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Bovine mastitis is an inflammation of the mammary gland, and it is the most common infectious disease in dairy cattle. Mastitis reduces milk yield and quality, costing dairy farmers millions of dollars each year. The aim of this study was to develop a point-of-need test for identifying mastitis pathogens that is field portable, cost-effective and can be used with minimal training. Using a proprietary polymer-based milk sample preparation method to rapidly extract pathogen DNA in milk samples, we demonstrated quantitative Polymerase Chain Reaction (qPCR) assays for six common bovine bacterial mastitis pathogens: Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Mycoplasma bovis and Escherichia coli. We also implemented this sample preparation method on a prototype point-of-need system in a proof-of-concept field trial to evaluate user experience. Importantly, the protype system enabled a sample-to-result turnaround time of within 70 min to quantitatively detect all six target pathogens. The key advantage of our point-of-need prototype system is being culture-independent yet providing automated milk sample preparation for molecular identification of key mastitis pathogens by non-expert users. Our point-of-need prototype system showed a good correlation to laboratory-based qPCR for target pathogen detection outcomes, thus potentially removing the need for milk samples to be transported off-site for laboratory testing. Above all, we successfully achieved our objective of developing a point-of-need biosensor technology for mastitis and increased its readiness level with industry partners towards technology commercialization., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

This study aimed to identify the pathogens isolated from the milk of cows with clinical mastitis in the subtropical region of Australia and to determine the antimicrobial susceptibility of these bacteria. Thirty dairy herds in the subtropical dairy region were asked to submit milk samples for the first 5 cases of clinical mastitis each month for 12 mo. Samples underwent aerobic culture, and isolates were identified via MALDI-TOF mass spectrometry. Antimicrobial susceptibility was determined for Escherichia coli, Enterococcus spp., Streptococcus agalactiae, Streptococcus uberis, Streptococcus dysgalactiae, Staphylococcus aureus, and non-aureus staphylococci and mammaliicocci (NASM). Between March 2021 and July 2022, 1,230 milk samples were collected. A positive culture result was recorded for 812 (66%) of the milk samples; from these samples, 909 isolates were obtained, including 49 isolates where no identification was possible. The remaining samples were classified as having no growth (16.8%) or as being contaminated (17.2%). The most common isolates with a MALDI-TOF diagnosis (n = 909) were Strep. uberis (23.6%), followed by the NASM group (15.0%). Farms enrolled in the study were in 3 distinct locations within the subtropical dairy region: North Queensland, Southeast Queensland, and Northern New South Wales. Some variation in isolate prevalence occurred between these 3 locations. We found lower odds of a sample being positive for E. coli in North Queensland (odds ratio [OR]: 0.25; 95% confidence interval [CI]: 0.07-0.87) and higher odds in Southeast Queensland (OR: 4.01; 95% CI: 1.96-8.20) compared with the reference, Northern New South Wales. We further found higher odds of Strep. dysgalactiae in North Queensland (OR: 5.69; 95% CI: 1.85-17.54) and Southeast Queensland compared with Northern New South Wales (OR: 3.99; 95% CI: 1.73-9.22). Although some seasonal patterns were observed, season was not significant for any of the analyzed isolates. Farm-level differences in pathogen profiles were obvious. Overall, clinical mastitis pathogens had low levels of resistance to the antimicrobials tested. This research demonstrates that Strep. uberis and the NASM bacterial group are the most common pathogens causing clinical mastitis in the subtropical dairy region. It highlights the importance of understanding pathogenic causes of mastitis at the farm and regional level for targeted control and therapy., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

A 5-year retrospective study was conducted to describe the mastitis-causing organisms isolated from bovine milk samples submitted to four veterinary diagnostic laboratories in Australia. The aim of this study was to identify temporal, geographical, and seasonal patterns of occurrence for the organisms and report the in vitro susceptibility of the most common mastitis-causing pathogens. In total, 22,102 milk samples were submitted between 2015 and 2019. The results were reported as positive growth for at least one significant organism ( n = 11,407; 51.6%), no growth ( n = 5,782; 26.2%), and mixed/contaminated growth ( n = 4,913; 22.2%). Culture results for no growth, gram-negative bacteria, and eukaryotic organisms were combined for each region, and they were accounted for between 23 and 46% of submissions. These results represent a subset of mastitis cases for which the antibiotic treatment may not be warranted. A total of 11,907 isolates were cultured from 11,407 milk samples. The most common isolated organisms were Streptococcus uberis [41.3%; 95% confidence interval (CI): 40.4-42.1%] and Staphylococcus aureus (23.6%; 95% CI: 22.8-24.3%). For S. uberis and S. aureus , there was an association between a positive culture result and the dairy region. All regions except for the Sub-tropical Dairy region were more likely to culture S. uberis compared to the reference, Dairy NSW ( P < 0.001). Similarly, for S. aureus , a positive culture result was more likely in all other dairy regions compared to Dairy NSW ( P < 0.001). The LISA cluster analysis identified differences between High-High (hotspot) postcodes for S . aureus and S . uberis throughout all the analyzed dairy regions. These results highlight the need for further investigations into specific risk factors, such as environmental factors and herd-level predictors, which may have influenced the observed regional variations. Common mastitis-causing pathogens showed overall good susceptibility to a range of antimicrobials used in the treatment of mastitis. On-going surveillance of mastitis-causing pathogens and their antimicrobial susceptibilities will facilitate targeted mastitis control and treatment programs., Competing Interests: MH is employed by The Milk Road. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Langhorne, Gupta, Horsman, Wood, Wood, Barker, Deutscher, Price, McGowan, Humphris, Ranjbar, Henning and Gibson.)

Herpesviruses have been reported in several Australian marsupial species, with an overt, sometimes fatal disease described in macropods. Our study identifies a gammaherpesvirus in northern brown bandicoots (Isoodon macrourus) and provides virus prevalence data for bandicoots in southeast Queensland, Australia. Herpesvirus DNA was detected using pan-Herpesviridae family primers in a nested PCR format. Samples from 35 northern brown bandicoots were screened, including whole blood (n=29), oropharyngeal swabs (n=34), urine (n=22), and feces (n=23). Combining all sample types, herpesvirus DNA was detected at a total prevalence of 51% (18/35). Whole blood and oropharyngeal swabs proved to be the optimal samples for detection of this virus, with prevalences of 34% and 38%, respectively. Herpesvirus DNA was detected in 4.5% (1/22) of urine samples and not at all in fecal samples. Detection of herpesvirus was more likely in males than females. Animals were trapped at eight different locations, and at all but one location at least one herpesvirus positive animal was detected. This study indicates a high prevalence of the virus within northern brown bandicoot populations in southeast Queensland. Further research is required to understand the clinical manifestations, if any, of herpesvirus infection in this species and how this may affect populations in the face of stressors such as land clearing and habitat fragmentation., (© Wildlife Disease Association 2021.)

The impetus of this study was the imperative to establish blood biomarker values for clinically healthy mahogany gliders ( Petaurus gracilis ) in order to monitor the health status of eight captive individuals during their movement to a new facility. The study established ranges for 18 hematologic and 21 biochemical blood biomarkers for healthy individuals in a captive environment. The reported values are consistent with those published for other Australian glider and possum species. No statistically significant differences were found between the sexes, but significant age effects were observed. Specifically, subadult animals reported significantly higher total white cell counts, lymphocyte counts, alkaline phosphatase, creatine kinase, and glucose and chloride levels, compared to adult animals. Although there were no clinically significant changes in blood biomarkers associated with the relocation, many of the hematologic and biochemical biomarkers demonstrated the expected changes associated with the physiological stress of relocation. Specifically, triglycerides, glucose, globulins, creatinine kinase, aspartate transferase (AST), total protein, urea, phosphorous, potassium, sodium, chloride, neutrophils, and hematocrit showed changes with the large environmental change. The majority of the blood biomarkers returned to baseline levels 5 wk postrelocation, with all but one aged animal showing no signs of chronic health derangements following the relocation. The abnormal blood biomarker profiles of two geriatric individuals, one male diagnosed with pericloacal and adrenal gland tumors at the beginning of the study, and one female diagnosed with chronic urinary tract infections and suspected bone marrow disease following the relocation, are presented. The findings of this study inform the health monitoring of native gliders in captivity, rehabilitation, and clinical research scenarios. These findings also provide useful baseline data to aid in the health assessment of captive-bred individuals during their reintroduction into free-living populations.

There is growing evidence that pathogens play a role in population declines and species extinctions. For small populations, disease-induced extinction may be especially probable. We estimated the susceptibility of two amphibian species of conservation concern (the dusky gopher frog [Lithobates sevosus] and boreal toad [Anaxyrus boreas boreas]) to an emerging pathogen (ranavirus) using laboratory challenge experiments, and combined these data with published demographic parameter estimates to simulate the potential effects of ranavirus exposure on extinction risk. We included effects of life stage during pathogen exposure, pathogen exposure interval, hydroperiod of breeding habitat, population carrying capacity, and immigration in simulations. We found that both species were highly susceptible to ranavirus when exposed to the pathogen in water at environmentally relevant concentrations. Dusky gopher frogs experienced 100 % mortality in four of six life stages tested. Boreal toads experienced 100 % mortality when exposed as tadpoles or metamorphs, which were the only life stages tested. Simulations showed population declines, greater extinction probability, and faster times to extinction with ranavirus exposure. These effects were more evident with more frequent pathogen exposure intervals and lower carrying capacity. Immigration at natural rates did little to mitigate effects of ranavirus exposure unless immigration occurred every 2 years. Our results demonstrate that disease-induced extinction by emerging pathogens, such as ranavirus, is possible, and that threat may be especially high for species with small population sizes. For the species in this study, conservation organizations should incorporate ranavirus surveillance into monitoring programs and devise intervention strategies in the event that disease outbreaks occur.

Declines of the southern Rocky Mountain population of boreal toad (Anaxyrus boreas boreas) have led to the establishment of a captive assurance population and reintroduction program, in an attempt to preserve and propagate this geographically isolated population. One of the unique adaptations of this species is its ability to survive in cold environments by undergoing long periods of hibernation. In captivity, hibernation can be avoided altogether, decreasing morbidity caused by compromised immune systems. However, it is not entirely clear how essential hibernation is to reproductive success. In this study, the effects of hibernation versus nonhibernation, and exogenous hormones on oviposition, were examined in boreal toad females in the absence of males. In the summers of 2011 and 2012, 20 females housed at Mississippi State University were treated with a double priming dose of hCG and various ovulatory doses of hCG and LH-releasing hormone analog but denied hibernation. Exogenous hormones, in the absence of hibernation, could not induce oviposition over two breeding seasons (2011-2012). In contrast, during the summer of 2012 and 2013, 17 of 22 females (77%) housed at the Native Aquatic Species Restoration Facility (Alamosa, CO, USA) oviposited after they were treated with two priming doses of hCG (3.7 IU/g each) and a single ovulation dose of hCG (13.5 IU/g) and LH-releasing hormone analog (0.4 μg/g) after hibernation. There was a significant difference in oviposition between females that were hibernated and received hormones (2012, P < 0.05 and 2013, P < 0.01) compared to hibernated control females. In 2013, 12 of 16 remaining Mississippi State University females from the same group used in 2011 and 2012 were hibernated for 1, 3, and 6 months, respectively and then treated with the same hormone regimen administered to females at the Native Aquatic Species Restoration Facility. Together, hibernation and hormone treatments significantly increased oviposition (P < 0.05), with 33% of females ovipositing. These results suggest that (1) hibernation is a key factor influencing oviposition that cannot be exclusively circumvented by exogenous hormones; (2) females do not require the presence of a male to oviposit after hormone treatments; and (3) longer hibernation periods are not beneficial for oviposition. The hormonal induction of oviposition in the absence of males and shorter hibernation periods could have important captive management implications for the boreal toad. Furthermore, the production of viable offspring by IVF where natural mating is limited could become an important tool for genetic management of this boreal toad captive population., (Copyright © 2015 Elsevier Inc. All rights reserved.)

A pathophysiological study of the pulmonary vasculature in 10 patients with hypoxic cor pulmonale and severe airways obstruction (five treated and five untreated with long-term oxygen) is presented. The media of muscular pulmonary arteries was normal or atrophic but, in the intima, there was active deposition of longitudinal muscle, fibrosis and elastosis. In the arterioles a medical coat of circular smooth muscle bounded by a new internal elastic lamina had developed, while there was deposition of longitudinal muscle and fibrosis in the intima. In five cases the lumen was subdivided into parallel tubes, found by serial section to lead into alveolar capillaries. These features are distinctive of hypoxaemia and obstructive airways disease. Changes continued until death. The conspicuous longitudinal muscle may be attributable to stretching of vessels round distorted terminal airways; further exploration into mechanisms is required. The hypothesis that vascular changes follow hypoxic vasoconstriction is no longer tenable. No correlations were found between quantitative pathological findings and arterial blood gas tensions, pulmonary artery pressure or haematocrit. There were no differences between patients treated or not treated with oxygen which might suggest that it arrests pathological changes. Thus, once a patient is given oxygen, survival probably depends as much on progressive mechanical changes in the lung as on continuing hypoxaemia.