**Practical Application of Urinary Zearalenone Monitoring System for Feed Hygiene Management of a Japanese Black Cattle Breeding Herd—The Relationship between Monthly Anti-Müllerian Hormone and Serum Amyloid A Concentrations**

**Oky Setyo Widodo 1,2 , Makoto Etoh <sup>3</sup> , Emiko Kokushi <sup>4</sup> , Seiichi Uno <sup>4</sup> , Osamu Yamato <sup>5</sup> , Dhidhi Pambudi <sup>6</sup> , Hiroaki Okawa <sup>7</sup> , Masayasu Taniguchi 1,8, Mirni Lamid <sup>2</sup> and Mitsuhiro Takagi 1,2,8,\***


**Abstract:** This study addresses an advantageous application of a urinary zearalenone (ZEN) monitoring system not only for surveillance of ZEN exposure at the production site of breeding cows but also for follow-up monitoring after improvement of feeds provided to the herd. As biomarkers of effect, serum levels of the anti-Müllerian hormone (AMH) and serum amyloid A (SAA) concentrations were used. Based on the results of urinary ZEN measurement, two cows from one herd had urinary ZEN concentrations which were two orders of magnitude higher (ZEN: 1.34 mg/kg, sterigmatocystin (STC): 0.08 mg/kg in roughages) than the levels of all cows from three other herds (ZEN: not detected, STC: not detected in roughages). For the follow-up monitoring of the herd with positive ZEN and STC exposure, urine, blood, and roughage samples were collected from five cows monthly for one year. A monitoring series in the breeding cattle herd indicated that feed concentrations were not necessarily reflected in urinary concentrations; urinary monitoring assay by ELISA may be a simple and accurate method that reflects the exposure/absorption of ZEN. Additionally, although the ZEN exposure level appeared not to be critical compared with the Japanese ZEN limitation in dietary feeds, a negative regression trend between the ZEN and AMH concentrations was observed, indicating that only at extremely universal mycotoxin exposure levels, ZEN exposure may affect the number of antral follicles in cattle. A negative regression trend between the ZEN and SAA concentrations could also be demonstrated, possibly indicating the innate immune suppression caused by low-level chronic ZEN exposure. Finally, significant differences (*p* = 0.0487) in calving intervals between pre-ZEN monitoring (mean ± SEM: 439.0 ± 41.2) and post-ZEN monitoring (349.9 ± 6.9) periods were observed in the monitored five cows. These preliminary results indicate that the urinary ZEN monitoring system may be a useful practical tool not only for detecting contaminated herds under field conditions but also provides an initial look at the effects of long-term chronic ZEN/STC (or other co-existing mycotoxins) exposure on herd productivity and fertility.

**Keywords:** AMH; cattle; long-term monitoring; sub-clinical contamination; SAA; urine; zearalenone

**Citation:** Widodo, O.S.; Etoh, M.; Kokushi, E.; Uno, S.; Yamato, O.; Pambudi, D.; Okawa, H.; Taniguchi, M.; Lamid, M.; Takagi, M. Practical Application of Urinary Zearalenone Monitoring System for Feed Hygiene Management of a Japanese Black Cattle Breeding Herd—The Relationship between Monthly Anti-Müllerian Hormone and Serum Amyloid A Concentrations. *Toxins* **2022**, *14*, 143. https://doi.org/ 10.3390/toxins14020143

Received: 19 January 2022 Accepted: 11 February 2022 Published: 16 February 2022

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**Key Contribution:** The urinary ZEN monitoring system is an important practical tool for revealing the effects of long-term chronic ZEN/STC (or other co-existing mycotoxins) exposure on cows productivity and fertility. The long-term use of the ZEN-contaminated feed affected the calving interval.
