----start---- repro 9.14 we were talking about BSE before... we were working on the semen evaluation component sperm morphology is a very important parameter. morphology is characterized as normal, or primary or secondary abnormalities, based on where they occur - primary during spermatogenesis in the testicle (although there is no real/true evidence for this) so we changed the terminology to major/minor abnormalities - major is that which we know causes fertility problems, and minor may not be involved with fertility as far as we know. most important thing: percentage of normal sperm. ** normal sperm % is related to fertility we talked about motility before - the thing with that is it has to be progressive motility. the progressively motile sperm are a subset of motile sperm. we want normal morphology and progressive motility to apply to at least 60% of the sperm. slides of sperm - mostly normal, nice and straight, one has abnormal acrosome, one has bent nucleus, one has hairpin fold in tail. these are major abnormalities. head defects are most important - double headed sperm is rare and bad. slide: WBC in ejaculate. these are abnormal findings. you're "allowed" to have 3 or 4 WBC/HPF but really any are abnormal slide: phase contrast picture of abnormal sperm with frayed midpiece behind head. another one with a bent midpiece, very abnormal, another with a proximal cytoplasmic droplet (minor) head morphology: now using automated morphometry to evaluate these suckers. also fluorescent acrosomal stain is sometimes used. acrosome is absolutely essential for fertility. Testicular size and volume: we look at this very critically when we do BSE, b/c larger testicles make more sperm. size may vary w/season (stallion, ram) the volume of parenchyma correlates to daily production of sperm. we use the volume to estimate the daily production of sperm - look for discrepancy. stallion: scrotal width > 8.0 cm; testicular volume - caliper, U/S, measure height, width, length of each testicle and calculate volume with this formula: 4/3 ¼ x (h/2 x w/2 x l/2) (that's 4/3 pi...) this is more accurate than just scrotal width. bull/ram: scrotal circumference, breed/age specific cutoff values; approximately >32 cm. use special scrotal measuring tape if you want :) in bull, breeds have different shaped testicles. also different timelines for maturity. one breed has long cylindrical testicle and circumference isn't as good a measure. slide: measuring scrotal circumference on a ram.pull scrotum out behind animal, and prior to measuring palpate the testicles, epididymis, spermatic cord, inguinal rings, ductus deferens, any abnormalities....also ensure testicles are freely mobile within the scrotum - important for temperature regulation. then place scrotal tape around circumference of scrotum at widest point. hold scrotum on the sides, without spreading testes apart. another way of measuring testicular size - used in human med alot! - uses beads of different sizes - compare size of testicle to the beads estimating daily sperm output: stallion gold standard: measure it every day - semen collection sid/bid x 7-10 days, take average of your totals, and that is an approximation of what horse makes daily. this technique stabilizes extragonadal sperm reserves, and is truest approximation of daily sperm production. but this is expensive and soemtimes dangerous... so, meausre h w and l of each testicle and put it into formula to get a predicted daily sperm output (DSO) number. compare to actual count. 4/3 ¼ x (h/2 x w/2 x l/2) = a [.024 x a] - 0.76 = predicted DSO compare the predicted DSO to actual sperm count. case 1: 2 yr old stallion, black and white with white strip/snip. novice breeder, good libido, normal mating behavior vol: 55 ml 40 ml total #: 12.1 6.2 % mot: 85/70 75/70 % normal: 47 51 #MNPM 4.6 2.2 he's only 2, so it's ok to have under 60% passing = minimum 1 billion MNPM in second ejaculate % normal cells a little low, but he's young. he passes but should recheck in a year. microbial cultures - done routinely, very important, done in stallions and bulls, not other spp. stallion: venereal pathogens prewash: fossa glandis, penis, prepuce preejaculate: urethra postejaculate: urethra then on second ejaculate, before it you take external cultures again. this is a post washing culture. we're most interested in pre and post urethral samples - to see if pure growth of bacteria comes from urethra post ejaculation. you expect flora to be the same pre and post, but if there is a change, there is probably an accessory gland infection. most animals do not have pathogenic bacteria in there. pseudomonas, klebsiella, and e.coli in pure growth are pathogenic in stallions. bull: only done when indicated preputial swabs or washes - diamond's medium, clark's medium. stick swab up sheath prior to erection, and either swab it or flush it, and culture. only interesting when bull will be in herd situation doing natural breeding, not for AI donors. major organisms: tritrichomonas fetus (bull = asympt carrier) and campylobacter fetus (aka vibrio fetus; bull = asympt carrier). need two different media - trich is protozoal and needs diamond's medium. infertility workup: semen evaluation daily sperm output rectal exam cultures ultrasonography endoscopy (not in bulls)(fairly often but not routine in stallions) - would use if there were blood or pus in stallion ejaculate, or history of stones in urethra or something. advanced diagnostics: automated motility analysis, endocrinology (FSH, LH, testosterone, estradiol, inhibin); karyotype, DNA index; Y chromosome fragments (useful with intersex conditions, chromosomal chimerism); sperm chromatin structure (highly correlated with fertility); electron microscopy; fluorescence microscopy (acrosomal proteins and membrane structures). generally speaking, there are not many hormonal problems in stallions. most cases of infertility will not be dxed with hormone analysis. final assessment and conclusion: classify your individual as satisfactory, questionable, or unsatisfactory for stallions; for bulls: satisfactory, unsatisfactory, or classification deferred (food animals are culled if they can't stand up to fertility - if bull, boar, ram, other can't be found fertile, he'll be culled. so a male's genes aren't as important as in horse business.]. stallion: amusingly, horses with highest stud fees tend to be subfertile...that's why stud fees are so high. to be satisfactory, you need a minimum of 1 billion MNPM (morphologically normal, progressively motile) sperm in second ejaculate; ability to breed a book of reproductively normal mares (40 by natural cover or 120 by AI) -(should get at least 70% pregnancy rate). what if he's bred to 80 mares? Some horses do not do as well if they have to breed more mares. they are changing the classification, now, to 80 instead of 40, actually... in handout - stuff we're not talking about - read but do not study. estimation of stallion's book - when stallions are classed as questionable - under a billion MNPM - you have to change # of mares. we're not going to discuss this. 1982 arabian stallion bred 4 mares/yr x 8 yrs up til 1994 no pregnancies 1995 no evidence of accessory gland pathology semen evaluation - no sperm at all in either ejaculate, normal volume. well, is he making any sperm? how do you find out? testicular biopsy is method of last resort. another test is used to look for blockage of duct - alkaline phosphatase! if you run an alk phos on this horse, you will find it to be normal. the normal level is >10,000. if you do not get that much (say, less than 100 u) there is probably a blockage in the duct system. this horse did have high alk phos, though - so fluid in semen was coming all the way from epididymis (which is where alk phos is made). so, you could do epididymal aspirate - but if you tear the artery or rupture the epididymis, you create big problems. other things to do include hormone assays - testosterone, FSH, LH. ultimately, testicular biopsy showed he had totally shut down sperm production. case 3 1994 scotch highland bull 2 yrs old - very slow to mature breed, so could be immature as well as having a problem. hx infertility and dysuria 0 pregnancy out of 8 cows discomfort during urination per owner electroejaculation: 2 ml, 75/70 motility, 65% NM, 15% WBC; scrotal circumference 37 cm (ok) so, he'd probably pass BSE, but he isn't getting anyone pregnant and has WBC in semen. turned out to have urinary infection of some kind. treated that for about a month with TMP-sulfa 1996 breeding season - 5/12 heifers pregnant and still breeding ---break---- note: strep also a stallion pathogenic bacteria note: stuff in back of handout is just for your info Preputial/penile problems 1. hematoma of the penis, aka "broken penis" swelling cranial to scrotum bulls: thrusting injury stallions: trauma/kick (rarely occurs) it's caused by rupture of tunica albuginea treatment: benign neglect or surgical debridement and lavage followed by rest. slide: huge swelling cranial to bull scrotum. takes months to resolve - if bull isn't valuable, usually will be culled. sequelae - preputial/penile abscess, prolapse 2. prolapsed penis/prepuce -sequela to hematoma or trauma -bulls mainly, esp bos indicus breeds which have long prepuces -predisposes to paraphimosis (dependent edema pulls penis out, can't retract it) 3. adhesions: sequelae to above in bulls, rams, stallions. -trauma or breeding injury (slide: tear in preputial fornix. ouch. need to extrude penis and put ointment on to prevent adhesion formation) -sigmoid flexure usually site, sometimes glans penis. when sigmoid flexure is site, can't retract or extend penis often it's all together - hematoma predisposes to prolapse, which predisposes to trauma, and adhesions occur... reefing/circumcision - removes a swath of skin from the penis - used to get rid of the preputial tear described above.... 4. deviations (bulls) 5. persistent frenulum (bulls) - normally separates at puberty; can correct surgically (just snip with scissors) 6. spiral/corkscrew penis (bulls) - normal post-breeding, abnormal prior to intromission, tx surgical implant, fascia lata, other. slide: dangling corkscrew penis in bull slide: rare condition - diphallus - two penises on one bull! [crude joke not told.] paraphimosis: inability to retract penis - secondary to trauma, stenotic preputial orifice, phenothiazine tranquillizers in stallions (or geldings) - so do not use phenothiazines in male horses (although, everyone uses ace to clean a gelding's sheath...but it isn't wise) phimosis: inability to extend penis - trauma, constriction of preputial opening, short retractor penis muscle or short penis. paraphimosis in stallions: tx fairly routine prevent dependent edema! apply ointment to reduce adhesions replace penis, place pursestring suture of umbilical tape or heavy suture around prepuce - use local anesth. preserve ability to urinate freely! +/- open and cleanse daily hydrotherapy slides: paraphimosis in horses. note with paraphimosis the penis is flaccid. another condition called priapism is persistent erection. slide: priapism secondary to acepromazine in gelding. males cycle through erections around the clock. this is true. geldings/castrated males still get erections now and then, but much less frequently than intact males. this poor guy got an erection while under influence of acepromazine. owners left it for 3 days before bringing him in. so blood in corpus cavernosum clotted. requires surgery. pharmacological tx for priapism can be used within 3 hrs of onset (alpha adrenergic agonist), but after 3-5 hrs you end up with real problems. this horse they did surgical irrigation of corpus cavernosum, penile bandages - it was a horrible mess. owners had to do a lot of bandage changes, stall rest x 6 weeks...then they planned a Bolz procedure - penile retraction procedure - so they went in and pulled the penis in from above. other option was penile amputation. the retraction procedure went beautifully but horse broke his leg in recovery and shattered his tibia and had to be put down. it was pretty heartbreaking. treatment of penis/prepuce lesions 1. support dependent structures - usually slings, sheets used, "jock straps" 2. hydrotherapy 3. antibiotics/ointments locally to prevent adhesions 4. systemic antibiotics to prevent infection with wounds 5. sexual rest for weeks to many months when penis is bandaged, be sure to watch for urine scald and attempt to bandage appropriately to allow urination. penile amputation is a serious thing - very prone to stricture and urinary blockage. balanoposthitis - pizzle rot - rams, dogs, bulls, stallions - ammonia/urea scalding of glans. bacterial, venereal. in bulls - IBR-IPV virus, in stallions habronemiasis (summer sores- fly lays egg in urethra, migrates into tissue, causes extensive damage, very painful, stallion reluctant to ejaculate or urinate and may have bloody ejaculate - blood kills sperm). habronemiasis ddx squamous cell carcinoma. if you see this tx for habronemiasis first, if it won't go away probably SCC... balanitis - stsallions - due to coital exanthema (EHV3), poxlike and transmissible to mares; self limitied 10-14 days. painful pustules and erosions on penis or vulva. sarcoid: stallions- space-occupying lesion, any skin surface affected. neoplasia: transmissible fibropapilloma, most common tumor in young bulls, esp housed together. viral. can be very extensive. squamous cell carcinoma - most common tumor of stallions, not usually malignant ??, noticed by bleeding at breeding. penis should be amputated. or, cryotherapy is effective for discrete lesions. Scrotal problems: swelling - external problems: frostbite, skin necrosis, dermatitis, heat, trauma, castration related problem, dependent ventral edema swelling - internal problems: hematocoele, hydrocoele, hernia, peritonitis slides: stallions with big scrota. you have to ultrasound these to figure out what is going on, where the actual swelling is. orchitis, cord torsions make the testicle itself get large. testicular problems: cryptorchidism - unilateral most common. horse, pig, dog most often. in horse, inheritance unknown, in pig, single sex linked recessive gene; in goat, recessive, linked to poll gene; horse polygenic. equine cryptorchidism - ridgeling, rig, high flanker (where testicle is up near inguinal ring). most cases unilateral, left retained left testis most often abdominal retained right testis most often inguinal descent may occur up to two years estrogens may be diagnostic- abdominal testicle may make high amt of estrogen. testosterone is diagnostic - if animal has no external testicles but has testosterone level, he has a retained testicle. dx: testosterone levels do in AM b/c that's when the level should be lowest- gelding < 100 pg/ml, stallion > 500 pg/ml hCG challenge test: 5-10,000 IU IV, get baseline and 90 min sample; level should double in 90 min in animals with a testicle. hCG is similar to LH, is made by pregnant women, and it stimulates testosterone production from the testicle. cryptorchidism tx: surgical excision - inguinal approach or ventral laparotomy - inguinal if epididymis descended. U/S or rectal palpation may help to locate it. GnRH therapy, multiple dosing - minimal success reported, mostly anecdotal, no controlled studies, probably not that useful. but might be worth a try prior to surgery. try not to do these surgeries in the field, either, b/c it's hard to have enough time under injectable anesth to finish.... hydrocoele - fluid around testicle. here we see an asymmetrical scrotum, and lots of fluid visible on u/s. sometimes you can find a hernia and small intestine or something on u/s. inguinal hernia in foal - with young animals, some of these guys that are days or weeks old- you can push it in manually, bandage it, and it will close. sometimes, people take a tennis ball and put it against the scrotum and tape it there, and that will reduce the hernia. slide: bull scrotum - neck of scrotum should narrow. if it doesn't, there is probably a problem. this one has a hernia or a hematoma or abcess or something. - turned out to be a huge blood clot. asymmetry is always weird - may be hard to tell which side is abnormal,though. scrotal dermatitis can be sequela to frostbite, chemical burn...can shut down spermatogenesis due to heat. torsion/rotation: stallions, men, dogs - >180 degree impedes blood flow. can be up to 720 degree rotation! severe, acute pain, scrotal swelling. treatment - surgical correction or removal testicular arteritis - migrating strongyle larvae, EVA (viral arteritis) varicocoele - spermatic vein thrombosis, rams/stallions, adverse effects on thermoregulation, uncommon neoplasia - interstitial cell, sertoli cell, or germinal cell origin most are benign, space occupying lesions. seminoma in stallion may be metastatic sertoli cell tumor in dogs metastatizes bull leydic cell tumor- yellow on cut section. most common in bulls - interstitial cell tumor, seen in older animals, not seen much b/c most bulls culled prior to 7-10 yrs of age. stallions: teratomas and dermoid cysts in retained testicles; seminoma most common in stallion boar/ram - rare dog- sertoli tumor most common, usually in retained testicle, feminization occurs, atrophy of contralateral testicle ---end----