PhD in Veterinary Sciences for Animal Health and Food Safety

Contacts

Supervisor

Antonio D'angelo

Phd thesis

Cerebrospinal fluid as a tool to improve diagnosis and treatment in bovine neurology

Achieving an etiological diagnosis in bovine central nervous system (CNS) infection is challenging. Cerebrospinal fluid (CSF) analysis in infection shows changes that are not specific. The timely analysis of CSF is essential, yet it is subject to operator proficiency and requires specialized facilities. Furthermore, bacterial identification in CNS infection relies on CSF culture, which lacks sensitivity. To address these limitations, pioneering approaches might hold potential. Machine learning (ML) is revolutionizing disease diagnosis. Nextgeneration sequencing (NGS) could bypass the need for CSF culture. CSF lactate is recognized in humans as a CNS infection biomarker. Lastly, automated CSF analysis could be an alternative to labor-intensive manual analysis. Based on these premises, we developed 4 projects (P1, P2, P3, P4) exploiting these pioneering approaches applied to analysis of bovine CSF. P1 compared ML models to predict the likelihood of CNS infection in cattle. We created a ML based web app for INF diagnosis. ML methods were trained on demographics, clinical findings and CSF analysis. P2 exploited 16S rRNA sequencing to compare the CSF microbial composition in cattle with CNS infection and other CNS disorders. P3 established a Reference Interval (RI) for CSF lactate and assessed its potential as a biomarker for detecting CNS INF. Statistical analysis was performed to demonstrate an association between CSF lactate levels and interpretation of CSF in sick animals. P4 assessed the Idexx ProCyte Dx® analyzer's capability in analyzing CSF in cattle. We investigated the instrument's performance in delivering total (TNCC) and differential cell count compared to the gold standard laboratory analysis. P1: 98 cattle with CNS infection and 86 with other CNS disorders were included. All methods had high prediction accuracy (≥80%). Logistic regression had the highest accuracy (0.84) and was implemented in a web app. 8 P2: an infectious-inflammatory (n = 3) and a non infectious-inflammatory group (n = 3) were formed. There were no detectable differences in the CSF microbial composition of the samples from the two groups. Pseudomonas spp. (45%) was the most frequently expressed bacterial genus. P3: 27 healthy and 86 sick cattle with CNS disorders were included. The RI for CSF lactate was 1.1-2.4 mmol/L. The highest lactate level was found with neutrophilic pleocytosis (median = 7; p < .0001). P4: CSF was collected from 113 cattle, 76 with normal TNCC and 37 with increased TNCC. Procyte Dx® TNCC correlated well with the laboratory TNCC (rho = 0.89; p < .01) while could not provide a differential count. With the thesis presented here we investigated and developed several novel methodological approaches to improve CSF analysis in infectious and non-infectious CNS disorders in cattle. We investigated the potential of novel computational and technical modalities for diagnosis including a prediction web app using ML and identification of CSF lactate as an inflammatory biomarker. Challenges such as distinguishing microbial communities in CSF and performing a complete automated CSF analysis remain, and the work described here will form the basis for future studies.

Research activities

Articles on indexed reviews

 

• Ferrini S, Rollo C, Bellino C, Borriello G, Cagnotti G, Corona C, Di Muro G, Giacobini M, Iulini B, D’Angelo A. A novel machine learning-based web application for field identification of infectious and inflammatory disorders of the central nervous system in cattle.Journal of Veterinary Internal Medicine 2023. 37 (2), 766-773 Doi: 10.1111/jvim.16664

 

• Cagnotti G, Ferrini S, Di Muro G, Avilii E, Favole A, D'Angelo A. Duration of constant rate infusion with diazepam or propofol for canine cluster seizures and status epilepticus. Frontiers in Veterinary Science 2023. 10, 1247100. https://doi.org/10.3389/fvets.2023.1247100

 

• Borriello G, Valentini F, Rampinelli M, Ferrini S, Cagnotti G, D'angelo A, Bellino C. Ocular Ultrasonography in Healthy Calves with Different Transducers. Animals 2023. 13(4), 742. https://doi.org/10.3390/ani13040742

 

• Borriello G, Cagnotti G, Avedano E, Bergagna S, Iannello P, Di Muro G, Ferrini S, D'Angelo A, Bellino C. Qualitative and quantitative monitoring of antibiotics on dairy cattle farms in relation to animal welfare indicators. Italian Journal of Animal Science 2023. 22:1, 760-768, DOI: 10.1080/1828051X.2023.2241878

 

• Ferrini S, Grego E, Ala U, Cagnotti G, Valentini F, Di Muro G, Iulini B, Stella MC, Bellino C, D'Angelo A. Feasibility of 16S rRNA sequencing for cerebrospinal fluid microbiome analysis in cattle with neurological disorders: a pilot study. Veterinary Research Communication 2022. 47(2), 373–383. https://doi.org/10.1007/s11259-022-09949-w

 

• Cagnotti G, Ferrini S, Di Muro G, Borriello G, Corona C, Manassero L, Avilii E, Bellino C, D'Angelo A. Constant rate infusion of diazepam or propofol for the management of canine cluster seizures or status epilepticus. Frontiers in Veterinary Science 2022. 9, 1005948. https://doi.org/10.3389/fvets.2022.1005948

 

• Cagnotti G, Ferrini S, Ala U, Bellino C, Corona C, Dappiano E, Di Muro G, Iulini B, Pepe I, Roncone S, D'Angelo A. Analysis of Early Assessable Risk Factors for Poor Outcome in Dogs With Cluster Seizures and Status Epilepticus. Frontiers in Veterinary Science 2020. 7, 575551. https://doi.org/10.3389/fvets.2020.575551

 

Abstract of international congresses

 

• Ferrini S, Cagnotti G, Ala U, Bellino C, Biasibetti E, Borriello G, Corona C, Di Muro G, Iamone G, Iulini B, Pezzolato M, D’Angelo A. Diagnostic accuracy of cerebrospinal fluid lactate levels in cattle with neurological disorders. 34th ECVN-ESVN Symposium (Palma de Mallorca 23rd-24th September 2022) (Flash presentation)
• Ferrini S, Rollo C, Bellino C, Borriello G, Cagnotti G, Corona C, Di Muro G, Giacobini M, Iulini B, D’Angelo A. Development of a Machine Learning based Web-app for in field identification of central nervous system infectious-inflammatory disorders in cattle. 34th ECVN-ESVN Symposium (Palma de Mallorca 23rd-24th September 2022) (Flash presentation)

Abstract of National congresses

• Ferrini S, Cagnotti G, Ala U, Bellino C, Biasibetti E, Borriello G, Corona C, Di Muro G, Iamone G, Iulini B, Pezzolato M, D’Angelo A. Accuratezza diagnostica del lattato nel liquido cefalorachidiano in bovini affetti da patologie del sistema nervoso centrale. 54th Congresso Nazionale Società Italiana di Buiatria (6-7 ottobre 2022 Piacenza) (Oral presentation)
• Ferrini S, Rollo C, Bellino C, Borriello G, Cagnotti G, Corona C, Di Muro G, Giacobini M, Iulini B, D’Angelo A. Sviluppo di una web-app per l’identificazione dei disturbi infettivo-infiammatori a carico del sistema nervoso centrale nel bovino. 54th Congresso Nazionale Società Italiana di Buiatria (6-7 ottobre 2022 Piacenza) (Oral presentation)
• Ala U, Ferrini S, Bianchi P, Bertero A, Colitti B, Cagnotti G, Bertolotti L. Microbiome and biological interaction: can sterile districts really be considered as such? XX Congresso Nazionale A.I.B.G. (Roma, 23-24 settembre 2022) (Oral presentation)
• Ferrini S. Machine Learning algorithms as diagnostic tools for infectious-inflammatory central nervous system disorders in cattle. Game of Research Event (16th December 2021 Turin) (Oral presentation).