Rationale: Coronavirus disease 2019 (COVID-19) is a highly infectious disease due to the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV2). Vertical transmission and clinical presentation of COVID-19 in pregnancy is still obscure. Additionally, the potential hazard of COVID-19 in pregnancy on the fetus and post-delivery risk for the neonate remain under investigations. Patient concern: A young-aged Asian pregnant woman with 28 weeks of gestation presented with fever, dyspnoea, headache, and joint pain with decreased fetal movement for about one week. Diagnosis: The patient was diagnosed with COVID-19 pneumonia. Interventions: The patient was referred to the quarantine sector and was treated with chloroquine orally, intravenous fluid and other supportive treatment. Outcomes: After one week of treatment, the patient improved. Seventeen days after admission to hospital and at 30 weeks of gestation, she started vaginal preterm delivery of a viable healthy neonate with negative COVID-19 test for two occasions. Lessons: COVID-19 pneumonia during pregnancy presents with similar clinical presentation of non-pregnant women. COVID-19 pneumonia during pregnancy increases the risk of preterm labour without evidence of vertical transmission.

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Objective: To test the effects of extenders and packaging methods on morphological and functional characteristics of frozen thawed Ossimi ram semen. Methods: Ram semen was pooled, diluted in 3 different extenders: Tris-egg yolk (TEY), Tris-soybean lecithin (TSBL), and Tris- butylatedhydroxytoluene (TBHT), equilibrated at 5 °C for 4 h, and packaged in straws or pellets for freezing. Semen was evaluated for sperm progressive motility, viability, abnormality, and membrane integrity after dilution, equilibration and thawing. The percentages of viable, early apoptotic, apoptotic, and necrotic spermatozoa as well as comet assay parameters were determined in post-thawed semen. Total antioxidants capacity, malondialdehyde and lactic dehydrogenase were assayed in thawed seminal plasma. Results: After equilibration, only sperm membrane integrity was significantly higher (P < 0.05) in TEY and TSBL than in TBHT. After thawing, TEY or TSBL in straws significantly improved sperm progressive motility and vitality (P < 0.05). In thawed seminal plasma, TBHT or TSBL in straws and TBHT in pellets significantly reduced malondialdehyde, and TBHT in pellets significantly increased lactic dehydrogenase (P < 0.05). TEY in straws increased viable sperm, while significantly decreased early apoptotic and apoptotic sperm (P < 0.05). DNA damage was significantly decreased (P < 0.05) in straws with TEY and TSBL, and tail moment decreased in straws with all extenders (P < 0.05). Conclusions: Despite the disadvantages of dilution of cryopreserved semen with egg yolk, ram semen cryopreserved with TEY gives the best physical, morphological and functional characteristics in straws compared with pellets, followed by semen diluted with TSBL. However, semen diluted with TBHT or TSBL, regardless of packaging method, showed the highest impact on antioxidant status of cyopreserved ram semen.

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Objective: To investigate the effects of Azadirachta (A.) indica methanol seed extract on canine male fertility in vitro. Methods: The effect of the A. indica methanol seed extract (2.5, 5.0, 10.0 and 20.0 mg/mL, respectively) on canine spermatozoa was evaluated in vitro at 1.0, 2.5, 5.0 and 10.0 min post-treatment. The two controls were comprised of sperm sample in phosphate- buffered saline, and sperm sample in phosphate-buffered saline and 1% dimethyl sulfoxide, respectively. The experiment was repeated (n=3) in duplicates with semen collected from 3 adult Basenjis. The effect of A. indica methanol seed extract treatments on canine spermatozoal total and progressive motility (phase-contrast microscopy), sperm vitality (eosin-nigrosin vital staining method) and total abnormalities (phase contrast microscopy, eosin-nigrosin staining and Papanicolaou staining methods) were determined. Results: A. indica methanol seed extract treatment caused a concentration and time-dependent decrease in sperm percentage total motility, progressive motility and vitality. The absence of sperm revival post-treatment provided evidence of a permanent or irreversible incapacitation of sperm by A. indica methanol seed extract. Conclusions: The inhibition of sperm motility and significant spermicidal effect observed post-treatment suggest that A. indica methanol seed extract may produce a potent antifertility or contraceptive effect in dogs if applied as an intra-vaginal cream pre- coitus.

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Objective: To compare clinical pregnancy rates following sequential day-3 and day-5 embryo transfer with double or sequential cleavage-stage transfers. Methods: This study enrolled 242 patients undergoing gonadotropin-releasing hormone antagonist protocol and fresh embryo transfer. Basal follicle stimulating hormone, luteinizing hormone, serum estradiol and anti-Müllerian hormone levels and controlled ovarian stimulation outcomes were noted. Of 242 women, 135 underwent double embryo transfer on day 2 or day 3 (the double group), 54 women underwent sequential embryo transfer on day 2 and day 3 (the D2/D3 group), and 53 underwent sequential embryo transfer on day 3 and day 5 (the D3/D5 group). Clinical pregnancy rates were compared among the groups. Results: Female age, body mass index, basal follicle stimulating hormone, luteinizing hormone and estradiol levels were similar among the groups (P>0.05). The D3/D5 group had a significantly higher number of metaphase II oocytes, fertilized oocytes and good quality embryos on day 3 compared with the double group and the D2/D3 group (P < 0.001). Clinical pregnancy rates in the double, D2/D3 and D3/D5 groups were 26.6% (36/135), 16.6% (9/54) and 37.7% (20/53), respectively. There was no significant difference in clinical pregnancy rates between the double group and the D2/D3 group (P =0.204) or the D3/D5 group (P =0.188). The D3/D5 group had significantly higher clinical pregnancy rates compared with the D2/D3 group (P =0.025). Conclusions: Sequential cleavage-stage transfer (D2/D3) or cleavage stage and blastocyst transfer (D3/D5) does not improve clinical pregnancy rates compared with double cleavage-stage embryo transfer. Although sequential transfer seems to be an effective option in certain patient populations, routine application of this technique might not be a suitable approach in an unselected population to improve assisted reproductive technology outcomes.

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Objective: To compare the effectiveness of blastocyst elective single embryo transfer (eSET) and double embryo transfer (DET) in reducing low birth weight, preterm birth, and perinatal mortality in in vitro fertilization (IVF) cycles of Indonesian women. Methods: A retrospective observational study was conducted at Morula IVF Clinic, Jakarta, Indonesia. A total of 179 women who underwent either eSET or DET and had met the eligibility criteria were included. Seventy-six women underwent eSET while 103 underwent DET in their IVF cycles. Low birth-weight rate, preterm birth rate, and perinatal mortality rate of both groups were measured as the primary study outcomes. Neonatal intensive care unit (NICU) admission rate, Apgar score, multiple pregnancy, and maternal complications during pregnancy were also evaluated. Results: The risk of low birth weight [odds ratio (OR)=0.21, 95% confidential interval (CI): 0.10-0.45, P<0.001] and preterm birth (0R=0.25, 95% CI: 0.13-0.49, P<0.001) was significantly lower in the eSET group compared with the DET group. Furthermore, eSET efficiently reduced the incidence of NICU admission and multiple pregnancy (P=0.01 and P<0.001, respectively). No significant difference was observed in terms of perinatal mortality rate, Apgar score, and maternal complications including gestational diabetes, preeclampsia as well as pregnancy-induced hypertension (P>0.05). However, a lower incidence of antepartum hemorrhage was noticed in the eSET group than in the DET group (P=0.03). Conclusions: Compared with DET, infants conceived through IVF cycles with eSET have a significantly lower risk of low birth weight, preterm birth, and NICU admissions. Moreover, eSET is shown to reduce multiple pregnancy rate, yet no significant differences are observed in the perinatal mortality rates, Apgar score and maternal complications (except for the incidence of antepartum hemorrhage) between both groups.

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Objective: To evaluate whether addition of symbiotic to clindamycin could reduce Gardnerella vaginalis, Atopobium vaginae, and Megasphaera phylotype I in pregnant women with bacterial vaginosis. Methods: This randomized controlled trial (RSUP Dr. Wahidin Sudirohusodo Makassar clinical trial registry UH17010021) included 61 samples. The intervention group was given clindamycin and synbiotic while the control group was given clindamycin and placebo (without synbiotic). Wilcoxon test and hypothesis test of two independent samples were used to compare the treatment efficacy. Results: This study showed a significant difference in Nugent score before and after treatment in each group. But there was no difference in Nugent score between the intervention group and the control group after treatment or in Nugent scores reduction in both groups. The most common type of bacteria found was Megasphaera phylotype I. There were no significant differences in the three types (Gardnerella vaginalis, Atopobium vaginae, and Megasphaera phylotype I) of bacteria after treatment between both groups. Additionally, there was no difference in therapeutic effect between the intervention group and the control group. Conclusions: Clindamycin along with synbiotics is no more effective for treated bacterial vaginosis than clindamycin without synbiotics. Megaesphaera is the most commonly found bacteria, which cannot be eradicated with clindamycin.

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Objective: To evaluate whether the use of vascular endothelial growth factor (VEGF) with alginate increases oocyte maturation following in vitro maturation. Methods: This experimental study was performed on 150 immature oocytes (germinal vesicle oocytes) from females who were candidates for assisted reproductive technology. The germinal vesicle oocytes were randomly placed in the control, alginate, and VEGF plus alginate groups. The basic culture medium for oocytes culture (tissue culture medium 199, follicle-stimulating hormone 0.075 IU/mL, and fetal bovine serum 10%) was used in the control, alginate, and VEGF plus alginate groups. For the treatment groups (alginate, and VEGF plus alginate groups), alginate (8%) and VEGF (5 ng/mL) were added to the basic culture medium. After culture, immature oocytes were considered as oocytes unchanged in the nucleus whereas oocytes with a polar body were considered as mature oocytes (metaphase II stage). The mature oocytes in each group were fertilized by intracytoplasmic sperm injection and formed embryos were evaluated by reverse microscope. Results: The oocyte maturation rate (metaphase II) significantly (P < 0.05) increased in the alginate plus VEGF group as compared with the alginate alone and control groups during in vitro maturation. On day 2, the cleavage rates were significantly different in the matured oocytes between the treatment groups and the control group. The percentage of the two-cell stage, four-cell stage and eight-cell embryos was significantly higher in the treatment groups compared with the control group (P <0.05). Conclusions: Supplementation of VEGF with alginate can improve oocyte maturation in culture media. VEGF with alginate may promote the quality of nuclear and cytoplasmic maturation of human oocytes in vitro.

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Objective: To study the protective effect of ascorbic acid and curcumin against the abnormal estrous cycle and morphological changes in cells induced by repeated ultraviolet B (UVB) radiation in female Wistar rats. Methods: Sixteen female sexually mature Wistar rats weighing 130-150 g and aged 12-16 weeks were randomly divided into four groups. The control group received normal food and water ad libitum. The UVB group was exposed to a dose of 280 nm of UVB radiation for 2 h daily. The UVB+curcumin group received a dose of 280 nm of UVB radiation for 2 h daily and also an oral dose of curcumin (25 mg/kg body weight) daily. The UVB+ascorbic acid group received a dose of 280 nm of UVB radiation for 2 h daily and also an oral dose of ascorbic acid (250 mg/kg body weight) daily. All the treatments last for 15 consecutive days. Body and ovary weight and gonadosomatic index were measured. The stages (proestrus, estrus, metaestrus and diestrus) of the estrous cycle were determined by the cell types observed in the vaginal smear. Results: UVB radiation caused irregular alterations on the estrous cycle and morphological changes of the female Wistar rat as compared with the control group. Ascorbic acid and curcumin protected UVB-induced estrous phases and their cells. But curcumin showed greater protection than ascorbic acid. Conclusions: Ascorbic acid and curcumin at low doses can alleviate abnormal estrous cycle and morphological changes in cells induced by UVB radiations in female Wistar rats.

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