Fetoplacental pathology of equine abortion, premature birth, and neonatal loss due to Chlamydia psittaci.

This report describes the fetoplacental pathology of Chlamydia psittaci-associated abortion, premature birth, and neonatal loss in 46 of 442 equine abortion investigations between 2015 and 2019. Seven abortions, 26 premature births, and 13 neonatal deaths with positive C. psittaci polymerase chain reaction (PCR) were evaluated. In 83% of cases (38/46), C. psittaci infection was considered as the primary cause of loss based on quantitative PCR (qPCR) confirmation, pathological findings, and exclusion of other causes, and was supported by Chlamydia spp immunolabeling in fetoplacental lesions. Lymphohistiocytic placentitis with vasculitis (36/38) affected the amnion, umbilical cord, and chorioallantois at the umbilical vessel insertion and/or cervical pole. Lymphohistiocytic chorionitis in the subvillous stroma extended to the allantois mostly without villous destruction. Lymphohistiocytic amnionitis and funisitis occurred at the amniotic cord attachment. Lymphohistiocytic hepatitis was observed in 19/38 cases and pneumonia was identified in 26 cases. Chlamydia spp immunolabeled in placenta, lung, liver, or splenic tissue in the cases that were tested (14/38). C. psittaci infection was not the cause of loss in 2 cases with other diseases and of uncertain significance in 6 cases with no conclusive cause of loss. immunohistochemistry (IHC) was negative for 6 of these cases (6/8). The highest Chlamydia load was detected in pooled placental tissues by qPCR. qPCR and IHC had 83% congruence at a qPCR cut-off of 1 gene copy. IHC limits of detection corresponded to infections with 2 × 102 gene copies identified by qPCR. This study confirms the etiological role of C. psittaci as a cause of naturally occurring equine reproductive loss.

A Scoping Review of the Global Distribution of Causes and Syndromes Associated with Mid- to Late-Term Pregnancy Loss in Horses between 1960 and 2020.

Equine pregnancy loss is frustrating and costly for horse breeders. The reproductive efficiency of mares has significant implications for a breeding operation's economic success, and widespread losses can have a trickle-down effect on those communities that rely on equine breeding operations. Understanding the causes and risks of equine pregnancy loss is essential for developing prevention and management strategies to reduce the occurrence and impact on the horse breeding industry. This PRISMA-guided scoping review identified 514 records on equine pregnancy loss and described the global spatiotemporal distribution of reported causes and syndromes. The multiple correspondence analysis identified seven clusters that grouped causes, syndromes, locations and pathology. Reasons for clustering should be the focus of future research as they might indicate undescribed risk factors associated with equine pregnancy loss. People engaged in the equine breeding industry work closely with horses and encounter equine bodily fluids, placental membranes, aborted foetuses, and stillborn foals. This close contact increases the risk of zoonotic disease transmission. Based on this review, research is required on equine abortion caused by zoonotic bacteria, including Chlamydia psittaci, Coxiella burnetii and Leptospira spp., because of the severe illness that can occur in people who become infected.

Willingness to adopt personal biosecurity strategies on thoroughbred breeding farms: Findings from a multi-site pilot study in Australia's Hunter Valley.

There are almost 9,500 full-time employees in Australia's thoroughbred horse breeding industry. During foaling, they can be exposed to bodily fluids and mucous membranes which may present risks for zoonotic disease. These risks can be mitigated through personal biosecurity strategies. The aim of this study was to identify which personal biosecurity strategies were more or less likely to be adopted by workers. Seventeen participants representing 14 thoroughbred breeding farms and three equine veterinary practices in Australia's largest thoroughbred breeding region trialed up to 16 stakeholder-nominated personal biosecurity strategies over the 2021 foaling season. The strategies encompassed personal protective equipment (PPE), zoonotic disease awareness, policies and protocols, supportive environments, and leadership. Strategy adoption was monitored through three repeated self-audit surveys designed around the Transtheoretical Model of change (TTM) and findings were reviewed in exit interviews. For all survey waves in aggregate, 13 strategies were practiced by at least 50.0% of participants. Participants were most likely to use a ready-made foaling box (98.0%), communicate the message that PPE usage is a personal responsibility (94.1%) and use ready-made PPE kits (88.2%). However, 31.4% had no intention of doing practice sessions and/or dummy runs for PPE use and 27.5% had no intention of using a buddy system on farm/practice to check use of PPE. Whilst these rates indicate workers' willingness to adopt and maintain personal biosecurity strategies, they also indicate capacity for more practices to be implemented more often. Overall, the findings highlight the need for personal biosecurity interventions to be sensitive to the demands of the annual thoroughbred breeding calendar, the size of the breeding operation and the availability of skilled staff.

Real-time fluorometric and end-point colorimetric isothermal assays for detection of equine pathogens C. psittaci and equine herpes virus 1: validation, comparison and application at the point of care.

BACKGROUND: C. psittaci has recently emerged as an equine abortigenic pathogen causing significant losses to the Australian Thoroughbred industry, while Equine herpesvirus-1 (EHV-1) is a well-recognized abortigenic agent. Diagnosis of these agents is based on molecular assays in diagnostic laboratories. In this study, we validated C. psittaci and newly developed EHV-1 Loop Mediated Isothermal Amplification (LAMP) assays performed in a real-time fluorometer (rtLAMP) against the reference diagnostic assays. We also evaluated isothermal amplification using commercially available colorimetric mix (cLAMP), and SYBR Green DNA binding dye (sgLAMP) for "naked eye" end-point detection when testing 'real-world' clinical samples. Finally, we applied the C. psittaci LAMP assays in two pilot Point-of-Care (POC) studies in an equine hospital. RESULTS: The analytical sensitivity of C. psittaci and EHV-1 rt-, and colorimetric LAMPs was determined as one and 10 genome equivalents per reaction, respectively. Compared to reference diagnostic qPCR assays, the C. psittaci rtLAMP showed sensitivity of 100%, specificity of 97.5, and 98.86% agreement, while EHV-1 rtLAMP showed 86.96% sensitivity, 100% specificity, and 91.43% agreement. When testing rapidly processed clinical samples, all three C. psittaci rt-, c-, sg-LAMP assays were highly congruent with each other, with Kappa values of 0. 906 for sgLAMP and 0. 821 for cLAMP when compared to rtLAMP. EHV-1 testing also revealed high congruence between the assays, with Kappa values of 0.784 for cLAMP and 0.638 for sgLAMP when compared to rtLAMP. The congruence between LAMP assays and the C. psittaci or EHV-1 qPCR assays was high, with agreements ranging from 94.12 to 100% for C. psittaci, and 88.24 to 94.12% for EHV-1, respectively. At the POC, the C. psittaci rt- and c-LAMP assays using rapidly processed swabs were performed by technicians with no prior molecular experience, and the overall congruence between the POC C. psittaci LAMPs and the qPCR assays ranged between 90.91-100%. CONCLUSIONS: This study describes reliable POC options for the detection of the equine pathogens: C. psittaci and EHV-1. Testing 'real-world' samples in equine clinical setting, represents a proof-of-concept that POC isothermal diagnostics can be applied to rapid disease screening in the equine industry.

"Prevention is the biggest success": Barriers and enablers to personal biosecurity in the thoroughbred breeding industry.

Employees in the equine industry are at occupational risk of zoonoses such as Hendra virus and equine chlamydiosis through exposure to infected materials. This study aimed to gain a deeper understanding of the views and experiences of employees, and the key drivers of infection control and personal biosecurity (PB) practices in the Thoroughbred breeding industry. Methods: An exploratory qualitative study was conducted in 2018 in New South Wales, Australia using interviews (9) and small group discussions (7). The 29 participants included veterinarians, veterinary nurses, foaling staff, stud managers and laboratory personnel working in a range of equine medicine settings. Interviews and focus groups were recorded, transcribed and analysed manually by at least two members of the research team. An iterative approach was used to derive themes. Results: Five main themes emerged: (i) greater awareness of current and emerging infectious risks promotes use of Personal Protective Equipment (PPE); (ii) currently available PPE is not comfortable, practical or well-suited to equine reproductive work in Australia's hot climate; (iii) creating supportive environments for PB reduces risk of exposure to infectious materials; (iv) strong leadership is required to implement sustainable change in workplace culture and practices; and (v) policy and economic factors play an important role in adopting biosecurity (BS) and PB measures in the workplace. Personnel working in the Australian Thoroughbred breeding industry face unique zoonotic risks in a challenging physical environment. A qualitative approach provided rich insights into social and physical factors motivating BS and PB in this occupational group. There is an opportunity for collaboration between Public Health services and industry partners to develop and implement strategies most likely to be effective in ensuring consistent uptake of PB measures in the workplace.

Osteoarticular Infection in Three Young Thoroughbred Horses Caused by a Novel Gram Negative Cocco-Bacillus.

We describe three cases of osteoarticular infection (OAI) in young thoroughbred horses in which the causative organism was identified by MALDI-TOF as Kingella species. The pattern of OAI resembled that reported with Kingella infection in humans. Analysis by 16S rRNA PCR enabled construction of a phylogenetic tree that placed the isolates closer to Simonsiella and Alysiella species, rather than Kingella species. Average nucleotide identity (ANI) comparison between the new isolate and Kingella kingae and Alysiella crassa however revealed low probability that the new isolate belonged to either of these species. This preliminary analysis suggests the organism isolated is a previously unrecognised species.

Challenges in using serological methods to explore historical transmission risk of Chlamydia psittaci in a workforce with high exposure to equine chlamydiosis.

Introduction: This report describes the challenges encountered in using serological methods to study the historical transmission risk of C. psittaci from horses to humans. Methods: In 2017, serology and risk factor questionnaire data from a group of individuals, whose occupations involved close contact with horses, were collected to assess the seroprevalence of antibodies to C. psittaci and identify risk factors associated with previous exposure. Results: 147 participants were enrolled in the study, provided blood samples, and completed a questionnaire. On ELISA testing, antibodies to the Chlamydia genus were detected in samples from 17 participants but further specific species-specific MIF testing did not detect C. psittaci-specific antibodies in any of these samples. Conclusion: No serological evidence of past C. psittaci transmission from horses to humans was found in this study cohort. There are major challenges in using serological methods to determine the prevalence of C. psittaci exposure.

An epizootic of Chlamydia psittaci equine reproductive loss associated with suspected spillover from native Australian parrots.

Chlamydia psittaci is an avian pathogen capable of spill-over infections to humans. A parrot C. psittaci strain was recently detected in an equine reproductive loss case associated with a subsequent cluster of human C. psittaci infections. In this study, we screened for C. psittaci in cases of equine reproductive loss reported in regional New South Wales, Australia during the 2016 foaling season. C. psittaci specific-PCR screening of foetal and placental tissue samples from cases of equine abortion (n = 161) and foals with compromised health status (n = 38) revealed C. psittaci positivity of 21.1% and 23.7%, respectively. There was a statistically significant geographical clustering of cases ~170 km inland from the mid-coast of NSW (P < 0.001). Genomic analysis and molecular typing of C. psittaci positive samples from this study and the previous Australian equine index case revealed that the equine strains from different studs in regional NSW were clonal, while the phylogenetic analysis revealed that the C. psittaci strains from both Australian equine disease clusters belong to the parrot-associated 6BC clade, again indicative of spill-over of C. psittaci infections from native Australian parrots. The results of this work suggest that C. psittaci may be a more significant agent of equine reproductive loss than thought. A range of studies are now required to evaluate (a) the exact role that C. psittaci plays in equine reproductive loss; (b) the range of potential avian reservoirs and factors influencing infection spill-over; and