Risk factors associated with self-reported Q fever in Australian wildlife rehabilitators: Findings from an online survey.

Australian wildlife rehabilitators (AWR) are at increased risk of developing Q fever, a serious zoonotic disease caused by the intracellular bacterium Coxiella burnetii. Previous studies have suggested that Australian wildlife may be a potential C. burnetii infection source for humans. However, a recent serological survey of AWR found no association between C. burnetii exposure and direct contact with any wildlife species. To further explore the potential risk that wildlife may pose, this study aimed to identify associations between self-reported Q fever in AWR and risk factors for exposure to C. burnetii. An online cross-sectional survey was implemented in 2018 targeting AWR nationwide. Risk factors for self-reported Q fever were determined using multivariable logistic regression. Medically diagnosed Q fever was self-reported in 4.5% (13/287) of unvaccinated respondents. Rehabilitators who self-reported medically diagnosed Q fever were significantly more likely to: primarily rehabilitate wildlife at a veterinary clinic (OR 17.87, 95% CI: 3.09-110.92), have domestic ruminants residing on the property where they rehabilitate wildlife (OR 11.75, 95% CI: 2.91-57.42), have been educated at a High School/Technical and Further Education level (OR 10.29, 95% CI: 2.13-84.03) and be aged >50 years (OR 6.61, 95% CI: 1.60-38.35). No association was found between self-reported Q fever and direct contact with wildlife. These findings support previous work suggesting that AWR are at increased risk of C. burnetii infection and may develop Q fever potentially via exposure to traditional infection sources including livestock, other domestic animals, or contaminated environments, in association with their rehabilitation practices and lifestyle. Although Q fever vaccination is recommended for AWR, vaccine uptake is low in this population. Future studies should aim to determine the level of Q fever awareness and identify barriers to Q fever vaccination in this at-risk group. The difficulty in accessing the AWR population also highlights the need for a national centralized AWR database.

Factors associated with Q fever vaccination in Australian wildlife rehabilitators.

Australian wildlife rehabilitators (AWR) are at risk of contracting Q fever, a serious zoonotic disease caused by Coxiella burnetii. Despite Australian government recommendations for AWR to receive Q fever vaccination (QFV), and the availability of a safe and effective vaccine in Australia, shortfalls in vaccine uptake have been observed in AWR. This study aimed to determine factors associated with QFV status and describe AWR attitudes and potential barriers towards QFV. Data were obtained from a nationwide, online, cross-sectional survey of AWR undertaken in 2018. Approximately-three quarters (200/265; 75.5 %) of those that had heard of Q fever were also aware of the Q fever vaccine, and of those, 25.5 % (51/200) were vaccinated. Barriers to QFV, among unvaccinated respondents who had also heard of Q fever and the vaccine (149/200; 74.5 %), included concerns regarding the safety, efficacy, and importance of the Q fever vaccine. Complacency toward vaccination, convenience of vaccination, and a lack of Q fever knowledge were also notable barriers. Only 27.7 % (41/148) of respondents reported having had vaccination recommended to them. Multivariable logistic regression identified that vaccinated AWR were more likely to be aged ≤ 50 years (OR 4.51, 95 % CI: 2.14-10.11), have had a university level education (OR 2.78, 95 % CI: 1.39-5.73), have resided in New South Wales/Australian Capital Territory and Queensland than in other Australian jurisdictions (OR 2.9, 95 % CI: 1.10-8.83 and OR 4.82, 95 % CI: 1.64-16.00 respectively) and have attended an animal birth (OR 2.14, 95 % CI: 1.02-4.73). Knowledge gaps regarding Q fever and QFV in AWR demonstrated the need for interventions to raise the awareness of the potential health consequences of C. burnetii exposure and Q fever prevention. Education programs to allow AWR to develop an informed perspective of Q fever and QFV, coupled with improvements in vaccine affordability and the implementation of programs to enhance accessibility, may also increase vaccine uptake.

Serological Evidence of Exposure to Spotted Fever Group and Typhus Group Rickettsiae in Australian Wildlife Rehabilitators.

Rickettsioses are arthropod-borne zoonotic diseases, several of which occur in Australia. This study aimed to assess the exposure levels and risk factors for Rickettsia spp. among Australian wildlife rehabilitators (AWRs) using serology, PCR and a questionnaire. Antibody titres against Spotted Fever Group (SFG), Typhus Group (TG) and Scrub Typhus Group (STG) antigens were determined using an immunofluorescence assay. PCR targeting the gltA gene was performed on DNA extracts from whole blood and serum. Logistic regression was used to identify risk factors associated with seropositivity. Of the 27 (22.1%; 27/122) seropositive participants all were seropositive for SFG, with 5/27 (4.1%) also positive for TG. Of the 27 positive sera, 14.8% (4/27) were further classified as exposure to R. australis, 3.7% (1/27) to R. honei, 3.7% (1/27) to R. felis and 77.8% (21/27) were classified as 'indeterminate'-most of which (85.7%; 18/21) were indeterminate R. australis/R. honei exposures. Rickettsia DNA was not detected in whole blood or serum. Rehabilitators were more likely to be seropositive if more than one household member rehabilitated wildlife, were older than 50 years or had occupational animal contact. These findings suggest that AWRs are at increased risk of contracting Rickettsia-related illnesses, however the source of the increased seropositivity remains unclear.

Coxiella burnetii seroprevalence and Q fever in Australian wildlife rehabilitators.

Coxiella burnetii is the causative bacterium of the zoonotic disease Q fever, which is recognised as a public health concern globally. Macropods have been suggested as a potential source of C. burnetii infection for humans. The aim of this cross-sectional study was to determine the prevalence of C. burnetii exposure in a cohort of Australian wildlife rehabilitators (AWRs) and assess Q fever disease and vaccination status within this population. Blood samples were collected from adult participants attending the Australian Wildlife Rehabilitation Conference in Sydney in July 2018. Participants completed a questionnaire at the time of blood collection. Antibody titres (IgG, IgA and IgM) against phase I and phase II C. burnetii antigens as determined by immunofluorescence assay, revealed that of the unvaccinated participants, 6.1% (9/147) had evidence of exposure to C. burnetii. Of the total participants, 8.1% (13/160) had received Q fever vaccination, four of whom remained seropositive at the time of blood collection. Participants reporting occupational contact with ruminants, were eight times more likely to have been vaccinated against Q fever, than those reporting no occupational animal contact (OR 8.1; 95% CI 1.85-45.08). Three AWRs (2%) reported having had medically diagnosed Q fever, two of whom remained seropositive at the time of blood collection. Despite the lack of association between macropod contacts and C. burnetii seropositivity in this cohort, these findings suggest that AWRs are approximately twice as likely to be exposed to C. burnetii, compared with the general Australian population. This provides support for the recommendation of Q fever vaccination for this potentially 'at-risk' population. The role of macropods in human Q fever disease remains unclear, and further research into C. burnetii infection in macropods including: infection rate and transmission cycles between vectors, macropods as reservoirs, other animals and humans is required.

Cat fleas (Ctenocephalides felis clade 'Sydney') are dominant fleas on dogs and cats in New South Wales, Australia: Presence of flea-borne Rickettsia felis, Bartonella spp. but absence of Coxiella burnetii DNA.

The cat flea (Ctenocephalides felis) is the most common flea species parasitising both domestic cats and dogs globally. Fleas are known vectors of zoonotic pathogens such as vector-borne Rickettsia spp. and Bartonella spp. and could theoretically transmit Coxiella burnetii, the causative agent of Q fever. A total of 107 fleas were collected from 21 cats and 14 dogs in veterinary clinics, a feline rescue organisation and a grooming salon in New South Wales, Australia, to undergo PCR detection of Bartonella spp., Rickettsia spp. and C. burnetii DNA. Morphological identification confirmed that the cat flea (C. felis) is the most common flea in New South Wales, Australia, with only a single stick fast flea, Echidnophaga gallinacea recorded. The examined fleas (n = 35) at the cox1 locus revealed five closely related C. felis haplotypes (inter-haplotype distance < 0.5%). Multiplex TaqMan qPCR targeting the gltA (Rickettsia spp.) and ssrA (Bartonella spp.) genes was positive in 22.9% (95% CI: 11.8-39.3%) and 11.4% (95% CI: 3.9-26.6%) of samples, respectively. None of the DNA isolated from fleas was positive on TaqMan qPCRs targeting the C. burnetii IS1111, Com1 and htpAB genes. Co-infection of C. felis with Bartonella henselae and Bartonella clarridgeiae was demonstrated using gltA and ssrA Illumina next-generation amplicon sequencing. These findings reinforce the importance of flea control on domestic dogs and cats to effectively control the transmission of Rickettsia felis and Bartonella spp. The flea, however, is unlikely to be a vector of C. burnetii between companion animals and humans.

Molecular detection of Coxiella burnetii in raw meat intended for pet consumption.

The discovery of antibodies against Coxiella burnetii in cattery-confined breeding cats indicating prior or current exposure (Shapiro et al., 2015) prompted an investigation into possible sources of infection. One hypothesis was that raw meat diets containing reservoir species may provide a source of C. burnetii transmission. The aim of this pilot study was to determine whether C. burnetii DNA was present in raw meat sold exclusively for companion animal consumption. The sample population consisted of raw meat packages (n = 58) of primarily kangaroo origin, with three to four aliquots (50-120 mg) randomly selected from each package. Genomic DNA was extracted from whole tissue in each of these aliquots using a modified protocol. Three quantitative PCR assays were used for the detection of C. burnetii targeting the IS1111 gene, the heat shock operon htpAB and the C. burnetii outer membrane protein-coding gene, com1. Coxiella burnetii DNA was detected in 25/58 samples (43%) using the IS1111, htpAB and/or com1 PCR assays and confirmed by DNA sequencing. All samples amplifying a product in the com1 assay also amplified a product in the htpAB and IS1111 assays. A total of 17/58 (29%) packets were positive with all three genes, 4/58 (7%) were positive with two genes (IS1111 and htpAB) and 4/58 (7%) were positive with the IS1111 gene only. Coxiella burnetii DNA was five times more likely to be found in offal than skeletal muscle meat samples. All meat samples in which C. burnetii DNA was found were from kangaroo tissues, while samples labelled as non-kangaroo meat (n = 4) were negative. Multi-locus variable number of tandem repeat analysis (MLVA) identified three different genotypes of C. burnetii that have all been identified previously from Australian human clinical Q fever cases. Further investigations are required to determine the potential role of certain raw meats in the transmission of C. burnetii to cats and humans.

New insights on the epidemiology of Coxiella burnetii in pet dogs and cats from New South Wales, Australia.

Q fever is considered one of the most important zoonoses in Australia. Whilst ruminants are the primary reservoirs for Coxiella burnetii, and the major source of human infection, human cases have also been reported following contact with pet dogs and cats. This study aimed to estimate the prevalence of seropositivity to, and bacterial shedding of, C. burnetii by pet dogs and cats in a region with a high human Q fever incidence and explore risk factors for C. burnetii exposure. Samples (serum, whole blood, reproductive tissue, reproductive swabs) and questionnaires (completed by the pet's owner) were collected from dogs and cats from eight communities across remote New South Wales (NSW), Australia. Overall 86/330 dogs (26.1%, 95% CI 21.3-30.8%) and 19/145 cats (13.1%, 95% CI 7.6-18.6%) were seropositive to C. burnetii. Seroprevalence varied significantly between communities and was highest in communities within 150 km of a 2015 human Q fever outbreak. Feeding raw kangaroo was identified as a risk factor for seropositivity (adjusted OR 3.37, 95% CI 1.21-9.43). Coxiella burnetii DNA was not detected from any dog or cat whole blood, reproductive tissue or vaginal/preputial swab using qPCR targeting the IS1111 and com1 genes. Our findings suggest that companion animals are frequently exposed to C. burnetii in western NSW. Geographical variation in C. burnetii seroprevalence amongst companion animals - which corresponds with a human Q fever outbreak - suggests a shared environmental source of infection is likely with important consequences for public and animal health. The lack of detection of C. burnetii DNA from healthy companion animals suggests that pet dogs and cats are not an important reservoir for human Q fever infection outside a narrow periparturient window.

Draft Genome Sequence of a Red Basidiomycete Yeast, Symmetrospora coprosmae Strain UCD350, Isolated from Soil in Ireland.

Symmetrospora coprosmae is a red yeast from the subphylum Pucciniomycotina in the phylum Basidiomycota. Here, we present the first genome sequence of S. coprosmae strain UCD350, from an isolate collected from soil in Ireland. The genome size is 20.2 Mb.

Development of a loop-mediated isothermal amplification assay for the detection of Streptococcus agalactiae in bovine milk.

Streptococcus agalactiae is a well-characterized bovine mastitis pathogen that is known to be highly contagious and capable of spreading rapidly in affected dairy herds. Loop-mediated isothermal amplification (LAMP) is a novel molecular diagnostic method that has the capability to provide rapid, cost-effective screening for pathogens to support on-farm disease control and eradication programs. In the current study, a LAMP test was developed to detect S. agalactiae in milk. The assay was validated on a bank of existing clinical mastitis milk samples that had previously been identified as S. agalactiae positive via traditional microbiological culture techniques and PCR. The LAMP assay was conducted on bacterial colonies and DNA extracted from milk in tube- and plate-based formats using multiple detection platforms. The 1-h assay conducted at 64 °C exhibited repeatability (coefficient of variation) of 2.07% (tube) and 8.3% (plate), sensitivity to ~20 pg of extracted DNA/reaction, and specificity against a panel of known bacterial mastitis pathogens. Of the 109 known S. agalactiae isolates assessed by LAMP directly from bacterial cells in culture, 108 were identified as positive, in accordance with PCR analysis. The LAMP analysis from the corresponding milk samples indicated that 104 of these milks exhibited a positive amplification curve. Although exhibiting some limitations, this assay provides an opportunity for rapid screening of milk samples to facilitate on-farm management of this pathogen.

Activation of human dendritic cells by p-phenylenediamine.

Exposure to p-phenylenediamine (pPD), a primary intermediate in hair dye formulations, is often associated with the development of allergic contact dermatitis. Such reactions involve activation of the subject's immune system. The aim of these studies was to explore the relationship between pPD oxidation and functional maturation of human monocyte-derived dendritic cells in vitro. Dendritic cells were incubated with pPD and Bandrowski's base (BB) for 16 h, and expression of the costimulatory receptors CD40, CD80, CD83, CD86, and major histocompatibility complex class II intracellular glutathione levels and cell viability were measured. In certain experiments, glutathione (1 mM) was added to culture medium. Liquid chromatography-mass spectrometry (LC-MS) analysis and exhaustive solvent extraction were used to monitor the rate of [(14)C]pPD oxidation and the extent of pPD binding to cellular and serum protein, respectively. Proliferation of allogeneic lymphocytes was determined by incorporation of [(3)H]thymidine. Exposure of dendritic cells to pPD (5-50 microM), but not BB, was associated with an increase in CD40 and MHC class II expression and proliferation of allogeneic lymphocytes. Dendritic cell activation with pPD was not associated with apoptotic or necrotic cell death or depletion of glutathione. Neither pPD nor BB altered dendritic cell expression of CD80, CD83, or CD86. LC-MS analysis revealed pPD was rapidly oxidized in cell culture media to BB. Addition of glutathione inhibited BB formation but did not prevent covalent binding of pPD to dendritic cell protein or dendritic cell activation. Collectively, these studies show that pPD, but not BB, selectively activates human dendritic cells in vitro.

Sensory neurons from mice lacking the Brn-3b POU family transcription factor are resistant to death-inducing stimuli both in vitro and in vivo.

The critical role for programmed cell death in the normal development of the nervous system and the abnormal cell death that occurs in human neurodegenerative diseases renders it of vital importance to identify factors that enhance or reduce the levels of cell death in specific neuronal cells in the nervous system. We show that the Brn-3b transcription factor is essential for normal cell death responses in the peripheral nervous system and that in its absence neurons are resistant to a variety of death-inducing stimuli both in vitro and in vivo.