10 INFLUENCE OF ADMINISTRATION OF HUMAN CHORIONIC GONADOTROPIN ON DAY 4 AFTER FIXED-TIME AI ON CORPUS LUTEUM SIZE, FUNCTION, AND PREGNANCY RATE IN BEEF COWS

2014 ◽  
Vol 26 (1) ◽  
pp. 119
Author(s):  
J. B. S. Borges ◽  
D. X. Thedy

Successful establishment of pregnancy in the cow depends on early rise of progesterone (P4) to improve embryo development. The present study was carried out to determine the corpus luteum (CL) function and pregnancy rate following the administration of 1.500 IU of hCG in suckled beef cows on Day 4 after fixed-time AI (FTAI). Crossbred Angus cows (n = 429), 40 to 70 days postpartum, were synchronized using 2 mg of oestradiol benzoate (EB) i.m. and an intravaginal progesterone-releasing device containing 1 g of P4 for 7 days. At device removal, prostaglandin F2a (150 mg of D-cloprostenol) was injected i.m.; 24 h later, 1 mg of EB was given and the FTAI were done 30 to 36 h after EB using semen of just 1 bull. Cows were assigned alternatively to receive 1.500 IU of hCG i.m. (n = 200) or 1 mL of saline i.m. (n = 229) on Day 4 after FTAI. Ovaries were examined by transrectal ultrasonography (n = 71) on Days 4 and 7 to detect the presence and to measure the area of CL. Blood samples were collected on Days 4, 7, 10, 14, and 30 post-oestrus to determine P4 concentration by RIA. Pregnancy diagnoses were done 30 days after FTAI. The effects of treatments on CL area and serum P4 levels were analysed by ANOVA (Proc GLM, SAS Institute Inc., Cary, NC, USA) and pregnancy rates compared by chi-square (Proc CATMOT, SAS). The induction of accessory CL determined on Day 7 was 44% in cows treated with hCG and not observed in the control group. The area of the CL on Day 7 was significantly higher (P < 0.01; 3.37 ± 2.56 v. 2.56 ± 0.22 cm2) in the hCG group. Despite the increase in CL size, the mean levels of P4 on Days 4, 7, 10, 14, and 30 post-AI indicated no difference (P > 0.05) in CL function between control and hCG-treated cows. On Day 30, pregnancy rates were not different (P = 0.08), at 46.7 and 55% for control and hCG-treated cows, respectively. In conclusion, administration of 1.500 IU of hCG on Day 4 post-FTAI did not improve pregnancy rate in suckled beef cows.

2004 ◽  
Vol 16 (2) ◽  
pp. 212 ◽  
Author(s):  
T. Nishisouzu ◽  
M. Sugawara ◽  
S. Aoki ◽  
K. Kishida ◽  
M. Moriyoshi ◽  
...  

Treatments with GnRH and PGF2α for synchronization of ovulation has resulted in acceptable pregnancy rates after fixed-time artificial insemination in dairy cows without estrus detection. The objective of the present study was to evaluate the practicability of ovulation synchronization (Ovsynch, Pursley JR et al. 1995 Theriogenology 44, 915–923) in dairy cattle using GnRH and PGF2α for the embryo transfer recipients. Dairy cattle (cows; n=100, heifers; n=88) were randomly allocated to one of two groups. The control group (cows; n=45, heifers; n=37) was composed of cows in natural estrus. The ovulation synchronization group (cows; n=55, heifers; n=51) was treated with an intramuscular injection of 100μg of GnRH at a random stage of the estrous cycle. Seven days later, the cattle received PGF2α (Cows; 25–30mg) or PGF2α analog (Heifers; 0.5mg) in order to regress the corpora lutea (CL). Forty-eight hours later, cows and heifers received a second injection of 100μg GnRH. Embryo transfer was carried out 7 days after the second injection of GnRH in the ovsynch group and 7 days after estrus in the control group. The cattle judged to have CL 17mm were classified as acceptable recipients. The size of the follicles and the CL were determined to be of estrus stage and embryo transfer by means of ultrasonography. The mean numbers of follicles and CL were analyzed by ANOVA, while pregnancy rates were analyzed by chi-square test. The results are presented in the Table. The proportion of cows and heifers determined to be acceptable embryo transfers was not different between the control group and the ovsynch group. There were no differences in the proportion of acceptable embryo transfers between the control group and the ovsynch group. Follicle diameter at the time of estrus in the control group (cows; 20.7±0.7mm, heifers; 16.8±0.5mm) were significantly larger than that of the ovsynch group (cows; 18.0±1.0mm, heifers; 14.7±0.2mm) (P&lt;0.05). Although CL diameter at the time of embryo transfer in heifers showed no differences between the control group and the ovsynch group (25.0±1.0mm v. 22.8±1.5mm), The CL diameter of the control cow group was larger than that of the ovsynch group (29.8±0.7mm v. 26.1±1.0mm, P&lt;0.05). However, no differences in pregnancy rate were seen between the control group and the ovsynch group. These results suggest that ovsynch can be effectively applied in an embryo transfer program for cattle. Table 1 Proportion of acceptable embryo transfer recipients and pregnancy rate in dairy cattle in the control ovsynch groups


2010 ◽  
Vol 39 (10) ◽  
pp. 2182-2186
Author(s):  
Cristiane Reinher ◽  
Júlio Otávio Jardim Barcellos ◽  
Vanessa Peripolli ◽  
Ênio Rosa Prates ◽  
Maria Eugênia Andrighetto Canozzi

It was evaluated in this work the effect of calving sub-season on the pregnancy rate of 7,726 multiparous Hereford beef cows grazing on natural pastures in southern Brazil, from 1994 to 2007. Calving sub-season periods were divided in 20-day intervals from August 12th to 31st; from September 1st to 20th; from September 21st to October 10th; from October 11th to 31st; from November 1st to 20th. Calving in each sub-season and pregnancy rate (PR) were analyzed by the Chi-square test. Overall means of each sub-season were 92.7% (from August 12th to 31st), 90.6% (from September 1st to 20th), 82.1% (from September 21st to October 10th), 77.7% (from October 11th to 31st), and 70.6% (from November 1st to 20th). Calving sub-season significantly influenced pregnancy rate. Cows which calved in the initial sub-season showed higher pregnancy rates than cows which calved from the fourth calving sub-season. However this difference did not occur in 1999 and in 2007 since calving rates did not statistically differ among the five sub-seasons. In 1998 and 2004, calving rates were lower and the effects of calving sub-season on pregnancy rates were higher, possibly due to climate variations. In general, pregnancy rate decreases as calving occurs later in the year, particularly in years when the weather adversely affects pasture growth.


2011 ◽  
Vol 23 (1) ◽  
pp. 163 ◽  
Author(s):  
R. Núñez ◽  
T. de Castro ◽  
L. Cutaia ◽  
G. Bó ◽  
A. Menchaca

An important proportion of embryonic deaths in cattle occur during the critical period of embryo development and maternal recognition of pregnancy. High serum progesterone concentrations during this period are associated to higher embryo survival, and the use of eCG could enhance luteal activity. Two experiments were carried out to evaluate the effect of administering eCG at the time of removal of a progesterone-releasing device and 14 days after fixed-time artificial insemination (FTAI) in beef cattle. Multiparous suckled cows between 60 and 90 days postpartum (n = 297; Exp. 1), and two-year old prepuberal heifers (n = 689; Exp. 2) with a body condition score (BCS) between 3.5 and 5 (1 to 8 scale) were used. At the beginning of the experiments, none of females used had a corpus luteum detected by ultrasonography. Females received an intravaginal device containing 0.5 g progesterone (DIB 0.5, Syntex, Buenos Aires, Argentina) and 2 mg oestradiol benzoate i.m. (Benzoato de Oestradiol, Syntex). Eight days later, 500 μg of cloprostenol (Ciclase DL, Syntex) and 0.5 mg oestradiol cypionate (Cipiosyn, Syntex) were given i.m. and devices were removed; FTAI was performed 52 to 56 h after DIB removal. At the time of device removal, the animals were randomly allocated to 4 groups, to receive 400 IU of eCG i.m. (Novormon, Syntex) or placebo at device removal and/or 14 days after FTAI, in a 2 × 2 factorial design. Control group did not receive eCG. In Exp. 1, nose plates were placed on calves on the day of DIB insertion and left for 10 days in order to prevent suckling. Pregnancy diagnosis was performed by ultrasonography at 30 and 60 days after FTAI. Data were analysed by logistic regression. Pregnancy rate at 30 days after FTAI was affected by treatment (P < 0.05; Table 1). Late embryonic/fetal loss rates (non-pregnant females at 60 days after FTAI over those pregnant at 30 days) were 1.6% (3/182) for cows in Exp. 1 and 4.8% (14/292) for heifers in Exp. 2; no differences were detected among groups. In conclusion, administration of 400 IU of eCG at device removal increases pregnancy rate, both in anestrus cows and heifers. However, the administration of a second dose of eCG 14 days after FTAI suggests a possible effect on embryo survival within the first 30 days of pregnancy in cows but not in heifers. Further studies are required to confirm these results. Table 1.Pregnancy rates after administration of eCG at time of progesterone device removal and/or 14 days after insemination in beef cattle Supported by Syntex.


2007 ◽  
Vol 19 (1) ◽  
pp. 197
Author(s):  
A. Guzeloglu ◽  
H. Erdem

We previously reported that administration of flunixin meglumine two times at a critical stage approaching pregnancy recognition associated with corpus luteum maintenance increased early embryo survival and pregnancy rate via an additive antiluteolytic effect with the conceptus (Guzeloglu et al. 2006 Reprod. Fertil. Dev. 18, 183). In this study, the objective was to determine if a single administration of meloxicam, a non-steroid anti-inflammatory drug (NSAID) with a longer half-life, could be used instead of flunixin meglumine to avoid repeated injections in heifers at a critical stage following insemination to increase pregnancy rate due to its inhibitory effect on PGF2� synthesis. Eighty-five 15-18-month-old Holstein heifers were synchronized; following insemination (Day 0), the heifers were randomly assigned to receive a subcutaneous meloxicam injection (0.5 mg kg-1; n = 37) in the afternoon of Day 15 or left untreated as controls (n = 48). Pregnancy rates were defined as the percentage of heifers diagnosed pregnant by ultrasound between Days 31 and 38 after AI. The effect of treatment on pregnancy rates was analyzed by chi-square test. Meloxicam treatment on Day 15 after insemination dramatically decreased the pregnancy rates in the heifers (25 of 48 (52%) in the control group vs. 9 of 37 (24.3%) in the meloxicam-treated group; P &lt; 0.01). This result indicates that administration of meloxicam around a time associated with pregnancy recognition and corpus luteum maintenance was harmful to the pregnancy even though the drug is considered to be safe during pregnancy in cattle. It is concluded that direct or indirect effects of meloxicam on the uterine environment and/or embryo are not favorable at least during early pregnancy in cattle. How meloxicam exerted its detrimental effect on pregnancy warrants further investigation.


2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 281-282
Author(s):  
Nicole N Noga ◽  
Jeremy G Powell ◽  
Brittni P Littlejohn ◽  
Whitney L Rook ◽  
Martin Mussard ◽  
...  

Abstract Preliminary data indicate that shortening the progesterone exposure with CIDR can improve conception rates compared to the established 7-day CO-Synch protocols. The objective was to determine the effects of shortening progesterone exposure from 7 days to 6 days among crossbred beef cows and fixed-time artificial insemination (FTAI) with sex-sorted semen. A total of 147 fall calving (September-November) crossbred beef cows from one location were randomized to one of four treatments: 1 and 4) 7-d CO-Synch + CIDR protocol with administration of gonadotropin-releasing hormone (GnRH) and a CIDR insert on day 0, prostaglandin F2α (PGF) at CIDR removal on day 7, and FTAI occurring 66 h (Treatment 1) or 74 h (Treatment 4) later; 2 and 3) 6-d CO-Synch + CIDR protocol with administration of GnRH and CIDR insert on day 0, prostaglandin F2α (PGF) at CIDR removal on day 6, and FTAI occurring 72 h (Treatment 2) or 80 h (Treatment 3) later. Sex-sorted semen from Angus sires was used for AI. Pregnancy was determined by ultrasound 40 days after AI. Pregnancy rate was analyzed using the MIXED procedure of SAS with CIDR and AI service time as fixed effects. Chi-square analysis was used to assess the number of pregnant and open cows in each treatment using the FREQ procedure of SAS. There was no difference (P = 0.78) in pregnancy rates between 6-d CIDR (0.59 ± 0.05%) and 7 d CIDR (0.57 ± 0.05%) protocols. Cattle bred a shorter period after CIDR removal (0.64 ± 0.05%) had numerically (P = 0.11) greater pregnancy rates compared to those with a longer period after CIDR removal (0.51 ± 0.05%). There was no difference in the number of pregnant and open cows between treatment groups. These data suggest that using a 6-day or 7-day protocol, regardless of breeding time, results in similar conception rates.


2020 ◽  
Vol 98 (Supplement_2) ◽  
pp. 1-1
Author(s):  
Jennifer McMurray ◽  
Andrea N DeCarlo ◽  
Nathan Long ◽  
Scott Pratt

Abstract A study was conducted over three years to determine if differences in pregnancy rates existed between the use of commercially available sex-selected (SEXED) or conventional semen (STD) coupled to fixed-time artificial insemination (TAI). A total of 240 (2016: n = 80; 2017: n = 95; 2018: n = 65) primi- or multiparous Angus or Angus-cross cows being at 45 days post-partum or greater were blocked by weight and BCS and assigned to be inseminated using SEXED semen or STD semen. Artificial insemination was performed by the same AI technicians (n = 3) across all years. All cows were subjected to estrous synchronization and insemination using the SEXED semen or STD semen with TAI between 63 and 66 h post-CIDR removal. Fourteen days post-TAI all cows were exposed to bulls for natural service. Cows were subjected to transrectal ultrasonography at day 30 and day 45 post-TAI to determine pregnancy rates and calving dates in the subsequent calving seasons were recorded to determine calving rates to TAI. Pregnancy rates were compared using a Chi-square test of two proportions. All statistical calculations were performed using JMP software and statistical significance was based on p-values less than 0.05. Pregnancy rates at day 30 and day 45 were 39.67% and 48.76% for SEXED and 47.90% and 54.62% for STD, respectively (P &gt; 0.05). The percentage of calves born to TAI pregnancies for sex-selected semen and conventional semen were 22.31% and 40.34%, respectively (P &lt; 0.05). These data indicate that the use of sex-selected semen can give similar pregnancy results to conventional semen in beef cow operations using TAI at day 30 of gestation; however, there was a reduction in the number of offspring to TAI comparing calving dates to animals pregnant at day 30.


2020 ◽  
Vol 48 ◽  
Author(s):  
Caio Cezar Da Silva ◽  
Hélton Aparecido Garcia Gregianini ◽  
Jennifer Teodoro Ferreira Gregianini ◽  
José Antônio Dell’Aqua Junior ◽  
Jefferson Viana Alves Diniz ◽  
...  

Background: In vitro embryo production (IVEP) allows the spread of superior animal genetics, but pregnancy rates show a high variability with this biotechnique. In the initial stage of pregnancy, progesterone plays a fundamental role in uterine preparation, acting on embryonic growth, implantation, and development. However, on the day of the IVEP transfer to the recipients, progesterone levels may be lower than that expected, influencing the uterine environment and, consequently, the pregnancy rate. Therefore, the objective of this study was to evaluate the pregnancy rate in heifers after the administration of injectable progesterone (P4) in the fixed-time embryo transfer (FTET) protocol.Materials, Methods & Results: The experiment was conducted inside a rural property near the city of Rio Branco, Acre, Brazil. The experimental group consisted of 232 animals, including 78 zebuine (Bos indicus) and 154 mixed (½ blood B. indicus and ½ blood B. taurus) animals, aged between 16 and 24 months, with a mean weight of 300 and 330 kg for zebuine and mixed animals, respectively. The selected animals were previously synchronized using the progesterone-estrogen-prostaglandin-estrogen protocol. Embryo transfer was performed on day 18 of the protocol, which was 9 days after the removal of intravaginal progesterone implant. On day 15 of the protocol, that is, 144 h (6 days) after the device removal, the animals were randomly distributed into two experimental groups: Control Group (CG; 0.5 mL of 0.9% saline solution, intramuscular) and Treated Group (P4G; 0.5 mL of injectable P4, 150 mg, intramuscular). Chi-square test was used for the statistical analysis of the pregnancy rate at a 5% probability. After 23 days of embryo transfer, pregnancy was diagnosed by ultrasonography. The general pregnancy rate, considering all groups (CG and P4G) and breeds included, was 55.17% (128/232). The pregnancy rates of the P4G and CG groups, regardless of breeds, were 55.08% (65/118) and 55.26% (63/114), respectively, with no statistical difference (P = 0.8344). Angus animals presented a similar pregnancy rate in the P4G and CG groups of 54.93% (39/71) and 57.83% (48/83), respectively. Furthermore, similar results were found for the Nellore breed, with no difference in pregnancy rate between the CG (55.81%, 24/43) and P4G (48.57%, 17/35) groups.Discussion: The strategy of using P4 to increase the pregnancy rate is very widespread in fixed-time artificial insemination (FTAI) protocols, although differences still exist. The literature presents zero or negative effects, as in this study, when exogenous P4 was used to increase endogenous P4 concentrations and, consequently, the pregnancy rate. However, some studies demonstrated the beneficial effects of increasing blood P4 concentrations, and that the increased fertility depends on the method and time of P4 supplementation and the animal’s physiological state. In this context, the fact that the injectable progesterone supplementation did not increase the pregnancy rate in this study is justified mainly by factors such as the animal category used (heifers), time of P4 supplementation (day 4 after ovulation), biotechnology used (FTET), and the heterogeneous characteristics related to phases of the estrous cycle of heifers. Supplementation with 150 mg of injectable long-acting progesterone intramuscularly did not interfere in the pregnancy rate of Angus and Nellore heifers receiving bovine embryos.


2006 ◽  
Vol 18 (2) ◽  
pp. 183
Author(s):  
A. Guzeloglu ◽  
H. Erdem ◽  
M. K. Saribay ◽  
W. W. Thatcher ◽  
T. Tekeli

The objective was to determine if administration of flunixin meglumine (FM) to heifers following insemination would increase pregnancy rate due to its inhibitory effect on prostaglandin F2� (PGF2�) synthesis. Fifty-two 15-month-old Holstein heifers were synchronized with single or double injections of PGF2� followed by an injection of gonadotropin-releasing hormone (GnRH) 48 h later and a timed artificial insemination at 12-14 h after injection of GnRH (Day 0). Heifers randomly assigned to the treatment group (FMG) were injected twice with FM (1.1 mg/kg BW; i.m.), given 12 h apart on the evening of Day 15 and the morning of Day 16. The control group (CG) was not treated. Pregnancy rates were defined as the percentage of heifers diagnosed pregnant by ultrasound at Days 29 and 65 after AI. Effects of treatment on pregnancy rates and pregnancy losses were analyzed by chi-square test and logistic regression analysis. Pregnancy rates in the heifers treated with FM were higher at Day 29 (76.9%; 20/26 [FMG] vs. 50%; 13/26 [CG]; P < 0.04) and tended to be higher at Day 65 (69.2%; 18/26 [FMG] vs. 46.2%; 12/26 [CG]; P < 0.09). Administration of FM two times at a critical stage leading up to pregnancy recognition, associated with corpus luteum maintenance, increased both early embryo survival and pregnancy rate via an additive antiluteolytic effect due to a delay in the luteolytic secretion of PGF2�; this provided conceptuses extra time to develop the capability to inhibit the luteolytic process.


2016 ◽  
Vol 28 (2) ◽  
pp. 134
Author(s):  
J. B. S. Borges ◽  
H. L. D. Neri ◽  
M. R. Almeida ◽  
E. P. Silva ◽  
A. Bilha

This study aimed to compare the effects of 2 inducers of new follicular wave [oestradiol benzoate (EB) or hCG] administrated at onset of resynchronization protocols initiated 22 days after the first timed AI (TAI) of beef heifers. We evaluated the effect of treatments on corpus luteum (CL) development, serum progesterone (P4) concentrations, and pregnancy rates. A total of 467 grazing Brangus heifers, with 24- to 26-months, 320 ± 10 kg of body weight, and 3.2 ± 0.5 body condition score, were utilised. The resynchronization protocol began 22 days after the first TAI (Day 22). Heifers were randomly allocated into 1 of 3 groups: EB (n = 199), hCG (n = 101), or control untreated (n = 167). The EB- and hCG-treated heifers received an intravaginal P4 device (Procliclar, 0.75 g of P4; Hertape, Brazil) and an IM treatment with 1 mg of EB (Benzoato HC, Hertape) or 1000 IU of hCG (Vetecor 5000, Hertape), respectively. On Day 30, P4 device was removed, and the pregnancy diagnosis was performed. At this time heifers had the CL area (cm2) determined by ultrasound and seric P4 concentration evaluated by chemiluminescence assay. The assay sensitivity was 0.1 ng mL–1, and the intra- and interassay coefficients of variation were 7.9 and 10.6%, respectively. Nonpregnant heifers after the first TAI (EB = 97, hCG = 36) received intramuscularly 150 μg of sodium cloprostenol (Veteglan Luteolítico, Hertape) and 1 mg of oestradiol cypionate (Cipionato HC, Hertape). The second TAI was performed 48 to 52 h after the P4 device removal. On Day 62, heifers were submitted to a second pregnancy diagnosis. No effect of the body condition score (P = 0.28) was observed on the first and second pregnancy per AI. Pregnancy rates after the first TAI were higher (P = 0.03) on hCG-treated heifers (64.4%) than EB (51.3%) or the control group (58.7%). The EB group determined greater (P = 0.04) pregnancy rate after the second TAI (49.5%) than the hCG group (22.2%). The cumulative pregnancy rates were similar (P = 0.46) between groups [EB (72.4%) and hCG (68.3%)]; however, both were greater (P < 0.05) than the control group (58.6%). Heifers treated with hCG (3.42 ± 0.76) presented greater CL area at Day 30 (P < 0.05) than the EB (2.44 ± 0.57) and control (2.61 ± 0.61) groups. The treatment with hCG 22 days after TAI determined greater (12.43 ± 3.48; P < 0.05) serum P4 concentrations on Day 30 than the EB (6.92 ± 3.04) and control (7.29 ± 2.45) groups. Therefore, the use of hCG at the beginning of resynchronization protocol 22 days after the TAI increased pregnancy rate following the first TAI. However, the use of 1 mg of EB determined greater pregnancy rate after the second TAI, and no detrimental effects on CL development, P4 concentration, and on first TAI pre-established pregnancy.


2013 ◽  
Vol 25 (1) ◽  
pp. 159
Author(s):  
J. P. Detomini ◽  
R. R. Tirloni ◽  
C. V. Araujo ◽  
L. R. Martins

The aim of this study was to establish a comparison between heterospermic and monospermic semen on the pregnancy rate of fixed-time AI Bos indicus cows. The experiment was conducted in the state of Mato Grosso, Brazil, during December 2010 and February 2011. Multiparous (n = 173) and primiparous (n = 204) Nelore cows, 45 ± 15 days postpartum, were evaluated on body condition score (BCS; score 1 to 9), and those that presented BCS between 5 and 8 were selected. Two protocols of estrous cycle synchronization were performed (PeCG and Pshang). Synchronization of the estrous cycle was achieved with the aid of the following protocol: on Day 0, all animals were treated with a progesterone vaginal implant with 1.9 g of progesterone combined with an IM injection of 2 mg of estradiol benzoate. On Day 7, an IM injection of 12.5 mg of dinoprost was administered. On Day 9, implants were withdrawn and the animals received an IM injection of 0.5 mg of estradiol cypionate. On Day 9, part of the animals (n = 222) received an IM injection of 300 IU mg of eCG (PeCG) and the other part (n = 155) had their calves removed for 48 hours (Pshang). All animals were inseminated 46 to 52 h after the end of the protocol. Cows were allocated randomly in 4 groups according to the donor male sample in the semen straw and groups were designated as MA (monospermic straw from bull A; n = 110), MB (monospermic straw from bull B; n = 102), MC (monospermic straw from bull C; n = 108), and HT (heterospermic sample from bulls A, B, and C; n = 57). Commercially available semen straws from Aberdeen Angus donors were used (ABS Pecplan, Brazil). Pregnancy diagnosis was performed 35 days after AI using real-time ultrasonography. Results are summarized in Table 1. There was no statistical difference between MA, MB, MC, and HT in any group according to the chi-square test (P > 0.05). However, there was a remarkable difference in pregnancy rates among the 3 bulls tested considering both protocols, probably due to the variation in the ovulation moment. Table 1.Pregnancy rates of Nelore cows submitted to FTAI protocol inseminated with monospermic (bull A, B, or C) or heterospermic (HT) semen


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