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.

Peritoneal fluid analysis in equine post-partum emergencies admitted to a referral hospital: A retrospective study of 110 cases.

BACKGROUND: Peritoneal fluid analysis has both diagnostic and prognostic value in colic but is little reported in the post-partum mare. Multiple conditions may present similarly in this period, and peritoneal fluid findings may aid a prompt diagnosis. OBJECTIVES: To describe the peritoneal fluid findings and their association with diagnosis in mares presenting to a single referral hospital for treatment of post-partum emergencies. STUDY DESIGN: A retrospective clinical study. METHODS: Clinical records of 110 Thoroughbred mares were reviewed. Details of peritoneal fluid analysis from samples obtained at admission were recorded, in addition to history, physical examination, presenting clinicopathological data. Cases were classified by their primary diagnosis into groups of gastrointestinal tract (GIT), urogenital trauma (UGT) and post parturient haemorrhage (PPH). Univariable analysis was performed to compare findings between groups, using one-way ANOVA and post hoc Tukey/Kruskal-Wallis, as appropriate. A multinomial logistic regression was performed for variables significant in the univariable analysis. RESULTS: When separated into their diagnostic categories, 33/110 (30%) mares were classified as GIT, 55/110 (50%) UGT and 22/110 (20%) PPH. Peritoneal fluid packed cell volume (PCV), nucleated cell count (WBCC) and cytological findings were significantly different between diagnostic categories. The likelihood of diagnosis of PPH increased with an increase in peritoneal fluid PCV, the absence of degenerate neutrophils on peritoneal fluid cytology and a decrease in the peritoneal fluid WBCC. Overall survival to discharge was 55%. MAIN LIMITATIONS: The study is referral hospital-based and retrospective in nature. Missing data reduced the power of analysis for several variables. CONCLUSIONS: Peritoneal fluid analysis may guide diagnosis in post-partum emergencies, but no one factor is uniformly diagnostic. Mares with PPH presented with a non-septic peritonitis with higher peritoneal PCV.

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.

Metagenomic investigation of potential abortigenic pathogens in foetal tissues from Australian horses.

BACKGROUND: Abortion in horses leads to economic and welfare losses to the equine industry. Most cases of equine abortions are sporadic, and the cause is often unknown. This study aimed to detect potential abortigenic pathogens in equine abortion cases in Australia using metagenomic deep sequencing methods. RESULTS: After sequencing and analysis, a total of 68 and 86 phyla were detected in the material originating from 49 equine abortion samples and 8 samples from normal deliveries, respectively. Most phyla were present in both groups, with the exception of Chlamydiae that were only present in abortion samples. Around 2886 genera were present in the abortion samples and samples from normal deliveries at a cut off value of 0.001% of relative abundance. Significant differences in species diversity between aborted and normal tissues was observed. Several potential abortigenic pathogens were identified at a high level of relative abundance in a number of the abortion cases, including Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Streptococcus equi subspecies zooepidemicus, Pantoea agglomerans, Acinetobacter lwoffii, Acinetobacter calcoaceticus and Chlamydia psittaci. CONCLUSIONS: This work revealed the presence of several potentially abortigenic pathogens in aborted specimens. No novel potential abortigenic agents were detected. The ability to screen samples for multiple pathogens that may not have been specifically targeted broadens the frontiers of diagnostic potential. The future use of metagenomic approaches for diagnostic purposes is likely to be facilitated by further improvements in deep sequencing technologies.

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.

Factors associated with survival and racing performance of 114 Thoroughbred foals with septic arthritis compared with maternal siblings (2009-2015).

BACKGROUND: There is little consensus on factors associated with survival in foals with septic arthritis and limited data on long-term racing performance of Thoroughbred foals treated for septic arthritis. A more thorough understanding of short- and long-term outcome is necessary to help inform owners, and subsequently guide treatment. OBJECTIVES: To investigate factors associated with survival, and to analyse racing performance of foals with septic arthritis compared with their maternal siblings. STUDY DESIGN: Retrospective cohort and a case-control study. METHODS: Veterinary clinical records of Thoroughbred foals ≤180 days old that underwent arthroscopic, cannulae or through-and-through needle lavage for the treatment of septic arthritis between 2009 and 2015 were reviewed. Data included signalment, and clinicopathological information. The dam's foaling records were reviewed and the lifetime racing records were obtained for affected foals and two of their maternal siblings. Logistic regression analysis was used to determine factors associated with survival to discharge or racing. Comparisons between treated foals and their maternal siblings were made. RESULTS: Ninety (78%) of 115 foals diagnosed with septic arthritis were discharged alive. Foals <26 days old at the time of admission were five times less likely (P = .003) and foals with concurrent multisystemic disease were six times less likely (P = .02) to be discharged alive. Sixty (67%) foals discharged alive started in ≥1 race, and there was no difference in the proportion of foals that started in a race or racing performance between foals treated for septic arthritis and their maternal siblings. MAIN LIMITATIONS: Retrospective study design, limited number of foals with multiple joint involvement and failure to accurately record duration of clinical signs. CONCLUSIONS: Foals treated for septic arthritis at the Scone Equine Hospital, New South Wales, Australia had a good prognosis for survival, and for this cohort, foals that survived to discharge had a similar ability to race as their maternal siblings.

"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.

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

Peripheral blood leukocytes.

Assessment of peripheral blood leukocytes is the most common clinical pathology test in equine practice. It is always a part of complete body function assessment and frequently is used alone, or in conjunction with, fibrinogen to monitor response to treatment.