% of its known and predicted habitat that was already under pressure from fragmentation. After the fires, we found evidence of eight individual dunnarts in the digestive tract of seven feral cats, out of the 86 collected in remaining unburnt refugia; thus demonstrating the need of immediate risk management efforts after large-scale stochastic events.

Introduction The spread of artemisinin resistant Plasmodium falciparum parasites is of global concern and highlights the need to identify new antimalarials for future treatments. Azithromycin, a macrolide antibiotic used clinically against malaria, kills parasites via two mechanisms: ‘delayed death’ by inhibiting the bacterium-like ribosomes of the apicoplast, and ‘quick-killing’ that kills rapidly across the entire blood stage development. Methods Here, 22 azithromycin analogues were explored for delayed death and quick-killing activities against P. falciparum (the most virulent human malaria) and P. knowlesi (a monkey parasite that frequently infects humans). Results Seventeen analogues showed improved quick-killing against both Plasmodium species, with up to 38 to 20-fold higher potency over azithromycin after less than 48 or 28 hours of treatment for P. falciparum and P. knowlesi , respectively. Quick-killing analogues maintained activity throughout the blood stage lifecycle, including ring stages of P. falciparum parasites (<12 hrs treatment) and were >5-fold more selective against P. falciparum than human cells. Isopentenyl pyrophosphate supplemented parasites that lacked an apicoplast were equally sensitive to quick-killing analogues, confirming that the quick killing activity of these drugs was not directed at the apicoplast. Further, activity against the related apicoplast containing parasite Toxoplasma gondii and the gram-positive bacterium Streptococcus pneumoniae did not show improvement over azithromycin, highlighting the specific improvement in antimalarial quick-killing activity. Metabolomic profiling of parasites subjected to the most potent compound showed a build-up of non-haemoglobin derived peptides that was similar to chloroquine, while also exhibiting accumulation of haemoglobin-derived peptides that was absent for chloroquine treatment. Discussion The azithromycin analogues characterised in this study expand the structural diversity over previously reported quick-killing compounds and provide new starting points to develop azithromycin analogues with quick-killing antimalarial activity.

The role of infectious diseases including coxiellosis in causing poorer reproductive performance of primiparous ewes are not well studied. The aims of this study were to determine if natural exposure to Coxiella burnetii is widespread in breeding ewes and whether seropositivity is associated with poor reproductive performance of primiparous ewes. Seropositivity to Coxiella burnetii was 0.08% (CI95% 0.01, 0.36) in primiparous ewes and 0.36% (CI95% 0.07, 1.14) in mature ewes. Coxiella burnetii was not detected in aborted or stillborn lambs using qPCR. These findings suggest C. burnetii infection was unlikely to be an important contributor to abortion and perinatal mortalities observed for primiparous ewe flocks, and exposure to C. burnetii was not widespread in ewes on farms located over wide geographical region of southern Australia. Whilst ewes on these farms were not an important reservoir for C. burnetii, sporadic zoonotic transmission from sheep is reported and has public health implications.

Recent concerns have arisen in Australia regarding detections of the exotic bacterium Ehrlichia canis which has resulted in ehrlichiosis outbreaks. In Australia, it is spread by the tropical brown dog tick Rhipicephalus linnaei, formerly Rhipicephalus sanguineus sensu lato tropical lineage. Previously, the tick has been recorded in South Australia in the Coober Pedy and the Oodnadatta areas. This study, which includes historical specimens data held in historical Australian arthropod collections, along with 10 sampled remote communities, confirms the wide distribution range of this species within the State. E. canis was detected by PCR in the ticks. The percentage of dogs hosting PCR‐positive ticks increased from 2.8% (95% confidence interval [CI]: 0.3 to 9.7) in November–December 2020 to 62.9% (95% CI: 44.9 to 78.5) end of February 2021, initially in two then in seven Anangu Pitjantjatjara Yankunytjatjara lands communities in the far northern regions of South Australia. Our results suggest a rapid spread of the pathogen. No evidence of E. canis was found in nine regional communities. The extended tropical brown dog tick distribution indicates a greater area where E. canis may occur and may require management to minimise the impacts of ehrlichiosis outbreaks. Without the implementation of effective detection and control programs, this extended distribution of R. linnaei is likely to result in the spread of the bacterium to other regions.

Objective: To review anthelmintic resistance globally in goats including the effect of location, mode of application and dosage on anthelmintic efficacy (assessed using Faecal Egg Count Reduction). Specifically, resistance of the three major classes of anthelmintics - Benzimidazole and Probenzimidazole (BP); Anti-cholinergics (AC); and Macrocyclic Lactone (ML) was investigated. Design/procedure: A PRISMA Framework was followed in order to conduct a thorough assessment of the literature on anthelmintic resistance in goats. A single factor ANOVA test was conducted in Microsoft Excel (2009) to test for the significance of the effect of location, mode of application and dosage on resistance. Three meta-analyses were also conducted in Microsoft Excel (2009) to quantify global resistance levels of the three major anthelmintic classes. Results: Of the 461 publications screened, 105 studies were included in the systematic review and 101 studies were included in the meta-analyses. Anthelmintic class as well as anthelmintic active principle selection in the BP and ML classes did have a significant effect on resistance (p < 0.05). Combination treatment groups had a lower amount of resistance than groups where anthelmintic classes were used alone. Mode of application of the treatment had a significant effect on resistance (p < 0.05), whilst the correlation of dosage with efficacy was low (r < 0.1). The effect of location (by continent) also had a significant influence on resistance for the AC anthelmintic class (p < 0.05). All GIN species assessed with the exception of Chabertia spp. exhibited anthelmintic resistance. Conclusions: Anthelmintic resistance is a substantial global issue in the goat industry. More research needs to be conducted into anthelmintic resistance in regard to effective ways to use anthelmintics and minimise resistance.

From four focused compound libraries based on the known anticoccidial agent robenidine, 44 compounds total were synthesised and screened for antigiardial activity. All active compounds were counter-screened for antibiotic and cytotoxic action. Of the analogues examined, 21 displayed IC50 <5 μM, seven with IC50 <1.0 μM. Most active were 2,2'-bis{[4-(trifluoromethoxy)phenyl]methylene}carbonimidic dihydrazide hydrochloride (30), 2,2'-bis{[4-(trifluoromethylsulfanyl)phenyl]methylene}carbonimidic dihydrazide hydrochloride (32), and 2,2'-bis[(2-bromo-4,5-dimethoxyphenyl)methylene]carbonimidic dihydrazide hydrochloride (41) with IC50 =0.2 μM. The maximal observed activity was a 5 h IC50 value of 0.2 μM for 41. The clinically used metronidazole was inactive at this timepoint at a concentration of 25 μM. Robenidine off-target effects at bacteria and cell line toxicity were removed. Analogue 41 was well tolerated in mice treated orally (100 mg/kg). Following 5 h treatment with 41, no Giardia regrowth was noted after 48 h.

The contribution of abortions to the overall mortality of lambs born to maiden (primiparous) ewes in Australia remains unclear. This cohort study aimed to quantify abortion and lamb mortality for ewe lambs and maiden Merino two-tooth ewes. Lamb mortality from pregnancy scanning to marking were determined for 19 ewe lamb and 11 Merino two-tooth ewe flocks across southern Australia. Average lamb mortality from scanning to marking was 35.8% (range 14.3–71.1%) for the ewe lambs and 29.4% (range 19.7–52.7%) for the two-tooth ewes. Mid-pregnancy abortion was detected in 5.7% of ewes (range 0–50%) in the ewe lamb flocks and 0.9% of ewes (range 0–4.4%) in the two-tooth ewe flocks. Mid-pregnancy abortion affecting ≥2% of ewes was observed in 6/19 ewe lamb flocks and 2/11 two-tooth ewe flocks. Lamb mortality from birth to marking represented the greatest contributor to foetal and lamb mortality after scanning, but mid-pregnancy abortion was an important contributor to lamb mortality in some ewe lamb flocks. Variability between the flocks indicates scope to improve the overall reproductive performance for maiden ewes by reducing foetal and lamb losses. Addressing mid-pregnancy abortion may improve the reproductive performance in some flocks.

Toxoplasma gondii is a protozoan parasite with a complex life cycle and a cosmopolitan host range. The asexual part of its life cycle can be perpetually sustained in a variety of intermediate hosts through a combination of carnivory and vertical transmission. However, T. gondii produces gametes only in felids after the predation of infected intermediate hosts. The parasite changes the behavior of its intermediate hosts by reducing their innate fear to cat odors and thereby plausibly increasing the probability that the definitive host will devour the infected host. Here, we provide a short description of such parasitic behavioral manipulation in laboratory rodents infected with T. gondii , along with a bird’s eye view of underpinning biological changes in the host. We also summarize critical gaps and opportunities for future research in this exciting research area with broad implications in the transdisciplinary study of host–parasite relationships. Graphical abstract