How FTAI and FTET impact reproductive efficiency of Brazilian dairy herds

Background: Throughout dairy cows evolution, milk production was always the key point to select the superior animal. Currently, several evidences has shown that high milk production have intensively contributed to the decline of dairy cattle fertility. Beyond milk production, dairy cows have their reproductive performance impaired by another factors, heat stress and repeat-breeding. Methods like fixed time artificial insemination and embryo transfer were developed to minimize the effects of these factors, and improve dairy herds profitability. This review aims to show some key-point experiments conducted to improve the efficiency of the self-appointed protocols for artificial insemination and embryo transfer in Brazil, overcoming several reproductive problems. Our goal is to develop cheap and easy self-appointed programs that facilitate animal handling and maximize their reproductive outcomes all over the year. Review: Failure in estrus detection is the mainly limiting factor for the use of artificial insemination in high-production dairy herd. An excellent alternative to overcome the need of estrus detection is fixed time artificial insemination. Many protocols with and without the use of estradiol have been developed to that end. Among the protocols for fixed time artificial insemination without estradiol, DoubleOvsynch has been extensively used recently in American dairy herds. In Brazil, similar pregnancy rate was obtained compared to progesterone-estradiol based protocols for fixed time artificial insemination. Particularities of progesterone-estradiol based protocols as (1) new progesterone device or devices previously used for eight days; (2) different doses of eCG; and (3) the use of estradiol cypionate for fixed time artificial insemination have been studied in Brazil. The use of self-appointed artificial insemination also enabled the reduction of the interval calving-conception compared to cows inseminated following the standing estrus. Regarding the low fertility of repeat breeders and the effect of heat stress at early pregnancy, other methods like embryo transfer became important tools to enhance reproductive efficiency of Brazilian dairy herds. Protocols were also developed to allow fixed time embryo transfer, eliminating the need of estrus detection and improving the reproductive efficiency of lactating recipients. As well as described for fixed time artificial insemination treatments, there is a large variety of hormone combination for fixed time embryo transfer (with and without estradiol). An experiment conducted in Brazil demonstrated that protocols for fixed time embryo transfer without estradiol can be as good as with estradiol to synchronize high-producing Holstein recipients, essentially during summer. Particularities related to embryos cryopreservation, synchronization of the estrus cycle of donors and recipients and the site of embryo release into the uterine horn were also investigated. Greater conception rates were achieved when fresh embryos were transferred compared to frozen-thawed ones. Also, the tight synchronization between donor and recipient (same day of estrus) resulted more pregnancies than when recipients were one day later or in advantage in relation to donors. Moreover, the site of embryo release into the uterine horn (ipsilateral to the corpus luteum) had no effect on pregnancy rates after in vivo produced embryo transfer. Conclusion: Both fixed time artificial insemination and fixed time embryo transfer are important tools to improve reproductive efficiency of high-producing dairy cows. These biotechnologies help bypassing some of the greatest challenges of dairy cattle reproduction: the difficulties of estrus detection, and the low fertility associated to heat stress and repeat breeding.

Doppler ultrasonography principles and methods of evaluation of the reproductive tract in mares

Background: Doppler ultrasonography is a non-invasive real time pulse-wave technique recently used for the transrectal study of the reproductive system hemodynamics in large animals. This technic is based in the Doppler Effect Principle that proposes the change in frequency of a wave for an observer (red blood cells) moving relative to the source of the respective wave (ultrasonic transducer). This method had showed to be effective and useful for the evaluation of the in vivo equine reproductive tract increasing the diagnostic, monitoring, and predictive capabilities of theriogenology in mares. However, an accurate and truthful ultrasonic exam requires the previous knowledge of the Doppler ultrasonography principles. Review: In recent years, the capabilities of ultrasound flow imaging have increased enormously. The current Doppler ultrasound machines offer three methods of evaluation that may be used simultaneously (triplex mode). In B-mode ultrasound, a linear array of transducers simultaneously scans a plane through the tissue that can be viewed as a two-dimensional gray-scale image on screen. This mode is primarily used to identify anatomically a structure for its posterior evaluation using colored ultrasound modes (Color or Spectral modes). Colored ultrasound images of flow, whether Color or Spectral modes, are essentially obtained from measurements of moving red cells. In Color mode, velocity information is presented as a color coded overlay on top of a B-mode image, while Pulsed Wave Doppler provides a measure of the changing velocity throughout the cardiac cycle and the distribution of velocities in the sample volume represented by a spectral graphic. Color images conception varies according to the Doppler Frequency that is the difference between the frequency of received echoes by moving blood red cells and wave frequency transmitted by the transducer. To produce an adequate spectral graphic it is important determine the position and size of the simple gate. Furthermore, blood flow velocity measurement is influence by the intersection angle between ultrasonic pulses and the direction of moving blood-red cells (Doppler angle). Objectively colored ultrasound exam may be done on large arteries of the reproductive tract, as uterine and ovary arteries, or directly on the target tissue (follicle, for example). Mesovarium and mesometrium attachment arteries also can be used for spectral evaluation of the equine reproductive system. Subjectively analysis of the ovarian and uterine vascular perfusion must be done directly on the corpus luteum, follicular wall and uterus (endometrium and myometrium associated), respectively. Power-flow imaging has greater sensitivity to weak blood flow and independent of the Doppler angle, improving the evaluation of vessels with small diameters and slow blood flow. Conclusion: Doppler ultrasonography principles, methods of evaluation and reproductive system anatomy have been described. This knowledge is essential for the competent equipment acquisition and precise collection and analysis of colored ultrasound images. Otherwise, the reporting of inconsistent and not reproducible findings may result in the discredit of Doppler technology ahead of the scientific veterinary community.

Acurácia da histerossonografia versus ultrassonografia transvaginal em mulheres inférteis candidatas às técnicas de reprodução assistida

PURPOSE: To compare the diagnostic accuracy of sonohysterography (HSN) and conventional transvaginal ultrasound (USG) in assessing the uterine cavity of infertile women candidate to assisted reproduction techniques (ART). METHODS: Comparative cross-sectional study with 120 infertile women candidate to ART, assisted at Centro de Reprodução Assistida (CRA) of Hospital Regional da Asa Sul (HRAS), Brasília-DF, from August 2009 to November 2010. Sonohysterography was performed with saline solution infusion in a close system. The sonohysterography finding was compared to previous USG results. The uterine cavity was considered abnormal whenthe endometrium was found to be thicker than expectedduring the menstrual cycle and whenan endometrial polyp, a submucous myoma and an abnormal shape of the uterine cavity were observed. The statistical analysis was doneusing absolute frequencies, percentage values and the χ 2, with the level of significance set at5%. RESULTS:HSN revealed that 92 (76.7%) infertile women candidate to ARThad anormal uterine cavity, while 28 (23.3%) had the following abnormalities: 15 polyps (12.5%).

Appendiceal endometriosis as a rare cause of abdominal pain - a case report and literature review

Endometriosis is an estrogen-dependent inflammatory disease, common in young women, characterized by the presence of endometrial tissue outside the uterine cavity. This ectopic endometrial tissue is most commonly found in the ovaries, peritoneum, uterosacral ligaments and rectovaginal cul-de-sac, with extremely rare involvement of the appendix. The main symptom is chronic abdominal pain, and the diagnosis is often made later, after the result of the histopathological examination. This study reports a 34-year-old patient complaining of chronic pelvic pain refractory to medical treatment, having undergone diagnostic laparotomy. During the surgery, we observed the presence of endometrioma fixed to the uterine wall, and the appendix was enlarged, but without evidence of inflammation. Endometrioma resection and appendectomy were performed, with good postoperative recovery. The anatomopathological exam showed endometriosis in the cecal appendix.

Prolapso da glândula da terceira pálpebra em cão -relato de caso

The updating of the of the prolapse of the third eyelid gland treatment in the literature is pertinent, due to the different surgical techniques for his resolution, with advantages and disadvantages. The work aimed to report the Moore technique in surgical treatment of the bilateral prolapse of the third eyelid gland in a female dog with bilateral entropion, six months, Shar-Pei breed, attended in the Veterinary Hospital School, Mozambique. The Moore technique showed good results and if the bilateral entropion was not a spastic type, was able to do the both surgeries in same day. The surgery using the Moore technique continues easy to learn, more simple to do and does not affect the third eyelid mobility.

Glicemia, proteinograma e perfil de alguns componentes bioquímicos séricos de cabritos da raça bôer

The newborn goat kids suffer several adjustments to life outside the uterus that affect different body functions that they need to produce heat, muscle activity and start searching for food. All these events lead to changes in several blood constituents such as proteins and serum biochemical parameters. Other studies have shown these variations, but not extend beyond the neonatal period to phase in young animals. The aim was to test the hypothesis that there is variation of the protein profile, and biochemical components of blood glucose in goat kids from birth to 75 days of life in terms of adaptation to extra uterine life. To achieve these objectives have been collected blood samples from 25 goats born by normal delivery, regardless of their sex. The variables serum total protein (TP), albumin, α-globulin, β-globulin, γ-globulin, which includes the immunoglobulin G (IgG), aspartate (AST), alkaline phosphatase (ALP), gamma-glutamyltransferase (GGT), creatinine and urea, and glucose were determined at times zero (immediately after birth), two, seven, 15, 30 and 75 days old. We observed significant differences in all variables between times, but only the creatinine concentration was higher than those of other moments in time zero and two days old, probably due to immaturity of renal function in newborn animals. The blood constituents of the kids had variations in the study period, related to physiological and nutritional causes.

Histomorfometria do corno uterino de gatas (Felis Catus) submetidas à ovariosalpingohisterectomia

Histomorphometric data were obtained from cats uterine horns: either nulliparous (n = 6), primiparous (n = 6), multiparous (n = 6) and treated with contraceptive (n = 6). The material was collected after surgery, fixed in paraformaldehyde and embedded in paraplast® resin to be sliced in a microtome. The obtained sections were stained with hematoxylin - eosin and measured under a light microscope: uterine wall total thickness (μm), endometrium total height, endometrial glands diameter and glandular epithelia height, total myometrium, internal and external myometrium and vascular layer thickness. It was concluded that: 1 - contraceptive use and number of pregnancies altered the uterine structure, 2 - one pregnancy does not appear to affect the uterine lining structures as occurs in multiparous cats, 3 - there was no variation on the evaluated structures between nulliparous and primiparous cats except for inner myometrium, 4 - the total myometrium hight and the endometrium hight showed similar variations except for the contraceptive treated cats group, 5 - the outer myometrium showed marked changes in the treated cats, 6 - the less marked morphological variations were for the endometrial glands diameter and glandular epithelium hight, 7 - the presence of dilated endometrial glands was found only in treated cats.

Reproductive biology of the guitarfish Rhinobatos percellens (Chondrichthyes, Rhinobatidae) from the São Paulo Coast, Brazil, western South Atlantic Ocean

The reproductive biology of the guitarfish Rhinobatos percellens was studied from 751 specimens caught by bottom pair trawlers off the coast of São Paulo, Brazil, between c. 24° 00′ S; 45° 15′ W and c. 25° 10′ S; 47° 52′ W, from September 2007 to August 2009. The total length (LT) and total mass (MT) relationship for males and females combined was MT = 1·29E-06 LT 3·15 (r = 0·99, n = 751). The mean LT of sexually mature specimens was 548 mm for males and 583 mm for females. Clasper growth was allometric and showed three distinct phases. Most claspers were calcified in specimens of c. 550 mm LT. The mean diameter of the largest oocyte was 29·8 mm, the mean ovarian fecundity was seven oocytes and ovulation occurred between August and November. Uterine fecundity ranged from two to 13 embryos (mean of five embryos). Larger females had higher litter sizes and larger embryos; the size-at-birth was c. 200 mm LT. The hepato-somatic index oscillated seasonally for males and females; the gonado-somatic index had little variation in males, but varied seasonally in females. The presence of many non-pregnant adult females and of encapsulated eggs during two consecutive seasons suggests a resting period between gestations and the possibility of diapause. © 2012 The Authors. Journal of Fish Biology © 2012 The Fisheries Society of the British Isles.

Treatments for invasive carcinoma of the cervix: What are their impacts on the pelvic floor functions?

Aims: Describe the impact of surgery, radiotherapy and chemoradiation in the pelvic floor functions in cervical cancer patients. Materials and Methods: A prospective study with women submitted to radical hysterectomy (RH) (n = 20),exclusive radiotherapy (RT) (n= 20)or chemoradiation (CT/RT)(n = 20)for invasive cervical cancer. Urinary, intestinal and sexual function, as well as vaginal length and pelvic floor musclecontraction were evaluated. Comparisons between groups were performed by Kruskal-Wallis and Chi-square tests (p < 0.05). Results: The groups were similar in stress urinary incontinence incidence (p = 0.56), urinary urgency (p = 0.44),urge incontinence (p = 0.54),nocturia(p = 0.53), incomplete bowel emptying (p = 0.76),bowel urgency(p = 0.12)and soilage(p = 0.43). The CT/ RT group presented a higher urinary frequency(p < 0.001)and diarrhea(p = 0.025). Patients in the RH group were more sexually active(p = 0.01) and experienced less dyspareunia (p = 0.021). Vaginal length was shorter in RT group (5.5 ± 1.9 cm) and CT/ RT(.3 ± 1.5 cm) than in the RH group (7.4 ± 1.1 cm) (p < 0.001). Pelvic floor muscle contraction was similar (p = 0.302). Conclusions: RT and CT/RT treatment for cervical carcinoma are more associated to sexual and intestinal dysfunctions.

Hormonal treatment of dairy goats affected by hydrometra associated or not with ovarian follicular cyst

Hydrometra is considered a very important pathological condition, because it represents one of the main causes of temporary infertility in dairy goats. The objective was (i) to evaluate a protocol for the treatment of hydrometra associated (n=2) or not (n=17) with follicular ovarian cyst in 19 dairy goats and (ii) to assess its reproductive efficiency after treatment. For this purpose, 10. mg of dinoprost (PGF) divided in two equal doses were administered to all animals intravulvosubmucosally on Days 0 and 10. In addition, 500. IU hCG were administered on Day 7. Ultrasound exams were performed in all females from Days 0 to 3, 7 and 10 to 13 of treatment, in order to evaluate uterus drainage after each treatment. Goats were monitored for estrus after both treatments and mated after the second dose of PGF. Blood samples were collected from 11 goats to determine plasma progesterone concentrations before, during and after treatment. Of the 19 goats treated, 16 lost weight after the first dose, probably due to uterine discharge. Complete drainage of uterine fluid was observed in 11/19 (57.9%) and 17/19 (89.5%) after the first and second doses, respectively. Afterwards, we diagnosed 2 more goats with follicular cysts, for a total of 21.1% (4/19) of animals exhibiting hydrometra and ovarian cyst concomitantly. In one doe the diagnosis was on Day 2 and in the other on Day 11. All does showed progesterone concentrations superior to 1. ng/mL at Day 0, with an average of 10.6 ± 1.4. ng/mL. Out of the 10 goats mated, only two became pregnant after treatment, corresponding to 10.5% of the total (2/19). Although prostaglandin was effective to drain the uterine fluid and led to the onset of estrus, it did not improve the pregnancy rate. The use of hCG in female goats was not effective in luteinizing the cysts. It can be concluded that hydrometra alone or associated with ovarian follicular cyst may adversely affect goat reproductive performance. © 2012 Elsevier B.V.

Timing of prostaglandin F2α treatment in an estrogen-based protocol for timed artificial insemination or timed embryo transfer in lactating dairy cows

Objectives were to investigate progesterone concentrations and fertility comparing 2 different intervals from PGF2α treatment and induced ovulation in an estrogen-based ovulation synchronization protocol for timed artificial insemination (TAI) or timed embryo transfer (TET) in lactating dairy cows. A total of 1,058 lactating Holstein cows [primiparous (n=371) and multiparous (n=687)], yielding 34.1±0.33 kg of milk/d at various days in milk were randomly assigned to receive treatment with PGF2α on either d 7 or 8 of the following protocol: d 0: 2mg of estradiol benzoate + controlled internal drug release device; d 8: controlled internal drug release device removal + 1.0mg of estradiol cypionate; d 10: TAI or d 17: TET. Only cows with a corpus luteum at d 17 received an embryo and all cows received GnRH at TET. Pregnancy diagnoses were performed by detection (transrectal ultrasonography) of an embryo on d 28 or a fetus on d 60. Fertility [pregnancy per artificial insemination (P/AI) or pregnancy per embryo transfer (P/ET)] was affected by breeding technique (AI vs. ET) and time of PGF2α treatment (d 7 vs. 8) at the 28-d pregnancy diagnosis for TAI [32.9% (238) vs. 20.6% (168)] and TET cows [47% (243) vs. 40.7% (244)] and at the 60-d pregnancy diagnosis for TAI [30% (238) vs. 19.2% (168)] and TET cows [37.9% (243) vs. 33.5% (244)]. The progesterone (P4) concentration at d 10 altered fertility in TAI cows, with higher P/AI in cows with P4 concentration <0.1 ng/mL compared with cows with P4 concentration ≥0.1 ng/mL, and in ET cows, with higher P/ET in cows with P4 concentration <0.22 ng/mL compared with cows with P4 concentration ≥0.22 ng/mL. Prostaglandin F2α treatment at d 7 increased the percentage of cows with P4 <0.1 ng/mL on d 10 [39.4 (85) vs. 23.2 (54)]. Reducing the period between PGF2α and TAI from 72 to 48h in dairy cows resulted in a clear reduction in fertility in cows bred by TAI and a subtle negative effect in cows that received TET. The earlier PGF2α treatment benefits are most likely mediated through gamete transport, fertilization, or early embryo development and a more subtle effect of earlier PGF2α treatment that may be mediated through changes in the uterine or hormonal environment that manifests itself after ET on d 7. © 2013 American Dairy Science Association.

Fetal-Maternal Interactions in the Synepitheliochorial Placenta Using the eGFP Cloned Cattle Model

Background: To investigate mechanisms of fetal-maternal cell interactions in the bovine placenta, we developed a model of transgenic enhanced Green Fluorescent Protein (t-eGFP) expressing bovine embryos produced by nuclear transfer (NT) to assess the distribution of fetal-derived products in the bovine placenta. In addition, we searched for male specific DNA in the blood of females carrying in vitro produced male embryos. Our hypothesis is that the bovine placenta is more permeable to fetal-derived products than described elsewhere. Methodology/Principal Findings: Samples of placentomes, chorion, endometrium, maternal peripheral blood leukocytes and blood plasma were collected during early gestation and processed for nested-PCR for eGFP and testis-specific Y-encoded protein (TSPY), western blotting and immunohistochemistry for eGFP detection, as well as transmission electron microscopy to verify the level of interaction between maternal and fetal cells. TSPY and eGFP DNA were present in the blood of cows carrying male pregnancies at day 60 of pregnancy. Protein and mRNA of eGFP were observed in the trophoblast and uterine tissues. In the placentomes, the protein expression was weak in the syncytial regions, but intense in neighboring cells on both sides of the fetal-maternal interface. Ultrastructurally, our samples from t-eGFP expressing NT pregnancies showed to be normal, such as the presence of interdigitating structures between fetal and maternal cells. In addition, channels-like structures were present in the trophoblast cells. Conclusions/Significance: Data suggested that there is a delivery of fetal contents to the maternal system on both systemic and local levels that involved nuclear acids and proteins. It not clear the mechanisms involved in the transfer of fetal-derived molecules to the maternal system. This delivery may occur through nonclassical protein secretion; throughout transtrophoblastic-like channels and/or by apoptotic processes previously described. In conclusion, the bovine synepitheliochorial placenta displays an intimate fetal-maternal interaction, similar to other placental types for instance human and mouse. © 2013 Pereira et al.

Nucleotide and phylogenetic analysis of human papillomavirus types 6 and 11 isolated from recurrent respiratory papillomatosis in Brazil

There are few studies about the distribution of natural molecular variants of low-risk HPVs. Our aim was to evaluate the E6 early gene variability among HPV-6 and HPV-11 isolates detected in recurrent respiratory papillomatosis (RRP) samples obtained in a cohort of Brazilian patients. We also performed a phylogenetic analysis in order to compare nucleotide sequences identified in our study with previously reported isolates from different anatomic sites (laryngeal papillomas, genital warts, cervical cancer and anal swabs) obtained from other parts of the world to determine the phylogenetic relationships of variants detected in Brazil. The complete coding region of the E6 gene of 25 samples was cloned and sequenced: 18 isolates of HPV-6 (72%) and 7 isolates of HPV-11 (28%). A total of four different HPV-6 genomic variants and two HPV-11 genomic variants was identified. It was not possible to correlate specific variants with disease severity. Phylogenetic trees for both HPV types were constructed enclosing both E6 sequences detected in our study and formerly published sequences. In both phylogenetic trees, the sequences from Brazil did not group together. We could not establish a geographical association between HPV-6 or HPV-11 variants, unlike HPV-16 and HPV-18. © 2013 Elsevier B.V.

Melatonin and ethanol intake exert opposite effects on circulating estradiol and progesterone and differentially regulate sex steroid receptors in the ovaries, oviducts, and uteri of adult rats

Chronic ethanol intake is associated with sex hormone disturbances, and it is well known that melatonin plays a key role in regulating several reproductive processes. We report the effects of ethanol intake and melatonin treatment (at doses of 100. μg/100. g. BW/day) on sex hormones and steroid receptors in the ovaries, oviducts and uteri of ethanol-preferring rats. After 150 days of treatment, animals were euthanized, and tissue samples were harvested to evaluate androgen, estrogen, progesterone and melatonin receptor subunits (AR, ER-α and ER-β, PRA, PRB and MT1R, respectively). Melatonin decreased estradiol (E2) and increased progesterone (P4) and 6-sulfatoxymelatonin (6-STM), while an ethanol-melatonin combination reduced both P4 and E2. Ovarian AR was not influenced by either treatment, and oviduct AR was reduced after ethanol-melatonin combination. Oviduct ER-α, ER-β and uterine ER-β were down-regulated by either ethanol or melatonin. Conversely, ovarian PRA and PRB were positively regulated by ethanol and ethanol-melatonin combination, whereas PRA was down-regulated in the uterus and oviduct after ethanol consumption. MT1R was increased in ovaries and uteri of melatonin-treated rats. Ethanol and melatonin exert opposite effects on E2 and P4, and they differentially regulate the expression of sex steroid receptors in female reproductive tissues. © 2013 Elsevier Inc.

Excess androgen during perinatal life alters steroid receptor expression, apoptosis, and cell proliferation in the uteri of the offspring

Exposure to environmental chemicals may contribute to reproductive disorders, especially when it occurs in critical periods of development. The female reproductive system can be a target for androgens derived from environmental contaminants or pathological conditions. The purpose of this study was to assess the long-term effects of androgens on uterine tissue after maternal exposure limited to the time of gestation and lactation. Pregnant Wistar rats were treated with testosterone propionate (TP) at 0.05. mg/kg, 0.1. mg/kg, 0.2. mg/kg or corn oil (vehicle), s.c., from gestational day 12 until the end of lactation. The results show changes in the pattern of expression of receptors for estrogen, progesterone, and androgen at all doses tested, and decreases in both apoptosis and cell proliferation indices at 0.1 and 0.2. mg/kg. We conclude that early TP exposure, under these experimental conditions, causes changes in cellular and molecular parameters that are essential for normal uterine function in the adult. © 2013 Elsevier Inc.