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Objective: The aim of this study was to assess ovarian blood flow characteristics by applying transabdominal color Doppler ultrasonography during the course of the estrous cycle in gilts., Materials and Methods: Estrus and ovulation were synchronized in 15 pubertal gilts. During their consecutive spontaneous estrous cycle the gilts were sonographically scanned daily and ovarian blood flow was determined by imaging perfused areas of the ovarian hilus and the ovarian tissue. Recorded video clips were then analyzed for the parameters perfused area (Amix), blood flow velocity (vmix) as well as resistance (RIvmix) and pulsatility index (PIvmix) using the software PixelFlux ® ., Results: All blood flow parameters showed an estrous cycle dependent course, with Amix and vmix being highest in diestrus, followed by proestrus, while RIvmix and PIvmix patterns were inversely proportional., Conclusion and Clinical Relevance: Transabdominal color Doppler ultrasonography proved feasible to study ovarian perfusion characteristics in pubertal gilts. The ovarian blood flow was dependent on the stage of estrous cycle and highest in diestrus. The results of this study encourage the use of color Doppler ultrasonography to also investigate reasons of ovary-caused infertility including corpus luteum insufficiency or seasonal effects on ovary function., Competing Interests: None of the authors have any conflict of interest to declare., (© Georg Thieme Verlag KG Stuttgart · New York.)

The aim of this study was to investigate the relationship of progesterone (P) and luteinizing hormone (LH) during recognition and establishment of pregnancy in the gilt. Therefore, the effects of eliminating episodic LH pulses on P patterns were determined during early pregnancy. To this end, a slow-release GnRH implant deslorelin was used for GnRH down-regulation. A group of gilts (GnRHa, n = 8) was implanted with the GnRH-agonist on Day 11 of pregnancy, while a control group (C, n = 5) was treated with a non-impregnated placebo implant. Blood was collected via a vena cava caudalis catheter at 10-min intervals for 8 hr on Day 16 and 21 of pregnancy. As expected, the GnRH implant reduced LH secretion (p < 0.01) and abolished LH pulses completely at Day 16 and Day 21 of pregnancy. On Day 16, there was no difference in P levels between the treatments. However, on Day 21, the GnRH-agonist treatment led to significantly increased P concentrations (p < 0.01) compared with the control gilts. Progesterone was secreted in a pulsatile manner in both treatment groups and no relationship between LH pulsatility and P pulsatility was observed. In conclusion, abolishment of LH pulsatility did not affect the pulsatile pattern of P secretion but led to an unexpected overall increase in P on Day 21 of pregnancy; this effect was delayed and occurred 10 days after commencing treatment with the GnRH depot agonist. The elevation of P on Day 21 of pregnancy in the GnRHa group suggests either a reduced negative feedback effect or an increased autocrine response by the corpora lutea., (© 2018 Blackwell Verlag GmbH.)

Background: Interaction of spermatozoa and Chlamydiae spp. might contribute to reduced fertility in cattle. To proof this hypothesis, bovine semen was incubated with viable or heat inactivated Chlamydia (C.) abortus or psittaci (Multiplicity of infection = 1) and sperm motility was monitored with a computer-assisted sperm analyzer over 24 h. Additionally, the interaction with the spermatozoa was further investigated by means of light and transmission electron microscopy (TEM)., Results: Only viable Chlamydiae of both species decreased sperm motility and this only after about 9 h. Taking binding rates into account, the loss of sperm motility after about 9 h could likely be a consequence of Chlamydiae attachment to the spermatozoa. About two thirds of the Chlamydiae elementary bodies were bound to the front third of the sperm, the acrosomal region. No inclusions of Chlamydiae in spermatozoa were observed in TEM after 2 h co-incubation., Conclusions: As initial motility was not affected following co-incubation of viable Chlamydiae and bovine sperm, it seems likely that sperm could serve as a carrier/vehicle for Chlamydiae facilitating cervical passage of Chlamydiae spp. in cattle. Additionally, our results suggest that spermatozoa carrying Chlamydiae may have no initial disadvantage in reaching the oviduct, but are immotile at the time of ovulation what might have an impact on fertilization capacities of the individual sperm. Consequently, high concentrations of the investigated Chlamydiae in the seminal plasma or female genital tract might play a role in reduced fertility in cattle.

Competing Interests: Disclosure The authors report no conflicts of interest in this work.

Objective: To test for the feasibility of transabdominal Doppler sonography (color, power, pulse wave) to define uterine perfusion characteristics throughout the estrous cycle in gilts., Materials and Methods: A total of 15 gilts were synchronized for estrus and scanned in their following spontaneous cycle while being restrained in a purpose-designed mobile crate. To define uterine perfusion characteristics, vessels in between and within uterine cross-sections were imaged and recorded as video sequences to be analyzed by PixelFlux® software for perfused area (Amix), blood flow velocity (vmix) and intensity (Imix) as well as resistance (RIvmix) and pulsatility index (PIvmix)., Results: Color Doppler sonography proved to be the only feasible technique, as it was less affected by animal movements than power and pulse wave sonography. As determined by color Doppler sonography, all five parameters determined showed specific patterns through the estrous cycle, i. e. Amix, vmix, Imix were high in proestrus, decreased in estrus and remained low in midestrus and most parts of diestrus; RIvmix and PIvmix with inversely paralleled patterns., Conclusion and Clinical Relevance: This study has demonstrated that transabdominal color Doppler but not power and pulse wave Doppler sonography is feasible to be performed in crate-restrained gilts for studying uterine perfusion characteristics during the estrous cycle, and that changes of uterine perfusion over the course of the estrous cycle can be clearly followed by color Doppler sonography. Results encourage the use of color Doppler sonography for studying i. e. uterine capacity or uterus related infertility such as for cases of clinically unapparent endometritis., Competing Interests: The authors confirm that they do not have any conflict of interest., (Schattauer GmbH.)

Background: Maintaining optimal reproductive and litter performance is essential for meeting economic targets in commercial pig production. Treatment with exogenous gonadotropins in sows after weaning or in gilts after altrenogest treatment has been used to stimulate follicular development leading to more piglets born and eventually higher birth weights. The effect of peforelin on litter performance was investigated in 212 gilts, primi- and pluriparous sows in three herds. Animals were randomly allocated to three treatments 24 h after weaning: peforelin (P group), eCG (E group), and physiological saline solution (C group). Numbers of total, liveborn and stillborn piglets and mortality rate during lactation were recorded. Birth weights and coefficient of variation in weights within litter were assessed. All parameters were compared among treatments., Results: Over all parities, no difference was found among treatments in litter size nor mortality rate, but birth weights were significantly lower in the E group. Stillbirth numbers in pluriparous sows were 2.2, 0.9 and 1.4 for P, E and C groups, respectively ( p  = 0.04). Piglets in the P group had significantly higher live born birth weights in gilts, compared to the E group (1.36, 1.26, 1.32 kg ( p  < 0.02) for P, E and C group, respectively). No significant differences were found for the other investigated parameters., Conclusions: Peforelin treatment showed no improvement of litter performance compared to no treatment.

Background: Cobalamin (vitamin B12 ) and folate (vitamin B9 ) are important for the amino acid metabolism and nucleic acid synthesis. Immunoassays for the measurement of both vitamins in serum are routinely used in people, cats, and dogs, serving as indicators for clinical disorders including cobalamin and/or folate deficiency, small intestinal dysbiosis, or inadequate dietary supply of these vitamins. The analysis of these analytes may also be clinically useful in pigs., Objective: The purpose of this study was to analytically validate immunoassays for the measurement of cobalamin and folate concentrations in porcine serum, and to determine serum cobalamin and folate concentrations in healthy newborn pigs pre- and postweaning., Methods: Assay validation for both vitamins included the determination of linearity, accuracy, and intra- and inter-assay variability using serum samples from 10 pigs. Also, serum cobalamin and folate concentrations were compared in piglets between pre- and postweaning., Results: For both vitamins, observed-to-expected ratios for linearity and accuracy were 93.2 ± 14.3% and 100.3 ± 8.1% (mean ± standard deviation), respectively. Intra- and inter-assay coefficients of variation for serum were ≤ 8.7% and ≤12.5%, respectively. Significantly higher serum cobalamin and lower folate concentrations were observed in piglets at the time of weaning than at postweaning (P < .0061; P < .0001, respectively)., Conclusions: Both immunoassays are linear, accurate, precise, and reproducible for measurement of porcine serum cobalamin and folate concentrations. Piglets differing in age by only 12 days had significantly different serum cobalamin and folate concentrations. The implications of differing serum cobalamin and folate concentrations in pigs at different stages of life should be further investigated., (© 2016 American Society for Veterinary Clinical Pathology.)

Miniature pet pigs are becoming ever more common among the patients of small animal practices. Due to an advanced age attained in these animals in comparison with conventional pigs in pork production, miniature pet pigs develop age-related diseases that are infrequently observed in farm pigs, including neoplastic alterations of the uterus. We describe two cases of a uterine adenocarcinoma and a leiomyoma, respectively, in two miniature pet pig sows. Options and limits of diagnostic measures (unspecific clinical symptoms, x-ray, sonography and computed tomography) and therapy within the legal limits as well as the risk of an incorrect diagnosis based upon the more common differential diagnoses in swine medicine, including obstipation, cystitis and endometritis, are discussed.

The ability of peforelin (l-GnRH-III) to stimulate follicular growth, FSH release, and estrus in gilts after altrenogest treatment and in sows after weaning was investigated. In three farrow-to-wean herds, with at least 600 sows and average production performance, 216 gilts, 335 primiparous, and 1299 pluriparous sows were randomly allocated to three treatments: peforelin (M group: Maprelin), eCG (F group: Folligon), and physiological saline solution (C group). Animals were treated 48 hours after their last altrenogest treatment (gilts) or 24 hours after weaning (sows). The weaning-to-estrus interval, estrus duration, estrus rate (ER), pregnancy rate, and total born (TB), live born, and stillborn (SB) numbers were recorded and compared between treatments for the different parity groups (gilts and primiparous and pluriparous sows). Follicle sizes were measured in representative animals from each group on the occasion of their last altrenogest treatment or at weaning, and also on the occasions of their first (FS1) and second (FS2) attempted inseminations. Blood samples were taken to determine FSH concentrations at weaning and 2 hours after injection, and progesterone concentrations 10 days after the first insemination attempt. The relative change in FSH concentrations was calculated. Significant differences were found for ER within 7 days of weaning in pluriparous sows (95%, 91%, and 90% for the M, F, and C groups, respectively, P = 0.005). Gilts in the F-group had high TB numbers, and pluriparous sows in the M group had high SB numbers (TB gilts = 13.6, 15.4, and 14.9 [P = 0.02] and SB pluriparous sows = 1.8, 1.4, and 1.7 [P = 0.05] for the M, F, and C groups, respectively). The M group had the highest FS1 (for gilts) and FS2 (for pluriparous sows) values: FS1 = 5.4, 4.9, and 4.9 mm [P = 0.02] and FS2 = 6.8, 5.3, and 6.3 mm [P = 0.03] for the M, F, and C groups, respectively. There were no significant differences between the different treatments within each parity group with respect to any of the other variables. Overall, peforelin treatment had small but positive effects on the ER and follicle growth in certain parity groups but did not seem to affect litter sizes or FSH and progesterone levels in sows on the occasions of the corresponding examinations., (Copyright © 2013 Elsevier Inc. All rights reserved.)

In cattle, elevated blood serum concentrations of haptoglobin, an acute phase protein, have been demonstrated in association with several diseases, but not with lameness. Serum haptoglobin was measured in 60 Holstein dairy cattle diagnosed with lameness due to four claw disorders, pododermatitis septica (PS; n=41), pododermatitis circumscripta (PC; n=8), interdigital necrobacillosis (IN; n=7), papillomatous digital dermatitis (PDD; n=4). Haptoglobin was measured on day 1 (0-3 days after lameness was observed but before treatment) and on days 3 and 5. A total of 10 healthy cows served as controls (haptoglobin values <1.0 mg/dL). Each of the claw disorders was associated with elevated haptoglobin on day 1 (PS, PC, IN and PDD: 65.9%, 37.5%, 71.4% and 25.0%, respectively). Trimming and antibiotic treatment led to a reduction in the number of PS and IN cows with increased haptoglobin concentrations, respectively (P<0.05), but trimming did not lead to any reduction in cows with PC. The study showed that lameness due to claw disorders can be associated with a systemic acute phase response and elevated serum haptoglobin in dairy cattle. Based on the course of haptoglobin, treatments seemed effective for all claw disorders except for PC., (Published by Elsevier Ltd.)

Immunization against GnRH has been proven effective for boar taint removal, and long-term treatment with GnRH analogues has been shown to suppress GnRH dependent reproductive processes in several species. This study was conducted to treat boars (n = 5) with Suprelorin, i.e., an implant that contains 4.7 mg of the long-acting GnRH analogue deslorelin, and to test the effects on sexual function. Insertion of the implant occurred at the age of 5 weeks and animals were observed until market age at 26-27 weeks. Surgically castrated (n = 4) and intact boars (n = 3) served as controls. Testes growth was markedly reduced and steroidogenesis (testosterone, estrone, estrone sulphate, estradiol 17beta) as well as spermatogenesis suppressed in 4 of 5 GnRH treated boars, respectively. The remaining fifth boar resumed testes growth after week 17 of age and had high hormone concentrations when tested at weeks 26 and 27. Restoration of spermatogenesis was observed at 34 weeks of age. There were no effects of treatment on general health, nor were there local inflammatory reactions. Results indicate that suppression of sexual functions in boars due to long-term treatment with the GnRH agonist deslorelin through an implant such as Suprelorin is possible and can last for several months up to market age; thus it has potential as an alternative to other methods used for boar taint removal. Because the maximum duration of suppression seems to vary between boars, further studies are necessary to refine the treatment., (Copyright 2010 Elsevier Inc. All rights reserved.)

This study was conducted to determine uterine echogenicity by grey-scale analysis (GSA) and transcutaneous ultrasonography in pregnant sows (P-sows; n=16) and gilts (P-gilts; n=13) vs. cyclic gilts (C-gilts; n=9) between days 8 and 16 post ovulation (po) with the aims of testing for feasibility of uterine GSA and of gathering reference data. Estruses and ovulations were hormonally synchronized and the animals artificially inseminated. Ovulation was monitored by ultrasound. The equipment used was a HS 2000 ultrasound unit and a 5 MHz linear probe. Unit settings were standardized for all GSA scanning sessions and the animals crated during scanning. For GSA, cross-sections of the uterine horns were imaged, entirely defined as regions of interest, and pixel analyses done. A total of 342 scanning sessions were performed, 341 GSA accomplished, and 1-13 cross-sections analyzed per session. Comparison of coefficients of variation suggests that analysis of two cross-sections per session is sufficient for a reliable GSA per animal. P-sows and P-gilts were similar in their echogenicity course, but differed from C-gilts. Most noticeable, echogenicity declined in pregnant animals on day 12 po, while it increased in cyclic gilts. In conclusion, the study demonstrates that GSA using transcutaneous ultrasound is a feasible procedure for the determination of uterine echogenicity in the pig, and that pregnant and cyclic pigs differ in the uterine echogenicity, particularly during the time when maternal recognition of pregnancy occurs.

The urinary bladders of sows (n=10) without urinary tract infection (UTI) were longitudinally transrectally scanned after emptying and refilling with 200, 400, 600 and 800mL saline, and a volume dependence was found for bladder depth (BD), dorsal (dWT) and ventral wall thicknesses (vWT), wall regularity (WR) and mucosal wall surface (mWS). When another 31 sows without and 15 with UTI (as defined on the basis of high bacterial count and macroscopic/biochemical urine abnormalities) were compared for these parameters using BD as volume equivalent, no differences were found. Sows with UTI more often had moderate to high amounts of sediment than animals without UTI. Ultrasonographic assessment of dWT, vWT, WR and mWS of the urinary bladder of sows requires knowledge of bladder volume, and BD may be used as a volume equivalent. However, the parameters are inappropriate for the diagnosis of UTI as defined in this study, while moderate/high amounts of sediment seem to be indicative. Sediment can be visualized by transrectal scanning, but this is also possible using the transcutaneous route., (Copyright 2008 Elsevier Ltd. All rights reserved.)

Hysterosalpingo-contrast sonography is an accurate first-line method used to test for tubal patency in human gynecology. Tubal pathology occurs in dairy cattle and is a reason for infertility, but easy and accurate methods to test for tubal patency are not available in the living cow. In this study it was thus investigated if contrast sonography (CS) using Echovist-200 as the echo-contrast medium is a feasible procedure to test for tubal patency in dairy cattle. In eight repeat breeder Holstein Frisian cows, all of them being in behavioral estrus, Echovist-200 was given into the uterus and its exit into the abdominal cavity then imaged by transrectal conventional B-mode ultrasound, and used as indicator for tubal patency. Animals were slaughtered one day later and the genital tracts subjected to gross morphology and histology in order to confirm the results of CS. In two cows, CS was prematurely terminated after examination of one oviduct because of rectal bleeding, while CS completely failed in another cow because of poor image quality. In five cows, both oviducts could be examined by CS, respectively. A total of five oviducts were found patent by CS and confirmed by post mortem examination. Two out of five oviducts diagnosed as occluded by CS were morphologically intact and thus misdiagnosed. Of the three non-patent oviducts, two were occluded because of a hydrosalpings, respectively, while the third was inflamed. In conclusion, CS has been shown a feasible procedure to test for tubal patency in dairy cattle. Further studies with more animals are however recommended to warrant this result.

The mycotoxin zearalenone (ZEA) is a selective estrogen receptor modulator that can contaminate cereal feeds and lead to reproductive disorders. To determine effects of perinatal ZEA exposure on uterine expression of genes associated with endometrial development in the neonatal gilt, pregnant sows were fed ZEA (1500 microg ZEA/kg of feed/day) or vehicle from 14 days before farrowing through postnatal day (PND) 20-21, when neonatal uterine tissues were collected. At birth, gilts were cross-fostered to generate four ZEA exposure groups (n= 5-6/group): unexposed controls or exposures limited to prenatal, postnatal, or pre- and postnatal (continuous) periods. Results showed that at PND 20-21, uterine Wnt7a, Hoxa10, estrogen receptor alpha, and RXFP2 mRNA levels were decreased in neonates exposed continuously to ZEA (P < 0.05). Uterine RXFP1 transcripts were decreased in postnatally and continuously exposed groups (P < 0.05). Neonatal uterine Wnt4 mRNA levels were unchanged.

Porcine neonatal uterine relaxin receptor (RXFP1) expression is tissue compartment specific and estrogen sensitive. Here, procedures were established for laser microdissection, tissue capture, and quantification of the effects of perinatal exposure (14 days pre- to 21 days postnatal) to a selective estrogen receptor modulator of environmental origin, zearalenone (ZEA), on endometrial RXFP1 expression. Total RNA from captured endometrium was used to generate cDNA for quantitative reverse transcription-PCR. Cycle threshold values indicated that ZEA reduced (P < 0.06) endometrial RXFP1 expression on postnatal days 20-21.

A technology that allows for manipulating of oestrus and ovulation, and would then also allow for fixed-time insemination, can be of great benefit for swine farms that operate using sow batch management due, at least in part, to savings in labour and the production of large batches of evenly developed pigs. Thanks to the current knowledge on endocrine regulation of follicle development and ovulation, and the availability of numerous reproductively active substances such a technology is now available. It covers procedures for synchronising oestrus based on the use of altrenogest in gilts and of batch-wise weaning in sows, for stimulating follicle development using eCG and for inducing of ovulation using hCG or LH as well as GnRH analogues. While the procedures for oestrus synchronisation stand alone, other procedures require additional treatments. If fixed-time insemination is the goal, oestrus needs to be synchronised and follicular development and ovulation induced by the use of GnRH analogues and hCG with ovulation occurring within 36-42 hrs. It is a general recommendation to inseminate those animals twice, i.e. 24 and 40 hrs after ovulation induction. However, the aforementioned technology requires healthy animals and a solid management and cannot be used to compensate for poor management.

Although there are indications for venereal transmission of chlamydiae in cattle, epidemiological data on the presence of these bacteria in bulls and bull semen in particular is still incomplete. We investigated semen (n=120), preputial washing samples (n=121) and faeces (n=122) of bulls from six bull studs located within five Federal States of Germany for the presence of chlamydiae using omp1-PCR and partial omp1 sequencing. Blood serum was examined for chlamydial antibodies using an indirect ELISA (n=122). Chlamydiae were found in 11 (9.2%), 13 (10.7%) and 22 (18.0%) of the semen, preputial washing and faecal samples, respectively. Among individual chlamydial species identified, Chlamydophila (Cp.) psittaci predominated in semen and preputial washing samples, and Cp. pecorum in faeces. Cp. abortus was the third frequently observed species. Chlamydial antibodies were detected in a total of 62 (50.8%) bulls. Bull studs differed in regard to the number of bulls found chlamydia-positive in faeces and serologically positive. No correlation was observed between serological data and PCR of semen, preputial washing samples or faeces. Standard ejaculate parameters did not differ between bulls that were chlamydia-positive and -negative in semen. In conclusion, detection of chlamydiae in semen of bulls suggests a potential for venereal transmission. Chlamydiae appear to be widespread within the bull population in Germany. Serological testing failed to identify bulls shedding chlamydiae in their semen.

A field study was conducted to investigate the effectiveness of a treatment with altrenogest, eCG and hCG or the GnRH-analogue D-Phe(6)-LHRH to synchronize estrus and ovulation of sows diagnosed as non-pregnant in order to reintegrate them back into a scheduled fixed-time insemination program. Sows (n=531) diagnosed as non-pregnant by ultrasonography on days 21-35 after insemination were subjected to one of three treatments: (1) 16 mg altrenogest/day/animal orally for 15 days to block follicular growth, followed by injection of 1000 IU eCG intramuscularly (i.m.) 24h after withdrawal of altrenogest to stimulate follicular growth and 500 IU hCG i.m. 78-80 h after eCG to induce ovulation; (2) similar to (1) except that 20mg altrenogest and 800 IU eCG were used and (3) similar to (2) except that 50 microg D-Phe(6)-LHRH was used to induce ovulation. Females were artificially inseminated (AI) twice at 24 and 40 h, respectively, after hCG/D-Phe(6)-LHRH. Success of treatments was checked by ultrasonography of the ovaries. Rates of conception and farrowing (CR, FR), and number of total and live born piglets (TB, LB) were recorded and compared to those of synchronized first served sows. Females had differing ovarian structures prior to treatment. Altrenogest effectively blocked follicular growth in >80% of the females irrespective of dosage, but 16 mg increased the development of polycystic ovarian degeneration. Four to 18% of the females still had corpora lutea after altrenogest. Most females ovulated either between both inseminations or thereafter (P<0.05). Females treated with D-Phe(6)-LHRH tended to ovulate earlier than those injected with hCG. The CR and FR were up to 25% lower for sows diagnosed as non-pregnant than for sows after first service (P<0.05). Among sows diagnosed as non-pregnant the CR was higher in females treated with D-Phe(6)-LHRH (P<0.05). No differences were found in regard to numbers of TB and LB. In conclusion, a treatment with 20mg altrenogest per day per animal, followed by 800 IU eCG and 50 microg the GnRH-analogue D-Phe(6)-LHRH is appropriate to synchronize estrus and ovulation of sows diagnosed as non-pregnant. Whether there might be a need to feed altrenogest for a longer interval of 18 days has to be investigated.

Chlamydial infections of the genital organs cause reproductive failure in female pigs, and the uterus is recognized a target tissue for an infection. In contrast, information on the effect of chlamydiae on the porcine oviduct is patchily and inconclusive, although the bacteria are known to cause severe tubal defects in humans and laboratory animals. The aim of this study was to examine the segments ampulla (A), isthmus (I) and utero-tubal junction of the left (n=20) or both (n=22) oviducts, and uteri (U) from 42 culled repeat breeder pigs for chlamydiae using ompA-PCR, partial ompA gene sequencing, immunohistochemistry (IHC) and microscopy of tissue specimens for histopathology. As revealed by PCR, among a total of 26 chlamydia-positive females, 19 were tested positive in one or more segments of one or both oviducts, 14 were found positive in the uterus, and concomitant infections of both organs were observed in 7 of them. Sequencing of 33 PCR products revealed the following chlamydial species: Chlamydophila (Cp.) psittaci (n=18), Cp. abortus (n=2), Chlamydia (C.) suis (n=10), and C. trachomatis (n=3). Immunopositive staining was observed within the surface epithelium (in A, I, U), stromal tissue (in I, U) and muscular layer (in A, I, U). A total of 24 females had inflamed oviductal segments (in A and/or I) and 36 inflamed uteri. However, there was no relationship between histopathology and results of PCR or IHC. In conclusion, chlamydiae were found to infect oviducts and uteri of pigs. Further studies are required to clarify whether chlamydial infection causes specific histopathology and alters tubal function.

Although there are indications for venereal transmission of chlamydiae in pigs, direct diagnostic evidence on the presence of these bacteria in boars and boar semen in particular is still incomplete. We investigated boars from two studs (A, B) in semen (A: n = 174; B: n = 100) and faeces (A: n = 174; B: n = 24) for chlamydiae using ompA-PCR and partial ompA gene sequencing. Additionally, blood serum was examined for chlamydial antibodies using an indirect ELISA (A: n = 171; B: n = 62). Chlamydiae were found in 9 (5.2%) and 24 (24.0%) semen specimens, and in 71 (40.1%) and 2 (8.3%) faecal samples from boars of stud A and B, respectively. Regarding individual chlamydial species, Chlamydophila psittaci and Chlamydia suis were identified most frequently, with the former predominating in semen (in 23 out of 33 positive samples) and the latter in faeces (68/73). In contrast, Chlamydophila pecorum was found only sporadically. Chlamydial antibodies were detected in 80 (46.8%) and 6 (9.7%) boars of stud A and B, respectively. No correlation was observed between the data from serology and PCR of semen or faeces in either of the studs. In conclusion, detection of chlamydiae in semen of boars suggests a potential for venereal transmission. Whether the high overall prevalence of chlamydial infections reflects a general situation in boars needs to be investigated. Serological testing failed to identify boars shedding chlamydiae in their semen.

Oviduct epithelial cells are important for the nourishment and survival of ovulated oocytes and early embryos, and they respond to the steroid hormones estrogen and progesterone. Endocrine-disrupting polyhalogenated aromatic hydrocarbons (PHAH) are environmental toxins that act in part through the ligand-activated transcription factor arylhydrocarbon receptor (AhR; dioxin receptor), and exposure to PHAH has been shown to decrease fertility. To investigate effects of PHAHs on the oviduct epithelium as a potential target tissue of dioxin-type endocrine disruptors, we have established a novel telomerase-immortalized oviduct porcine epithelial cell line (TERT-OPEC). TERT-OPEC exhibited active telomerase and the immunoreactive epithelial marker cytokeratin but lacked the stromal marker vimentin. TERT-OPEC contained functional estrogen receptor (ER)-alpha and AhR, as determined by the detection of ER-alpha- and AhR-specific target molecules. Treatment of TERT-OPEC with the AhR ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) resulted in a significant increase in the production of the cytochrome P-450 microsomal enzyme CYP1A1. Activated AhR caused a downregulation of ER nuclear protein fraction and significantly decreased ER-signaling in TERT-OPEC as determined by ERE-luciferase transient transfection assays. In summary, the TCDD-induced and AhR-mediated anti-estrogenic responses by TERT-OPEC suggest that PHAH affect the predominantly estrogen-dependent differentiation of the oviduct epithelium within the fallopian tube. This action then alters the local endocrine milieu, potentially resulting in a largely unexplored cause of impaired embryonic development and female infertility.

This study was conducted to define the characteristics of the uterus of reproductively failed pigs by transcutaneous ultrasonography (SONO) in order to investigate the appropriateness of SONO to diagnose presumptive uterine disorders. Zearalenone (ZEA) is known to affect uterine function and causes endometrial liquid accumulation and was also determined. In 33 sows and 14 gilts, of unknown reproductive stages and culled for failing to conceive, the uterus was scanned transcutaneously and the females slaughtered on the same day or the day after scanning. Parameters determined by SONO were uterine echotexture (UET; graded 1 for homogeneous to 4 for highly heterogeneous), uterine size (US; expressed as the mean sectional area of two to three cross-sections of the uterine horns given in cm2) and intrauterine content. Post mortem, the ovarian structures were assessed and females grouped accordingly into those in estrus (n=2), early diestrus (n=14), diestrus (n=15), late diestrus (n=1), anestrus (n=10) and having polycystic ovarian degeneration (n=5). The uterine weight (UW) was recorded and uterine specimens microscopically evaluated for an endometrial oedema (EO; grades 1 for none, to 4, if an oedema was clearly evident) and for immune cells to assess endometritis. Total ZEA was analysed in bile and females with >or=50 ng/ml judged as positive. The uterus could be examined in all animals. UET, US, UW and EO was found to be different between groups, and a positive correlation (P<0.001) established for US and UET (r=0.71), US and EO (r=0.51), UET and EO (r=0.57), US and UW (UW=357.6 x US(0.801); r=0.88). One female had intrauterine fluid and an acute-chronic endometritis diagnosed. Almost all females had a chronic endometritis and a majority found ZEA positive. No differences were observed between groups and a relationship between ZEA or chronic endometritis and UET, US, UW and EO were not established. In conclusion, transcutaneous SONO is appropriate to examine the uterus in reproductively failed pigs on farms, and the estimation of UET and US gives information on EO and UW. Intrauterine fluid is indicative for a severe uterine inflammation. Since groups differed in UET and US, but were equally ZEA positive and the uteri chronically inflamed, an UET and US specifically associated with ZEA or chronic endometritis is questioned.

This study investigated the viability of embryos from non-stimulated 2-3-month-old calves generated in vitro using oocytes from follicles of defined size in terms of their ability to produce full-term pregnancies. Ablation of follicles>or=4 mm was used to induce the emergence of a new follicular wave, and calves (n=3) were laparoscopically punctured three times at 7-day-intervals to recover cumulus-oocyte-complexes (COCs) from follicles>8 (group A) and between 4-8 mm (group B). Calves were aged 49, 56, and 80 days, respectively, at first recovery. Morphologically intact COCs were subjected to in vitro maturation, fertilization, and embryo culture, and compact morulae/blastocysts were transferred on day 7 post-insemination to synchronized virgin heifers. Blood typing was used for maternity analysis. A total of 29 COCs were recovered, 21 cultured, yielding 11 cleaved embryos (52.4%) and 6 compact morulae/blastocysts (28.6%). No differences were observed between groups. Transfer of the 6 embryos to 5 recipients resulted in three pregnancies (one from group A and two from group B). Two normal male offspring (both from group B), with birth weights of 44 and 51 kg, were born, and two donor calves, aged 56 and 59 days, were identified as the dams. In conclusion, the results demonstrate that embryos generated in vitro from oocytes from non-stimulated calves at an age younger than two months are viable in terms of their ability to produce full-term pregnancies, and suggest that the viability of calf embryos is not related to follicle size.

This study investigated the relationship between follicle size (FS) and developmental competence of calf oocytes. Cumulus-oocyte-complexes (COCs) from follicles>8 (L-COCs; n=19), 4-8 (M-COCs; n=54), and 2-3 mm (S-COCs; n=155) were recovered from non-stimulated 1-4 months old dairy calves post mortem and ex vivo (laparoscopy), and in parallel from slaughtered adult cows from follicles of identical size categories [>8 (n=91); 4-8 (n=138); 2-3 mm (n=193)]. Morphologically intact COCs were subjected to in vitro maturation, fertilization, and embryo culture. Cleavage rate (CR; 46 h post-insemination=p.i.), rate of morulae/blastocysts (M/Bl; day 7 p.i.), and blastocysts (Bl; day 9 p.i.) were recorded. FS had no effect on the CR in calves. However, calf L-COCs yielded the highest rates of M/Bl and Bl compared with the two other size categories (P<0.05). In contrast, calf S- and M-COCs gave similar rates of M/Bl, whereas the proportion of Bl was lowest for S-COCs (P<0.05). This was almost identical to findings in cows, except that the CR was highest for L-COCs and M/Bl yields were lowest for S-COCs (P<0.05). There were no differences between calf and cows with regard to CR for the respective FS categories. L-COCs from calves and cows yielded similar rates of M/Bl and Bl, whereas calf S- and M-COCs yielded lower rates of Bl than S- and M-COCs from cows and a lower rate of M/Bl when S-and M-COCs were analyzed as one group (P<0.05). Whereas the CR was similar in calves and cows, calf COCs yielded lower rates of M/Bl and Bl (P<0.05). In conclusion, the results show that the developmental competence of calf oocytes is higher in those derived from follicles larger than 8 mm, and thus are almost equally as competent as cow oocytes derived from follicles of identical size. This suggests that calf oocytes acquire developmental competence within the large follicle, potentially due to a process similar to prematuration of the oocyte in the adult cow. It is proposed that procedures that facilitate prematuration, such as "coasting" following a preceding superstimulation, might increase the developmental competence of calf oocytes.

In two experiments (EXP), 44 and 52 crossbred gilts (mean age+/-S.D. and weight+/-S.D.: 204+/-22 and 203+/-9 days, 114+/-13 and 127+/-12 kg, respectively, in EXP 1 and 2) from four farms were examined by means of transcutaneous ultrasonography (US) to define the characteristics of the ovaries and the uterus (echotexture, size) and to investigate the appropriateness of US to determine sexual maturity. Gilts were judged as prepubertal [PRE; follicles 2-5 mm (F2-5) only] or pubertal [PUB; F7-8, corpora lutea (CL), corpora haemorrhagica (CH)] at the first (PUB-1; EXP 1) or a subsequent estrous cycle [PUB-2; additionally corpora albicantia (CA); EXP 1] by US, and results were verified by postmortem examination (EXP 1), or progesterone analysis and detection of estrous signs (EXP 2). Accuracy of US was 100% for PRE and PUB (both EXP) and 77.3% for PUB-1 and PUB-2 (EXP 1). PRE and PUB with CL/CH had uteri of homogeneous, PUB with F7-8 of heterogeneous echotexture. The size was expressed as the mean sectional area (SAsono) of 2-5 cross-sections of the uterine horns (calculated by multiplication of 1/2 the maximum x the minimum dimension of the cross-sections x pi). SAsono corresponded with the sectional area of postmortem dissected transverse uterine segments relatively with r=0.92 (P<0.0001; EXP 1). Mean SAsono (both EXP) and mean uterine weight (EXP 1) were PRE

This study was aimed firstly, to examine the ovaries in non-pregnant first served sows and gilts by transcutaneous ultrasonography and secondly, to evaluate the suitability for this procedure to be performed routinely on farms. Two thousand five hundred and twenty-three females on a 1250-sow unit, synchronized with Regumate (gilts only) and/or gonadotropins (sows and gilts) not detected as returned to estrus by daily boar contact prior to scanning were ultrasonographically tested for pregnancy between days 20 and 114 postinsemination (p.i.). Of 256 sows (S) and 130 gilts (G) found to be non-pregnant the ovaries were visualized in 87.1 and 80.0% of them, respectively. Ovarian findings were: corpora lutea (CL); follicles of 2-6mm (F(2-6)); peri-ovulatory ovarian structures (POS; comprising follicles of 7-8mm and corpora haemorrhagica); single cysts (SC); oligocystic ovarian degeneration (OOD) and polycystic ovarian degeneration (POD). Their incidence was: CL>F2-6>POS>POD ( P<0.05 ) in both S and G. POD and SC plus OOD were more frequently in S ( P<0.05 ). The ovarian findings were related to the intervals of regular (days 18-25 p.i. (R1), 38-46 p.i. (R2)) and irregular service returns (days 26-37 p.i. (IR1), 47-114 p.i. (IR2)). Comparison within intervals: CL tended to be more frequently with P<0.05 only at IR2 in S. Comparison among intervals (R1 to IR2): The percentage of females (1) with CL tended to increase (S and G) and (2) with F2-6 plus POS decreased significantly (S; P<0.05 ) or tendentiously (G). SC plus OOD was higher before R2, POD after IR1 (S and G; P<0.05 ). In conclusion, the results indicate a high heterogeneity of ovarian structures in non-pregnant first served sows and gilts up to day 114 after service and suggest CL as an important cause for a delayed and, rather than POD, a failed service return. The results further demonstrate that transcutaneous ultrasonography is an appropriate and recommended method for examining the ovaries on farm in female pigs with reproductive failures.

We employed molecular and immunological techniques to investigate the expression of INSL3, a member of the insulin-like superfamily, in prepubertal testis, postpubertal testes exhibiting normal and disturbed spermatogenesis, and cryptorchid testes of male horses. In addition, the partial cDNA coding sequences of the equine homologue of the human relaxin/INSL3-receptor Lgr8 were determined. Nonradioactive in-situ hybridization with a cRNA probe for equine Insl3 and immunohistochemistry with a specific rabbit INSL3 antiserum localized Insl3 transcripts and immunoreactive INSL3 ligand to Leydig cells in all types of testes investigated. Quantitative polymerase chain reaction analysis revealed a down-regulation of Insl3 and an up-regulation of the relaxin/INSL3-receptor expression in unilateral cryptorchid versus descended testes. Western blot analysis of protein extracts from adult normal and cryptorchid testes and prepubertal testes showed a single immunoreactive band at 14.5 kDa, which correlates with the predicted size of equine proINSL3. Densitometric analysis of Western blot data of adult normal testes revealed significantly stronger expression of immunoreactive proINSL3 as compared to extracts derived from cryptorchid or prepubertal testes. Thus, decreased expression of immunoreactive INSL3 in cryptorchid and prepubertal equine testis is transcriptionally regulated. The detection of transcripts for equine Lgr8 in the testis has identified the testis as a potential target of INSL3.

Employing postpubertal testicular tissue, we determined the cDNA coding sequence of a truncated canine relaxin-like factor (RLF) consisting of a signal peptide of 28 amino acids (aa), a B-domain of 23 aa, a truncated C-domain of 34 aa, and an A domain of 26 aa, respectively. Within the B-domain of canine RLF, the putative relaxin receptor binding motif contained a single substitution with the C-terminal arginine replaced by a serine residue, and the putative RLF receptor binding motif was truncated. Leydig cells specifically expressed RLF in the normal postpubertal and cryptochid testis as well as in testicular Leydig cell adenoma. The epididymis was an additional source of RLF in the dog. In the female reproductive tract, expression of immunoreactive RLF and relaxin were compared. Within the ovary, RLF, but not relaxin, was detected in follicular theca interna and granulosa cells and the corpus luteum. In the nonpregnant uterus, luminal and glandular epithelium coexpressed RLF and relaxin. Uteroplacental tissue at early stages of gestation revealed RLF expression in the proliferative fetal villous cytotrophoblast and in maternal uterine cells. In the mature canine placenta, the trophoblast surrounding the maternal blood vessels and the hemophagous cytotrophoblast of the paraplacental zone expressed RLF. Canine relaxin was absent in the paraplacental areas. Western analysis of placental tissue extracts revealed the presence of specific immunoreactive bands likely resembling unprocessed and enzymatically cleaved RLF. Differential expression of RLF and relaxin appears to reflect distinct autocrine and paracrine functions of RLF in canine reproductive tissues.

The relaxin-like factor (RLF) is one of the insulin-like molecules, which also includes insulin, insulin-like growth factor I and II, placentin, and relaxin. Employing RT- and RACE-PCR on RNA isolated from goat testicular tissue, we report the cloning and nucleic acid sequence of goat RLF. The caprine RLF cDNA coding sequence consisted of 396 base pairs encoding a peptide of 131 amino acids. Caprine RLF showed the highest homology in nucleic acid and amino acid sequence with bovine and sheep RLF, suggesting conservation of the RLF gene among ruminants. Northern blot analysis revealed a single 0.9 kb RLF transcript expressed in the goat testis but not in the epididymis, liver, or muscle tissue. Only a single goat RLF gene is present in the goat genome as determined by Southern blot analysis. Employing nonradioactive in situ hybridization for goat RLF mRNA and immunohistochemistry for 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and 17alpha-hydroxylase, we identified the Leydig cells as the sole source of RLF mRNA in the goat testis.

The cat placenta is known to secrete large amounts of relaxin. We employed uteroplacental tissue at approximately Day 35 of gestation to determine the nucleic acid sequence of feline preprorelaxin using reverse transcription- and rapid amplification of cDNA ends-polymerase chain reaction. Feline preprorelaxin cDNA was found to consist of 540 base pairs encoding a protein of 180 amino acids (aa). We identified a signal peptide of 25 aa, a B domain of 33 aa, a C domain of 98 aa, and an A domain of 24 aa. The putative receptor binding region in the N'-terminal part of the B domain contained one substitution from the classical GRELVR motif (L-->F). Feline preprorelaxin shared highest homology with porcine and equine preprorelaxin. Northern analysis revealed a specific 1-kilobase transcript present in total RNA of feline uteroplacental tissue but not of liver tissue. Nonradioactive in situ hybridization was used to localize relaxin mRNA, and immunohistochemistry was used to localize the relaxin hormone and cytokeratin, in tissues of the feto-maternal interface recovered from two queens at Day 35 of gestation. Specific hybridization signals for relaxin mRNA were exclusively detected in cells located in the lamellar placental labyrinth but were absent from other placental and nonplacental uterine parts. The cells expressing relaxin mRNA also displayed immunoreactivity for cytokeratin and were, therefore, identified as trophoblast cells. Immunoreactive relaxin colocalized in those placental areas expressing relaxin mRNA. Trophoblast cells located at the villous chorioallantoic tips invading the endometrium and extravillous trophoblast cells in the junctional placental zone were devoid of relaxin.

This paper presents the results of 16,592 pregnancy diagnoses in 4086 gilts and 12,506 sows. The animals were scanned transcutaneously between days 20 and 110 after mating or insemination (m/i). Animals being obviously pregnant or nonpregnant were scanned once, whereas sows which could not be recognized definitely as pregnant or not were characterized as animals with equivocal status and examined once again seven to 14 days later. Between days 20 to 23 after m/i pregnant sows were obviously identified to 99.5% (3336/3352). Nonpregnant sows were clearly recognized to 97% (3145/3243) between days 20 and 28 after m/i. Beginning on days 26 and 32, respectively, and later in gestation, pregnant and nonpregnant sows could be diagnosed to 100%. Confirmed through the re-examination, altogether nine (of 16 ambiguous pregnant sows) and 19 (of 97 ambiguous nonpregnant sows) animals were diagnosed false pregnant and nonpregnant, respectively. To avoid incorrect diagnoses or assessments, numerous phenomena should be attended. On days 20 to 23 after m/i, sows which did not establish pregnancy often show middle- or nonechogenic areas in the uterus and/or between the uterus loops. Since this can hardly be distinguished from pregnant uterine structures, it is recommended to examine the ovaries as well. Pregnant animals with signs of embryonic death should be grouped separately and examined twice, because it is unpredictable whether the pregnancy will stay intact or not. In conclusion, the ultrasonography is well suitable to detect pregnant and nonpregnant sows and can be applied very early in gestation.