Capacity and needs assessment of veterinary services in Vietnam in biosecurity, biosafety and One Health.

The Asia-Pacific region is recognised as an epicentre of emerging infectious diseases (EIDs), of which 75% are zoonotic in nature. Vietnam is recognised as a potential hotspot for zoonotic EIDs. There is a growing recognition that progress towards global health security requires greater focus on collaboration between the human health and animal health sectors to control diseases at their animal source and prevent against human health impacts. Assessment of veterinary epidemiology capacity in Vietnam is paramount to strengthening the health security of Asia-Pacific. This study aims to evaluate the national capacity and needs of veterinary services in Vietnam in biosecurity, biosafety and One Health. A cross-sectional, convergent mixed-methods study was conducted between November 2020 and April 2021. An online questionnaire was administered to government-employed field veterinarians. Descriptive analyses and logistic regression models were performed using survey responses to understand capacity in the field. Semi-structured interviews were also conducted with stakeholders in veterinary services including government, academia, research institutes, non-profit and international organisations. Coding and thematic analysis using a deductive approach was used for data collected from interviews to contextualise findings from the survey and understand institutional capacity. In total, 178 field veterinarians completed the online survey and 25 stakeholders were interviewed. The majority of participants had reported receiving training in biosecurity and biosafety, including use of personal protective equipment. Most respondents reported practicing good biosecurity measures (92%) and good biosafety measures (88%). Physical and socioeconomic barriers to practicing biosecurity were reported to be prevalent for smallholder farmers, which may suggest a gap in the capacity of veterinary services to provide cost-effective and practical biosecurity strategies. Seventy five percent of participants had never or rarely participated in One Health approaches in the field in the last 12 months and 69% reported further training as a high priority. There was a knowledge gap reported amongst district and commune-level veterinary staff about the need for, and awareness of multisectoral collaboration. Respondents that completed postgraduate qualifications in epidemiology or Field Epidemiology Training Programs (adjusted OR: 3.06; 95% CI: 1.01, 9.23, p = 0.046) and had longer job tenure between 10-12 years (OR: 10.38; 95% CI: 3.06, 35.15, p = <0.001) were more likely to have higher levels of experience in One Health. This study identified gaps in knowledge, attitudes and adoption of practices related to biosecurity, biosafety and One Health specifically in lower-level or less experienced veterinary staff without further training opportunities in epidemiology. These findings enable prioritisation of training, policy, and planning activities to further enhance the national capacity of veterinary services in Vietnam.

Impact of bushfires on Australian livestock health, welfare and carcase quality.

The 2019/2020 Australian bushfires were unprecedented in terms of total area burned and impact on livestock and wildlife populations. However, there is currently limited literature available relating the consequences of bushfire or smoke exposure to livestock health, welfare, and carcase quality. A retrospective cross-sectional study was conducted using historical monitoring data from a Meat Standards Australia (MSA) accredited abattoir located on the mid-north coast of New South Wales, Australia. The spatiotemporal association between exposure to bushfire smoke (specifically the duration of the bushfires, distance from closest bushfire, annual bushfire season, proportion of the property of origin burned during the bushfires and time frame since exposure to bushfires) and effects on carcase meat quality metrics and pathology were measured, by building linear, generalised linear and cumulative link mixed models. Our findings indicate that hot carcase weight increased as the distance between the property of origin and the closest bushfire became greater and decreased with exposure to bushfires of longer duration or when greater proportions of the property of origin were burnt during bushfires. Subcutaneous rib fat of carcases also increased with an increasing distance of properties from the closest recorded bushfire and decreased with exposure to bushfires during the 2019/2020 season. Higher meat colour scores (darker meat colour) were associated with exposure to bushfires during the 2019/2020 season and exposure to bushfires of longer durations. There was only a weak association between increasing distance to the closest bushfire and higher marbling scores. Evidence of pneumonia in carcases was associated with exposure to bushfires of longer duration, specifically increasing risk of pneumonia was associated with fires of longer durations. Greater periods of time since exposure (i.e., >6 months) to bushfires were also associated with a higher risk of evidence of pneumonia at the time of processing. With increasing incidence of bushfires in Australia forecasted as a result of climate change, there is an urgent need to understand the impact of bushfires on livestock, to limit the effects on livestock health and mitigate the risk of significant socioeconomic impacts to the livestock industry. By providing a greater understanding of the impact of bushfires, the findings of this study can support producers to make informed decisions to mitigate the effects of bushfires on livestock health and carcase meat quality.

A retrospective study on antibacterial treatments for koalas infected with Chlamydia pecorum.

Chlamydiosis remains the leading infectious disease and is one of the key factors responsible for the dramatic reduction of koala populations in South-East Queensland (SEQ) and New South Wales (NSW) regions of Australia. Possible infection outcomes include blindness, infertility, painful cystitis, and death if left untreated. Studies have reported the treatment efficacy of chloramphenicol and doxycycline, which are the two most commonly administered treatments in diseased koalas, in clinical settings. However, none have directly compared the treatment efficacy of these antibacterials on koala survival. A retrospective study was essential to identify any relationships between the demographical information, and the animals' responses to the current treatment regimens. Associations were explored between six explanatory (sex; maturity; location; clinical signs, treatment; treatment duration) and two outcome variables (survival; post-treatment PCR). Results showed that female koalas had a statistical trend of lower odds of surviving when compared to males (OR = 0.36, p = 0.05). Koalas treated with chloramphenicol for ≥ 28 days had greater odds of surviving than when treated for < 28 days (OR = 8.8, p = 0.02), and those koalas administered doxycycline had greater odds of testing PCR negative when compared to chloramphenicol treatments (OR = 5.45, p = 0.008). There was no difference between the antibacterial treatments (chloramphenicol, doxycycline, and mixed/other) and the survival of koalas. Female koalas had greater odds of exhibiting UGT signs only (OR = 4.86, p < 0.001), and also greater odds of having both ocular and UGT clinical signs (OR = 5.29, p < 0.001) when compared to males. Of the koalas, 28.5% initially had no clinical signs but were PCR positive for C. pecorum. This study enables further understanding of the complex nature between chlamydial infection and response to antibacterial treatment.

Isolated paediatric orbital fractures: a case series and review of management at a major trauma centre in the UK.

PURPOSE: Paediatric orbital fractures are rare. Existing literature demonstrates wide variation in estimates of incidence, aetiology, management protocols and outcomes. Despite this, it is generally acknowledged that orbital fractures with entrapment of the extraocular muscles constitute a surgical emergency due to the potential for persistent diplopia secondary to muscle ischaemia and necrosis. METHODS: This retrospective study was conducted to determine the characteristics and outcomes of management of orbital fractures amongst the paediatric population. It involved patients presenting to a major trauma unit in London between 2010 and 2020. RESULTS: Thirteen patients with isolated orbital fractures presented to our unit in this period. The average age was 13 years. Surprisingly the predominant aetiology was interpersonal violence. The most common fracture pattern involved the orbital floor and medial wall. One medial wall fracture case was missed in the emergency department. Eight patients required surgical intervention due to diplopia caused by muscular entrapment of extraocular muscles; the final patient had a large defect resulting in enophthalmos requiring a large titanium plate. A transconjuctival approach was preferred for surgical access and resorbable sheet was used in the remaining cases. Five patients had nausea, vomiting or bradycardia associated with the oculocardiac reflex. Surgical intervention occurred within 24-48 h of injury in 6 cases. Resolution of diplopia occurred in 7 patients within 6 months. CONCLUSION: Paediatric patients with orbital fractures should be assessed on the day of injury by a maxillofacial surgeon. Due to the risk of persistent diplopia, urgent surgical intervention in patients with entrapment of extraocular muscles should occur as soon as possible.

A randomised control trial to evaluate the effectiveness of a commercial vaccine for pinkeye in Australian beef cattle.

Pinkeye (a generic term to describe infectious bovine keratoconjunctivitis) is a significant disease of cattle worldwide, impacting productivity and animal welfare. One commercial pinkeye vaccine, a systematically administered Moraxella bovis bacterin, has been available in Australia since 2007. This is the first field trial of the effectiveness of this vaccine for the prevention of naturally occurring disease in Australia. Extensively run beef herds in southwest Queensland that regularly experienced pinkeye were enrolled in the trial and animals were randomly allocated to vaccinated and control groups in different proportions in each herd. The subsequent incidence of clinical pinkeye between the two groups was compared for animals less than one-year-old. Data were analysed from 649 cattle from five herds over two pinkeye seasons: three herds of 390 calves from 1st November 2019 to 20th January 2020 and two herds of 259 calves from 23rd September 2020 to 21st April 2021. Pinkeye was common with 24% of all calves (156/649) contracting the disease during the trial. Univariable and multivariable binary logistic mixed-effect models were fitted to account for clustered data and potential residual confounding due to sex, weight, breed, coat colour, and periocular pigmentation. The incidence of pinkeye was not significantly different between vaccinated and control groups, both alone (p = 0.67) and after adjusting for sex and weight differences (p = 0.69). The vaccine was not protective against naturally occurring pinkeye under the field conditions.

A survey to understand farmers' perceptions and risk factors for hoof diseases including footrot in sheep in New South Wales, Australia.

The aims of this study were to develop an understanding of farmers' perceptions and risk factors for footrot, including its less severe forms, and other hoof diseases in sheep in New South Wales (NSW). A questionnaire was developed and administered to sheep farmers in Local Land Services (LLS) regions across NSW. LLS staff selected sheep farmers who met the inclusion criteria which included farmers with a minimum of 100 sheep, a history of having had foot problems in their flock or having expressed an interest in improving sheep health and production. Farmers completed the questionnaire either by telephone or via the REDCap online survey platform. Descriptive analyses and multivariable logistic regression models were created. The survey was completed by 43 sheep farmers with a median farm size of 1,500 Ha and flock size of 2,300; footrot was present on 39% of farms while 75.6% had other hoof diseases. A flock of >3,000 sheep were more likely to have footrot than a smaller flock (OR = 11.99, 90% CI = 3.02-63.92, P-value = 0.005) and footrot was less likely to be present on farms when an Animal Health Statement was requested while purchasing sheep (OR = 0.10, 90% CI = 0.01-0.56, P-value = 0.04). Hoof conditions other than footrot were likely to be present in flocks when foot inspections were conducted at a time other than weekly inspections (OR = 0.13, 90% CI = 0.01-0.68, P-value = 0.04) and flocks kept on undulating ground were more likely to have diseases other than footrot compared to those kept on flat ground (OR = 3.72, 90% CI = 1.02-15.80, P-value = 0.09). Most farmers agreed that footrot including its less severe forms can cause production losses and negatively affect animal health and welfare. Limitations of the study were the sample size and dry environmental conditions prior to and during study period in many regions of NSW which limited the expression of footrot.

Design and Evaluation of the Veterinary Epidemiology Teaching Skills (VETS) Workshop: Building Capacity in the Asia-Pacific Region.

Building workforce capacity in epidemiology skills for veterinarians in the Asia-Pacific region is crucial to health security. However, successful implementation of these programs requires a supply of trained veterinary epidemiology teachers and mentors. We sought to design and evaluate delivery of a 4-day Veterinary Epidemiology Teaching Skills (VETS) workshop as part of a larger project to strengthen field veterinary epidemiology capacity. Thirty-five veterinarians were selected to participate in the 4-day VETS workshop, consisting of nine modules delivered synchronously online. Participants were formatively assessed and given feedback from peers and facilitators on all activities. Data were collected with pre- and post-course questionnaires. Numeric values were categorized to convert into an ordinal scale with four categories. Qualitative data were analyzed using thematic analysis. Thirty-four veterinary epidemiologists from eight countries of the Asia-Pacific completed the workshop. Participants felt able to achieve most key learning outcomes through provision of succinct literature, teaching frameworks, and active participation in small groups, with multiple opportunities to give and receive feedback. Although the online workshop provided flexibility, participants felt the addition of face-to-face sessions would enrich their experience. Additionally, protected time from work duties would have improved their ability to fully engage in the workshop. The VETS workshop granted an effective online framework for veterinary epidemiologists to develop and practice skills in teaching, facilitation, assessment, feedback, case-based learning, program evaluation, and mentorship. A challenge will be ensuring provision of local teaching and mentoring opportunities to reinforce learning outcomes and build workforce capacity.

Fat Grafting as Regenerative Surgery: A Current Review.

SUMMARY: Autologous fat grafting has long been regarded as an ideal filler, lauded for its ability to restore soft-tissue contour. In recent times, fat grafting has exhibited regenerative capacity, largely secondary to the action of adipose-derived stem cells and preadipocytes in the stromal vascular fraction of adipose tissue. This has birthed a new field of regenerative surgery with benefits germane to scarring, fibrosis, atrophy, burns, neuropathic pain, and autoimmune disease. The broad clinical applications of regenerative fat grafting have the potential to improve quality of life through functional and aesthetic improvement. Fat grafting has the potential to serve as a regenerative option for difficult clinical problems that cannot be treated effectively at the present time. Fat grafting also exhibits angiogenic and immunomodulatory properties in the context of autoimmune disease. The broad clinical applications of regenerative fat grafting have the potential to improve quality of life both functionally and aesthetically. It may present a less invasive avenue for clinical issues that today necessitate conventional surgical techniques. However, regenerative fat grafting is still in its infancy; further research is required to ascertain evidence-based protocols for the various clinical indications and better understand the precise regenerative mechanisms after fat grafting.

Perceptions of Australian cattle farmers regarding the impact of pinkeye on farm productivity and animal welfare.

Pinkeye (infectious bovine keratoconjunctivitis, IBK) is an important disease of cattle worldwide. It has a substantial negative impact on farm productivity and is a major cost burden, but specific data on losses are lacking. This study was conducted to understand farmers' perceptions of the impact of pinkeye on farm productivity and animal welfare, and factors influencing the money farmers estimated spending on pinkeye in 2018. Data were collected by the first Australia-wide online survey on pinkeye. There were 1035 suitable responses analysed for impact on farm productivity. From these 82% of respondents represented farms in southern Australia, 58% reported cattle breeding as their main enterprise, and 89% bred animals on farm. Farmers were more likely to rank the impact of pinkeye on farm productivity as high if they had younger cattle, treated cattle with pinkeye more frequently, and as their herd size increased. Fewer farmers chose pinkeye as an animal welfare concern than as an economic and farm management issue, but overall animal welfare was rated by the greatest number of farmers as a high severity concern (n = 691), followed by decreased sale value and farm profits (n = 561). This suggests a shift in the equipoise between the economics of food animal production and animal welfare expectations. The median amount reportedly spent on pinkeye in 2018 by Australian farmers (n = 779) was $250.00 per farm. Farmers reported spending more money on pinkeye as herd size and number of cattle affected by pinkeye increased, their perception of pinkeye impact on farm productivity and animal welfare increased, if they treated pinkeye more frequently, reported higher fly worry, if their herds contained Angus cattle, if they bred on farm, and if they were located in southern Australian regions. Study findings should be used to better understand pinkeye, target expenditure, and improve outcomes for cattle and farmers.

Perceptions and practices of Australian cattle farmers for the treatment of pinkeye (infectious bovine keratoconjunctivitis).

Pinkeye or infectious bovine keratoconjunctivitis is a globally significant disease and occurs in every state of Australia. Economic loss due to pinkeye can be considerable and it is a major welfare concern, but not all cattle with the disease are treated by farmers. This study was conducted to understand the perceptions and practices of Australian farmers regarding the treatment of pinkeye: factors influencing when farmers treat pinkeye, treatments used and considered effective, and reasons for not treating. Data were gathered using a custom designed online questionnaire. Farmer responses suitable for assessment (n = 985) were analysed using descriptive analyses along with univariable and multivariable ordinal logistic regression models to evaluate the association of 15 explanatory variables with the outcome "when do you treat pinkeye?". Results revealed three variables, farm size, times yarded and ranking of the pain caused by pinkeye, were significantly associated with the frequency of pinkeye treatment. Specifically, farmers with smaller farm sizes were more likely to treat their cattle for pinkeye more frequently. So too were those who yarded their cattle more, and those that rated pinkeye as highly painful. The most used treatments for pinkeye in Australia were pinkeye ointments (n = 861), followed by eye patches (n = 637), pinkeye spray (n = 623), fly control (n = 507), and pinkeye powder (n = 408). Over half of those who had used subconjunctival injection, pinkeye ointments, eye patches, injectable antibiotics and veterinarians rated them as highly effective treatments. Kerosene is still used by some farmers (n = 106). The most common reason for not treating pinkeye was that it was 'too difficult to treat individuals'. These findings provide insight into the attitudes of Australian farmers to the treatment of pinkeye and should be used to improve pinkeye outcomes in cattle.

Risk factors associated with pinkeye in Australian cattle.

Pinkeye is the most important bovine ocular disease worldwide and a major welfare and economic concern to the Australian cattle industry. Pinkeye can occur in epidemic proportions, but severity and susceptibility vary within and between herds, indicating that the disease is multifactorial. This study was conducted to identify the on-farm risk factors associated with pinkeye disease in Australian cattle. Data were gathered from cattle farmers using a custom designed online questionnaire. Farmer responses suitable for assessment (n = 999) were analysed with descriptive, univariable and multivariable logistic regression to evaluate the association of 26 explanatory variables with pinkeye within-herd prevalence. Results revealed that farm location, farm grazing area, farmer-reported dust levels, fly levels, rain levels, animal zebu content and cattle age were significantly associated with pinkeye prevalence. More specifically, having a farm located in southern Australia, of smaller grazing area with cattle ≤ 2 years of age, was associated with a higher pinkeye prevalence. Pinkeye prevalence was also greater if respondents ranked their farms as having high fly levels compared to moderate and low fly levels, respectively and on farms ranked low for rainfall compared to moderate and high rainfall, respectively. Those that ranked their farms as having high dust levels had more pinkeye compared to moderate and low dust levels, but moderate dust levels were protective compared to low dust levels. The results confirm that pinkeye disease is multifactorial and is associated with a range of host and environmental factors. These findings should be used to assist in the control of the disease and improve pinkeye outcomes in Australian cattle.

Factors influencing the effectiveness of the Gudair vaccine for controlling Johne's disease in sheep flocks in Australia.

Ovine Johne's disease is a chronic debilitating disease of sheep caused by Mycobacterium avium subsp. paratuberculosis (Mptb) which results in diarrhoea, emaciation and mortalities in infected animals. Vaccination with Gudair® has been a key strategy for controlling the disease in Australia since its approval in 2002. Previous research conducted in Australia has demonstrated that the vaccine is quite effective in reducing sheep mortalities. While some farms have also been successful in reducing the prevalence of the disease in their flocks to undetectable levels, sheep in other flocks continue to shed Mptb in faeces even after an ongoing vaccination program . This study was conducted to investigate management, husbandry and biosecurity factors associated with paratuberculosis infection in Gudair® vaccinated sheep flocks in Australia. We enrolled 64 sheep farmers and interviewed them to obtain information about their management and biosecurity practices. Pooled faecal samples were collected from sheep at each farm and cultured to create two outcome variables: Mptb positive (yes/no) and disease prevalence level (nil, < 1 %, ≥ 1 %). Binary and ordinal logistic regression analyses were conducted to evaluate the association of management, husbandry and biosecurity factors with these outcome variables. Farms were more likely to have Mptb positive sheep and a higher disease prevalence in their flocks if they: (a) provided supplementary feed on the ground (instead of in a trough); (b) had a greater number of neighbours with sheep; and (c) had introduced rams from a greater number of sources. The results suggest the effectiveness of Gudair® vaccination to control OJD can be improved if sheep producers maintain other risk management strategies and biosecurity practices. Extension agencies should advise farmers not to relax their biosecurity practices and to purchase rams from only low-risk sources, even if they are continuing to vaccinate their flocks.

Demographics and management of paediatric dental-facial trauma in the 'lockdown' period: A UK perspective.

BACKGROUND: The United Kingdom entered 'lockdown' on the 23 March 2020 due to the COVID-19 pandemic. This resulted in school closures forcing children to remain at home. Dental-facial trauma was still likely to be common place due to falls and injuries exercising. The aim of this study was to explore the impact of the COVID-19 pandemic on clinical practice in a paediatric population in a tertiary care hospital and a linked Dental Institute. METHOD: A proforma was designed to capture the demographics, presenting complaints, type of dental-facial injury, treatment need and the treatment received for all paediatric patients presenting face to face with dental-facial trauma to King's College Hospital during the 'lockdown' period (23 March- 14 June 2020). RESULTS: Four hundred and twenty calls were triaged, converting to 102 patients seen face-to-face for dental-facial trauma. The remainder were able to be triaged 'virtually'. Injuries seen included 56 (54.9%) dentoalveolar injuries, 37 (36.2%) lacerations, five (4.9%) suspected facial fractures and four (3.9%) dog bites. Males and females were equally affected. The majority of incidents occurred in the home environment (n = 60, 58.8%), with the remainder (n = 42, 41.2%) occurring outside the home environment. The main causes of dental-facial trauma were falls (n = 47, 46.1%) and bicycles/scooters (n = 29, 28.4%). The most common type of dentoalveolar injury was lateral luxation (n = 15, 26.7%), followed by avulsion (n = 12, 21.4%). Only one child required treatment under general anaesthesia (GA). CONCLUSION: The demographic, presenting complaints and treatment needs of patients who presented during the lockdown period with dental-facial trauma were unusual. The overwhelming majority were able to be treated without the use of GA. The attendance protocol in a tertiary care setting and the use of 'teledentistry' ensured only the most severe trauma cases were seen. This highlights how more complex trauma can still occur during 'lockdown' and requires immediate management.

Smoking and Physical Activity Significantly Influence Stromal Vascular Fraction Cell Yield and Viability.

BACKGROUND: Subcutaneous tissue is an abundant source of adipose-derived regenerative cells. It is readily available and easy to extract by means of liposuction, making it one of the most popular sources for tissue engineering and regenerative medical applications. METHODS: The stromal vascular fraction (SVF) cell yield and viability of the lipoaspirate obtained from 43 patients undergoing elective liposuction were examined in correlation with their age, gender, body mass index, smoking status, and physical activity. The lipoaspirate was processed with the Celution® 800/CRS system to isolate the SVF and a few drops of the obtained pellet were used for cell counting with NecleoCounter® NC-100TM. RESULTS: Twenty-eight (65.1%) were men and 15 (34.9%) were women with an average age of 40.7 ± 10.4 years (women) and 38.9 ± 11.8 years (men). Viable SVF cells/g fat was significantly correlated with smoking level (negative correlation, ρ= - 0.312, P < 0.05) and with marginal significance with female gender. Cell viability showed a significant negative correlation with physical activity level (ρ = - 0.432, P < 0.01); borderline significance for correlation of this parameter with smoking level should not be neglected. Other parameters did not influence the cell yield nor the viability of the stromal vascular fraction. CONCLUSION: Many factors may influence SVF cell yield and viability. Our findings indicate that age and smoking significantly influenced SVF cell yield, age positively while smoking negatively. Increased physical activity had a negative correlation with SVF cell viability. LEVEL OF EVIDENCE N/A: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Vocal production in postpartum dairy cows: Temporal organization and association with maternal and stress behaviors.

Mammalian vocalizations can encode contextual information in both the spectrographic components of their individual vocal units and in their temporal organization. Here we observed 23 Holstein-Friesian dairy cows immediately after birth during interactions with their calf and when their calf was separated to the other side of a fence line. We investigated whether the vocalizations emitted in these postpartum contexts would vary temporally. We also described the maternal and stress behaviors preceding and following postpartum vocal production using kinematic diagrams and characterized call sequence structure. The kinematic diagrams highlight the disruption of maternal responses caused by calf separation and show that behavioral and vocal patterns varied according to the cows' emotional states and proximity to the calf in both contexts. During calf interactions, cows mainly produced closed-mouth calls simultaneous to licking their calf, whereas an escalation of stress responses was observed during calf separation, with the cows approaching the fence line, becoming alert to the calf, and emitting more mixed and open-mouth calls. Call sequences were similarly structured across contexts, mostly containing repetitions of a single call type, with a mean interval of 0.57 s between calls and a greater cumulative vocalization duration, attributed to an increased number of vocal units per sequence. Overall, calf separation was associated with a greater proportion of calls emitted as a sequence (inverse of single isolated calls), a shorter interval between separate call sequences, and a greater number of vocal units per sequence, compared with calf interactions. These temporal vocal features varied predictably with the high stress expression from cows during calf separation and may represent temporal modulations of emotional expression. Despite the noisy farm soundscape, empirical call type and temporal vocal features were easy to measure; thus, findings could be applied to future cattle studies wishing to analyze vocalizations for on-farm welfare assessments.

Kangaroos display gazing and gaze alternations during an unsolvable problem task.

Domestication is generally assumed to have resulted in enhanced communication abilities between non-primate mammals and humans, although the number of species studied is very limited (e.g. cats, Felis catus; dogs, Canis familiaris; wolves, Canis lupus; goats, Capra hircus; horses, Equus caballus). In species without hands for pointing, gazing at humans when dealing with inaccessible food during an unsolvable task, and in particular gaze alternations between a human and the unsolvable task (considered forms of showing), are often interpreted as attempts at referential intentional communication. We report that kangaroos, marsupial mammals that have never been domesticated, actively gazed at an experimenter during an unsolvable problem task (10/11 kangaroos tested), thus challenging the notion that this behaviour results from domestication. Nine of the 10 kangaroos additionally showed gaze alternations between the unsolvable task and experimenter. We propose that the potential occurrence of these behaviours displayed towards humans has been underestimated, owing to a narrow focus on domestic animals, as well as a more general eutherian research bias.

A 3-step Mechanical Digestion Method to Harvest Adipose-derived Stromal Vascular Fraction.

BACKGROUND: Adipose stromal vascular fraction (SVF) isolation with enzymatic digestion is the gold standard, but is expensive, having practical and legal concerns. The alternative mechanical SVF isolation methods provide lower cell yields as they employ either centrifugation, emulsification, or digestion steps alone. We combined mechanical processing with buffer incubation and centrifugation steps into an isolation method called "mechanical digestion" and compared the cell yields with that of enzymatic digestion. METHODS: A total of 40-mL lipoaspirate was harvested from 35 women undergoing liposuction and was submitted to conventional enzymatic digestion for SVF isolation or mechanical digestion using a closed unit harnessing 3 ports with blades, followed by buffer incubation and centrifugation. Culture of the SVFs and flow cytometry were performed. RESULTS: The SVF cell yield obtained by enzymatic digestion was significantly higher 3.38 × 106/mL (±3.63; n = 35) than that obtained by mechanical digestion 1.34 × 106/mL (±1.69; n = 35), P = 0.015. The average cell viability was 82.86% ± 10.68 after enzymatic digestion versus 85.86% ± 5.74 after mechanical digestion, which was not significant. Mechanical digested SVF expressed 2-fold higher stem cell surface markers compared with enzymatically digested SVF. Mechanical digestion was less time consuming, cost effective, and did not require a specific laboratory environment. CONCLUSIONS: Mechanically digested SVF was comparable to enzymatically digested SVF in terms of stromal cell composition and viability. With mechanical digestion, we can isolate 30%-50% SVF cells of that isolated with enzymatic digestion. Further studies are warranted to determine the clinical outcomes.

Vocal individuality of Holstein-Friesian cattle is maintained across putatively positive and negative farming contexts.

Cattle mother-offspring contact calls encode individual-identity information; however, it is unknown whether cattle are able to maintain individuality when vocalising to familiar conspecifics over other positively and negatively valenced farming contexts. Accordingly, we recorded 333 high-frequency vocalisations from 13 Holstein-Friesian heifers during oestrus and anticipation of feed (putatively positive), as well as denied feed access and upon both physical and physical & visual isolation from conspecifics (putatively negative). We measured 21 source-related and nonlinear vocal parameters and stepwise discriminant function analyses (DFA) were performed. Calls were divided into positive (n = 170) and negative valence (n = 163) with each valence acting as a 'training set' to classify calls in the oppositely valenced 'test set'. Furthermore, MANOVAs were conducted to determine which vocal parameters were implicated in individual distinctiveness. Within the putatively positive 'training set', the cross-validated DFA correctly classified 68.2% of the putatively positive calls and 52.1% of the putatively negative calls to the correct individual, respectively. Within the putatively negative 'training set', the cross-validated DFA correctly assigned 60.1% of putatively negative calls and 49.4% of putatively positive calls to the correct individual, respectively. All DFAs exceeded chance expectations indicating that vocal individuality of high-frequency calls is maintained across putatively positive and negative valence, with all vocal parameters except subharmonics responsible for this individual distinctiveness. This study shows that cattle vocal individuality of high-frequency calls is stable across different emotionally loaded farming contexts. Individual distinctiveness is likely to attract social support from conspecifics, and knowledge of these individuality cues could assist farmers in detecting individual cattle for welfare or production purposes.

An Enzyme-free Method for Isolation and Expansion of Human Adipose-derived Mesenchymal Stem Cells.

Mesenchymal stem cells (MSCs) are a population of multipotent cells that can be isolated from various adult and fetal tissues, including adipose tissue. As a clinically relevant cell type, optimal methods are needed to isolate and expand these cells in vitro. Most methods to isolate adipose-derived MSCs (ADSCs) rely on harsh enzymes, such as collagenase, to digest the adipose tissue. However, while effective at breaking down the adipose tissue and yielding a high ADSC recovery, these enzymes are expensive and can have detrimental effect on the ADSCs - including the risks of using xenogeneic components in clinical applications. This protocol details a method to isolate ADSCs from fresh lipoaspirate and abdominoplasty samples without enzymes. Briefly, this method relies on mechanical disassociation of any bulk tissue followed by an explant-type culture system. The ADSCs are permitted to migrate out of tissue and onto the tissue culture plate, after which the ADSCs can be cultured and expanded in vitro for any number of research and/or clinical applications.