----start---- reproduction in the male dog dr ward her two loves: endocrinology and sperm :) The same ground rules apply here - you're not responsible for anything not in handout or for anything in the little 8 page thing at the end she wrote for Iams, that's for your own information. You're only responsible for what is in the handout. a few cases will be discussed today; she'll try to tell you some stuff about doing repro work in practice. usually most of our time is spent working with the female, but there is a little bit of stuff to know about the male, too :) Repro in small animal (private) practice: -one reason MDs go into infertility work is you have nothing to lose. worst case, you end up with no pregnancy - it's not like they get worse,generally. if you get them pregnant, you're a hero, if you don't, you haven't lost a lot. -easy -small amount of equipment -involves mostly common sense and logic -provides a "hard to find" service: no one does this, not sure why. maybe because people do not like to deal with the breeders. but you can help these people and their animals. working up infertility in a male dog is easier than regulating a diabetic, too -can be a practice builder! -breeders are grateful -you end up with puppies which grow into dogs and come back to you male anatomy: penis and prepuce penis easily extruded from prepuce normal things to look for - for each male dog that comes in for a physical, make sure you can extrude penis, that it all looks normal. males have an os peniss, and a bulbus glandis which is one of the sex organs that swells during breeding. penis should be nice light glistening pink color. Testicles and epididymides: dogs have huge epididymides - can feel all the parts, so that might help you localize disease. testicles should be firm, symmetrical, and descended by six months of age. don't dx cryptorchidism til 6 mos. prostate: accessory sex gland that you can palpate rectally. dogs get BPH like men do, all older intact male dogs will have some degree of this. physiology: puberty: in male about 10-12 mos with range of 6-24 mos. defined as animal with normal libido and motile, mature sperm in ejaculate endocrinology: GnRH, FSH, LH secreted in pulsatile manner - this is important b/c it means a baseline measurement isn't helpful. you need the normal pulse pattern. also tx for lack of these hormones isn't just giving one shot - you need multiple shots, or some kind of pump. testosterone is 50-100 times higher in the testes than in the plasma - this is important b/c if you have an animal with possible testosterone deficiency, plasma testosterone level isn't so useful. you need testicular testosterone level. GnRH - hypothalamus -> FSH and LH - pituitary --> testis. FSH trophic to sertoli cells, stimulates gametogenesis and inhibin production. inhibin --> neg feedback, reduced FSH secretion LH --> from pituitary to testis under influence of GnRH --> trophic to leydig cells, promotes testosterone secretion. testosterone acts locally on seminiferous tubules, involved in spermatogenesis. spermatogenesis is really really cool, and this is a reason why most animals are so reproductively successful - b/c this happens all the time, and this saves our butts. if there is testicular damage, normal sperm production can return if you just wait. it's a continuous process, and takes about 60 days in the dog. least differentiated cell is near membrane, most mature in lumen of tubule. negative influences on spermatogenesis: elevated testicular temperature nutritional deficiency toxins - big in human med right now epididymal maturation: takes 15 days in dog. so, 75 total days for spermatogenic cycle. don't expect normal sperm for 75 days following testicular insult. that's 2-3 mos, ok?. sperm mature in caput and corpus and are stored in cauda. then they wait for ejaculation. there is a storage area. if they sit there too long,t hey can degenerate a bit. so a first ejaculate after a long rest can be sort of crappy. Blood testis barrier: this is important. tight junctions b/w sertoli cells near basal lamina, steroids can pass through; protects germ cells from noxiou agents and prevents antigenic sperm products from entering circulation and causing massive inflammatory response. so you must be very careful sticking needle into testis or taking biopsy. disorders of penis and prepuce: congenital: hypospadia (David Holt discovered this in cat), deformity of os penis, persistent frenulum. these are all rare. balanoposthitis - inflammation of penis/prepuce - relatively common - opportunistic invasion of normal flora. in normal dogs, small amount of d/c normal. sometimes in morning is more copious than other times. but can get opportunistic invasion. Phimosis: inability to protrude the penis from the prepuce. congenital or more commonly acquired due to scarring, neoplasia, hair ring. Paraphimosis: inability to withdraw penis back into the prepuce. gross and painful. penis dries out, necrosis, urethral obstruction, etc. people may breed a male and put back in cage and not ensure penis is back in prepuce and this can occur. may require surgical intervention. shrink it down using some dextrose to draw fluid out first. insert urinary cath, put in purse string suture to hold penis in. so be careful post breeding to ensure penis is in place. Trauma: penis is very vascular. if you stick it, it bleeds a lot. Neoplasia: squamous cell carcinoma, transmissible venereal tumor TVT (tx vincristine), papillomas, mast cell tumors. Testicular/epidydimal disorders: 1. cryptorchidism: lack of descent of testicles by 6 mos of age. most common congenital testicular abnormality. (note: breeders often bring in puppies at 4 mos and want to know if testes will descend. you can say probably not, but can't say for sure til 6 mos). can be unilateral or bilateral. testes can be in inguinal canal or abdomen. might be single autosomal recessive diseae. purebreds, toys, unilateral most common. These animals are fertile but should not be bred or shown. torsion and neoplasia (sertoli cell tumors) of retained testicles are common. unilateral descended testicle is normal and fertile. You should *always* neuter these animals. 2. orchitis/epididymitis: inflammation in testicle or epididymus - usually can't tell them apart. trauma, infection, autoimmune disease can cause this. brucella canis is a big cause and you should worry about that. do brucella titers on these animals b/c it is zoonotic. note that scrotal skin is very sensitive and betadine can cause scrotal inflammation. signs: enlarged, painful, inflamed testes or epididymides; erythematous scrotum, if chronic problem testes are small and firm. there may be systemic illness. slide: sick dog with huge red swollen scrotum and multiple petechiae all over body. guarded prognosis for fertility - inflammation raises temperature in testis, may be permanently damaged. neuter these animals, and check for brucella canis. 3. testicular tumors: common in older dogs, usually not a problem. unilateral or bilateral. sertoli cell tumors: most commonly intra-abdominal and can get large. slide: tumor size of watermelon on a spaniel of some sort. these are the ones that are problems, b/c of estrogen production causing secondary feminization, attraction of male dogs, bone marrow suppression, paraneoplastic syndrome...gyecomastia, alopecia, aplastic anemia, male dogs hanging around. often the unaffected testicle atrophies. neuter these dogs. leydig cell tumor: incidental findings, no big deal. seminomas: also usually incidental, arise from germ cells, mets uncommon Disorders of the Prostate: her viewpoint on things. this is part of the repro system and you need to know about it b/c it affects fertility Benign prostatic hyperplasia: a normal age related change that is associated with some problems -- it's hyperplasia and hypertrophy, btw. most dogs do not develop signs but if they do, clinical signs are: -tenesmus (prostate compressing rectum) -urethral d/c (may be bloody) -hematuria On PE you feel: -enlarged prostate - sometimes feel this abdominally as well. bilaterally symmetrical and nonpainful. definitely feel it on rectal exam which you should do on all your patients! dx: biopsy, PE, u/s, prostatic fluid analysis tx: *castration*; finasteride (Proscar) - 5a-reductase inhibitor (anti androgen; 1 mg/kg/day) - use that really just in animal to palliate while preparing to breed him - breed him and then neuter him; estrogen (DES) and megestrol acetate (Ovaban) - bad side effects of aplastic anemia, diabetes. Prostatitis, prostatic abscesses - acute or chronic, usually ascending infection, 10% have recurrent UTI. the bad thing about these is they can get septic, and often there may be endocarditis and death, argh. you need to jump right on this and treat. usually e.coli, sometimes brucella. signs: usually sick dog, febrile, stilted gait, caudal abdominal gain (may be referred, may mimic pancreatitis), urethral d/c. on PE, rectal exam is very painful, they will try to kill you. sometimes the chronic cases come in for refusal to breed, reduced libido. or infertile - sperm don't handle influx of RBC and WBC into semen well. with chronic cases, signs can be intermittent, sometimes they come in with history of waxing/waning illnes, recurrent UTI. clinpath: neutrophilia with left shift. hematogenous spread to heart, joints. could present with lameness. dx: hematology, u/s, u/a, cytology/culture of urine, semen, prostatic fluid. could do u/s guided prostatic aspirate. biopsy rarely needed since advent of ultrasound. usually cytology or prostatic wash is inflammtory, and culture reveal >10,000 CFU/ml. is semen sterile? no. so here, if you're trying to decide if you have an infectious process in some area semen passes through, you can't just culture it. you need to look at number of colonies and also look for inflammatory cells. if there's infection there are >10,000 CFU and there are WBCs. slide: normal prostatic cells from prostatic wash - if you see these clumping cells, that doesn't mean there is a tumor. these are healthy normal cells, all same size. slide: PMNs, bacteria - infection from prostate. --break-- treatment for prostatitis/abscess of prostate: -antibiotics: 4-6 weeks with lipid soluble drugs; baytril, TMS, chloramphenicol. need to recheck culture - urine or prostatic wash. -castration for refractory cases - usually you neuter them -surgical drainage may be required for abscesses prognosis: acute disease - resolves quickly (but may recur); chronic will recur. prostatic neoplasia: rare. seen in intact and neutered males. types: adenocarcinoma, transitional cell carcinoma, bad tumors in general. neutering doesn't prevent these. signs: tenesmus, dysuria, hemorrhagic urethral d/c - similar to BPH PE: nonpainful, asymmetrically enlarged prostate. should be nonpainful, however, with neoplasia there is increased risk of infection. if inflamed or infected will be painful. sublumbar LNs may be enlarged (may be with infxn also). dx: prostatic fluid cytology; prostatic tissue aspirate, biopsy mets: local, bladder, bones, kidney, lung slides: low and high mag of a transitional cell CA of prostate. these epithelial cells are not the same size - different sizes, large nuclear: cytoplasmic ratio. inflammatory cells also present. tx: radiation, chemo, prostatectomy not easy to do; px bad b/c usually mets are present. Prostatic wash: a good clinical tool; lots of people do not like them, Dr W does. it's a way to get prostatic fluid w/o semen collection. purpose: collect sample for cytology/culture. procedure: awake or light sedation; bladder empty of urine; infuse sterile saline to remove residual urine. you don't want urine in your prostatic fluid sample if possible. if you can't do the rinsing, it's ok. just empty the bladder. palpate prostate rectally. withdraw catheter until at prostatic level - you will feel it. inject about 5 mL sterile saline, while massging the prostate. then suck out the fluid. do this a couple of times. then do culture/cytology. look at it yourself, too. Artificial Insemination: easy to do! low tech. why? -semen extension to help sperm quality -combination of several ejaculates to improve sperm quality -prevention of injury during breeding -chilled or frozen semen allows you do to this w/o stud being present equipment needed: -artificial vagina (these are cheap and easy) (ha!) and semen collecting equipment -insemination pipette -just a long tube that isn't glass -20 cc syringe Fresh semen: collect from stud put vaginally in bitch (+/- holding her butt in the air) best conception rates - sperm love semen extender so you can add that too Chilled semen: collect extend refrigerate vaginal insemination within 24-48 hrs conception rate is still pretty good, better than frozen; canine semen is pretty hardy as long a you extend it well. frozen semen: worst conception rates collect, extend with cryoprotectant added freeze in straws or pellets in liquid nitrogen vaginal or intrauterine insemination a few groups report better insemination with intrauterine insemination one woman in NZ gets into uterus with insemination pipette - no one else can, though. hmm. everyone else is doing surgery. if cells are extended wel, probably get pretty good rates with vaginal insemination. still not as good as fresh or chilled, though. in practice, you'll probably use only vaginal insemination. Brucellosis: brucella canis: zoonotic diseae, transmission is via ingestion, inhalation, or venereal transmission signs: itises and ilitie - orchitis, epididymitis, infertility, discospondylitis. dx: serology - need 3 neg results 1 month apart to prove neg rapid slide agglutination test: sensitive but not specific - you get false positives. do not condemn a positive tester - send for serology tube aagglutination test: same probs as RSAT - gives a titer result AGID test: gold standard - very specific; not sensitive. positive long after anmial is abacteremic. if inhouse test is positive, send off for AGID. definitive dx: bacterial culture. very time consuming, so we don't do this. tx: abx not curative. neuter affected animals; consider euthanasia due to zoonotic potential, spread to other dogs. Male cat: cats almost never show up here for repro stuff... we have an underdeveloped repro dept at VHUP. anatomy and physiology: puberty 8-12 mos two accessory glands - bulbourethral and prostate penile spines appear at 6-7 mos; androgen dependent; disappear after castration. male torties, calico cats: "I have a male calico cat - do you want it for research??" black and orange are on X can have normal male orange or black true tortie or calico means you need to have two X chromosome. these cats are sterile we do NOT want them for research :) cryptorchidism: less common than in dogs; testicles usually inguinal. if spines on penis, no testicles - cryptorchid. urine spraying: common reason for abandonment of cats. tx: castration, behavioral modification, drugs castration - decrease odor; decrease spraying behavior but not later on... drugs - megestrol acetate (Ovaban) but may cause permanent diabetes mellitus; valium; tricyclic antidepressants behavior modifications Case: this dog came in to VHUP 6 mos ago signalment: 12 yr old M intact lab "Carob" vomiting, anorexia, HL lameness, not doing well last normal 5 days ago hx: lethary progessing over 5 days, weak HL/limping x 5 days, inappetance progressing to anorexia, vomiting stool, urine normal no c/s vacc 5 mos ago no meds no garbage or toxin exposure leash walks only lives in Philadelphia area PE: T 103.3, P 175, RR 32, hydration poor, mm tacky, CRT >2 sec, scleral injection, lungs harsh esp ventrally, poor pulses, no murmur, painful abdomen, lymphadenopathy noted on rectal, testicles palpate normally, popliteal LN enlarged, joint effusion right hock, painful hocks bilaterally problems: vomiting, anorexia, dehydration, limping, harsh lung sounds, abdominal pain, sublumbar and popliteal lymphadenopathy, joint effusion, fever, scleral injection major rule outs for acute abdomen ddx: sepsis, pancreatitis, foreign body in GI tract with perforation, pyelonephritis, neoplasia, abscess, prostatitis, peritonitis rule outs for lymphadenopathy: lymphosarcoma (though usually multicentric, huge, firm) other neoplasia infection inflammation rule outs for joint effusion: sepsis, trauma, immune mediated disease... plan: cbc/chem/u/a; rads or u/s tx: fluids right away - IV Normosol R bolus, then 4 ml/kg/hr monitor pulses, temp, rr/effort results: PCV 50, TS 7.8, otherwise ok u/a: bili 4+, sg 1.044, blood 2+, protein 3+; sediment RBCs many, WBC >20 note: male dogs normally have bilirubin in urine; any febrile animal may be bilirubinuric. urine culture sample was lost CBC: neutrophilia w/left shift amylase and lipase - under 1000 so not too worrisome chem: creat 1, Na and CL low, total bili 1.0, alt/ast high, not in renal failure sample was hemolyzed abd rads: soft tissue mass - may be bladder, mareked prostatomegaly abd u/s: big hypoechoic area in prostate aspirated prostate - pus came out chest rads normal joint rads: effusion w/o erosion joint tap: healthy PMNs, healthy lymphocytes - non-infectious inflammatory dz this is probably an immune mediated response to the sepsis, not an additional autoimmune disease on top of prostatic abscess. meanwhile back in the cage: T 103.8, P 175, RR pant, pulse quality sucks, BG 65, scleral injection. he's getting more septic. ok, give abx - baytril IV, maybe add some ampicillin exploratory drained abscess took biopsy, cytology castration cytology: suppurative inflammation 1880 Rittenhouse recovery was slow - this is not unusual culture showed no growth but it was infection pathology - prostatitis, no neoplasia, bilateral interstitial cell tumors in testes - incidental. c&s - beta hemolytic strep, sensitive to TMS kept on abx 4-6 wks return for wash and culture in a week or two. ---break--- this is for fun not testable material ACVIM lecture: clinical approach to infertility in the stud dog dog comes in infertile - what do you do? the point of fertility is to make babies like matthew and theodore, here :) why is dog brought in? -unproven, no pregnancies ever -proven, but decreased pregancy rates -proven, but decreased litter sizes these appointments take some time get a super history logical thinking, 1/2 hr of time - you will dx most dogs on history! schedule for 1/2 hr, charge double exam fee! need to know about: breeding management - number one cause of infertility reproductive history on stud: libido (LH or testosterone problem?), breeding use (every day? what?), # pups/litter general health: any past illnesses? remember 75 days for sperm cycle. urination, defecation patterns toxin exposure - any disease in the family? any other infertility? :) environmental change trauma medication history - hormones, glucocorticoids (can suppress repro axis), including topical meds! borough's solution? family history PE: do a thorough one! systemic disease endocrinopathies (endocrine alopecia is a key sign) skeletal diseases prepuce - d/c, penile extrusion penis - color, trauma, neoplasia do good full pe feel vasa deferentia, do a good rectal, palpate prostate, LNs, testes then do semen collection: allows you to evalute libido, mounting ability, intromission semen evaluation semen collection/evaluation used to check infertile dogs, dogs with prostate dz. how to do it: relaxed atmosphere - no stress secure footing required teaser bitch - estrus bitch, or anestrus bitch with estrus vaginal secretions frozen on 4x4 gauze pads applied to perineal area. use artificial vagina - very helpful but not required clear, conical tube with side vent clean container with sharp edges wrapped water based lubricant without stuff in it things sperm hate: antiseptics, lubes, water, blood, urine, alcohol have someone holding the teaser bitch. let male sniff her let him get excited and mount get prepuce pulled back behind bulbus glandis stick penis into AV and watch for ejaculation ejaculate: three fractions: presperm - clear, small volume - drops to mLs sperm - rich, milky collect those together prostatic - collect this separately - esp if you think there is prostatic disease. usually there is a break before this comes out so you can put a separate container on. handler has hand behind bulbus glandis. can massage if needed to keep ejaculate going. always ensure prepuce is back over the penis before stud returns to cage. semen evaluation: microscope, slides, coverslips, blood cell dilution kit, hemocytometer, diffquik or wright's stain, transfer pipette should look milky,white. red = blood, yellow = urine. evaluation: look at color llook at cells motility morphology number rbc/wbc/bacteria/neoplastic cells? motility: total, progressive; examine several areas. look at it first b/c light, coldness will affect it. make wet mount, look at it. look at various areas of the slide b/c one area may be crappy and the rest fine. sperm agglutination - brucella causes this. other immune dz too sperm count - estimate volume from conical tube; for cell count use a unopette blood dilution kit and a hemocytometer. could dilute in water, too. count like you would blood cells. abnormal motility: consider contamination as most likely cause - water, antiseptic, lube - somethign got in there and decreased the motility. other causes - infection, inflammation, congenital problem. 99% of the time, contamination problem. morphology - stained, air dried smear, diffquick stain. look for primary and secondary abnormalities. her take is - they should look normal and if they do not, that's not good. primary - may be due to faulty spermatogenesis secondary - may be due to faulty maturation or bad handling head/tail separation - can be handling artifict leukocytes/erythrocytes in sample indicate a problem. cytology and culture of third ejaculate fraction normals - >70 to 80% progressively motile; 200-300,000 sperm/ejaculate GSD or bigger dog 500-600,000 75% or more should be morphologically normal if there are 60% normal, he may still get bitch pregnant,b ut it isn't normal. abnormal semen evaluation causes: overuse underuse (sperm sitting in storage area) inexperienced/frightened always repeat when evaluating sample, big factor is - is sperm present. if no sperm - aspermia - could be ejaculatory failure due to nerves or something. repeat collection after 48 hr rest. use comfy conditions and a real estrus bitch. you may have to go to his house. another cause of aspermia is retrograde ejaculation - hard to dx actual azoospermia - ejaculates but no sperm. a true ejaculate contains alk phos >5000 u/l, and carnitine azoospermia can be congenital (chromosomal sex disorders xxy, xx) or acquired - obstructive: inflammation, neoplasia, sperm granuloma, spermatocoele, aplasia failure of spermatogenesis: steroids - topical, injectable, oral; toxins, infections, immune mediated dz, endocrinopathy (cushings) teratozoospermia - morphologically abnormal sperm asthenozoospermia - lack of progressively motile sperm agglutinatoin: brucella, other - antisperm antibodies are present, can happne with any severe inflammation or immune mediated process. other rule out UTI, prostatitis semen culture - breeders want to do this. not that useful since semen isn't sterile. look for >10^6 bacteria to be significant. mycoplasma/ureaplasma - everyone said this was a problem and cultured everyone. bottom line- it has been proven that these are normal, resident flora in repro tract. nothing to do with infertility. breeder will ask you to do this test though. you can do it but results are not significant. other tests: u/s, epididymal aspiration (be careful about BTB), testicular biopsy (same thing), sperm function testing - new tests coming out, so far nothing useful; yearly brucella screening, biochemical database, karyotype. endocrine testing - this is too much for me. i am just not writing any more down. oh, ok. a few things. treatment: do nothing - wait 3-6 mos stop aall meds change environment a course of abx treat systemic illnesses/endocrinopathie hormonal therapy - frequent injections of GnRH/FSH/LH - or pumps, sustained release preps?? excellent breeding management is key - optimize the bitch if male is subfertile. use combined ejaculates with extenders. ----end----