Canine infertility. Alfort National Veterinary College Paris - France. Alain Fontbonne. DVM, PhD, Dipl. ECAR

Transcription

1 Canine infertility Alain Fontbonne DVM, PhD, Dipl. ECAR Alfort National Veterinary College Paris - France

2 Introduction Nowadays: vets are more often asked to manage fertility problems in the dog: increased popularity of purebred dogs increased technicity in human medicine sentimental or financial reasons

3 Introduction (2): Many different problems may lead to infertility: - hormonal problems - infectious diseases - congenital or acquired defects of the genital tract to manage infertility, it is necessary for the veterinarian to follow a very strict diagnosis procedure

4 A bitch is pregnant if One spermatozoon + One oocyte = One pup

5 A breeder is happy if Many spermatozoa + Many oocytes = Several puppies

6 1. Mistimed breeding

7 According to the vast majority of publications: mistimed mating represents the commonest cause of infertility or hypofertility

8 Therefore, the first step in an infertile bitch is performing ovulation timing!

9 Oocytes

10 First phase of heat = pro-oestrus Oestrogens Sexual hormone Follicular growth Vulvar bleeding Changes of the vaginal mucosa

11 Around the end of the pro-oestrous period LH peak * * LH : Luteinizing Hormone Acceptance of mating Start of progesterone rise Specificity of the bitch

12 Second phase of heat = oestrus 2 days after the LH peak Occurrence of ovulation

13 IN MOST MAMMALS Oocyte maturation in pre-ovulatory follicles OVULATION Mature oocyte BITCH: Oocyte maturation In the oviduct OVULATION Min. 48 hours Pre-mature oocyte Mature oocyte

14 Thus, in the bitch The time of ovulation is not the time of fertilisation

15 After 2 to 3 days Ovulation Maturation LH 48h 48h 48h-72h the oocytes degenerate

16 Oocytes can be fertilised during 2 (or 3) days 2 or 3 days

17 Spermatozoa

18 Spermatozoa Rapid transfer to the anterior part of the uterus increase of uterine contractions around ovulation Sperm may fertilise oocytes 7 to 8 days after mating: for many stud dogs when using chilled or frozen semen it may not survive more than 2 days

19 2 or 3 days 2 to 8 days

20 Matings after ovulation In case of natural mating AI with fresh semen: if 2 matings: 1 day and 3 days after ovulation is ideal if a single mating is performed: ideally 1 or 2 days after ovulation In case of AI with chilled or frozen semen ideally 2 or even 3 days after ovulation

21 1st case Ov LH Maturation 48h 48h 48h-72h Mating too early Not enough sperm Not pregnant or small litter

22 2nd case Ov LH Maturation 48h 48h 48h-72h Not pregnant Mating too late

23 3rd case Ov LH Maturation 48h 48h 48h-72h PREGNANCY Mating 1 to 4 days after ovulation

24 How can we determine the optimal mating time?

25 Duration of heat

26 Fertile bitches are not always mated between the 10 th and the 13 th day of their heats Mating date of 745 pregnant bitches ( Alfort National Veterinary College - Paris)

27 Also the optimal breeding period may vary among successive cycles in the same bitch

28 Variation in the optimal time of breeding 44% (16/36) bitches showed significant variations in successive cycles (Badinand et al. 1993)

29 Electrical resistivity of the vaginal mucus

30 A recent study (Clero UMES) Ovulstart (Nature et compléments France)

31 Results Ovulation

32 Results This technique: - Is much more precise than vaginal smears to detect ovulation - In 69 to 76% of the cases, ovulation may be detected with an accuracy of ± 24 hours - The maximal difference between real ovulation and ovulation detected Ovulation by the system is 60 hours

33 Endoscopic appearance of the vaginal folds

34 Flat and thin vaginal wall Anoestrus

35 Proestrus

36 Oestrus

37 End of heats = beginning of dioestrus

38 Progesterone assays

39 Progesterone and LH blood levels (ng/ml) 2 main events in progesterone profile Ovulation 5 to 10 ng/ml (15 to 30 nmol/l) 10 8 Pg LH 6 LH peak h00 30/10 21h00 9h00 31/10 15h30 02/11 13h45 03/11 20h00 04/11 15h00 05/11 20h00 9h00 06/11 14h30 20h30 9h30 07/11 15h00 19h00 17h00 08/11 20h00 10h30 09/11 Dates and time of blood samples

40 Progesterone ng/ml Blood progesterone concentration at ovulation is fairly constant Ovulation Days

41 Progesterone ng/ml Progesterone blood concentration varies a lot at the optimal fertilisation period Ovulation Days

42 Ultrasonography of the ovaries

43 Position of the bitch?

44 Position

45 Pre-ovulatory period (end of pro-oestrus beginning of oestrus)

46 Pre-ovulatory period

47 Pre-ovulatory period

48 Ovulation

49 Ovulatory period

50 Ovulatory period

51 Ovulatory

52 Ovulatory period

53 Ovulatory period

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56 Marseloo et al Ovulation can be accurately determined using ovarian ultrasonography in the bitch Compared with progesterone assays: ovarian ultrasound examination improves the detection of ovulation in around 10 to 15 % of bitches, depending on the breed

57 In practice?

58 Progesterone and LH blood levels (ng/ml) In practice Begin with progesterone assays: when blood progesterone begins to rise eventually: begin ovarian ultrasound examination Pg LH h00 21h00 30/10 9h00 15h30 13h45 20h00 15h00 20h00 31/10 02/11 03/11 04/11 05/11 9h00 14h30 20h30 9h30 15h00 19h00 17h00 20h00 10h30 06/11 07/11 08/11 09/11 Dates and time of blood samples

59 Interests in infertile bitches Hypofertile or old bitch: helps to estimate the number of follicles per ovary (?) Ovulation problems: veterinarians may see in situ what is happening: easier diagnosis of ovulation problems detection of ovarian cysts it may help to use appropriate treatments if necessary

60 2. Male infertility

61 Male infertility «After mistimed breeding, male infertility is the next common cause of conception failure in bitches presented with infertility» (Johnston et al. 1994) Photo: Arthus-Bertrand

62 Introduction High value of reproduction in genetically selected male dogs Very little is known about male infertility: in humans, the cause remains unknown in more than 70% of the cases: use of IVF or ICSI in dogs: fertility is restored in only 10% of the cases (Johnston et al. 2001) Veterinarians must have total comprehension concerning the investigation of the male reproductive function

63 Examination of semen The first and essential clinical approach to infertility in the male dog Veterinarians have to perform two different things: collect the semen evaluate the semen

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69 What causes low quality semen?

70 Prostatic problems

71 Prostatic problems

72 Prostatic problems Important role of prostatic diseases in canine infertility

73 Epididymal or testicular problems

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75 Urinary or urethral problems

76 Hormonal problems

77 Infectious diseases

78 Orchitis/Epididymitis acute orchitis: failure to mate chronic orchitis: alteration of spermogram - decreased progressive motility (asthenozoospermia) - increased secondary morphologic abnormalities or oligo-teratozoospermia (Hastings et al. 1987)

79 Miscellaneous problems

80 3. Other causes of infertility

81 3.1. Anoestrus / Infertility with prolonged interestrous intervals

82 No puberty?

83 No puberty? Sexual ambiguity = pseudo-hermaphrodism

84 No puberty? Silent heats: 18% of anoestrus cases (Arbeiter et al. 1972)

85 In the adult bitch, which has already cycled

86 Progesterone induced problem Progesterone secreted by the ovaries: Silent heat +++ Ovarian cyst or tumour

87 «Grace» Collie bitch, 4 years old Anoestrus for 14 months

88 «Grace» Collie bitch, 4 years old Pg = 18.1 ng/ml

89 Heats induction protocols

90 Heats induction GnRH and GnRH agonists: discontinuous administration works badly in a near future: SC agonists implants? i.e. Deslorelin (Suprelorin ) (Kützler et al. 2003) oestrus induced in 2 to 9 days in 8/8 bitches more synchronised if sub-mucosal vulval implantation

91 Gonadotropins Potential use of: PMSG (ecg) However: Low quality of the corpora lutea (England 1999) Frequent luteal failure (Kützler ) Alltogether: rather moderate pregnancy rates 0 to 60%

92 Antiprolactin drugs: Heats induction dopaminergic products mainly (cabergoline) interest in case of prolonged anoestrus Buff et al.1996: 0.1 ml/kg/day 3 weeks 72% of heats induction in the following month (versus 6 % in non-treated bitches) works badly to shorten interoestrous intervals

93 Underlying illness

94 Incidence of hypothyroidism on infertility Hericher 2004 / Segalini et al : 102 infertile or hypofertile dogs and bitches from 5 breeds (Great Dane, Dogues de Bordeaux, English mastiffs, Leonbergers, Golden Retrievers) Only 3 dogs showed a low T4 and high TSH

95 3.2. Infertility with shortened interestrous intervals

96 Need of endometrial repair Approx. 130 to 150 days for endometrial involution and repair (Johnston et al. 2001) cycles intervals less than 4 months are often infertile

97 Shortened interestrous intervals Pathological factors: ovarian «hyperstimulation»: follicular cysts, GC tumors premature decline of progesterone during diestrus

98 Ovarian follicular cysts or tumors

99 Follicular cysts Follicular cysts:

100 Follicular cysts Potential clinical signs: shortened interestrous intervals prolonged estrus (or proestrus) anovulation alopecia aregenerative anemia (medullar aplasia) CEH/pyometra

101 Diagnosis: ovarian ultrasonography Size > 1cm = cyst

102 Follicular cysts Hormonal treatments (Johnston et al. 2001): From our point of view: high danger of induced pyometra Surgical treatments: Fine needle aspiration under ultrasound Surgical removal of the cyst

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104 «Teresa s friend» greyhound 3 years old

105 History Interoestrous intervals: every 5 months Two previous matings: ovulation detected by progesterone assys no puppies produced

106 Ovarian ultrasound

107 Ultrasound emptying of cyst failed Decision of surgical removal of cyst Next heats: 3 pups

108 Surgical removal of the cyst

109 Premature decline in progesterone during diestrus

110 Premature decline in progesterone Anovulatory cycles: 1% of the cycles (Arbeiter 1993) role of stress? / shipping of the bitch? (Cain 2001) Always re-assay progesterone: at the end of the heat period

111 Premature decline in progesterone Luteal failure: lack of progesterone secretion during pregnancy influence of the breed: Rottweilers, German shepherds Newfoundlands, Bernese

112 During pregnancy Perform progesterone assays once (Grundy et al. 2002) or twice a week At least: assay progesterone in infertile bitches at the time of the early pregnancy diagnosis

113 3.3. Infertility with normal interestrous intervals

114 3.3.1 Hormonal problems

115 Infectious diseases

116 How do infectious diseases cause infertility?

117 Vaginal infection may result in: spermicidal activity (Jones and Joshua 1982, Burke 1986) penetration of infectious agents into the uterus during prooestrus and oestrus (Van Duijkeren 1992)

118 Uterine infection may result in: hostile environment for the sperm and eggs (Linde-Forsberg and Bölske 1995) early embryonic death (Freshman 1991) inflammatory process endometritis placentitis or placental infection (Anvik 1991) (Van Duijkeren 1992)

119 Viral infections

120 Canine Herpes virus

121 Other viral infections that may play a Canine distemper role on fertility Canine minute virus (CPV1): resorption in 3/8 pregnant bitches (Carmichael et al. 1991) one clinical case described in Germany (Tryuen et al. 1996) role on fœtal anasarcoid?

122 Bacterial infections

123 Specific bacterial diseases acting on infertility Potential role of: Brucella canis Mycoplasms / Ureaplasms Others: Campylobacter Salmonella sp. / Listeria sp. Leptospira Chlamydia etc

124 Usual vaginal bacteria

125 In the vagina

126 The incidence of vaginal infection on infertility There is a high evidence that vaginal infection leads to infertility (Fayrer-Hosken et al. 1994) Any bacteria may become pathogen: when it has gained competitive advantage and has overgrown the normal bacteria (Caudle et al. 1994)

127 Quantitative variations in case of vaginal infection Often, a pure culture is obtained (Johnson 1991)

128 Symptoms in case of vaginitis Often very subtle: licking of the vulva / sticky vulvar hair very subtle vulvar discharge attraction of the males urinary problems infertility or abortion

129 Therefore, genital infection can be suspected if (Grundy et al. 2002): in the same infertile bitch: growth of bacteria from the deep vagina vaginal mucosal inflammation +/- peripheral leukocytosis if possible: uterine cytology should be performed

130 How to diagnose vaginitis?

131 Vaginal smears

132 Vaginoscopy: an alternative technique?

133 Vestibulitis

134 Vestibular inflammation

135 Vaginitis

136 Chronic vaginitis

137 Chronic vaginitis

138 Cervicitis?

139 Cervicitis

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141 3 clinical cases

142 Vanda, cocker spaniel, 2 years old Frenetic vulvar licking Clinical exam: Serous vulvar swelling +++ Vaginal swab: anoestrus, few PMN P4 low

143 Vanda, cocker spaniel, 2 years old

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145 Vanda, cocker spaniel 2 years old

146 Vanda, cocker spaniel 2 years old Vaginal Bacteriology: E.coli in heavy growth

147 Very Glamour, Colley 2 years old Vulvar discharge and attract males Clinical exam: Slight serous vulvar swelling Vaginal swab: RAS

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149 Very Glamour, Colley 2 years old Vaginal bacteriology: Proteus mirabilis +++ Resistant to cefalexin Streptococcus dysgalactiae +

150 Reine, Landseer, 5 years old Infertility

151 Reine, Landseer, 5 years old Vaginal swab

152 Reine, Landseer, 5 years old Vaginal bacteriology: Pasteurella multocida in heavy growth Resistant to most ATBQ Intermédiaire: fluoroquinolones

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154 Diagnosis of vaginal inflammation Interests in infertile bitches: Correlation between vaginal infection and infertility / neonatal diseases (Fayrer-Hosken et al. 1994, Münnich and Lübke- Becker 2003) Subsequent risk of endometritis?

155 Anatomical abnormalities of the vulva, vestibule or vagina

156 Anatomical malformations

157 Anatomical defects of the vagina Leads to infertility due to: bitches that refuse to be mated impossibility of the tie Always make a good genital investigation of the vagina at the begining of the heats

158 Vaginal septum

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160 Ectopic ureter Lulich 2006 Pechereau 2003

161 Stenosis Vagino-vestibular Circumferancial stricture and vaginitis Severe vaginal stenosis

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163 Occlusion of the vulva

164 Vaginal hyperplasia 2ème degré 1er degré 3ème degré

165 Vestibular and vaginal masses /tumours

166 Vaginal masses/tumours

167 Uterine pathology

168 Endometritis Etiology: Frequent role of bacterial infection: Role of mycoplasms unclear Role of biochemical substances: i.e.: tris-egg yolk buffer (Coelho Ribeiro et al. 2006) Role of endometrial trauma

169 Endometritis in the bitch Symptoms: Infertility: Failure of implantation Placentitis: embryonic or fetal resorption Abortion Vulvar discharge not systematic

170 Principle: Diagnosis of endometritis: bacteriology + cytology Guiding a catheter through the cervix Injecting sterile saline solution Re-aspirating the injected fluid for

171 Fontaine et al /26 bitches showed endometritis

172 Fontaine et al Five bitches were further bred after treatment and all became pregnant

173 Diagnosis of endometritis: uterine biopsy Downs et al. 1994

174 Cystic endometrial hyperplasia Often leads to pyometra (CEH)

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178 Abnormal sexual behavior

179 Abnormal sexual behavior very dominant bitches with subordinate males sexual preference standing oestrus disconnected to ovulation perform A.I.s

180 Conclusion: the infertile bitch in practice

181 In practice: During the heat period: Make a careful examination of the genital tract Follow carefully the ovulation process: Progesterone assays are esential Ovarian ultrasound may be useful Check the semen of the male Check infectious problems: Brucellosis testing Swabbing of the anterior vagina for bacterial identification

182 In practice: During the pregnancy period: Vaccinate against CHV Assay progesterone: ideally once a week at least: around 30 days of pregnancy Perform early pregnancy diagnosis: if possible by ultrasound

183 Thank you for your attention!