Clinical evaluation of reproductive disorders in buffaloes under field conditions using ultrasonography and its implications for on-farm reproductive management

Introduction

Buffaloes (Bubalus bubalis) constitute a critical component of dairy-based livelihood systems in South Asia and contribute substantially to India's national milk production, thereby functioning as a primary income buffer for smallholder farmers (FAO, 2023). In such production systems, reproductive efficiency is not merely a biological trait but a central economic determinant governing calving interval, lactation continuity, replacement rate, and lifetime productivity. Even marginal impairment in fertility extends days open, increases insemination frequency, and amplifies cumulative economic losses at the household level (Noakes et al., 2018) [8] . Field-level reproductive inefficiency in buffaloes commonly presents as anoestrus, repeat breeding, uterine infections (metritis and endometritis), ovarian cystic conditions, hydrometra, retained fetal membranes, abortion, and early embryonic death (EED). Among these, early embryonic mortality represents a biologically silent yet economically significant cause of reproductive wastage, frequently occurring before clinical recognition and often misinterpreted as repeat breeding (Sahu et al., 2025) [9] . Such diagnostic ambiguity leads to repeated inseminations, irrational hormonal use, delayed treatment, and erosion of farmer confidence in reproductive interventions. Additionally, postpartum disorders including genital prolapse and fetal mummification, though relatively less frequent, reflect underlying reproductive management deficiencies and further compromise herd productivity (Sahu et al., 2024a; Sahu et al., 2024b) [12,13] . Per-rectal (PR) palpation remains the most widely practiced reproductive examination method under field conditions because of its low cost and minimal equipment requirement. However, PR examination has intrinsic limitations in accurately differentiating uterine pathologies, detecting early pregnancy, assessing embryonic viability, and characterizing ovarian structures with precision (Noakes et al., 2018) [8] . Subtle intrauterine fluid accumulation, early conceptus development, and luteal morphology are particularly difficult to evaluate reliably through palpation alone. These diagnostic constraints often translate into delayed therapeutic decision-making and avoidable economic loss. Real-time transrectal ultrasonography (USG) has transformed bovine reproductive diagnostics by enabling direct visualization of ovarian follicular dynamics, corpus luteum morphology, uterine lumen characteristics, embryonic vesicles, and fetal heartbeat (Ginther, 2014) [4] . Early pregnancy detection is feasible as early as 25-30 days post-insemination, substantially improving management precision compared with palpation-based diagnosis (Noakes et al., 2018) [8] . Furthermore, Doppler ultrasonography has expanded diagnostic capacity beyond structural imaging by allowing assessment of uterine artery hemodynamics and tissue perfusion, thereby offering functional insight into estrous cyclicity, uterine inflammation, and recovery dynamics (Sahu et al., 2026a; Sahu et al., 2026b) [10,11] . This integration of structural and vascular assessment strengthens evidence-based reproductive management. Despite its established diagnostic superiority in organized dairy systems, the systematic deployment of portable ultrasonography as a structured field extension intervention in buffalo-dominated smallholder systems remains inadequately documented. Importantly, ultrasonography functions not only as a diagnostic modality but also as a visual communication tool. Real-time visualization of fetal heartbeat, uterine pathology, or ovarian activity enhances farmer understanding, builds trust in veterinary recommendations, and improves compliance with reproductive management protocols. Thus, its value extends beyond diagnostic accuracy into the domains of extension effectiveness and income protection. Therefore, the present study was undertaken to evaluate ultrasonography-led field extension interventions in buffalo herds under rural conditions. Specifically, the objectives were to (i) compare the diagnostic performance of ultrasonography with conventional per-rectal examination for common reproductive disorders, and (ii) assess changes in farmers' perception following ultrasonographic demonstration. By integrating diagnostic precision with participatory extension outcomes, this work positions portable ultrasonography not merely as a reproductive imaging tool, but as a scalable intervention capable of strengthening reproductive efficiency and enhancing income resilience in buffalo-based dairy systems.

Materials and Methods

Study Area and Duration

The study was conducted in Bareilly and adjoining districts of western Uttar Pradesh, India, under field conditions typical of buffalo-dominated smallholder dairy systems (Fig. 1). The intervention was implemented over a period of three years through organized reproductive health camps conducted in collaboration with field extension units (Fig. 2).

Study Population

A total of 21 field reproductive health camps were organized during the study period. Across these camps, 368 buffaloes presented with reproductive complaints were examined. Animals included both cyclic and non-cyclic females of varying parity brought by farmers for infertility, suspected pregnancy, repeat breeding, anoestrus, postpartum complications, or other reproductive concerns.

Diagnostic Protocol

Each animal underwent a two-step diagnostic procedure during the same visit. Initially, conventional per-rectal (PR) examination was performed following standard large-animal reproductive evaluation procedures as described in established veterinary texts (Noakes et al., 2018) [8] . Uterine size, tone, horn symmetry, cervical status, ovarian structures, and presumptive pregnancy indicators were assessed manually. Following PR examination, transrectal ultrasonography (USG) was performed using a portable ultrasound unit equipped with a linear-array transducer appropriate for bovine reproductive imaging. Ultrasonography was conducted according to standard reproductive imaging criteria, allowing visualization of www.extensionjournal.com ovarian follicles, corpus luteum morphology, intrauterine fluid accumulation, embryonic vesicle presence, and fetal heartbeat where applicable (Ginther, 2014) [4] . Diagnostic categorization was based on ultrasonographic visualization of reproductive structures and pathological changes. Diagnostic outcomes recorded through PR and USG were documented separately for comparative evaluation.

Farmer Perception Assessment

To evaluate the extension impact of ultrasonography-based reproductive examination, farmers attending the camps were assessed using a structured Likert-scale questionnaire administered before and after ultrasonographic demonstration. The perception survey evaluated farmer understanding of reproductive status, confidence in diagnosis, satisfaction with examination, willingness to use ultrasonography in future, and likelihood of recommending the method to other farmers. Responses were recorded on a standardized five-point Likert scale. Pre-and postdemonstration perception scores were compared to determine changes attributable to visual diagnostic confirmation.

Statistical Analysis

Diagnostic agreement between PR and USG findings was assessed using McNemar's chi-square test for paired nominal data. Differences in detection rates between the two diagnostic modalities were expressed as frequency and percentage. Changes in farmer perception scores before and after ultrasonographic demonstration were analyzed using paired t-test. Data were analyzed using GraphPad Prism 10.1.2. Statistical significance was considered at p < 0.05.

Results

A total of n= 368 buffaloes examined across 21 field camps were evaluated using both per-rectal (PR) examination and transrectal ultrasonography (USG). The comparative diagnostic findings are summarized in Table 1 and representative images are shown in Fig. 3. Farmer Perception Analysis Farmer perception outcomes comparing per-rectal (PR) examination and ultrasonography (USG) are presented (Fig. 4, Fig. 5). Ultrasonography significantly enhanced farmer understanding of reproductive status, trust in diagnosis, and confidence in reproductive decision-making (p < 0.01) (Fig. 4). Perception regarding differentiation between pregnancy and reproductive disorders did not differ significantly, likely reflecting moderate baseline awareness among farmers. Preference patterns clearly favored ultrasonography over conventional palpation (p < 0.01) (Fig. 5). Farmers reported greater perceived economic benefit, improved clarity of demonstration, and a higher likelihood of recommending USG-based examination to others (p < 0.01). However, willingness to spend on treatment did not differ significantly between methods, indicating that increased diagnostic clarity strengthened confidence but did not immediately alter cost-related decisions. Overall satisfaction levels were substantially higher following ultrasonographic evaluation compared with PR examination (Fig. 6), underscoring the extension value of visual diagnostic confirmation under field conditions. The overall findings indicate that ultrasonography significantly enhanced detection rates of early pregnancy, uterine inflammatory conditions, ovarian cysts, hydrometra, and early embryonic death under field conditions. In parallel, ultrasonographic demonstration positively influenced farmer perception, diagnostic trust, and willingness to adopt reproductive management interventions.

Comparative Diagnostic Performance

Discussion

Accurate reproductive diagnosis under field conditions remains one of the most decisive determinants of fertility management efficiency in buffalo-based smallholder systems. Conventional per-rectal palpation, while economical and widely practiced, is inherently limited by its dependence on tactile interpretation and operator experience. Subtle intrauterine fluid accumulation, early embryonic structures, and differentiation between follicular and luteal ovarian states often fall below the resolution threshold of manual assessment (Noakes et al., 2018) [8] . In contrast, real-time ultrasonography provides direct visualization of reproductive structures, thereby reducing diagnostic ambiguity and enabling physiologically informed www.extensionjournal.com decision-making. The enhanced diagnostic capacity of ultrasonography observed in this study is biologically consistent with established reproductive physiology. Early gestation detection through visualization of embryonic vesicles and cardiac activity allows confirmation of pregnancy at stages where uterine asymmetry may not yet be appreciable by palpation (Ginther, 2014) [4] . Similarly, differentiation of uterine inflammatory conditions depends on identification of echogenic versus anechoic intrauterine contents, endometrial changes, and structural integrity parameters that cannot be reliably evaluated through tactile examination alone. This capacity for structural differentiation has long been recognized as a defining advantage of ultrasonographic imaging in bovine reproduction (Noakes et al., 2018) [8] . Beyond structural imaging, contemporary research has expanded the role of ultrasonography into functional assessment. Doppler-based evaluation of uterine hemodynamics has demonstrated measurable variations in blood flow associated with estrous cyclicity, inflammation, and luteal activity, offering insights into reproductive physiology that extend beyond gross morphology (Sahu et al., 2026a; Sahu et al., 2026b) [10,11] . Such developments reinforce the conceptual shift from purely anatomical diagnosis toward integrated structural-functional reproductive assessment. Importantly, the significance of ultrasonography in this study extends beyond diagnostic sensitivity. The observed improvement in farmer perception and trust following ultrasonographic demonstration highlights the role of visualization in extension science. In smallholder systems, where reproductive disorders are often interpreted through experiential heuristics, the ability to visually demonstrate fetal heartbeat, ovarian dynamics, or uterine pathology transforms abstract explanations into observable evidence. Behavioral adoption theory consistently emphasizes that visibility, perceived reliability, and experiential validation strongly influence technology acceptance (Neethirajan, 2021) [7] . Thus, ultrasonography functions not merely as a diagnostic tool but as an extension catalyst that strengthens veterinarian-farmer interaction and informed reproductive decision-making. Early embryonic mortality and subclinical uterine pathology represent major hidden contributors to infertility in bovines (Sahu et al., 2025; Andrade et al., 2024) [9,1] . Without imaging support, such conditions are frequently misclassified as repeat breeding, leading to repeated inseminations and inappropriate hormonal interventions. By enabling earlier and more accurate differentiation between embryonic loss, luteal insufficiency, and uterine pathology, ultrasonography supports targeted therapeutic strategies and rational reproductive management. Despite these advantages, several practical considerations merit attention. Ultrasonographic accuracy remains operator-dependent, requiring structured training and calibration to ensure reproducibility. Equipment cost, maintenance, and portability may also influence scalability in resource-limited settings. Furthermore, while visual confirmation enhances confidence, economic constraints may still limit treatment uptake, underscoring the need for integrated financial and advisory support mechanisms within extension programs. Collectively, the present findings align with the growing body of literature supporting ultrasonography as a cornerstone of modern bovine reproductive management (Fricke, 2022) [3] . When deployed through structured field extension camps, portable ultrasonography bridges the gap between advanced reproductive diagnostics and rural livestock production systems. Its dual role as a diagnostic and educational instrument positions it as a scalable intervention capable of strengthening reproductive efficiency and, ultimately, income resilience in buffalobased dairy enterprises.

Statements and Declarations Ethics Approval and Consent to Participate

The clinical examinations performed in this study involved non-invasive reproductive evaluation procedures routinely conducted under veterinary field practice. No experimental interventions were performed beyond standard diagnostic procedures. All animal handling complied with accepted animal welfare guidelines for large-animal clinical examination. Verbal informed consent was obtained from livestock owners prior to examination and inclusion of their animals in study.

Consent for Publication

All participating farmers provided consent for the use of anonymized clinical data for research and publication purposes.

Acknowledgement

AcknowledgementsThe authors acknowledge the cooperation of participating livestock farmers and the support of field extension staff involved in organizing reproductive health camps.

References

1. Andrade MF, et al. Embryonic and fetal mortality in dairy cows: Incidence, diagnosis, and prevention strategies. Animals. 2024;14(3):40-40. doi:10.3390/ani14030040.
2. Food and Agriculture Organization of the United Nations. FAOSTAT statistical database. Rome: FAO; 2023. https://www.fao.org/faostat/
3. Fricke PM. Reproductive management of dairy cows in the precision dairy era. Journal of Dairy Science. 2022;105(4):3535-3550. doi:10.3168/jds.2021-21245.
4. Ginther OJ. How ultrasound technologies have expanded and revolutionized research in reproduction in large animals. Theriogenology. 2014;81(1):112-125. doi:10.1016/j.theriogenology.2013.09.007.
5. Government of India. Basic animal husbandry statistics 2023. Department of Animal Husbandry and Dairying, www.extensionjournal.com Ministry of Fisheries, Animal Husbandry and Dairying; 2023.
6. Government of Tamil Nadu. Animal husbandry policy note 2024-2025. Department of Animal Husbandry; 2024.
7. Neethirajan S. Digital livestock farming: Opportunities and challenges. Animal Frontiers. 2021;11(1):50-57. doi:10.1093/af/vfaa054.
8. Noakes DE, Parkinson TJ, England GCW, editors. Arthur's veterinary reproduction and obstetrics. 10th ed. Elsevier Health Sciences; 2018.
9. Sahu UK, Kumar A, Kumar B, Patra MK, Khan M. Early embryonic mortality in bovines: Current insights and interventions. Animal Reproduction Update. 2025;5(2):1-20. doi:10.48165/aru.2025.5.2.1.
10. Sahu UK, Kumar B, Khan MH, Warghat C, Singh M, Chauhan VS, et al. Spectral Doppler assessment of middle uterine artery hemodynamics across the estrous cycle in Tharparkar cows. Frontiers in Animal Science. 2026;7:1740299-1740299. doi:10.3389/fanim.2026.1740299.
11. Sahu UK, Kumar P, Kumar B, Khan MH, Baghel MS, Vadera V, et al. Transrectal Doppler characterization of uterine vascular dynamics in buffaloes with clinical metritis and subsequent recovery. International Journal of Bio-Resource and Stress Management. 2026;17(1):17. doi:10.23910/1.2026.6759.
12. Sahu UK, Singh K, Pandiyan N, Kumar B, Khan MH. Fetal mummification in a doe: A case report. The Indian Journal of Animal Reproduction. 2024;45(1):107-108.
13. Sahu UK, Singh M, Warghat C, Khan MH. Post-partum prolapse of genitalia in a cow. The Indian Journal of Animal Reproduction. 2024;45(2):93-95.