Mathematical modelling of human corneal oxygenation and cell kinetics in corneal epithelial wound healing

Healthy transparent cornea depends upon the regulation of fluid, nutrient and oxygen transport through the tissue to sustain cell metabolism and other critical processes for normal functioning. This research considers the corneal geometry and investigates oxygen distribution using a two-dimensional Monod kinetic model, showing that previous studies make assumptions that lead to predictions of near-anoxic levels of oxygen tension in the limbal regions of the cornea. It also considers the comparison of experimental spatial and temporal data with the predictions of novel mathematical models with respect to distributed mitotic rates during corneal epithelial wound healing.

Evaluation and histological examination of a Campylobacter fetus subsp. venerealis small animal infection model

Bovine genital campylobacteriosis (BGC), caused by Campylobacter fetus subsp. venerealis, is associated with production losses in cattle worldwide. This study aimed to develop a reliable BGC guinea pig model to facilitate future studies of pathogenicity, abortion mechanisms and vaccine efficacy. Seven groups of five pregnant guinea pigs (1 control per group) were inoculated with one of three strains via intra-peritoneal (IP) or intra-vaginal routes. Samples were examined using culture, PCR and histology. Abortions ranged from 0 to 100 and re-isolation of causative bacteria from sampled sites varied with strain, dose of bacteria and time to abortion. Histology indicated metritis and placentitis, suggesting that the bacteria induce inflammation, placental detachment and subsequent abortion. Variation of virulence between strains was observed and determined by culture and abortion rates. IP administration of C. fetus subsp. venerealis to pregnant guinea pigs is a promising small animal model for the investigation of BGC abortion.

Molecular evidence of Ebola Reston virus infection in Philippine bats

Background: In 2008-09, evidence of Reston ebolavirus (RESTV) infection was found in domestic pigs and pig workers in the Philippines. With species of bats having been shown to be the cryptic reservoir of filoviruses elsewhere, the Philippine government, in conjunction with the Food and Agriculture Organization of the United Nations, assembled a multi-disciplinary and multi-institutional team to investigate Philippine bats as the possible reservoir of RESTV. Methods: The team undertook surveillance of bat populations at multiple locations during 2010 using both serology and molecular assays. Results: A total of 464 bats from 21 species were sampled. We found both molecular and serologic evidence of RESTV infection in multiple bat species. RNA was detected with quantitative PCR (qPCR) in oropharyngeal swabs taken from Miniopterus schreibersii, with three samples yielding a product on conventional hemi-nested PCR whose sequences differed from a Philippine pig isolate by a single nucleotide. Uncorroborated qPCR detections may indicate RESTV nucleic acid in several additional bat species (M. australis, C. brachyotis and Ch. plicata). We also detected anti-RESTV antibodies in three bats (Acerodon jubatus) using both Western blot and ELISA. Conclusions: The findings suggest that ebolavirus infection is taxonomically widespread in Philippine bats, but the evident low prevalence and low viral load warrants expanded surveillance to elaborate the findings, and more broadly, to determine the taxonomic and geographic occurrence of ebolaviruses in bats in the region. © 2015 Jayme et al.

Placental infection with Ureaplasma species is associated with histologic chorioamnionitis and adverse outcomes in moderately preterm and late-preterm infants

Objective The human Ureaplasma species are the microbes most frequently isolated from placentae of women who deliver preterm. The role of Ureaplasma species has been investigated in pregnancies at <32 weeks of gestation, but currently no studies have determined the prevalence of ureaplasmas in moderately preterm and late-preterm (hereafter, “moderate/late preterm”) infants, the largest cohort of preterm infants. Methods Women delivering moderate/late preterm infants (n = 477) and their infants/placentae (n = 535) were recruited, and swab specimens of chorioamnion tissue, chorioamnion tissue specimens, and cord blood specimens were obtained at delivery. Swab and tissue specimens were cultured and analyzed by 16S ribosomal RNA polymerase chain reaction (PCR) for the presence of microorganisms, while cord blood specimens were analyzed for the presence of cytokines, chemokines, and growth factors. Results We detected microorganisms in 10.6% of 535 placentae (443 were delivered late preterm and 92 were delivered at term). Significantly, Ureaplasma species were the most prevalent microorganisms, and their presence alone was associated with histologically confirmed chorioamnionitis in moderate/late preterm and term placentae (P < .001). The presence of ureaplasmas in the chorioamnion was also associated with elevated levels of granulocyte colony-stimulating factor (P = .02). Conclusions These findings have important implications for infection and adverse pregnancy outcomes throughout gestation and should be of major consideration for obstetricians and neonatologists.

Preliminary investigation of some physiological responses of Bos indicus heifers to surgical spaying

Objective To determine the value of peripheral blood concentrations of cortisol, creatine phosphokinase (CPK), aspartate aminotransferase (AST), non-esterified fatty acids (NEFAs) and haptoglobin as indicators of welfare in Brahman heifers spayed by either the Willis dropped ovary technique (WDOT) or the flank laparotomy method. Design A total of 24, 2-year-old Brahman heifers were allocated to: crush (head-bail) restraint alone (Control, n = 5); crush restraint and ear-punch (Ear-punch, n = 5); crush restraint, WDOT spay and ear-punch (WDOT, n = 9); or crush restraint, elecrtoimmobilisation, flank spay and ear-punch (Flank; n = 5). Cattle were blood sampled frequently to 8 h, and then daily to day 4 and were monitored to 42 days post-procedure. Peripheral blood concentrations of bound and unbound cortisol, CPK, AST, NEFAs and haptoglobin were determined. Results Concentrations of plasma bound cortisol peaked in the spayed heifers 3-4 h post-procedure; values in the Flank (1603 nmol/L) and WDOT (1290 nmol/L) groups were similar and significantly greater (P < 0.05) than in the Controls (519 nmol/L). Flank heifers had elevated plasma haptoglobin levels to day 4 postprocedure. Liveweights were significantly lower in the spayed compared with the Control heifers at 21 and 42 days post-procedure, with liveweight gains also significantly reduced at day 21. Conclusions Bound cortisol responses in spayed heifers were elevated to 6 h post-procedure and similar in WDOT- and flank-spayed animals, indicating comparable levels of pain and stress. An inflammatory response, indicated by haptoglobin concentrations, was sustained for longer in Flank than in WDOT spayed heifers, suggesting longer-lasting adverse effects on welfare from flank spaying than WDOT spaying. © 2011 The State of Queensland (Department of Employment, Economic Development and Innovation). Australian Veterinary Journal © 2011 Australian Veterinary Association.

Welfare outcomes for 3- and 6-month-old beef calves in a tropical environment castrated surgically or by applying rubber rings

Castration of cattle using rubber rings is becoming increasingly popular due to the perceived ease of the procedure and greater operator safety when compared with surgical castration. Few comparative studies have investigated the effects of different castration methods and calf age on welfare outcomes, particularly in a tropical environment. Thirty Belmont Red (a tropically adapted breed), 3-month-old (liveweight 71–119 kg) and 30, 6-month-old (liveweight 141–189 kg) calves were assigned to a two age × three castration (surgical, ring and sham) treatment factorial study (Surg3, Surg6, Ring3, Ring6, Sham3 and Sham6, n = 10 for each treatment group). Welfare outcomes were assessed post-castration using: behaviour for 2 weeks; blood parameters (cortisol and haptoglobin concentrations) to 4 weeks; wound healing to 5 weeks; and liveweights to 6 weeks. More Surg calves struggled during castration compared with Sham and Ring (P < 0.05, 90 ± 7% vs. 20 ± 9% and 24 ± 10%) and performed more struggles (1.9 ± 0.2, 1.1 ± 0.3 and 1.1 ± 0.3 for Surg, Sham and Ring, respectively), suggesting that surgical castration caused most pain during performance of the procedure. A significant (P < 0.05) time × castration method × age interaction for plasma cortisol revealed that concentrations decreased most rapidly in Sham; the Ring6 calves failed to show reduced cortisol concentrations at 2 h post-castration, unlike other treatment groups. By 7 h post-castration, all treatment groups had similar concentrations. A significant (P < 0.01) interaction between time and castration method showed that haptoglobin concentrations increased slightly to 0.89 and 0.84 mg/mL for Surg and Ring, respectively over the first 3 days post-castration. Concentrations for Surg then decreased to levels similar to Sham by day 21 and, although concentrations for Ring decreased on day 7 to 0.76 mg/mL, they increased significantly on day 14 to 0.97 mg/mL before reducing to concentrations similar to the other groups (0.66 mg/mL) by day 21. Significantly (P < 0.05) more of the wounds of the 3-month compared with the 6-month calves scored as ‘healed’ at day 7 (74% vs. 39%), while more (P = 0.062) of the Surg than Ring scored as ‘healed’ at day 21 (60% vs. 29%). At day 14 there were significantly (P < 0.05) fewer healed wounds in Ring6 compared with other treatment groups (13% vs. 40–60%). Liveweight gain was significantly (P < 0.05) greater in 3-month (0.53 kg/day) than in 6-month calves (0.44 kg/day) and in Sham calves (P < 0.001, 0.54 kg/day), than in Ring (0.44 kg/day) and Surg (0.48 kg/day) calves. Overall, welfare outcomes were slightly better for Surg than Ring calves due to reduced inflammation and faster wound healing, with little difference between age groups.

Welfare outcomes for 3- and 6-month-old beef calves in a tropical environment castrated surgically or by applying rubber rings

Castration of cattle using rubber rings is becoming increasingly popular due to the perceived ease of the procedure and greater operator safety when compared with surgical castration. Few comparative studies have investigated the effects of different castration methods and calf age on welfare outcomes, particularly in a tropical environment. Thirty Belmont Red (a tropically adapted breed), 3-month-old (liveweight 71–119 kg) and 30, 6-month-old (liveweight 141–189 kg) calves were assigned to a two age × three castration (surgical, ring and sham) treatment factorial study (Surg3, Surg6, Ring3, Ring6, Sham3 and Sham6, n = 10 for each treatment group). Welfare outcomes were assessed post-castration using: behaviour for 2 weeks; blood parameters (cortisol and haptoglobin concentrations) to 4 weeks; wound healing to 5 weeks; and liveweights to 6 weeks. More Surg calves struggled during castration compared with Sham and Ring (P < 0.05, 90 ± 7% vs. 20 ± 9% and 24 ± 10%) and performed more struggles (1.9 ± 0.2, 1.1 ± 0.3 and 1.1 ± 0.3 for Surg, Sham and Ring, respectively), suggesting that surgical castration caused most pain during performance of the procedure. A significant (P < 0.05) time × castration method × age interaction for plasma cortisol revealed that concentrations decreased most rapidly in Sham; the Ring6 calves failed to show reduced cortisol concentrations at 2 h post-castration, unlike other treatment groups. By 7 h post-castration, all treatment groups had similar concentrations. A significant (P < 0.01) interaction between time and castration method showed that haptoglobin concentrations increased slightly to 0.89 and 0.84 mg/mL for Surg and Ring, respectively over the first 3 days post-castration. Concentrations for Surg then decreased to levels similar to Sham by day 21 and, although concentrations for Ring decreased on day 7 to 0.76 mg/mL, they increased significantly on day 14 to 0.97 mg/mL before reducing to concentrations similar to the other groups (0.66 mg/mL) by day 21. Significantly (P < 0.05) more of the wounds of the 3-month compared with the 6-month calves scored as ‘healed’ at day 7 (74% vs. 39%), while more (P = 0.062) of the Surg than Ring scored as ‘healed’ at day 21 (60% vs. 29%). At day 14 there were significantly (P < 0.05) fewer healed wounds in Ring6 compared with other treatment groups (13% vs. 40–60%). Liveweight gain was significantly (P < 0.05) greater in 3-month (0.53 kg/day) than in 6-month calves (0.44 kg/day) and in Sham calves (P < 0.001, 0.54 kg/day), than in Ring (0.44 kg/day) and Surg (0.48 kg/day) calves. Overall, welfare outcomes were slightly better for Surg than Ring calves due to reduced inflammation and faster wound healing, with little difference between age groups.

Infection and pathology in Queensland grouper, Epinephelus lanceolatus, (Bloch), caused by exposure to Streptococcus agalactiae via different routes

Since 2007, 96 wild Queensland groupers, Epinephelus lanceolatus, (Bloch), have been found dead in NE Australia. In some cases, Streptococcus agalactiae (Group B Streptococcus, GBS) was isolated. At present, a GBS isolate from a wild grouper case was employed in experimental challenge trials in hatchery-reared Queensland grouper by different routes of exposure. Injection resulted in rapid development of clinical signs including bilateral exophthalmia, hyperaemic skin or fins and abnormal swimming. Death occurred in, and GBS was re-isolated from, 98% fish injected and was detected by PCR in brain, head kidney and spleen from all fish, regardless of challenge dose. Challenge by immersion resulted in lower morbidity with a clear dose response. Whilst infection was established via oral challenge by admixture with feed, no mortality occurred. Histology showed pathology consistent with GBS infection in organs examined from all injected fish, from fish challenged with medium and high doses by immersion, and from high-dose oral challenge. These experimental challenges demonstrated that GBS isolated from wild Queensland grouper reproduced disease in experimentally challenged fish and resulted in pathology that was consistent with that seen in wild Queensland grouper infected with S. agalactiae.

Natural Hendra Virus Infection in Flying-Foxes - Tissue Tropism and Risk Factors

Hendra virus (HeV) is a lethal zoonotic agent that emerged in 1994 in Australia. Pteropid bats (flying-foxes) are the natural reservoir. To date, HeV has spilled over from flying-foxes to horses on 51 known occasions, and from infected horses to close-contact humans on seven occasions. We undertook screening of archived bat tissues for HeV by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Tissues were tested from 310 bats including 295 Pteropodiformes and 15 Vespertilioniformes. HeV was detected in 20 individual flying-foxes (6.4%) from various tissues including spleen, kidney, liver, lung, placenta and blood components. Detection was significantly higher in Pteropus Alecto and Pconspicillatus, identifying species as a risk factor for infection. Further, our findings indicate that HeV has a predilection for the spleen, suggesting this organ plays an important role in HeV infection. The lack of detections in the foetal tissues of HeV-positive females suggests that vertical transmission is not a regular mode of transmission in naturally infected flying-foxes, and that placental and foetal tissues are not a major source of infection for horses. A better understanding of HeV tissue tropism will strengthen management of the risk of spillover from flying-foxes to horses and ultimately humans.

Spatiotemporal aspects of Hendra Virus infection in Pteropid Bats (Flying-Foxes) in Eastern Australia

Hendra virus (HeV) causes highly lethal disease in horses and humans in the eastern Australian states of Queensland (QLD) and New South Wales (NSW), with multiple equine cases now reported on an annual basis. Infection and excretion dynamics in pteropid bats (flying-foxes), the recognised natural reservoir, are incompletely understood. We sought to identify key spatial and temporal factors associated with excretion in flying-foxes over a 2300 km latitudinal gradient from northern QLD to southern NSW which encompassed all known equine case locations. The aim was to strengthen knowledge of Hendra virus ecology in flying-foxes to improve spillover risk prediction and exposure risk mitigation strategies, and thus better protect horses and humans. Monthly pooled urine samples were collected from under roosting flying-foxes over a three-year period and screened for HeV RNA by quantitative RT-PCR. A generalised linear model was employed to investigate spatiotemporal associations with HeV detection in 13,968 samples from 27 roosts. There was a non-linear relationship between mean HeV excretion prevalence and five latitudinal regions, with excretion moderate in northern and central QLD, highest in southern QLD/northern NSW, moderate in central NSW, and negligible in southern NSW. Highest HeV positivity occurred where black or spectacled flying-foxes were present; nil or very low positivity rates occurred in exclusive grey-headed flying-fox roosts. Similarly, little red flying-foxes are evidently not a significant source of virus, as their periodic extreme increase in numbers at some roosts was not associated with any concurrent increase in HeV detection. There was a consistent, strong winter seasonality to excretion in the southern QLD/northern NSW and central NSW regions. This new information allows risk management strategies to be refined and targeted, mindful of the potential for spatial risk profiles to shift over time with changes in flying-fox species distribution.

Coronavirus Infection and Diversity in Bats in the Australasian Region

Following the SARS outbreak, extensive surveillance was undertaken globally to detect and identify coronavirus diversity in bats. This study sought to identify the diversity and prevalence of coronaviruses in bats in the Australasian region. We identified four different genotypes of coronavirus, three of which (an alphacoronavirus and two betacoronaviruses) are potentially new species, having less than 90% nucleotide sequence identity with the most closely related described viruses. We did not detect any SARS-like betacoronaviruses, despite targeting rhinolophid bats, the putative natural host taxa. Our findings support the virus-host co-evolution hypothesis, with the detection of Miniopterus bat coronavirus HKU8 (previously reported in Miniopterus species in China, Hong Kong and Bulgaria) in Australian Miniopterus species. Similarly, we detected a novel betacoronavirus genotype from Pteropus alecto which is most closely related to Bat coronavirus HKU9 identified in other pteropodid bats in China, Kenya and the Philippines. We also detected possible cross-species transmission of bat coronaviruses, and the apparent enteric tropism of these viruses. Thus, our findings are consistent with a scenario wherein the current diversity and host specificity of coronaviruses reflects co-evolution with the occasional host shift.

The Use of Australian Honey in Moist Wound Management

This project has investigated the correlation between antimicrobial activity and the non-peroxide activity of particular Leptospermum-derived honeys. Initial investigations tested the efficacy of these honeys against one particular bacteria (Staphylococcus aureus), while later studies assessed the effect of this honey on a range of food pathogens, animal pathogens (e.g. in mastitis) and human pathogens (e.g. golden Staph). Using honeys sourced from around Australia and screened against several reference microorganisms, the anti-microbial potency of one particular Australian honey (from Leptospermum polygalifolium) was shown to be comparable to NZ Manuka honey (from L.scoparium). The results of this screening of active honeys against pathogenic bacteria has supported the registration of honey by Capilano Honey Limited as a “Drug” with the Therapeutic Goods Administration based on its antimicrobial activity.

Prevention and management of foot problems in diabetes: A summary guidance for daily practice 2015, based on the IWGDF Guidance Documents

In this 'Summary Guidance for Daily Practice', we describe the basic principles of prevention and management of foot problems in persons with diabetes. This summary is based on the International Working Group on the Diabetic Foot (IWGDF) Guidance 2015. There are five key elements that underpin prevention of foot problems: (1) identification of the at-risk foot; (2) regular inspection and examination of the at-risk foot; (3) education of patient, family and healthcare providers; (4) routine wearing of appropriate footwear, and; (5) treatment of pre-ulcerative signs. Healthcare providers should follow a standardized and consistent strategy for evaluating a foot wound, as this will guide further evaluation and therapy. The following items must be addressed: type, cause, site and depth, and signs of infection. There are seven key elements that underpin ulcer treatment: (1) relief of pressure and protection of the ulcer; (2) restoration of skin perfusion; (3) treatment of infection; (4) metabolic control and treatment of co-morbidity; (5) local wound care; (6) education for patient and relatives, and; (7) prevention of recurrence. Finally, successful efforts to prevent and manage foot problems in diabetes depend upon a well-organized team, using a holistic approach in which the ulcer is seen as a sign of multi-organ disease, and integrating the various disciplines involved.

Assessment of signs of foot infection in diabetes patients using photographic foot imaging and infrared thermography

Background Patients with diabetic foot disease require frequent screening to prevent complications and may be helped through telemedical home monitoring. Within this context, the goal was to determine the validity and reliability of assessing diabetic foot infection using photographic foot imaging and infrared thermography. Subjects and Methods For 38 patients with diabetes who presented with a foot infection or were admitted to the hospital with a foot-related complication, photographs of the plantar foot surface using a photographic imaging device and temperature data from six plantar regions using an infrared thermometer were obtained. A temperature difference between feet of > 2.2 °C defined a ''hotspot.'' Two independent observers assessed each foot for presence of foot infection, both live (using the Perfusion-Extent-Depth- Infection-Sensation classification) and from photographs 2 and 4 weeks later (for presence of erythema and ulcers). Agreement in diagnosis between live assessment and (the combination of ) photographic assessment and temperature recordings was calculated. Results Diagnosis of infection from photographs was specific (> 85%) but not very sensitive (< 60%). Diagnosis based on hotspots present was sensitive (> 90%) but not very specific (<25%). Diagnosis based on the combination of photographic and temperature assessments was both sensitive (> 60%) and specific (> 79%). Intra-observer agreement between photographic assessments was good (Cohen's j = 0.77 and 0.52 for both observers). Conclusions Diagnosis of foot infection in patients with diabetes seems valid and reliable using photographic imaging in combination with infrared thermography. This supports the intended use of these modalities for the home monitoring of high-risk patients with diabetes to facilitate early diagnosis of signs of foot infection.