Changes in anti-Müllerian hormone concentrations in bitches throughout the oestrous cycle

Studies in female dogs proved the measurement of anti-Müllerian hormone (AMH) as a diagnostic tool to distinguish ovarioectomized from intact females or to diagnose ovarian remnant syndrome. Furthermore, its usability to diagnose a granulosa cell tumour and predict litter size was also investigated in female dogs. Although serum AMH was previously shown to increase during the transition from anoestrus to pro-oestrus in dogs, changes in AMH concentration over the entire oestrous cycle have not yet been determined. The aim of this study was to investigate the secretion pattern of AMH throughout the oestrous cycle in non-pregnant bitches. As a preliminary step, we tested our assay by measuring AMH concentrations in serum samples of 19 intact bitches (10 in anoestrus, 3 in pro-oestrus, 3 in oestrus, 3 in metoestrus) and 19 spayed females of different age and breed. For the main study, 20 healthy female dogs were examined (10 Beagles, and 10 Labrador crossbreeds, body weight 10-28 kg, age 1-6 years) during a normal oestrous cycle. Serum samples were collected in late anoestrus, at several times during pro-oestrus and oestrus, as well as 28 days and 4.5 months after ovulation. Blood collection was combined with a gynaecological examination including progesterone measurement. Serum concentration of AMH was determined using a chemiluminescence immunoassay validated for dog serum. In the preliminary test, intact bitches in various stages of the oestrous cycle had significantly higher serum AMH levels (0.19-1.45 ng/ml) than spayed females (0.01-0.06 ng/ml, P < 0.001). In cycling Beagle and Labrador crossbreed bitches participating in our main study, AMH concentrations were between 0.09 and 2.65 ng/ml with higher AMH concentrations in Beagles but overall high inter-individual and intra-individual variation. The highest AMH values were reached in the last three weeks before the onset of heat until six days before ovulation, and the lowest AMH concentrations were recorded during preovulatory oestrus in 6, metoestrus in 7 or anoestrus in 7 dogs. Serum AMH concentrations increased significantly from late anoestrus up to six days before ovulation and decreased significantly over the last three days before ovulation. A further significant decrease occurred from the last days of oestrus to metoestrus and mid anoestrus. This study shows that the used AMH assay can clearly distinguish between intact and spayed females and that the serum AMH pattern over the oestrous cycle is similar in all bitches, but with high variation among the breeds and among and within bitches. These changes in the AMH concentration are a challenge in timing serum sample collection and interpreting AMH values in bitches. Further studies are necessary to emphasize the factors influencing the AMH concentration.