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The Australian sea lion (Neophoca cinerea) experiences high pup mortality of seasonally alternating severity, partly attributed to endemic hookworm (Uncinaria sanguinis) infection. To further explore health outcomes of early hookworm elimination, a treatment trial was conducted at Seal Bay Conservation Park, South Australia, over consecutive lower and higher mortality breeding seasons (2019, 19.2%; 2020–1; 28.9%). Pups (n = 322) were stratified into two age cohorts (median 14 d and 24 d recruitment ages) and randomly assigned to treated (topical ivermectin 500 μg/kg) or control (untreated) groups. A younger prepatent cohort a posteriori. A seasonally independent growth benefit resulted from hookworm elimination across all age cohorts. The greatest relative improvements (bodyweight + 34.2%, standard length + 42.1%; p ≤ 0.001) occurred in the month post-treatment, in the youngest prepatent cohort. A significant benefit of lesser magnitude (bodyweight + 8.6–11.6%, standard length + 9.5–18.4%; p ≤ 0.033) persisted up to 3 months across all age cohorts – greatest in the youngest pups. Treatment resulted in immediate improvement in hematological measures of health – decreased anemia and inflammation severity (p ≤ 0.012). These results enhance our understanding of host–parasite–environment interactions within the context of hematological ontogenesis, confirm the seasonally independent benefits of hookworm disease intervention, and further inform conservation recommendations for this endangered species.

Reverse transcription polymerase chain reaction (RT-PCR) is currently the standard diagnostic method to detect symptomatic and asymptomatic individuals infected with Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, RT-PCR results are not immediate and may falsely be negative before an infected individual sheds viral particles in the upper airways where swabs are collected. Infected individuals emit volatile organic compounds in their breath and sweat that are detectable by trained dogs. Here, we evaluate the diagnostic accuracy of dog detection against SARS-CoV-2 infection. Fifteen dogs previously trained at two centres in Australia were presented to axillary sweat specimens collected from known SARS-CoV-2 human cases (n = 100) and non-cases (n = 414). The true infection status of the cases and non-cases were confirmed based on RT-PCR results as well as clinical presentation. Across dogs, the overall diagnostic sensitivity (DSe) was 95.3% (95%CI: 93.1-97.6%) and diagnostic specificity (DSp) was 97.1% (95%CI: 90.7-100.0%). The DSp decreased significantly when non-case specimens were collected over 1 min rather than 20 min (p value = .004). The location of evaluation did not impact the detection performances. The accuracy of detection varied across dogs and experienced dogs revealed a marginally better DSp (p value = .016). The potential and limitations of this alternative detection tool are discussed.

Objective To characterize and compare two intramuscular drug protocols using alfaxalone and alfaxalone–medetomidine combination for the field immobilization of free-ranging koalas. Study design Blinded, randomized, comparative field study. Animals A total of 66 free-ranging koalas from the Mount Lofty Ranges, South Australia. Methods Koalas were randomly allocated into two groups. Group A animals were given alfaxalone alone at 3.5 mg kg–1. Group AM animals were given alfaxalone 2 mg kg–1 and medetomidine 40 μg kg–1, reversed with atipamezole at 0.16 mg kg–1. Blinded operators recorded heart rate (HR), respiratory rate (fR), cloacal temperature, depth of sedation and times to: first effect, sedation suitable for clinical interventions, first arousal and full recovery. Data were analysed using independent t test, Mann–Whitney U test, chi-square analysis and log-rank test at 5% level of significance. Results Suitable immobilization for clinical examination and sample collection was achieved in all animals. In groups A and AM, median time to working depth was 6.5 minutes (range: 3.4–15) and 8.1 minutes (range: 4.3–24) and time to complete recovery was 66 minutes (range: 12–138) and 34 minutes (range: 4–84), respectively, following reversal. Time to first effect was significantly shorter in group A (p = 0.013), whereas time to full arousal was significantly shorter in group AM (p = 0.007) probably due to the administration of atipamezole. Maximum HR was 117 ± 28 beats minute–1 in group A, which was a significant increase from baseline values (p Conclusions and clinical relevance Both the protocols produced immobilization, enabling clinical examination and sample collection; however, protocol AM was more suitable for field work due to shorter recovery times.

Little is known about the global bat souvenir trade despite previous research efforts into bat harvest for bushmeat. We screened eBay listings of bats in Australia, Canada, Italy, Switzerland, United Kingdom and USA to assess the nature and extent of the online offers. A total of 237 listings were retrieved in between the 11th and 25th of May 2020 with a median price per item of US$38.50 (range: US$8.50–2,500.00). Items on offer were mostly taxidermy (61.2%) or skull (21.1%) specimens. Overall, 32 different species of bat were advertised, most of which (n = 28) are listed as “Least Concern” on the International Union for Conservation of Nature (IUCN) Red List. One species (Nycteris javanica) is classified as “Vulnerable” and one (Eidolon helvum) as “Near Threatened.” Pteropus spp. specimens were the most expensive specimens on offer and the conservations status of these species may range from “Critically Endangered” to “Data Deficient” by IUCN and the entire genus is listed in the Appendix II by the Convention on the International Trade in Endangered Species of Wild Fauna and Flora (CITES). However, the exact species concerned, and their respective conservation status, could not be confirmed based on the listings' photos. The sourcing of bat was restricted to mostly South-East Asian countries (a third of items sourced from Indonesia) and to two African countries. Our survey revealed that the online offer of bat products is diverse, abundant, and facilitated by worldwide sellers although most offered bats species are from South-East Asia. With a few exceptions, the species on offer were of little present conservation concern, however, many unknowns remain on the potential animal welfare, biosecurity, legal implications, and most importantly public health risks associated with this dark trade.

This study evaluated the diagnostic test accuracy of disc diffusion relative to broth-microdilution for clinical Staphylococcus pseudintermedius isolated from dogs in Australia (n = 614). Accuracy of disc diffusion and broth-microdilution for oxacillin relative to mecA real-time PCR was also assessed. Each isolate had paired minimum inhibitory concentration and zone diameter values for ten antimicrobial agents. Data was dichotomised using Clinical and Laboratory Standards Institute susceptible and resistant clinical breakpoints. Test accuracy was reported using relative diagnostic sensitivity (RSe), specificity (RSp), likelihood ratio pairs, diagnostic odds ratio, and area-under-the receiver-operating characteristic (ROC AUC) analysis. Disc diffusion was found to have high test accuracy for most antimicrobials (ROC AUC range: 0.96 – 0.99) except rifampicin (ROC AUC = 0.80). The RSp of disc diffusion was high for all antimicrobials (range, 97.1%–100%). However, RSe was considerably variable (range, 35.7%–98.8%), particularly for amoxicillin-clavulanic acid (51.5%, 95% CI, 38.9%, 64.0%), cefoxitin (35.7%, 95% CI, 12.8%, 64.9%), and cephalothin (43.6%, 95% CI, 27.8%, 60.4%). When disc diffusion and broth-microdilution were compared to mecA real-time PCR, the overall accuracy of both assays was similar (ROC AUC, 0.99 respectively). However, the RSe for broth-microdilution (96.1%, 95% CI, 88.9%, 99.2%) was significantly higher than for disc diffusion (86.8%, 95% CI, 77.1%, 93.5%) (McNemars mid-p value 0.01). Overall, these findings demonstrate that for most antimicrobials, disc diffusion performed according to CLSI guidelines can be used to differentiate clinical S. pseudintermedius isolates that might otherwise be assessed by broth-microdilution, provided consideration is given to the performance estimates reported here.

Many infectious pathogens can be transmitted by highly mobile species, like bats that can act as reservoir hosts for viruses such as henipaviruses, lyssaviruses and coronaviruses. In this study, we investigated the seroepidemiology of protein antigens to Severe acute respiratory syndrome virus (SARS-CoV-1) and Middle eastern respiratory syndrome virus (MERS-CoV) in Grey-headed flying foxes (Pteropus poliocephalus) in Adelaide, Australia sampled between September 2015 and February 2018. A total of 301 serum samples were collected and evaluated using a multiplex Luminex binding assay, and median fluorescence intensity thresholds were determined using finite-mixture modelling. We found evidence of antibodies reactive to SARS-CoV-1 or a related antigen with 42.5% (CI: 34.3%-51.2%) seroprevalence but insufficient evidence of reactivity to MERS-CoV antigen. This study provides evidence that the Grey-headed flying foxes sampled in Adelaide have been exposed to a SARS-like coronavirus.

Rodents comprise a major component of cat (Felis catus) diets in many ecosystems, and life cycle diagrams of Toxoplasma gondii typically depict small rodents as quintessential intermediate hosts. Counter-intuitively, small rodents often experience a lower T. gondii seroprevalence than do larger sympatric mammals. This observation has repeatedly caused confusion about the relative importance of small rodents to the ecology of T. gondii. To address this confusion, we modified the Reed-Frost epidemic model to develop a simple binomial equation to model T. gondii transmission from prey to feline predators. This equation takes into account variations in prey seroprevalence and the frequency with which they are consumed by felids. Even when T. gondii seroprevalence in prey is < 1%, computation reveals that the risk of feline exposure to T. gondii can easily exceed 50 % annually. For example, if cats eat an average of 1 mouse per day, a seroprevalence of 0.2 % (1/500) in mice will cause 51.9 % of cats to be exposed to T. gondii annually. Our simple equation demonstrates that both prey seroprevalence and the rate at which prey are consumed are of approximately equal importance to the ecology of T. gondii. When inferring the importance of various prey species to the ecology of T. gondii, researchers must consider the predation and dietary habits of felids from within their study system. Our simple binomial equation could also be used to predict T. gondii exposure rates of humans or other carnivorous animals from various dietary sources or be applied to other predator-prey parasite life cycles.

Background Coxiella burnetii causes Q fever, a zoonotic bacterial disease with a multi-host cycle and reservoirs in wild and domestic animal species. Q fever has a significant impact on the Australian public health and economy but its ecology and contributing reservoir species remain poorly understood. In Europe, rabbits (Oryctolagus cuniculus) were identified as a major reservoir of C. burnetii and it is possible that they play a similar role in Australia. In absence of commercial kit available for rabbit, the Thermo Fisher - PrioCHECK™ Ruminant Q fever Ab Plate Kit was adapted to successfully screen rabbits population in Europe. However, this assay is not accessible in Australia and we assessed the equivalency of two commercially available kits in Australia – IDEXX - CHEKIT Q Fever Antibody ELISA kit and IDVet - ID Screen® Q Fever Indirect Multi-species with the Thermo Fisher kit (reference kit). Results A total of 94 rabbit sera were screened by all three ELISA kits using the same confirmed positive and negative controls. While the IDEXX kit failed to agree the other two assays (concordance correlation coefficient, rb rb = 0.927). Conclusion IDvet kit provides the best alternative for Thermo Fisher in the detection of C. burnetii specific antibodies in rabbits in Australia. Further trials are required to confirm these preliminary results due to the low seroprevalence of Coxiella burnetii observed in the study sera.

Major health conditions in sheep contribute to substantial economic losses throughout the sheepmeat supply chain in Australia. A systematic review was undertaken to explore the measurable impact of six conditions: arthritis, sheep measles, pleurisy, pneumonia, grass seeds and rib fractures, on the production of lamb and mutton across the meat value chain. Peer-reviewed scientific literature from three databases and non-peer-reviewed articles and reports from Australian government and non-government websites were searched between 11 and 17 November 2019. Original articles, including studies conducted in Australia and New Zealand, that had measurable impacts on conditions of interest were included. The search yielded 16 articles and reports and were classified as producer impact and/or processor impact studies. Mortalities were quantified for pneumonia and arthritis, with pneumonia having the highest impact for producers. Grass seed infestation resulted in the highest impact on carcase and liveweight losses compared to arthritis and pneumonia. Arthritis had the highest trim weight losses for both lamb and mutton and the highest rate of carcase condemnation. Grass seed was the only condition where other impacts on the processor (chain speed and staff relocation to the boning room) were quantified. Although quantifiable production and processing losses were available for some conditions, this review has highlighted that limited quantifiable data based on scientifically sound research were not available for other conditions. The evidence for some conditions found in this review can be used to target future research activities and to further assist producers in making informed management decisions on prevention and control.

BackgroundThe Dachshund is a chondrodystrophic breed of dog predisposed to premature degeneration and calcification, and subsequent herniation, of intervertebral discs (IVDs). This condition is heritable in Dachshunds and breeding candidates are screened for radiographically detectable intervertebral disc calcification (RDIDC), a feature of advanced disc degeneration and a prognostic factor for clinical disease. RDIDC scoring has been previously shown to be consistent within scorers; however, strong scorer effect (subjectivity) was also reported. The aim of this study was to estimate the within- and between-scorer agreement (repeatability and reproducibility, respectively) of computed tomography (CT) scanning and magnetic resonance imaging (MRI) for scoring IVD calcification, and to compare these modalities with radiographic scoring.ResultsTwenty-one Dachshund dogs were screened for IVD calcification using the three imaging modalities. Three scorers scored each case twice, independently. Repeatability was highest for radiography (95.4%), and significantly higher than for CT (90.4%) but not MRI (93.8%). Reproducibility was also highest for radiography (92.9%), but not significantly higher than for CT or MRI (89.4% and 86.4%, respectively). Overall, CT scored IVDs differently than radiography and MRI (64.8% and 62.7% agreement, respectively), while radiography and MRI scored more similarly (85.7% agreement).ConclusionsDespite high precision for radiography, previous evidence of scorer subjectivity was confirmed, which was not generally observed with CT and MRI. The increased consistency of radiography may be related to prior scorer experience with the modality and RDIDC scoring. This study does not support replacing radiography with CT or MRI to screen for heritable IVD calcification in breeding Dachshunds; however, evaluation of dog-level precision and the accuracy of each modality is recommended.

OBJECTIVE To compare outcomes following trans-scleral cyclophotocoagulation (TSCP) and 350-mm2 Baerveldt implantation in the treatment of canine refractory glaucoma. DESIGN Retrospective case study. CASE SELECTION Client owned dogs undergoing surgical treatment of glaucoma within a veterinary referral hospital. PROCEDURES Eighty-six glaucoma surgeries were performed on 83 eyes (69 dogs) diagnosed with primary or secondary glaucoma. Medical records were retrieved, and baseline data, surgery, medications, intraocular pressures (IOPs), vision, and complications were extracted. RESULTS Fifty-four eyes (44 dogs) were treated with TSCP and placement of an anterior chamber suture shunt; 28 eyes (24 dogs) were implanted with a Baerveldt glaucoma drainage device (GDD); and four eyes (4 dogs) underwent GDD implantation after failure of TSCP to manage IOP. Following TSCP, IOP control (

Cat-borne parasites and their associated diseases have substantial impacts on human, livestock, and wildlife health worldwide. Despite this, large and detailed datasets that allow researchers to study broad-scale trends in the ecology of cat-borne diseases are either difficult to obtain or non-existent. One condition that is easily detected at slaughter is macroscopic sarcocystosis, a cat-borne parasitosis of sheep (Ovis aries). We conducted a cross-sectional study to describe the geographic distribution of sarcocystosis in sheep throughout South Australia and investigate ecosystem characteristics associated with the presence of disease. Data were obtained from two slaughterhouses which processed 3,865,608 sheep from 4,204 farms across 385,468 km2 of South Australia's land mass for the period 2007-2017. A Poisson point process model was developed to quantify environmental characteristics associated with higher densities of sarcocystosis-positive farms. Sarcocystosis was highly clustered on a large island off of the Australian coast and the density of sarcocystosis-positive farms increased in areas of low soil pH (intensity ratio: 0.86, 95% CI: 0.78, 0.95) and high clay content. We hypothesize that region was confounded by, and predominately acted as a proxy for, cat density. Our results have broader implications regarding the health, welfare, economic, and conservation impacts of other cat-borne parasitosis, such as toxoplasmosis.

Listed as near-threatened by the International Union for Conservation of Nature (IUCN), the southern hairy-nosed wombat (SHNW, Lasiorhinus latifrons) faces threats such as drought, habitat degradation and loss, disease, and persecution because of competition with agriculture. To assist with evaluation of wombat health, this study reports serum biochemical reference intervals (RIs) for wild-caught SHNW from South Australia established from 126 apparently healthy SHNW using a Beckman Coulter AU480® Automated Chemistry Analyzer using RefVal Advisor. Partitioning of RIs for male and female wombats and for the two methods of sampling was performed as appropriate, and additional significant differences (P < 0.05) in biochemical profiles were identified across age class and season examined. A number of differences were observed between male and female wombats, most notably higher creatinine, urea, and sodium in females. Subadult and juvenile wombats had significantly lower total protein, globulin, and ALT activity, and significantly higher ALP activity than adults. Wombats sampled in winter and spring had significantly higher total protein, albumin, potassium, bicarbonate, and enzyme activities (ALP, ALT, AST, GGT, GLDH, lipase), and significantly lower glucose and creatinine when compared to individuals sampled in summer and autumn. Differences in CK activity and anion gap observed between the two methods of sampling likely reflect delay and handling of animals between capture and blood collection. The serum biochemical RIs documented here are considered representative of a population of healthy SHNW, providing a tool for health assessment and monitoring of SHNW health in South Australia and elsewhere.

BACKGROUND: Canine heartworm (Dirofilaria immitis) was reported as endemic to South Australia over 25 years ago, but the current status of canine heartworm infection in South Australia is unknown. METHODS: We surveyed South Australian veterinary practices to collect evidence about canine heartworm cases, prevention, screening and treatment practices. RESULTS: In total, 141 veterinary practices operating in South Australia (92.3% response rate) completed a 12‐question telephone survey. Five confirmed cases of heartworm and one suspected false‐positive case were reported from separate practices over the 2016–17 financial year. All confirmed cases had recently relocated to South Australia from interstate. Incidence of canine heartworm was estimated at 0.00% (95% confidence interval (CI): 0.00–0.0006) and prevalence at 0.002% (95%CI: 0.00–0.004) using conservative estimates of the South Australian dog population. Attitudes and protocols regarding heartworm differed significantly between metropolitan and rural veterinary practices. All metropolitan practices recommend systematic medicated prevention against heartworm, while some rural practices (20.7%) recommend prevention only when travelling interstate (P < 0.001). CONCLUSION: Although there is a minor source of heartworm from introduction of infected dogs into South Australia, there is no evidence that the infection is transmitted to other dogs within the state. Therefore, it is unlikely that heartworm is currently endemic to South Australia as reported in the past. This new evidence provides a strong case to refine current prevention practices to reflect the current negligible risk of heartworm infection within South Australia.

Objective Improving antimicrobial stewardship in the livestock sector requires an understanding of the motivations for antimicrobial use and the quantities consumed. However,detailed information on antimicrobial use in livestock sectors is lacking. This cross‐sectional study aimed to better understand antimicrobial use in the beef feedlot sector in Australia. Design A self‐administered questionnaire asking about antimicrobial use and reasons for use was designed and mailed to beef feedlot operators in Australia. Respondents were asked to report the percentage of animals treated, purpose of use, and disease conditions targeted for 26antimicrobial agents. Results In total, 83 of 517 (16.1%) beef feedlot operators completed the survey. Monensin (61.0%of respondents) and virginiamycin (19.5%of respondents) were the most commonly reported in‐feed antimicrobials. In‐feed antimicrobial agents were most frequently used by respondents for treatment of gastrointestinal diseases (52.8%). Antimicrobials were used for growth promotion by 42.1% of respondents, with most (85.7%) reporting the use of ionophores(a group of compounds not used in human medicine). Short‐acting penicillin(69.1%), short‐acting oxytetracycline, and tulathromycin (both 57.3%) werethe most common injectable antimicrobial agents used. Injectable antimicrobials were most frequently used to treat respiratory (72.3%) and musculoskeletal (67.5%) conditions. Conclusion Overall,the use of antimicrobials was appropriate for the purpose indicated, and there was a strong preference for drugs of low‐importance in human medicine. The data described here stand to be a strong influence on the implementation of an antimicrobial stewardship program in the sector.

The assessment of antimicrobial resistance in bacteria derived from animals is often performed using the disc diffusion assay. However broth-microdilution is the preferred assay for national antimicrobial resistance surveillance programs. This study aimed to evaluate the accuracy of disc diffusion relative to broth-microdilution across a panel of 12 antimicrobials using data from a collection of 994 clinical Escherichia coli isolates from animals. Disc diffusion performance was evaluated by diagnostic sensitivity, specificity, likelihood ratio pairs and receive-operating characteristic (ROC) analysis. Data was dichotomised using CLSI susceptible and resistant clinical breakpoints. In addition, disc diffusion breakpoints produced using diffusion Breakpoint Estimation Testing Software (dBETS) were evaluated. Analysis revealed considerable variability in performance estimates for disc diffusion susceptible and resistant breakpoints (AUC ranges: 0.78-0.99 and 0.92-1.0, respectively) across the panel of antimicrobials. Ciprofloxacin, tetracycline, and ampicillin estimates were robust across both breakpoints, whereas estimates for several antimicrobials including amoxicillin-clavulanic acid, cefoxitin and gentamicin were less favourable using susceptible breakpoints. Overall performance estimates were moderately improved when dBETS susceptible breakpoints were applied. For most antimicrobials, disc diffusion was accurate at predicting resistance of clinical E. coli from animals that could otherwise be determined by broth-microdilution. While disc diffusion is suboptimal for assessing the proportion of fully susceptible isolates for some drugs, sensitivity and specificity estimates provided here allow for the use of standard formula to correct this. For this reason, disc diffusion has applicability in national surveillance provided the performance of the assay is taken into account.

Free range production systems are becoming a major source of egg production in Australia and worldwide. This study investigated shedding and ecology of Salmonella Typhimurium and Salmonella species in a free range layer flock, wild birds and foxes in the vicinity of the free range farm in different seasons. Shedding of Salmonella was significantly higher in summer. Within the shed, overall, Salmonella prevalence was highest in dust. Corticosterone level in faeces was highest in spring and lowest in winter. There was no direct association between the Salmonella shedding (MPN/gm) and corticosterone levels in faeces. Salmonella Typhimurium MLVA types isolated from fox and wild birds were similar to MLVA types isolated from layer flock and reported during human food borne illness. Wild birds and foxes appear to play an important role in S. Typhimurium ecology and food safety. Environmental factors could play a role in evolution of S. Typhimurium in free range environment.

Nineteen white rhinoceroses (Ceratotherium simum) were anesthetized with 4 mg of etorphine hydrochloride; 35-40 mg of midazolam; and 7,500 international units of hyaluronidase for dehorning purposes at a game ranch in South Africa, to investigate this anesthetic combination. Median time to recumbency was 548 sec (range 361-787 sec). Good muscle relaxation and no muscle rigidity or tremors were observed in 18 animals, and only 1 individual showed slight tremors. In addition, all animals received butorphanol i.v. 5 min after recumbency at the ratio of 10 mg of butorphanol per 1 mg of etorphine. Blood gas and selected physiologic parameters were measured in the recumbent animal, immediately before and 10 min after the administration of butorphanol. Statistically significant improvements were observed in blood gas physiologic and cardiopulmonary parameters 10 min after the administration of butorphanol, with a reduction in arterial partial pressure of carbon dioxide, systolic blood pressure, and heart rate and an increase in pH, arterial partial pressure of oxygen, oxygen saturation, and respiratory rate (all P < 0.005). After i.v. naltrexone reversal, recovery was uneventful, and median time to walking or running was 110 sec (range 71-247 sec). The results indicate etorphine and midazolam combination is an effective alternative anesthetic protocol and produces good muscle relaxation. Furthermore, i.v. butorphanol was associated with improved blood gas values and cardiopulmonary function for at least 10 min postinjection. Refereed/Peer-reviewed

Blood samples were collected from 69 'healthy' female alpacas aged ≥12 months from 11 properties in South Australia. The 10-90 percentile ranges of the 16/19 analytes measured in this sample population were within the published ranges of four healthy alpaca populations from other geographic locations. Marginal exceptions were glutamate dehydrogenase and bicarbonate. Potassium was notably elevated, probably because of haemolysis of some samples. The sample size was insufficient to provide the appropriate statistical power to define diagnostic references ranges according to international standards. The health status of the sample population of alpacas was presumptive based on a physical examination.

Diagnostic laboratories frequently select a subjective cutoff value for real-time amplification assays, above which a threshold cycle (Ct) value is deemed false. Commonly, higher Ct values are interpreted as amplification or fluorescence artifacts, or cross contaminations. Although the implementation of Ct cutoff might be reasonable, its justification and selection should be based on evidence. The current article reviewed evidence-based strategies to select Ct cutoffs grouped in analytical and epidemiologic approaches. Analytical strategies use criteria gathered during the assay development and include fluorescence threshold, reaction end-cycle, limit of detection, and artifact investigation. Variability in amplification efficacy across test runs may induce some instability in an intended Ct cutoff and requires some standardization or normalization procedures. Epidemiologic strategies use criteria based on either the probability or the cost of a false test result associated with a specified cutoff. Cutoffs, depending on the intended purpose of the test, can be selected graphically to minimize the probability of either false-positive or false-negative results by using two-graph receiver operating characteristics curves. The assay's diagnostic sensitivity and specificity may vary with the tested population, thus, the estimated two-graph receiver operating characteristics curve is population dependent and should be established for the targeted population. Although the selection of a cutoff based on misclassification cost depends on infection prevalence, the selection based on predictive values does not. To optimize the test average diagnostic performance, the Ct cutoff should be selected when diagnostic odds ratio is maximal. Epidemiologic approaches were illustrated by selecting Ct cutoffs for a real-time assay for Infectious salmon anemia virus.