Sutherland et al

…Numerous studies have shown that surgical castration causes a significant and marked increase in pigs’ cortisol concen- Infrared thermography trations (Prunier et al., 2005; Carroll et al., 2006; Sutherland et al., IRT…

…Numerous studies have shown that surgical castration causes a significant and marked increase in pigs’ cortisol concen- Infrared thermography trations (Prunier et al., 2005; Carroll et al., 2006; Sutherland et al., IRT…

population data (5, 6) in bushfire-affected regions of NSW and Victoria Research Team and Reflexivity indicate that there were 3.6 million cattle and 21 million sheep Three authors (BC, MW and MB-T) developed the semi- in bushfire-affected regions, although many would not have been structured interview guide independently of other authors close to fire within those regions because of the coarse scale of (see Supplementary Material). The interviews were conducted the population data (BC, unpublished data). The local impact by the lead author (BC). The analyses were first conducted on some individual farmers was very high. For example, in a by BC with subsequent assistance and commentary from all recent case control study, some farms suffered an impact of up co-authors. BC is a male veterinary epidemiologist (PhD, to $2 million (AUD) and deaths of all livestock on a farm (BC, FANZCVS) and beef producer who was from a bushfire affected unpublished data). farm. MB-T is a female medical epidemiologist (PhD) who Bushfires (wildfires) are increasing in frequency globally, has extensive experience in qualitative and mixed methods especially as a result of longer fire seasons in temperate or boreal epidemiology and strategically assisted in the project to ensure regions (7, 8). Little research has been conducted on the impacts methodologies were well-implemented. MW is a male veterinary of bushfires on livestock in any part of the world. For example, epidemiologist (Ph.D., FANZCVS) and has used qualitative a systematic literature review by co-authors (BC, AH and CP) methods in veterinary epidemiology for several years. revealed barely a dozen publications, mostly case studies in The interviewer (BC) established a new professional Australia (9–22). More specific published

fficulty, walk normally, on pasture, (p = 2.0 × 10–6) for appetite score, indicating treatment effects were and salivating) were analysed using linear models with fixed ef- changing over the study period. Model-based appetite score prob- fects for Treatment, Age, Breed and Sex. Due to the positive skew, abilities are displayed (Figure 3). Most control cattle had no appe- a loge(y + 1) transformation was applied. However, with the large tite with low scores over the duration of the study period (score of number of ‘zero’ durations, hypothesis testing was conducted with 1). However, the two treatment groups had moderate (MO) to high permutation tests rather than F tests, using the aovperm function in (TS) scores on Day 0, low on Day 3, then progressively increasing the permuco package (Frossardm & Renaud, 2019) in R. scores after that. After Day 0, there were no significant differences in scores between MO and TS-treated cattle (all p > .10). Neither Age (p = .810), Breed (p = .524), nor Sex (p = .324) had a significant effect 3 | R E S U LT S in appetite score. 3.1 | Lesion healing scores 3.3 | Lesion size There was a highly significant Treatment × Day interaction (p = 3.4 × 10–7), indicating differing lesion healing score time courses There was a highly significant Treatment × Day interaction (p < 2 × across the three treatment groups. Model-based lesion score prob- 10–16) for lesion size (Figure 4), indicating different rates of healing abilities are displayed (Figure 2). While control cattle maintained the across the three groups. Initially, there were no significant differ- presence of erosions/ulcers (a score of 1), recoveries were observed ences in mean wound sizes amongst the three groups (all p > .25). in the two treatment groups. From Day 9

0). After this decrease in an decrease in power, increase power, in power an increase at frequencies in power at frequencies>10>10HzHz waswas observed observed starting after starting after approximately 35approximately s until approximately 90 s after90t = 35 s until approximately 0 (‘late‘ s after response). t = 0 (‘late‘ Figure response). Figure22presents presents the DSAin the DSA of the change of power the change in power relative torelative to the baseline the baseline and highlights and highlights thetheearly earlyand and late late components observed following the interdigital pinch. components observed following the interdigital pinch. Figure 2. DSA of the Pooled Figure 2. DSAMedian Response of the Pooled Median(n = 44) to Response (n Interdigital Pinch = 44) to Interdigital Stimulation Pinch Stimulation over 90s. s. over 90 Areas with bold colours indicate the frequency and time regions with significant differences Areas with bold colours indicate the frequency and time regions with significant differences between pre- and poststimulus conditions. The colour bar indicates the change in the poststimulus between pre- and poststimulus powerconditions. compared with The colour barbaseline. the prestimulus indicates the change The stimulus inatthe started t = 0.poststimulus power Briefly, the interdigital compared with thepinch leads to an prestimulus ‘early‘ reduction baseline. in powerstarted The stimulus in the slow at t =frequencies 0. Briefly,below 10 Hz and a pinch the interdigital ‘late‘ activation of higher frequencies above 10 Hz starting approximately 35 s after the stimulation. leads to an ‘early‘ reduction in power in the slow frequencies below 10 Hz and a ‘late‘ activation of higher frequencies3.3. above 10 Hz starting

0). After this decrease in an decrease in power, increase power, in power an increase at frequencies in power at frequencies>10>10HzHz waswas observed observed starting after starting after approximately 35approximately s until approximately 90 s after90t = 35 s until approximately 0 (‘late‘ s after response). t = 0 (‘late‘ Figure response). Figure22presents presents the DSAin the DSA of the change of power the change in power relative torelative to the baseline the baseline and highlights and highlights thetheearly earlyand and late late components observed following the interdigital pinch. components observed following the interdigital pinch. Figure 2. DSA of the Pooled Figure 2. DSAMedian Response of the Pooled Median(n = 44) to Response (n Interdigital Pinch = 44) to Interdigital Stimulation Pinch Stimulation over 90s. s. over 90 Areas with bold colours indicate the frequency and time regions with significant differences Areas with bold colours indicate the frequency and time regions with significant differences between pre- and poststimulus conditions. The colour bar indicates the change in the poststimulus between pre- and poststimulus powerconditions. compared with The colour barbaseline. the prestimulus indicates the change The stimulus inatthe started t = 0.poststimulus power Briefly, the interdigital compared with thepinch leads to an prestimulus ‘early‘ reduction baseline. in powerstarted The stimulus in the slow at t =frequencies 0. Briefly,below 10 Hz and a pinch the interdigital ‘late‘ activation of higher frequencies above 10 Hz starting approximately 35 s after the stimulation. leads to an ‘early‘ reduction in power in the slow frequencies below 10 Hz and a ‘late‘ activation of higher frequencies3.3. above 10 Hz starting

e commer­ of orf virus. This was followed by a similar sampling of cial MagMAX™ Pathogen RNA/DNA kit (Thermo Fisher the lesions on days 1 (T1), 3 (T2) and 5 (T3) post- Scientific) and the automated magnetic particle processor treatment of all the lesions observed on the lambs. KingFisher Flex System (Thermo Fisher Scientific), fol­ Samples of 3mL of whole blood were also collected lowing the manufacturer’s instructions. Extracted DNA from the jugular vein through a vacutainer system into samples were stored at −80°C until end of sampling in EDTA tubes to perform haematology in all the animals order to evaluate all of them in a single run of real-time prior to (T0) and ten days (T4) following treatment, PCR quantification. Orf virus detection was performed For personal use only. respectively. using the commercial qPCR kit EXOone Contagious Ecthyma (Exopol, Spain) which targets the BL2 gene Treatment Application that encodes a major envelope viral antigen. The kit con­ Following confirmation of the presence of orf virus in the tains a quantified synthetic positive control, and also an lesions, a single spray of 1.5mL of TS was applied liber­ endogenous control to avoid false-negative results. ally with a spray gun to all orf lesions in Group A lambs, Amplification was performed on a FAST 7500 cycler with the Group B lambs remaining untreated. (Applied Biosystems), and a cut-off value for positive samples was established at cycle quantification (Cq) Clinical Progression values lower than 38. Clinical examination of all lambs was performed by obser­ vers blinded to treatment. This occurred daily for 11 days Virus Culture to determine the clinical progression of the lesions, with Sterile swabs collected at T0, T1, T2 and T3 were submitted data collected on the

R-Interval [ms] between baseline and during castration and (D) between baseline and the bar and standard deviation post is represented castration. The barbyindicates the whiskers. Boxplots the mean of (E) difference is displayed as the in topstandard of the bar deviation and standard rate (SDHR) of heartdeviation is [beats minute −1] between baseline and during castration and (F) between baseline and post represented by the whiskers. Boxplots of (E) difference in standard deviation of heart rate (SDHR) castration, (G) difference in standard deviation of RR-Interval (SDRR) [ms] between baseline and [beats minute−1 ] between baseline and during castration and (F) between baseline and post castra- during castration and (H) between baseline and post castration and (I) difference in low tion, (G) differencefrequency frequency/high in standard deviation (LF/HF) ratioofbetween RR-Interval (SDRR) baseline and[ms] between during baseline castration and and during (J) between baselineand castration and(H) postbetween baseline castration. and represent The boxes post castration the first and(I)third and lowthe quartileinand difference frequency/high whiskers range frequency (LF/HF)to from minimum ratio betweenThe maximum. baseline median and during castration is indicated by the bandand inside (J) between the box.baseline post andletters Different (a, b, c, d) castration. show The boxes significant representdifferences between the first and thirdthe experimental quartile and the groups whiskers 0.05).from (p <range The superscript minimum “e” differentiates to maximum. time periods/episodes(“ The median is indicated by the e”) from time points (MAP and respiratory rate) band inside the box. Different letters (a, b, c, d) show significant differences between the

R-Interval [ms] between baseline and during castration and (D) between baseline and the bar and standard deviation post is represented castration. The barbyindicates the whiskers. Boxplots the mean of (E) difference is displayed as the in topstandard of the bar deviation and standard rate (SDHR) of heartdeviation is [beats minute −1] between baseline and during castration and (F) between baseline and post represented by the whiskers. Boxplots of (E) difference in standard deviation of heart rate (SDHR) castration, (G) difference in standard deviation of RR-Interval (SDRR) [ms] between baseline and [beats minute−1 ] between baseline and during castration and (F) between baseline and post castra- during castration and (H) between baseline and post castration and (I) difference in low tion, (G) differencefrequency frequency/high in standard deviation (LF/HF) ratioofbetween RR-Interval (SDRR) baseline and[ms] between during baseline castration and and during (J) between baselineand castration and(H) postbetween baseline castration. and represent The boxes post castration the first and(I)third and lowthe quartileinand difference frequency/high whiskers range frequency (LF/HF)to from minimum ratio betweenThe maximum. baseline median and during castration is indicated by the bandand inside (J) between the box.baseline post andletters Different (a, b, c, d) castration. show The boxes significant representdifferences between the first and thirdthe experimental quartile and the groups whiskers 0.05).from (p <range The superscript minimum “e” differentiates to maximum. time periods/episodes(“ The median is indicated by the e”) from time points (MAP and respiratory rate) band inside the box. Different letters (a, b, c, d) show significant differences between the