Fertility testing of preserved epididymal sperm by microinjection: A model for the rescue and utilization of genetically superior animals.

Background: Epididymal sperm preservation is a simple conservation approach that can help prevent the loss of high genetic quality of farm animals. The chance of loss increases, especially during disease outbreaks or other interruptions to normal reproduction function. Aim: This study looked into the ability of preserved ram epididymal sperm to fertilize oocytes. Due to motility becoming an issue following sperm storage for fertilization, the sperm microinjection known as intracytoplasmic sperm injection approach was employed. Methods: The study was divided into two parts. First, involved the preservation of epididymal sperm at 5°C for 12 days. During preservation, sperm quality parameters namely motility, viability, intact membrane, acrosome, and Deoxyribonucleic acid (DNA) are evaluated every three days. For the fertility test in the second experiment, matured oocytes were injected with immotile sperm discovered in the last days of preservation. The presence of pronucleus development following in vitro culture is used as an indicator of sperm's ability to activate and fertilize oocytes. Results: All sperm quality parameters significantly (p < 0.05) declined during preservation time. On day 12, motility was discovered to be 0%, but viable sperm, sperm with intact membrane, acrosome, and DNA remained at 41.86% ± 9.30%, 31.18% ± 5.15%, 21.88% ± 1.93%, and 33.35% ± 8.74%, respectively. On the fertility test, we inject immotile sperm from day 12 of preservation, which has the lowest motility found, into matured oocytes. Those sperms are able to activate (52.05% ± 7.15%) and fertilize (31.37% ± 1.75%) the injected oocytes, but their fertilizing ability is significantly lower (p < 0.05) when compared to the sperm derived from the ejaculate. Conclusion: In this study, simple preservation of epididymal sperm reduces all sperm quality criteria, particularly motility. Using the microinjection approach preserved sperm which had no motility, still demonstrated its ability to activate and fertilize the oocytes. According to that, this study provides potential approaches and tools for using genetically superior animals that have lost their ability to execute regular fertilization, and also prolong reproduction function.

The effect of activated silicon dioxide and betaine supplementation on quails' growth and productivity.

BACKGROUND AND AIM: Silicon dioxide and betaine supplements are essential in the poultry diet to improve growth and laying performance. This study aimed to determine the effect of activated silicon dioxide (ASD) and betaine supplementation on quails' growth performance during the starter period and on follicular development and productive parameters at the onset of the laying period. MATERIALS AND METHODS: The research used 1064 quails, aged 7 days. Four treatments were employed: A basal diet (control), a diet supplemented with 200 ppm ASD, a diet supplemented with 0.12% betaine, and a diet supplemented with a combination of 200 ppm ASD and 0.12% betaine (ASD+Betaine). Each diet group included seven replicates of 38 quails. The observed parameters were the quails' growth performance during the starter period and follicular development and egg production during the onset of the laying period. RESULTS: The results showed that combined supplementation with ASD+Betaine improved the quails' growth performance during the starter period (p<0.05). However, these supplements, either as single feed additives or in combination, did not affect ovarian and follicular characteristics during the onset of the laying period. Still, ASD and betaine enhanced egg production and weight. The highest magnitude of change resulted from combined supplementation with ASD+Betaine (p<0.05). CONCLUSION: The ASD+Betaine could be productively applied to quails' diets during the starter period and at the onset of the laying period.

Deleterious effect of short-term gavage of an ethanol extract of cogon grass (Imperata cylindrica L.) roots on testis and epididymal sperm quality.

BACKGROUND AND AIM: Cogon grass (Imperata cylindrica L.) (CGG) is a herbal medicine that could be developed into a male antifertility agent. The present study aims to determine the effect of an ethanol extract of CGG roots on mice testicular activity, reproductive hormone levels, and epididymal sperm quality. MATERIALS AND METHODS: This study was designed as completely randomized with three different doses, such as an ethanol extract of CGG roots at 0 (control), 90, and 115 mg/kg body weight. In total, 21 male DDY mice strain were treated with the CGG extract (by gavage) for 14 days, followed by an evaluation of reproductive organs, epididymal sperm quality, testis histology, histomorphometry, and reproductive hormone assays. All quantitative data were analyzed by analysis of variance, followed by Tukey's post hoc test at α=0.05. RESULTS: The results showed that the administration of the CGG root ethanol extract disrupted the testis interstitial area and seminiferous tubules, resulting in decreased epididymal sperm quality as well as serum testosterone levels in a dose-dependent pattern. CONCLUSION: Oral administration of a CGG root ethanol extract induced testicular damage, decreased epididymal sperm quality, and impaired testosterone secretion.

Understanding divergent domestication traits from the whole-genome sequencing of swamp- and river-buffalo populations.

Domesticated buffaloes have been integral to rice-paddy agro-ecosystems for millennia, yet relatively little is known about the buffalo genomics. Here, we sequenced and assembled reference genomes for both swamp and river buffaloes and we re-sequenced 230 individuals (132 swamp buffaloes and 98 river buffaloes) sampled from across Asia and Europe. Beyond the many actionable insights that our study revealed about the domestication, basic physiology and breeding of buffalo, we made the striking discovery that the divergent domestication traits between swamp and river buffaloes can be explained with recent selections of genes on social behavior, digestion metabolism, strengths and milk production.

Intestinal and hepatic coccidiosis among rabbits in Yogyakarta, Indonesia.

BACKGROUND AND AIM: The attention to rabbit meat production in Indonesia is comparatively less to other farm animals such as cattle and poultry industries. However, future prospect of rabbit to be seriously industrialized seemed quite promising due to rabbit is highly productive and has short reproduction cycle as well as generation interval. One of the diseases infecting many rabbits is coccidiosis caused by protozoan parasite, Eimeria spp. The infectious stage of Eimeria spp. presents ubiquitously in the environment and increases the risk of parasite transmission. Preventive methods such as vaccination are not yet fully developed, while sporadic treatment is not efficiently reduce the cases. In this study, Eimeria spp. infecting rabbits in Yogyakarta Province, Indonesia, were investigated with the aim for precise diagnosis to determine targeted treatment and as a baseline epidemiological data from rabbit in Indonesia. MATERIALS AND METHODS: Sample collection was performed randomly for 3 months, from March 2017 to May 2017 and covered areas in Yogyakarta, Indonesia. A total of 750 samples were collected. Eimeria species identification was determined morphologically from the samples after sporulation in 2.5% potassium dichromate byCOCCIMORPH. RESULTS: Ten species of Eimeria spp. were identified in this study from the positive samples (527/750; 70.3%). Eimeria flavescens was present in 80% of the positive samples, Eimeria coeciola in 78%, Eimeria perforans in 61%, Eimeria exigua in 37%, Eimeria media in 33%, Eimeria stiedae in 31%, Eimeria irresidua in 12%, Eimeria magna in 11%, Eimeria intestinalis in 10%, and Eimeria piriformis in 10%. Coinfection as noted in 80% of the positive samples with 2-6 species in a specimen. E. flavescens and E. coeciola were the most prevalent among all Eimeria spp. (p≤0.0001). CONCLUSION: Eimeria spp. is detected in high prevalence among rabbit in Yogyakarta, Indonesia, with commonly occurs in mixed infections. In this paper, we describe Eimeria spp. that are circulating in Indonesia and present it as updated information to farmers and veterinarians. To the best of our knowledge, we provide the first information about rabbit coccidiosis in Indonesia.

Bovine coccidiosis cases of beef and dairy cattle in Indonesia.

The number of bovine coccidiosis cases is generally high and distributed worldwide. It can reach up to 100% in the early weeks of age in calves and has a corresponding impact on further cattle development and the economic outcome. For these reasons, this study was conducted to detect the presence of Eimeria spp. oocysts followed by morphologically species characterization in different breeds of beef and dairy cattle. These breeds were reared by rural farmers in 9 provinces in Indonesia from March to October 2017. A total of 2150 fecal samples were collected, along with farmer knowledge about coccidiosis through interviews. Results show that prevalence varied as indicated by the following regions: Riau (68.73%), South Sumatera (78.34%), Banten (63.73%), Special Region of Yogyakarta (54.69%), East Java (63.89%), Bali (83.34%), West Nusa Tenggara (78.43%), Central Kalimantan (72.46%), and South Sulawesi (85.07%). The overall prevalence of bovine coccidiosis in all provinces sampled was 72.07%. Eight species of Eimeria spp. were identified, namely, E. auburnensis (52.11%), E. bovis (81.89%), E. bukidnonensis (18.44%), E. canadensis (38.22%), E. cylindrica (31.67%), E. zuernii (60%), E. ellipsoidalis (10.11%), and E. alabamensis (11.67%). The highest prevalence was found in E. bovis compared with any other Eimerias (P ≤ .0001) followed by E. zuernii. Most of the interviewed farmers in this study did not know about coccidia, especially Eimeria spp. in cattle. They were more familiar with worm infections because of both their experience and knowledge. The distribution of cattle with ages <6 months, 6-18 months, and >18 months exhibited 97%, 81%, and 38% infection, respectively. Based on these results, the authors saw the necessity to inform farmers about bovine coccidiosis as well as the veterinarian regarding better preventive and curative treatments.

Floor space and betaine supplementation alter the nutrient digestibility and performance of Japanese quail in a tropical environment.

This study investigated the effects of floor space and betaine supplementation on the nutrient digestibility and performance of laying quail, at an advanced stage of production, in a tropical environment. In total, 408 laying quail (23 weeks old) were distributed in 3 × 2 factorial arrangements with four replicates. The first factor was floor space (250 cm2 [FS-1], 221 cm2 [FS-2], and 200 cm2 per bird [FS-3]) by allocating 15, 17, and 19 birds per cage (cage size: 3750 cm2). The second factor was betaine supplementation, administered at levels of 0 (Control) and 0.12% (Betaine). The birds were housed under a natural, tropical environment that was outside the predominant thermoneutral zone, indicating heat stress conditions. Interactions were found between floor space and betaine on crude fiber digestibility, egg production, and eggshell thickness, with the highest values being yielded in FS-2 group supplemented with betaine. Reducing the floor space to 200 cm2 negatively affected nutrient digestibility and performance. The birds kept in FS-3 had lower (P < 0.05) dry matter, crude protein, crude fat, and calcium digestibility values than those kept in FS-1 and FS-2. Birds housed in FS-2 exhibited higher egg weight than those in FS-3 (P < 0.05), although they did not differ from those in FS-1. Furthermore, a lower eggshell weight in FS-3, compared with FS-1 and FS-2, was observed (P < 0.01). Betaine supplementation enhanced the dry matter, crude protein, crude fat, crude ash, and calcium digestibility values (P < 0.05). Accordingly, betaine improved feed intake, egg weight, feed conversion, and protein and energy efficiency ratios (P < 0.01). Betaine also resulted in higher albumen, yolk, and eggshell weights (P < 0.01). In conclusion, a floor space of 221 cm2, combined with betaine supplementation at 0.12%, can be applied for raising quail under high environmental temperatures.

Correction to: Sulforaphane protects granulosa cells against oxidative stress via activation of NRF2-ARE pathway.

There is an error in the original publication of this paper. Figures 1-6 were shown in the wrong version, thus corrected figures provided were in this article.

Sulforaphane protects granulosa cells against oxidative stress via activation of NRF2-ARE pathway.

Sulforaphane (SFN) has been considered as an indirect antioxidant and potential inducer of the Nrf2-ARE pathway. This study was conducted to investigate the protective role of SFN against oxidative stress in bovine granulosa cells (GCs). GCs were collected from antral follicles (4-8 mm) and cultured according to the experimental design where group 1 = control, group 2 = treated with SFN, group 3 = treated with hydrogen peroxide (H2O2), group 4 = pretreated with SFN and then with H2O2 (protective) and group 5 = treated with H2O2 followed by SFN treatment (rescuing). Results showed that SFN pretreatment significantly increases cell viability and reduces cytotoxicity in GCs under oxidative stress. Following H2O2 exposure, expression of NRF2 was found to be significantly increased (p < 0.05) in SFN-pretreated cells, while no significant differences were observed between group 3 and group 5, although the expression was significantly increased compared to the control group. Moreover, the relative abundance of the NRF2 downstream target antioxidant genes (CAT, PRDX1, SOD1 and TXN1) were higher (fold change ranged from 7 to 14, p < 0.05) in sulforaphane pretreated GCs. Low level of ROS and lipid accumulation and higher mitochondrial activity were observed in GCs pretreated with SFN, whereas no such changes were observed in GCs treated with SFN after exposure to oxidative stress (group 5). Thus, we suggest that SFN pretreatment effectively protects GCs against oxidative damage through the activation of the NRF2-ARE pathway, whereas addition of SFN during oxidative insult failed to rescue GCs.

Chicken Coccidiosis in Central Java, Indonesia: A Recent Update.

Avian coccidiosis is a huge problem worldwide. Heavily infected animals that show severe clinical signs and coccidiostat resistance are causing important economic losses. The present study aimed to update the recent cases of coccidiosis in Central Java, Indonesia, and to show the importance of the disease in the region. A total of 699 samples were obtained from different chicken breed. Different Eimeria species were detected in 175 individuals (25.04%). Three different groups of chicken breed were considered: local chicken (autochthonous chickens of Sentul and Jawa), commercial broiler, and layer. Broiler chickens showed the highest prevalence of infection (34%), followed by layer (26.26%) and local chickens (10.45%). Mild to severe clinical signs of avian coccidiosis were observed in 42% of the infected animals, while 58% of the infected animals showed no clinical signs other than low feed conversion rates. Seven different Eimeria species were identified: E. tenella was the most prevalent (43.3%), followed by E. maxima (26.3%), E. necatrix (15.7%), E. acervulina (8%), E. praecox (3.1%), E. mitis (2.2%), and E. brunetti (1.3%). Coinfections with several Eimeria species were diagnosed. With this study we found massive usage of coccidiostat in the region even though its usage cannot guarantee coccidiosis-free chicken production.

Bovine embryo survival under oxidative-stress conditions is associated with activity of the NRF2-mediated oxidative-stress-response pathway.

In present study, we sought to examine the ability of preimplantation bovine embryos to activate the NF-E2-related factor 2 (NRF2)-mediated oxidative-stress response under an oxidative stress environment. In vitro 2-, 4-, 8-, 16-cell-, and blastocyst-stage embryos were cultured under low (5%) or high (20%) oxygen levels. The expression of NRF2, KEAP1 (NRF2 inhibitor), antioxidants downstream of NRF2, and genes associated with embryo metabolism were analyzed between the embryo groups using real-time quantitative PCR. NRF2 and KEAP1 protein abundance, mitochondrial activity, and accumulation of reactive oxygen species (ROS) were also investigated in blastocysts of varying competence that were derived from high- or low-oxygen levels. The expression levels of NRF2 and its downstream antioxidant genes were higher in 8-cell, 16-cell, and blastocyst stages under high oxygen tension, whereas KEAP1 expression was down-regulated under the same conditions. Higher expression of NRF2 and lower ROS levels were detected in early (competent) blastocysts compared to their late (noncompetent) counterparts in both oxygen-tension groups. Similarly, higher levels of active nuclear NRF2 protein were detected in competent blastocysts compared to their noncompetent counterparts. Thus, the survival and developmental competence of embryos cultured under oxidative stress are associated with activity of the NRF2-mediated oxidative stress response pathway during bovine pre-implantation embryo development.