First Report of Fatal Feline Pulmonary Mycobacteriosis Caused by the Emerging Zoonotic Pathogen Mycobacterium orygis in a cat from India

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Dhende, Akshay P. Hendre, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8387028/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 19 Feb, 2026 Read the published version in Veterinary Research Communications → Version 1 posted 11 You are reading this latest preprint version Abstract Feline mycobacteriosis, often involving members of the Mycobacterium tuberculosis complex (MTBC), presents ongoing diagnostic and public health challenges. Mycobacterium orygis , an emerging and genetically distinct MTBC member, is increasingly recognized for its wide host range, including humans, domestic livestock, and wildlife, particularly in South Asia. We report the first documented case of fatal pulmonary mycobacteriosis caused by M. orygis in a companion animal host. A 5-month-old stray domestic shorthair cat in India presented in profound shock with severe, rapidly progressive respiratory distress. Thoracic radiography showed a diffuse bronchointerstitial pattern. Diagnostic workup of bronchoalveolar lavage fluid showed abundant acid-fast bacilli on Ziehl–Neelsen staining. Definitive diagnosis was achieved through advanced molecular methods-MTBC-specific real-time PCR followed by sequencing of the ETR-D spacer and gyrB gene, which identified the species as M. orygis . The patient succumbed to the fulminant disease within 24 hours of presentation. This finding establishes the domestic cat as a previously unrecognized host for M. orygis and highlights the pathogen's capacity to cause severe primary pulmonary infection. Given the documented circulation of M. orygis among humans, livestock, and wildlife in India, this case underscores a critical and often-overlooked zoonotic risk, emphasizing the necessity of integrated One Health surveillance and rapid molecular diagnostics for species-level identification of mycobacteriosis. Mycobacterium orygis Feline mycobacteriosis MTBC ETR-D gyrB India Figures Figure 1 Figure 2 Figure 3 Figure 4 Background Mycobacterial infections in cats are increasingly recognized as a concern in veterinary and public health. Members of the Mycobacterium tuberculosis complex (MTBC), feline leprosy and non-tuberculous mycobacteria (NTM) have been linked to feline illnesses (Gunn-Moore 2014 ; Gunn-Moore et al. 2011 ). However, the most commonly identified feline mycobacteriosis is caused by Mycobacterium microti and Mycobacterium bovis , which are the members of the MTBC (Gunn-Moore et al. 2011 ; Snider et al. 1971 ). The symptoms of mycobacterial infections in cats depend on the mode of exposure. Infections resulting from skin contact, typically caused by bites from infected rodents or wound contamination, usually lead to skin and subcutaneous tubercles; inhalation of infectious material can lead to lung disease; and ingesting the contaminated material, such as infected raw meat or unpasteurized milk from affected cattle can cause gastrointestinal signs (Gunn-Moore 1997 ; Gunn-Moore 2014 ; Smith et al. 2009 ). Mycobacterium orygis , previously known as the Oryx bacillus, is a genetically distinct member of the MTBC. Initially discovered in wildlife, it is increasingly being reported from various animal species, especially domestic cattle in South Asia, and a growing number of human cases, which highlights its zoonotic potential (Duffy et al. 2020 ; Kock et al. 2021 ; Malone and Gordon 2017 ). In India, M. orygis has been reported in a diverse range of animal hosts including spotted deer, black buck, Indian bison, cattle and buffalo with cases confirmed across different states (Refaya et al. 2022 ; Sharma et al. 2023 ). Cases of human tuberculosis (TB) caused by M. orygis have also been reported from India (Duffy et al. 2020 ; Sumanth et al. 2023 ). The documented presence of M. orygis in humans, domestic livestock, and diverse wildlife across India highlights a critical and often-overlooked public health threat. Diagnosing mycobacterial infections has remained challenging primarily because the causative agents are slow-growing or even unculturable (such as, M. lepraemurium ) using standard laboratory techniques, thereby delaying or preventing definitive identification (Kassa et al. 2021 ). Clinical signs, like respiratory distress, non-healing skin nodules or abscesses, are non-specific and often mimic other common bacterial or fungal infections, while traditional diagnostic tests like the tuberculin skin test are unreliable in cats (O'Halloran et al. 2019 ). Traditional methods, such as Ziehl–Neelsen staining, provide supporting evidence but are limited in sensitivity and cannot differentiate between mycobacterial species. Tests such as interferon-gamma release assays (IGRAs) and advanced imaging techniques are also used to aid diagnosis, although their availability is limited (Major et al. 2015; Tobaiqi et al. 2025 ). This diagnostic difficulty is being overcome by molecular approaches, specifically Polymerase Chain Reaction (PCR). This technique rapidly amplifies and identifies minute amounts of mycobacterial deoxyribonucleic acid (DNA) directly from a clinical sample, bypassing the long culture period and allowing for species identification even when the organism is unculturable or present in low numbers. Molecular diagnostics, particularly PCR often complemented by sequencing, have thus significantly enhanced the ability to detect and distinguish mycobacterial species in clinical samples (Zhang et al. 2024 ), enabling targeted and timely treatment that significantly improves patient prognosis. Given the diagnostic challenges as well as zoonotic implications, case reports remain crucial in increasing the veterinary community’s knowledge of feline mycobacterial infections worldwide. In India, such reports are rare, with only a few cases documented, often identified retrospectively or through advanced molecular techniques. This underlines the necessity for more documented instances of feline TB in the Indian subcontinent, where the close contact among humans, domestic animals, and wildlife can promote the spread of Mycobacterium spp. and elevate the risk of zoonotic transmission. Case Presentation and Diagnostic Workup A 5-month-old female domestic shorthair (DSH) stray cat was presented with severe respiratory distress, marked by difficult breathing, lethargy, and decreased activity levels. The cat, found visibly ill and lying stationary in the same location for over 12 hours, was rescued and presented for evaluation by a concerned resident. Initial parameters were highly concerning with hypothermia (92ºF), tachypnoea (Respiratory Rate > 50/min), tachycardia (Heart Rate 230 beats per minute) and severe hypoxemia (SpO 2 ​ 65%). The patient was in profound shock, evidenced by unreadable blood pressure and an unpalpable pulse, along with severe hypoglycemia (Blood Glucose: 12mg/dL). Lung auscultation revealed abnormal sounds indicative of lower respiratory tract disease. An X-ray confirmed a diffuse bronchointerstitial pattern suggestive of bronchopneumonia (Fig. 1). There were no signs of lymph node swelling or skin lesions at the time of presentation. The combination of the clinical signs, physical exam findings, and the supportive bronchointerstitial X-ray pattern led to a working diagnosis of severe lower respiratory tract disease. General anaesthesia (Butorphanol, Midazolam, Propofol, Isoflurane) was initiated due to persistent apnea and hypoxemia despite being kept in the oxygen cage. The patient immediately received fluid boluses and dextrose for hypoglycemia. Due to continuous deterioration (SpO 2 dropping to 55–62%) while on Total Intravenous Anesthesia (TIVA), the subject was placed on mechanical ventilation with 100% oxygen. Ventilator support successfully achieved stability, normalizing SpO 2 (~ 95–100%) and end-tidal carbon dioxide (EtCO 2 ) (35–37 mmHg) within 90 minutes. Once the patient was stable, bronchoalveolar lavage (BAL) was obtained and submitted for cytology and culture. The blood workup comprised a complete blood count (CBC), liver function tests (LFT), and kidney function tests (KFT). Routine bacterial culture of the BAL fluid was negative, showing no bacterial growth after 48 hours of incubation at 37ºC. However, BAL cytology with Leishman stain revealed chronic active inflammatory changes with a high number of neutrophils, followed by macrophages and lymphocytes, alongside numerous intra- and extracellular unstained rod-shaped bacteria (Fig. 2a, 2b). To further characterize these bacteria, Ziehl–Neelsen staining was performed, which demonstrated numerous acid-fast bacilli scattered both extracellularly (Fig. 3 a) and intracellularly within macrophages (Fig. 3 b). Initial Complete Blood Count (CBC) diagnostics revealed severe neutrophilic leukocytosis with a left shift and anemia, consistent with chronic inflammatory disease. The KFT showed a creatinine level of 0.47 mg/dl, which was slightly below the normal range (0.5–1.9 mg/dl), while the LFT indicated a total bilirubin of 0.92 mg/dl, slightly above the reference range (0-0.4mg/dl). Other liver parameters, such as Alanine Transaminase (ALT) and Alkaline Phosphatase (ALP), were within normal limits. The combined findings from BAL cytology, positive Ziehl–Neelsen staining (confirming acid-fast bacilli), and relevant CBC/blood parameters significantly heightened our suspicion for mycobacterial lung infection. Consequently, the BAL sample was submitted for molecular identification of MTBC. DNA was extracted from the BAL fluid using QIAamp DNA blood mini kit (Qiagen, Hilden, Germany). A SYBR green-based real-time PCR targeting the MTBC insertion element IS 6110 was performed using TB Green Premix Ex Taq II (TaKaRa, Japan). The real-time PCR tested positive for MTBC with low cycle threshold (Ct) value of 23.2, strongly indicative of an active mycobacterial replication. The patient experienced subsequent episodes of refractory hypotension through the night and next day requiring continuous rate infusion of Norepinephrine starting from the next day early morning. The overall prognosis was considered poor. The cat died on the second day, within 24 hours of admission, before specific TB treatment could be initiated. Nevertheless, in an effort to identify the precise species of Mycobacterium involved in this case of pulmonary TB, the DNA isolated from the BAL was subjected to amplification and sequencing of the Exact Tandem Repeat D (ETR-D) spacer located between the sensor histidine kinase SenX3 gene upstream and the response regulator RegX3 gene downstream using a set of published primers (Djelouadji et al. 2008 ). Conventional PCR amplification was performed using CloneAmp HiFi PCR Premix (Takara, Japan), yielding a product ~ 700 base pairs (bp) in size. The specific band was excised from the agarose gel and purified using QIAquick PCR & Gel cleanup kit (Qiagen, Hilden, Germany). The gel purified product was then submitted to a commercial facility for A-tailing, cloning and sequencing. The resulting forward and reverse sequences were aligned and cropped to yield a final consensus sequence of 691 bp which when analyzed against the non-redundant nucleotide collection database (nr/nt) of NCBI using the BLASTn algorithm matched with M. orygis with 100 percent (%) identity. In silico alignment of 691bp sequence, obtained in this study, with sequences of other Mycobacterium spp. of MTBC group exhibited a newly identified G to A polymorphism (G 57 A) specific to M. orygis at position 57 in the intergenic spacer between SenX3 and RegX3 genes (Fig. 4 a). To reconfirm the identity of M. orygis , another set of amplification, cloning and sequencing was performed to detect a known synonymous G to A mutation specific to M. orygis at position 870 (G 870 A) of gyrB gene (Huard et al. 2006 ; Islam et al. 2023 ). BLASTn analysis and sequence alignment with other members of MTBC confirmed the presence of mutation (Fig. 4 b) Both ETR-D and gyrB sequences of M. orygis were submitted to NCBI GenBank and accession numbers (PX395407 and PX698743 respectively) were obtained. Discussion and Conclusions The present case report details a confirmed instance of fatal pulmonary mycobacteriosis in a young, stray DSH cat, attributed to the emerging MTBC member, M. orygis . To the best of our knowledge, this is the first documented case of feline tuberculosis caused by this specific, emerging member of the MTBC in India and globally, highlighting a critical and previously unrecognized host species in the zoonotic cycle. Feline TB is frequently reported as the skin form, presenting as localized cutaneous or subcutaneous granulomas often due to bite wounds from infected rodents, followed by respiratory and gastrointestinal forms (Gunn-Moore 2014 ; The Cat Group 2006 ). The cat in discussion however, had no cutaneous lesions and suffered from respiratory symptoms with presentation of severe respiratory distress, hypoxemia, and a bronchointerstitial radiographic pattern, strongly suggesting a primary pulmonary infection. Primary respiratory TB is a rare finding in cats; and a secondary spread from cutaneous sites, via hematogenous route, to the lungs is more common, resulting in diffuse and interstitial pulmonary lesions (Bennett et al. 2011 ; Gunn-Moore 2014 ; Mitchell and Gunn-Moore 2019 ). Clinical signs may take time to appear until the disease advances to bronchial infiltration, at which point dyspnea and a mild cough develop (Gunn-Moore et al. 2011 ; Mitchell and Gunn-Moore 2019 ; The Cat Group 2006 ). In the present case, the clinical course had already progressed to advance stage, resulting in profound shock and death within 24 hours despite intensive supportive care, consistent with severe, rapidly progressive nature of mycobacterial pneumonia, which reportedly carries a poor prognosis in cats (Lloret et al. 2013 ). The definitive diagnosis of mycobacterial infections is often complicated by the slow-growing nature of the causative agents, as reflected by our initial negative routine bacterial culture from the BAL fluid. However, the results of subsequent cytology and staining on the BAL fluid proved critical; Ziehl–Neelsen staining confirmed the presence of abundant acid-fast bacilli both free and intracellularly within macrophages. Clinical and microscopic findings were decisively complemented with molecular diagnostics, the positive real-time PCR for the MTBC, establishing a robust, culture-independent basis for a confirmed diagnosis of feline pulmonary TB. The subsequent sequencing of the ETR-D spacer and the detection of the characteristic G 870 A synonymous mutation in the gyrB gene were pivotal in precisely identifying the pathogen as M. orygis . M. orygis has been identified as a genetically distinct MTBC member with a growing host range, being increasingly reported in cattle, buffalo, and diverse wildlife across India (Rani et al. 2025 ; Refaya et al. 2022 ; Sharma et al. 2023 ). Furthermore, a number of human TB cases caused by M. orygis have been documented in the country (Duffy et al. 2020 ; Sumanth et al. 2023 ). The detection of M. orygis in this domestic cat, a known sentinel species for environmental pathogens, strongly suggests that the pathogen is circulating in the local environment or food chain (e.g., scavenging infected carcasses or raw milk exposure), thereby bridging the gap between wildlife, livestock, and the human population. Spillover of M. orygis from humans to animals has previously been reported by Dawson et al. ( 2012 ). This case serves as a crucial warning signal for the One Health framework in the Indian subcontinent. Given the close human-animal-wildlife interface characteristic of South Asia, the identification of M. orygis in a feline host elevates the perceived zoonotic risk of this pathogen and emphasizes the need for enhanced surveillance and species-level identification of mycobacteriosis in both veterinary and public health settings, in India and around the globe (Malone and Gordon 2017 ). This case also highlights the importance of considering mycobacterial infections in cats with chronic respiratory disease, particularly in endemic regions or households with immunocompromised individuals or history of TB. Despite the novel findings, this case is subject to crucial clinical and diagnostic limitations that warrant discussion. Primarily, the patient's fulminant clinical course led to death within 24 hours of admission, preceding both the confirmation of the mycobacterial etiology and the initiation of specific antituberculosis therapy. This rapid deterioration precluded necropsy, leaving the full extent of organ involvement, the pathological classification of the pulmonary lesions, and the definitive tissue-level burden of M. orygis uncharacterized. Furthermore, the molecular diagnosis could not be complemented by conventional microbiology because the small volume of the BAL sample was insufficient for parallel inoculation onto specialized mycobacterial culture media, thereby precluding confirmation via culture and subsequent traditional drug susceptibility testing. In conclusion, this case provides the first documented evidence of fatal pulmonary mycobacteriosis caused by the emerging MTBC member, M. orygis , in a DSH cat in India. This finding highlights the pathogen's increasing host range and its capacity to cause severe, rapidly progressive disease when presenting as primary pulmonary infection. The case also warns that the pathogen has successfully bridged the gap between its typical hosts and companion animals, thereby increasing the risk of transmission to the human population. Given the known circulation of M. orygis among livestock, wildlife, and humans in India, this feline case emphasizes the significant zoonotic potential and the necessity for a unified One Health approach to surveillance and management of tuberculosis in this highly interconnected environment. Declarations Competing Interests The authors have no relevant financial or non-financial interests to disclose. Ethics approval The cat was sampled for clinical reasons. No additional ethical approval was needed. Funding The authors declare that no funds, grants, or other support were received during the preparation of this manuscript. Author Contribution NM, HNP and AJ conceived and designed the study. SS and AR were attending clinicians and collected clinical data. AJ was pathologist on the case and collected pathological data and figures. ATB assisted with pathological investigation. NM and HNP wrote the first draft, supervised molecular investigation and analyzed sequencing data. AVD and APH carried out molecular testing and collected data. All authors read, reviewed and approved the final manuscript. Data Availability ETR-D and gyrB sequences of M. orygis were submitted to NCBI GenBank and accession numbers (PX395407 and PX698743 respectively) were obtained. References Bennett AD, Lalor S, Schwarz T, Gunn-Moore DA (2011) Radiographic findings in cats with mycobacterial infections. 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Front Public Health 12:1410672. https://doi.org/10.3389/fpubh.2024.1410672 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 19 Feb, 2026 Read the published version in Veterinary Research Communications → Version 1 posted Editorial decision: Revision requested 09 Jan, 2026 Reviews received at journal 09 Jan, 2026 Reviews received at journal 05 Jan, 2026 Reviewers agreed at journal 31 Dec, 2025 Reviewers agreed at journal 27 Dec, 2025 Reviews received at journal 26 Dec, 2025 Reviewers agreed at journal 22 Dec, 2025 Reviewers invited by journal 22 Dec, 2025 Editor assigned by journal 22 Dec, 2025 Submission checks completed at journal 22 Dec, 2025 First submitted to journal 17 Dec, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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1","display":"","copyAsset":false,"role":"figure","size":336748,"visible":true,"origin":"","legend":"\u003cp\u003eThoracic radiograph.\u003cstrong\u003e \u003c/strong\u003eNote the bronchointerstitial pattern\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8387028/v1/bdc55bd184bbd70c54a6cd59.png"},{"id":99316570,"identity":"148b2128-47d5-438f-986d-f8de4cb09d6c","added_by":"auto","created_at":"2025-12-31 16:28:36","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1381138,"visible":true,"origin":"","legend":"\u003cp\u003eBAL cytology a) Presence of abundant neutrophils followed by macrophages and lymphocytes. Note presence of negatively stained rod-shaped bacteria intracellularly (red arrow) as well as extracellularly (black arrow); Leishman stain (100X) b) Numerous negatively stained rod-shaped bacteria inside a macrophage (red arrow); Leishman stain (100X)\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8387028/v1/8f798accbc93ce3e9e5ba1b1.png"},{"id":99316528,"identity":"3b836e90-c808-41d8-98df-c18c4ad29ce6","added_by":"auto","created_at":"2025-12-31 16:28:34","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1320218,"visible":true,"origin":"","legend":"\u003cp\u003eZiehl-Neelsen staining on BAL fluid a) Extracellular pinkish-red acid-fast bacilli (black arrow) scattered in the smear b) Macrophage packed with multiple acid-fast bacilli (red arrow)\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8387028/v1/e8c4eea19d19bd3ff53628e7.png"},{"id":99192527,"identity":"18ae780a-7736-4d72-9105-c5866c074836","added_by":"auto","created_at":"2025-12-30 01:04:33","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":556353,"visible":true,"origin":"","legend":"\u003cp\u003eNucleotide sequence alignment of \u003cem\u003eM. orygis\u003c/em\u003e (obtained in this study) with other members of MTBC a) Nucleotide sequence alignment of \u003cem\u003eSenX3\u003c/em\u003e-\u003cem\u003eRegX3\u003c/em\u003e intergenic spacer showing specific G\u003csub\u003e57\u003c/sub\u003eA polymorphism in the vertical box b) Nucleotide sequence alignment of \u003cem\u003egyrB\u003c/em\u003e partial gene sequence showing specific G\u003csub\u003e870\u003c/sub\u003eA polymorphism in the vertical box\u003c/p\u003e\n\u003cp\u003e★Highlights the sequence of \u003cem\u003eM. orygis\u003c/em\u003e isolated in the present study.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-8387028/v1/d3d6191364fc776617609435.png"},{"id":103251231,"identity":"967f6b12-90e8-4b5e-9e15-7a0890f7f4ac","added_by":"auto","created_at":"2026-02-23 16:06:40","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4890349,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8387028/v1/64822059-0e48-4116-9cc0-5f343fee6537.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"First Report of Fatal Feline Pulmonary Mycobacteriosis Caused by the Emerging Zoonotic Pathogen Mycobacterium orygis in a cat from India","fulltext":[{"header":"Background","content":"\u003cp\u003eMycobacterial infections in cats are increasingly recognized as a concern in veterinary and public health. Members of the \u003cem\u003eMycobacterium tuberculosis\u003c/em\u003e complex (MTBC), feline leprosy and non-tuberculous mycobacteria (NTM) have been linked to feline illnesses (Gunn-Moore \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Gunn-Moore et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). However, the most commonly identified feline mycobacteriosis is caused by \u003cem\u003eMycobacterium microti\u003c/em\u003e and \u003cem\u003eMycobacterium bovis\u003c/em\u003e, which are the members of the MTBC (Gunn-Moore et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Snider et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1971\u003c/span\u003e). The symptoms of mycobacterial infections in cats depend on the mode of exposure. Infections resulting from skin contact, typically caused by bites from infected rodents or wound contamination, usually lead to skin and subcutaneous tubercles; inhalation of infectious material can lead to lung disease; and ingesting the contaminated material, such as infected raw meat or unpasteurized milk from affected cattle can cause gastrointestinal signs (Gunn-Moore \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1997\u003c/span\u003e; Gunn-Moore \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Smith et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eMycobacterium orygis\u003c/em\u003e, previously known as the Oryx bacillus, is a genetically distinct member of the MTBC. Initially discovered in wildlife, it is increasingly being reported from various animal species, especially domestic cattle in South Asia, and a growing number of human cases, which highlights its zoonotic potential (Duffy et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Kock et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Malone and Gordon \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). In India, \u003cem\u003eM. orygis\u003c/em\u003e has been reported in a diverse range of animal hosts including spotted deer, black buck, Indian bison, cattle and buffalo with cases confirmed across different states (Refaya et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Sharma et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Cases of human tuberculosis (TB) caused by \u003cem\u003eM. orygis\u003c/em\u003e have also been reported from India (Duffy et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Sumanth et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The documented presence of \u003cem\u003eM. orygis\u003c/em\u003e in humans, domestic livestock, and diverse wildlife across India highlights a critical and often-overlooked public health threat.\u003c/p\u003e \u003cp\u003eDiagnosing mycobacterial infections has remained challenging primarily because the causative agents are slow-growing or even unculturable (such as, \u003cem\u003eM. lepraemurium\u003c/em\u003e) using standard laboratory techniques, thereby delaying or preventing definitive identification (Kassa et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Clinical signs, like respiratory distress, non-healing skin nodules or abscesses, are non-specific and often mimic other common bacterial or fungal infections, while traditional diagnostic tests like the tuberculin skin test are unreliable in cats (O'Halloran et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Traditional methods, such as Ziehl\u0026ndash;Neelsen staining, provide supporting evidence but are limited in sensitivity and cannot differentiate between mycobacterial species. Tests such as interferon-gamma release assays (IGRAs) and advanced imaging techniques are also used to aid diagnosis, although their availability is limited (Major et al. 2015; Tobaiqi et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). This diagnostic difficulty is being overcome by molecular approaches, specifically Polymerase Chain Reaction (PCR). This technique rapidly amplifies and identifies minute amounts of mycobacterial deoxyribonucleic acid (DNA) directly from a clinical sample, bypassing the long culture period and allowing for species identification even when the organism is unculturable or present in low numbers. Molecular diagnostics, particularly PCR often complemented by sequencing, have thus significantly enhanced the ability to detect and distinguish mycobacterial species in clinical samples (Zhang et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), enabling targeted and timely treatment that significantly improves patient prognosis.\u003c/p\u003e \u003cp\u003eGiven the diagnostic challenges as well as zoonotic implications, case reports remain crucial in increasing the veterinary community\u0026rsquo;s knowledge of feline mycobacterial infections worldwide. In India, such reports are rare, with only a few cases documented, often identified retrospectively or through advanced molecular techniques. This underlines the necessity for more documented instances of feline TB in the Indian subcontinent, where the close contact among humans, domestic animals, and wildlife can promote the spread of \u003cem\u003eMycobacterium spp.\u003c/em\u003e and elevate the risk of zoonotic transmission.\u003c/p\u003e"},{"header":"Case Presentation and Diagnostic Workup","content":"\u003cp\u003eA 5-month-old female domestic shorthair (DSH) stray cat was presented with severe respiratory distress, marked by difficult breathing, lethargy, and decreased activity levels. The cat, found visibly ill and lying stationary in the same location for over 12 hours, was rescued and presented for evaluation by a concerned resident. Initial parameters were highly concerning with hypothermia (92\u0026ordm;F), tachypnoea (Respiratory Rate\u0026thinsp;\u0026gt;\u0026thinsp;50/min), tachycardia (Heart Rate 230 beats per minute) and severe hypoxemia (SpO\u003csub\u003e2\u003c/sub\u003e​ 65%). The patient was in profound shock, evidenced by unreadable blood pressure and an unpalpable pulse, along with severe hypoglycemia (Blood Glucose: 12mg/dL). Lung auscultation revealed abnormal sounds indicative of lower respiratory tract disease. An X-ray confirmed a diffuse bronchointerstitial pattern suggestive of bronchopneumonia (Fig.\u0026nbsp;1). There were no signs of lymph node swelling or skin lesions at the time of presentation. The combination of the clinical signs, physical exam findings, and the supportive bronchointerstitial X-ray pattern led to a working diagnosis of severe lower respiratory tract disease.\u003c/p\u003e \u003cp\u003eGeneral anaesthesia (Butorphanol, Midazolam, Propofol, Isoflurane) was initiated due to persistent apnea and hypoxemia despite being kept in the oxygen cage. The patient immediately received fluid boluses and dextrose for hypoglycemia. Due to continuous deterioration (SpO\u003csub\u003e2\u003c/sub\u003e dropping to 55\u0026ndash;62%) while on Total Intravenous Anesthesia (TIVA), the subject was placed on mechanical ventilation with 100% oxygen.\u003c/p\u003e \u003cp\u003eVentilator support successfully achieved stability, normalizing SpO\u003csub\u003e2\u003c/sub\u003e (~\u0026thinsp;95\u0026ndash;100%) and end-tidal carbon dioxide (EtCO\u003csub\u003e2\u003c/sub\u003e) (35\u0026ndash;37 mmHg) within 90 minutes. Once the patient was stable, bronchoalveolar lavage (BAL) was obtained and submitted for cytology and culture. The blood workup comprised a complete blood count (CBC), liver function tests (LFT), and kidney function tests (KFT).\u003c/p\u003e \u003cp\u003eRoutine bacterial culture of the BAL fluid was negative, showing no bacterial growth after 48 hours of incubation at 37\u0026ordm;C. However, BAL cytology with Leishman stain revealed chronic active inflammatory changes with a high number of neutrophils, followed by macrophages and lymphocytes, alongside numerous intra- and extracellular unstained rod-shaped bacteria (Fig.\u0026nbsp;2a, 2b). To further characterize these bacteria, Ziehl\u0026ndash;Neelsen staining was performed, which demonstrated numerous acid-fast bacilli scattered both extracellularly (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e3\u003c/span\u003ea) and intracellularly within macrophages (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e3\u003c/span\u003eb).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eInitial Complete Blood Count (CBC) diagnostics revealed severe neutrophilic leukocytosis with a left shift and anemia, consistent with chronic inflammatory disease. The KFT showed a creatinine level of 0.47 mg/dl, which was slightly below the normal range (0.5\u0026ndash;1.9 mg/dl), while the LFT indicated a total bilirubin of 0.92 mg/dl, slightly above the reference range (0-0.4mg/dl). Other liver parameters, such as Alanine Transaminase (ALT) and Alkaline Phosphatase (ALP), were within normal limits.\u003c/p\u003e \u003cp\u003eThe combined findings from BAL cytology, positive Ziehl\u0026ndash;Neelsen staining (confirming acid-fast bacilli), and relevant CBC/blood parameters significantly heightened our suspicion for mycobacterial lung infection. Consequently, the BAL sample was submitted for molecular identification of MTBC. DNA was extracted from the BAL fluid using QIAamp DNA blood mini kit (Qiagen, Hilden, Germany). A SYBR green-based real-time PCR targeting the MTBC insertion element IS\u003cem\u003e6110\u003c/em\u003e was performed using TB Green Premix Ex Taq II (TaKaRa, Japan). The real-time PCR tested positive for MTBC with low cycle threshold (Ct) value of 23.2, strongly indicative of an active mycobacterial replication.\u003c/p\u003e \u003cp\u003eThe patient experienced subsequent episodes of refractory hypotension through the night and next day requiring continuous rate infusion of Norepinephrine starting from the next day early morning. The overall prognosis was considered poor.\u003c/p\u003e \u003cp\u003eThe cat died on the second day, within 24 hours of admission, before specific TB treatment could be initiated. Nevertheless, in an effort to identify the precise species of \u003cem\u003eMycobacterium\u003c/em\u003e involved in this case of pulmonary TB, the DNA isolated from the BAL was subjected to amplification and sequencing of the Exact Tandem Repeat D (ETR-D) spacer located between the sensor histidine kinase \u003cem\u003eSenX3\u003c/em\u003e gene upstream and the response regulator \u003cem\u003eRegX3\u003c/em\u003e gene downstream using a set of published primers (Djelouadji et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Conventional PCR amplification was performed using CloneAmp HiFi PCR Premix (Takara, Japan), yielding a product\u0026thinsp;~\u0026thinsp;700 base pairs (bp) in size. The specific band was excised from the agarose gel and purified using QIAquick PCR \u0026amp; Gel cleanup kit (Qiagen, Hilden, Germany). The gel purified product was then submitted to a commercial facility for A-tailing, cloning and sequencing.\u003c/p\u003e \u003cp\u003eThe resulting forward and reverse sequences were aligned and cropped to yield a final consensus sequence of 691 bp which when analyzed against the non-redundant nucleotide collection database (nr/nt) of NCBI using the BLASTn algorithm matched with \u003cem\u003eM. orygis\u003c/em\u003e with 100 percent (%) identity. In silico alignment of 691bp sequence, obtained in this study, with sequences of other \u003cem\u003eMycobacterium spp.\u003c/em\u003e of MTBC group exhibited a newly identified G to A polymorphism (G\u003csub\u003e57\u003c/sub\u003eA) specific to \u003cem\u003eM. orygis\u003c/em\u003e at position 57 in the intergenic spacer between \u003cem\u003eSenX3\u003c/em\u003e and \u003cem\u003eRegX3\u003c/em\u003e genes (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e4\u003c/span\u003ea). To reconfirm the identity of \u003cem\u003eM. orygis\u003c/em\u003e, another set of amplification, cloning and sequencing was performed to detect a known synonymous G to A mutation specific to \u003cem\u003eM. orygis\u003c/em\u003e at position 870 (G\u003csub\u003e870\u003c/sub\u003eA) of \u003cem\u003egyrB\u003c/em\u003e gene (Huard et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Islam et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). BLASTn analysis and sequence alignment with other members of MTBC confirmed the presence of mutation (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e4\u003c/span\u003eb) Both ETR-D and \u003cem\u003egyrB\u003c/em\u003e sequences of \u003cem\u003eM. orygis\u003c/em\u003e were submitted to NCBI GenBank and accession numbers (PX395407 and PX698743 respectively) were obtained.\u003c/p\u003e"},{"header":"Discussion and Conclusions","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003cp\u003eThe present case report details a confirmed instance of fatal pulmonary mycobacteriosis in a young, stray DSH cat, attributed to the emerging MTBC member, \u003cem\u003eM. orygis\u003c/em\u003e. To the best of our knowledge, this is the first documented case of feline tuberculosis caused by this specific, emerging member of the MTBC in India and globally, highlighting a critical and previously unrecognized host species in the zoonotic cycle.\u003c/p\u003e \u003cp\u003eFeline TB is frequently reported as the skin form, presenting as localized cutaneous or subcutaneous granulomas often due to bite wounds from infected rodents, followed by respiratory and gastrointestinal forms (Gunn-Moore \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; The Cat Group \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). The cat in discussion however, had no cutaneous lesions and suffered from respiratory symptoms with presentation of severe respiratory distress, hypoxemia, and a bronchointerstitial radiographic pattern, strongly suggesting a primary pulmonary infection. Primary respiratory TB is a rare finding in cats; and a secondary spread from cutaneous sites, via hematogenous route, to the lungs is more common, resulting in diffuse and interstitial pulmonary lesions (Bennett et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Gunn-Moore \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Mitchell and Gunn-Moore \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Clinical signs may take time to appear until the disease advances to bronchial infiltration, at which point dyspnea and a mild cough develop (Gunn-Moore et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Mitchell and Gunn-Moore \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; The Cat Group \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). In the present case, the clinical course had already progressed to advance stage, resulting in profound shock and death within 24 hours despite intensive supportive care, consistent with severe, rapidly progressive nature of mycobacterial pneumonia, which reportedly carries a poor prognosis in cats (Lloret et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe definitive diagnosis of mycobacterial infections is often complicated by the slow-growing nature of the causative agents, as reflected by our initial negative routine bacterial culture from the BAL fluid. However, the results of subsequent cytology and staining on the BAL fluid proved critical; Ziehl\u0026ndash;Neelsen staining confirmed the presence of abundant acid-fast bacilli both free and intracellularly within macrophages. Clinical and microscopic findings were decisively complemented with molecular diagnostics, the positive real-time PCR for the MTBC, establishing a robust, culture-independent basis for a confirmed diagnosis of feline pulmonary TB.\u003c/p\u003e \u003cp\u003eThe subsequent sequencing of the ETR-D spacer and the detection of the characteristic G\u003csub\u003e870\u003c/sub\u003eA synonymous mutation in the \u003cem\u003egyrB\u003c/em\u003e gene were pivotal in precisely identifying the pathogen as \u003cem\u003eM. orygis\u003c/em\u003e. \u003cem\u003eM. orygis\u003c/em\u003e has been identified as a genetically distinct MTBC member with a growing host range, being increasingly reported in cattle, buffalo, and diverse wildlife across India (Rani et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Refaya et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Sharma et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Furthermore, a number of human TB cases caused by \u003cem\u003eM. orygis\u003c/em\u003e have been documented in the country (Duffy et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Sumanth et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe detection of \u003cem\u003eM. orygis\u003c/em\u003e in this domestic cat, a known sentinel species for environmental pathogens, strongly suggests that the pathogen is circulating in the local environment or food chain (e.g., scavenging infected carcasses or raw milk exposure), thereby bridging the gap between wildlife, livestock, and the human population. Spillover of \u003cem\u003eM. orygis\u003c/em\u003e from humans to animals has previously been reported by Dawson et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). This case serves as a crucial warning signal for the One Health framework in the Indian subcontinent. Given the close human-animal-wildlife interface characteristic of South Asia, the identification of \u003cem\u003eM. orygis\u003c/em\u003e in a feline host elevates the perceived zoonotic risk of this pathogen and emphasizes the need for enhanced surveillance and species-level identification of mycobacteriosis in both veterinary and public health settings, in India and around the globe (Malone and Gordon \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). This case also highlights the importance of considering mycobacterial infections in cats with chronic respiratory disease, particularly in endemic regions or households with immunocompromised individuals or history of TB.\u003c/p\u003e \u003cp\u003eDespite the novel findings, this case is subject to crucial clinical and diagnostic limitations that warrant discussion. Primarily, the patient's fulminant clinical course led to death within 24 hours of admission, preceding both the confirmation of the mycobacterial etiology and the initiation of specific antituberculosis therapy. This rapid deterioration precluded necropsy, leaving the full extent of organ involvement, the pathological classification of the pulmonary lesions, and the definitive tissue-level burden of \u003cem\u003eM. orygis\u003c/em\u003e uncharacterized. Furthermore, the molecular diagnosis could not be complemented by conventional microbiology because the small volume of the BAL sample was insufficient for parallel inoculation onto specialized mycobacterial culture media, thereby precluding confirmation via culture and subsequent traditional drug susceptibility testing.\u003c/p\u003e \u003cp\u003eIn conclusion, this case provides the first documented evidence of fatal pulmonary mycobacteriosis caused by the emerging MTBC member, \u003cem\u003eM. orygis\u003c/em\u003e, in a DSH cat in India. This finding highlights the pathogen's increasing host range and its capacity to cause severe, rapidly progressive disease when presenting as primary pulmonary infection. The case also warns that the pathogen has successfully bridged the gap between its typical hosts and companion animals, thereby increasing the risk of transmission to the human population. Given the known circulation of \u003cem\u003eM. orygis\u003c/em\u003e among livestock, wildlife, and humans in India, this feline case emphasizes the significant zoonotic potential and the necessity for a unified One Health approach to surveillance and management of tuberculosis in this highly interconnected environment.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003ch2\u003eCompeting Interests\u003c/h2\u003e \u003cp\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/p\u003e \u003ch2\u003eEthics approval\u003c/h2\u003e \u003cp\u003eThe cat was sampled for clinical reasons. No additional ethical approval was needed.\u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThe authors declare that no funds, grants, or other support were received during the preparation of this manuscript.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eNM, HNP and AJ conceived and designed the study. SS and AR were attending clinicians and collected clinical data. AJ was pathologist on the case and collected pathological data and figures. ATB assisted with pathological investigation. NM and HNP wrote the first draft, supervised molecular investigation and analyzed sequencing data. AVD and APH carried out molecular testing and collected data. All authors read, reviewed and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eETR-D and gyrB sequences of M. orygis were submitted to NCBI GenBank and accession numbers (PX395407 and PX698743 respectively) were obtained.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBennett AD, Lalor S, Schwarz T, Gunn-Moore DA (2011) Radiographic findings in cats with mycobacterial infections. J Feline Med Surg 13:718\u0026ndash;724. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jfms.2011.06\u003c/span\u003e\u003cspan address=\"10.1016/j.jfms.2011.06\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDawson KL, Bell A, Kawakami RP, Coley K, Yates G, Collins DM (2012) Transmission of \u003cem\u003eMycobacterium orygis\u003c/em\u003e (\u003cem\u003eM. tuberculosis\u003c/em\u003e Complex Species) from a tuberculosis patient to a dairy cow in New Zealand. 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Front Public Health 12:1410672. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpubh.2024.1410672\u003c/span\u003e\u003cspan address=\"10.3389/fpubh.2024.1410672\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"veterinary-research-communications","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"verc","sideBox":"Learn more about [Veterinary Research Communications](https://www.springer.com/journal/11259)","snPcode":"11259","submissionUrl":"https://submission.nature.com/new-submission/11259/3","title":"Veterinary Research Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Mycobacterium orygis, Feline mycobacteriosis, MTBC, ETR-D, gyrB, India","lastPublishedDoi":"10.21203/rs.3.rs-8387028/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8387028/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eFeline mycobacteriosis, often involving members of the \u003cem\u003eMycobacterium tuberculosis\u003c/em\u003e complex (MTBC), presents ongoing diagnostic and public health challenges. \u003cem\u003eMycobacterium orygis\u003c/em\u003e, an emerging and genetically distinct MTBC member, is increasingly recognized for its wide host range, including humans, domestic livestock, and wildlife, particularly in South Asia. We report the first documented case of fatal pulmonary mycobacteriosis caused by \u003cem\u003eM. orygis\u003c/em\u003e in a companion animal host. A 5-month-old stray domestic shorthair cat in India presented in profound shock with severe, rapidly progressive respiratory distress. Thoracic radiography showed a diffuse bronchointerstitial pattern. Diagnostic workup of bronchoalveolar lavage fluid showed abundant acid-fast bacilli on Ziehl\u0026ndash;Neelsen staining. Definitive diagnosis was achieved through advanced molecular methods-MTBC-specific real-time PCR followed by sequencing of the ETR-D spacer and \u003cem\u003egyrB\u003c/em\u003e gene, which identified the species as \u003cem\u003eM. orygis\u003c/em\u003e. The patient succumbed to the fulminant disease within 24 hours of presentation. This finding establishes the domestic cat as a previously unrecognized host for \u003cem\u003eM. orygis\u003c/em\u003e and highlights the pathogen's capacity to cause severe primary pulmonary infection. Given the documented circulation of \u003cem\u003eM. orygis\u003c/em\u003e among humans, livestock, and wildlife in India, this case underscores a critical and often-overlooked zoonotic risk, emphasizing the necessity of integrated One Health surveillance and rapid molecular diagnostics for species-level identification of mycobacteriosis.\u003c/p\u003e","manuscriptTitle":"First Report of Fatal Feline Pulmonary Mycobacteriosis Caused by the Emerging Zoonotic Pathogen Mycobacterium orygis in a cat from India","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-30 01:04:28","doi":"10.21203/rs.3.rs-8387028/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-01-09T12:04:21+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-09T12:00:22+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-05T14:21:32+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"298324500448880677091017108606185056792","date":"2025-12-31T07:15:53+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"27645281521816380771597869883197950762","date":"2025-12-27T16:50:47+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-26T12:22:57+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"134582418138941107690262040213605404076","date":"2025-12-22T11:25:03+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-12-22T08:54:26+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-12-22T08:25:37+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-12-22T08:22:19+00:00","index":"","fulltext":""},{"type":"submitted","content":"Veterinary Research Communications","date":"2025-12-17T14:38:42+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"veterinary-research-communications","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"verc","sideBox":"Learn more about [Veterinary Research Communications](https://www.springer.com/journal/11259)","snPcode":"11259","submissionUrl":"https://submission.nature.com/new-submission/11259/3","title":"Veterinary Research Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"9a89f416-ded4-44fb-8ba8-392eadc04d69","owner":[],"postedDate":"December 30th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-02-23T16:03:08+00:00","versionOfRecord":{"articleIdentity":"rs-8387028","link":"https://doi.org/10.1007/s11259-026-11115-5","journal":{"identity":"veterinary-research-communications","isVorOnly":false,"title":"Veterinary Research Communications"},"publishedOn":"2026-02-19 15:59:15","publishedOnDateReadable":"February 19th, 2026"},"versionCreatedAt":"2025-12-30 01:04:28","video":"","vorDoi":"10.1007/s11259-026-11115-5","vorDoiUrl":"https://doi.org/10.1007/s11259-026-11115-5","workflowStages":[]},"version":"v1","identity":"rs-8387028","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8387028","identity":"rs-8387028","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}