Infertility in the queen Alain Fontbonne, DVM , PhD, Ass. Pr., Dipl. ECAR Past-President of EVSSAR Alfort Veterinary College, Paris, France Elise Malandain, DVM, MSc R&D , ROYAL CANIN France Infertility in the queen Introduction Anestrus Hyperestrus Mating failure Ovulation failure Lack of fertilization Early embryonic death Introduction Nowadays Pure bred cats become more and more popular People are more professional: They ask veterinarians to provide more technical help It is necessary to have an accurate diagnosis procedure Begin with the most frequent causes Follow a chronological order Infertility in the queen Introduction Anestrus Hyperestrus Mating failure Ovulation failure Lack of fertilization Early embryonic death Anestrus Usual in northern hemisphere during the short days period Physiological anestrus is longer in long haired breeds False anestrus Silent heat: Normal hormonal events without obvious estrus behaviour Timid queens; low on social scale; crowding Differentiate from pseudopregnancy: vaginal cytology, serum progesterone Perform vaginal smears and hormonal assays More difficult to appreciate than in the bitch: Generally progesterone blood level does not increase except if a spontaneous ovulation occurs . Anestrus Primary anestrus Check eventual drugs administration: - Progestagens (given when the queen suffers from prurit) - Antifungic drugs (ketoconazole) Congenital problem - Clinical examination - Ultrasound examination Check that ovaries are present Beware of neutered queens Anestrus Primary anestrus Environmental problems Frequent travel, exhibition Overcrowding, temperature/ventilation Antagonistic social interactions - Social life (stress) - Lack of light Stress Transport Delayed puberty! Persians may reach puberty near 1214 months. Anestrus Secondary anestrus Racial variations Some persian queens may exhibit only one or two heat periods per year Changes in the living environment Light period Below 10 hours of light per day anoestrus Persistant corpus luteum Continuous progesterone secretion Normally <1ng/L Estrus induction Indications Medical indications Prolonged anestrus Zootechnical indications Try to breed queens at the non-breeding period Long-haired queens Problem due to the « group effect » Methods Light stimulation Hormonal treatments: gonadotropins: eCG=PMSC Light stimulation Interest Is a « physiological » method No adverse reactions Methods A 14 hours day light helps to get two monthly estrous periods Needs 3 weeks before the beginning of heats Avoid continuous light Light stimulation In practice Hurni makes the proposal of -one month of 9 hours of day light -followed by one month of 14 hours of day light. Programs using dis-continuous lighting programs 12 h-1h-1h-9h seem to give better results. In all cases, the best results are obtained when light stimulation is associated with a social stimulus (other queens + males). Infertility in the queen Introduction Anestrus Hyperestrus Mating failure Ovulation failure Lack of fertilization Early embryonic death Hyper-estrus Siamese cats Social factors Queens that are not in heat may accept a mating Aetiology : Successive phases of follicular growth Hormonal secreting follicular cysts: GnRH, hCG, follicular puncture, unilateral ovariectomy Granulosa Cells Tumor Drugs Infertility in the queen Introduction Anestrus Hyperestrus Mating failure Ovulation failure Lack of fertilization Early embryonic death Mating failure Before doing any complementory investigation, check that the mating has occured In practice, mating failure is THE big cause of infertility in the queen Mating failure Environnemental causes Transport of the queen leading to interruption of estrous behavior Time necessary for adaptation Non-experimented or scared male cat... …or agressive queen! -Organise transport -Help.... Mating failure Physical causes in the male Gingivitis or pain of the jaws Penis stricture due to hair Anatomical abnormalities of the genital tract Infection Retrograde ejaculation Urinari calculi Pain during ejaculation Mating failure Lack of libido of the male Hypothyroidism Hormonal treatments Age Lack of A vitamine, iodide, poly insaturated fatty acids Weight excess Low efficiency of usual treatments(testosterone IM) Try with another male cat AI Infertility in the queen Introduction Anestrus Hyperestrus Mating failure Ovulation failure Lack of fertilization Early embryonic death Ovulation failure Mounting Not repeated enough Too long period between two successive mountings 1 coitus leads to ovulation in only 50% of the cases Before going through many complementary investigations,the breeder has to know the frequency and intensity of mountings, and to check if the queen exhibits a normal post-coital reaction. Ovulation failure Progesterone secretion (ovarian tumors, luteal cysts) Some queens may have follicular maturation under progesterone secretion. This level can be sufficient to prevent ovulation, without preventing estrus: It is useful to assay progesteronemia before a mating: <1ng/L May induce false interpretation of the ovulation detection after mating To confirm ovulation: progesterone testing !! 35 progesterone ng/L 30 25 20 15 10 5 0 0 1 2 3 4 Days post ovulation 5 6 7 Ovulation failure Sedatives/anesthesic drugs Used in agressive queens May prevent the LH peak Stress A stressed queen may not ovulate, even when exhibiting a normal post-coital reaction Try eventually to bring the male and to allow him to mate the queen , after a period of adaptation Infertility in the queen Introduction Anestrus Hyperestrus Mating failure Ovulation failure Lack of fertilization Early embryonic death Lack of fertilization Diagnosis Ovulation has to be confirmed Mating has to be confirmed Negative early pregnancy diagnosis using ultrasonography Impossible to differenciate from early embryonic death Cause Spontaneous ovulation, before presenting the queen to the male Relatively rare MALE INFERTILITY Male infertility Genetical problems « 3 colors » male cat (tortoise shell) Infertility: Abnormalities of semen (Cf. AI) Treatments are of low efficiency (GnRH) Presence of spicles on the penis Under the control of androgens Male infertility Miscellaneous problems Urinary calculi, Increased temperature of the testis, Testicular tumors Inflammatory diseases ( check testis and prepucial cavity). Back pain Gingivitis!!! Male infertility Drugs Cortico-steroids, anabolic steroids, antifungic compounds, progestagens Age Too young or too old ! Male infertility: procedure History Previous treatments (ketoconazole, progesterone, anti-androgens) Previous matings Clinical examination External genitalia Check the urine Semen evaluation See AI Infertility in the queen Introduction Anestrus Hyperestrus Mating failure Ovulation failure Lack of fertilization Early embryonic death Cystic endometrial hyperplasia First stage of pyometra Due to hormonal disorders - Repeated pseudo pregnancies - Drugs (pills, injection) Secondary, infection and accumulation of fluid = pyometra Particularly frequent in Siamese queens Leads to infertility Ovulation occurs, without implantation No other clinical signs…. Infertility in the queen Introduction Anestrus Hyperestrus Mating failure Ovulation failure Lack of fertilization Early embryonic death Abortion/Pregnancy arrest • Early embryonic resorption and further abortion most often unnoticed in the queen: infertility is the main sign • Scientific knowledge on this topic is very limited (Schlafer 2003) Early embryonic death In practice Try to diagnose pregnancy as early as possible D 10 D 11 D 13 D 15 Hormonal problems Hormonal problems • Hypoluteoidism: Very rare in queens Progesterone therapy may be used be careful not to counteract with normal parturition Pregnancy Infectious diseases Infectious diseases • Viruses: FelV the most common cause: Testing and housing procedures FIV: Outdoors catteries Feline Herpes virus Feline calicivirus: Many different strains - Cases in vaccinated animals ? Rarely suspected (Van Vuuren et al. 1999) Feline panleucopenia virus: Beware of attenuated live vaccines injected in pregnant queens Feline coronavirus: Uncommon cause of abortion in cats (Ström-Holst 2002) Infectious diseases • Bacteria: Probably under-diagnosed E.coli, Staphylococcus sp., Streptococcus sp.: - potential causes of abortion (Root-Kustritz 2006) Chlamydophila felis ? Its role is unclear : experimentally: suppurative placentitis may occur (Schlafer 2003) Treatment: Doxyxyclin 28 days (Dean et al. 2005) Diagnosis is difficult Vaccination does not protect against infection Salmonella sp. Bartonella henselae (Guptill et al. 1998) • Parasites: Toxoplama gondii (Root-Kustritz 2006) Cytauxzoon felis ? (Weismann et al. 2007) Infectious causes of « abortion » In practice Test the queen for FelV-FIV Serology-PCR for Chlamydiosis, FHV Vaginal smear: many neutrophils ? Bacterial identification: vaginal culture with a urinary catheter : infuse a few drops of sterile fluid in the vagina, and aspirate them again Chromosomal problems Chromosomal defects • May cause abortion or fetal death 37, X0 Trisomies • If repeated abortions: karyotype the aborted fetuses (Romagnoli 2003) Nutritional causes Proteins and Taurine Proteins Higher level diet recommended for reproduction Taurine During gestation Essential amino acid selectively transported through the placenta A deficiency may induce early embryonic resorption (before 25 days) (Sturman 1991, Dieter 1993, Champion et al. 2005) - with or without an increase in relaxine plasma level Arachidonic acid Essential in cats Can not be synthesized in quantity (limiting action of desaturase δ 6) Not enough to cover high needs But adults cat can survive with a deficiency During pregnancy A deficiency IMPAIRS reproduction in queens Essential for gestation and viability of kittens! (Pawlosky1996) No effect on males reproduction , Copper (Fascetti 1998, 2000) • Role on reproduction: Has been particularly studied in cats Direct link between Copper and reproductive performances Some chemical forms are not assimilated - Copper oxide for example Under 10 mg/kg, progressive decrease of fertility, prolificity and vitality of kittens To ensure the best assimilation Some authors recommend to use chelated forms - Trace element link with an amino acid - Less competition for absorption Early embryonic death In practice Test the queen for FelV Vaginal smear: many neutrophils ? Bacterial identification: vaginal swabbing: Tom Cat Catheter : inect a few drops of sterile fluid in the vagina, and aspirate them again Infertility in the queen : summary The infertile queen 1. Before the heat period Conduct a good clinical and genital examination Progesterone assay: level has to be basal Vaginal smear: interoestrus Ultrasonography : the ovary should be homogeneous or should not show any « big » follicular structure; the uterus should be normal Check the history of the male (previous litters ? ) Test FeLV The infertile queen 2. During the heat period Vaginal smear : confirms that the queen is in estrus Try to observe the mating -Is there enough mating? Frequency? Ultrasonography: helps to check follicular maturation and ovulation In summary: the infertile queen 3. Four days after the last mating Progesterone assay : +/- 5ng/L 35 progesterone ng/L 30 25 20 15 10 5 0 0 1 2 3 4 Days post ovulation 5 6 7 In summary: the infertile queen 4. Early pregnancy diagnosis: 14 days after mating Progesterone assay : >10ng/L Check male’s fertility Check uterus Pyometra, CEH? Return to estrus in less than 18 days indicates failure to ovulate Return to estrus 35-45 days pseudopregnancy Elevated progesterone without confirmed pregnancy Causes: CEH, endometritis, early embryonic loss, male infertility Ultrasound exam at 14-21 days post-breeding In summary: the infertile queen 5. If the queen undergoes a resorption or abortion Investigate infectious diseases: FeLV, FIV, FPV, FHV-1, FCV, Toxoplasma, etc. Clinical examination - FeLV - Herpesvirosis Bacterial identification from the vagina - Specific Chlamydiosis - Non specific Progesterone assay Thank you for your attention !