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Department of Animal Sciences
The Robert H. Smith Faculty
of Agricultural, Food & Environment

The Hebrew University of Jerusalem.

Herzl 229, Rehovot 7610001, Israel
Phone: +972-(0)8-9489119;
Fax: +972-(0)8-9465763;
Yael Lewitus, Department's Secretary
e-mail: yaellew@savion.huji.ac.il

Publications

2021
Basavaraja, R. ; Drum, J. N. ; Sapuleni, J. ; Bibi, L. ; Friedlander, G. ; Kumar, S. ; Sartori, R. ; Meidan, R. . Downregulated Luteolytic Pathways In The Transcriptome Of Early Pregnancy Bovine Corpus Luteum Are Mimicked By Interferon-Tau In Vitro. BMC GENOMICS 2021, 22.Abstract
BackgroundMaintenance of the corpus luteum (CL) beyond the time of luteolysis is essential for establishing pregnancy. Identifying the distinct features of early pregnancy CL remains unresolved, hence we analyzed here the transcriptome of CL on day 18 pregnant (P) and non-pregnant (NP) cows using RNA-Seq. CL of P cows expressed ISGs, verifying exposure to the pregnancy recognition signal, interferon-tau (IFNT), whereas the CL of NP cows had elevated luteal progesterone levels, implying that luteolysis had not yet commenced.ResultsThe DEGs, IPA, and metascape canonical pathways, along with GSEA analysis, differed markedly in the CL of P cows from those of NP cows, at the same day of the cycle. Both metascape and IPA identified similar significantly enriched pathways such as interferon alpha/beta, sonic hedgehog pathway, TNFA, EDN1, TGFB1, and PDGF. However, type-1 interferon and sonic hedgehog pathways were positively enriched whereas most of the enriched pathways were downregulated in the P compared to NP samples. Thirty-four % of these pathways are known to be elevated by PGF2A during luteolysis. Notably, selective DEGs in luteinized granulosa cells were modulated by IFNT in vitro in a similar manner to their regulation in the CL of P cows.ConclusionThis study unraveled the unique transcriptomic signature of the IFNT-exposed, early pregnancy CL, highlighting the abundance of downregulated pathways known to be otherwise induced during luteolysis. These and IFNT-regulated in vitro pregnancy-specific DEGs suggest that IFNT contributes to the characteristics and maintenance of early pregnancy CL.
Roth, Z. . Heat Stress Reduces Maturation And Developmental Capacity In Bovine Oocytes. REPRODUCTION FERTILITY AND DEVELOPMENT 2021, 33, 66-75.Abstract
The ovarian pool of follicles, and their enclosed oocytes, is highly sensitive to hyperthermia. Heat-induced changes in small antral follicles can later manifest as impaired follicle development and compromised competence of the enclosed oocytes to undergo maturation, fertilisation and further development into an embryo. This review describes the main changes documented so far that underlie the oocyte damage. The review discusses some cellular and molecular mechanisms by which heat stress compromises oocyte developmental competence, such as impairment of nuclear and cytoplasmic maturation and mitochondrial function, changes in the expression of both nuclear and mitochondrial transcripts and the induction of apoptosis. The review emphasises that although the oocyte is exposed to heat stress, changes are also evident in the developed embryo. Moreover, the effect of heat stress is not limited to the summer; it carries over to the cold autumn, as manifest by impaired steroid production, low oocyte competence and reduced fertility. The spontaneous recovery of oocytes from the end of the summer through the autumn until the beginning of winter suggests that only subpopulations of follicles, rather than the entire ovarian reserve, are damaged upon heat exposure.
Bartman, J. ; Zaguri, S. ; Avital-Cohen, N. ; Dishon, L. ; Druyan, S. ; Gumulka, M. ; Rozenboim, I. . Targeted Differential Illumination Improves Reproductive Traits Of Broiler Breeder Males. POULTRY SCIENCE 2021, 100.Abstract
Artificial targeted illumination has a pivotal role in reproductive processes of poultry. The light-absorption mechanism in birds consists of 2 main components: the eye (retinal photoreceptors) and extraretinal photoreceptors located in the brain. Previous studies conducted on hens have shown that photostimulation of brain extraretinal photoreceptors elevates reproductive activity, whereas retinal photostimulation suppresses it. We tested the effect of targeted differential photostimulation (TDP) on reproductive activities of broiler breeder males. Fifty broiler breeder roosters (Ross), 21 wk of age, were divided into 5 environmentally controlled light-treatment rooms (n = 10) equipped with individual cages. Rooms 1 and 2 had 2 parallel lighting systems consisting of red light (630 nm) and green light (514 nm), and rooms 3 and 4 had parallel red and blue (456 nm) lighting systems. Room 5, illuminated with white light, served as the control. Birds of all groups were kept under short day (6L:18D) for 2 wk with both lighting systems in each treatment room turned on. At 23 wk of age, birds were photostimulated by gradually increasing one of the lighting systems to 14 h of light in each room, while the other lighting system was left on short day (6L:18D). Weekly semen samples were collected until 65 wk of age and analyzed for volume, motility, concentration and vitality. Monthly blood samples were drawn for plasma hormone assays. At 65 wk of age, roosters were euthanized and hypothalamus, pituitary gland, retina and testes samples were taken for mRNA expression analysis. TDP using long-day red light and short-day green light significantly increased reproductive performance, manifested by higher semen volume, motility and concentration, and testis weight; furthermore, this group had higher plasma testosterone levels, higher GnRH mRNA expression in the hypothalamus, lower levels of aromatase in the testes, and lower mRNA expression of hypothalamic serotonin transporter, and of pituitary prolactin and its receptors in the testes. This is the first study showing a positive effect of TDP on reproduction of broiler breeder roosters.
Komsky-Elbaz, A. ; Kalo, D. ; Roth, Z. . Carryover Effect Of Atrazine And Its Metabolite-From Treated Bovine Spermatozoa To The Embryo'S Transcriptome. BIOLOGY OF REPRODUCTION 2021, 104, 1162-1180.Abstract
Atrazine (ATZ) is an extensively used herbicide and ubiquitous environmental contaminant. ATZ and its metabolite, diaminochlorotriazine (DACT), cause several cellular and functional alterations in spermatozoa. We aimed to examine the effect of ATZ/DACT on spermatozoon DNA integrity, fertilization competence, embryonic development, and transcriptome profile of in vitro-produced embryos derived from fertilization with pre-exposed sperm. Bovine spermatozoa exposed to ATZ (0.1 or 1 mu M) or DACT (1 or 10 mu M) during in vitro capacitation were used for in vitro fertilization of untreated oocytes. Cleavage and blastocyst-formation rates were evaluated 42 h and 7 days postfertilization, respectively. The association between DNA fragmentation and apoptosis (annexin V kit) was determined. Fertilization competence of annexin-positive (AV+) and annexin-negative (AV-) spermatozoa was examined. Microarray analysis was performed for 7-day blastocysts. Intracytoplasmic sperm injection was performed with control (AV+, AV-) and DACT (AV+, AV-) spermatozoa. Cleavage rates did not differ between groups and blastocyst formation tended to be higher for AV- vs. AV+ in both control and DACT groups, suggesting that acrosome reaction, rather than DNA fragmentation, underlies the reduced cleavage. Transcriptomic analysis revealed 139 and 230 differentially expressed genes in blastocysts derived from ATZ- and DACT-exposed spermatozoa, respectively, relative to controls. Proteomic analysis shown differential expression of proteins in ATZ- or DACT-treated spermatozoa, in particular proteins related to cellular processes and biological pathways. Therefore, we assume that factors delivered by the spermatozoa, regardless of DNA fragmentation, are also involved. Overall, the current study reveals a deleterious carryover effect of ATZ/DACT from the spermatozoa to the developing embryo. Summary sentence Summary sentence: Atrazine and its metabolite, diaminochlorotriazine, induce multifactorial damage in sperm, which is carried over to the embryo's transcriptome.
Azaria, S. ; Nussinovitch, A. ; Nir, S. ; Mordechai, S. ; van Rijn, J. . Removal Of Geosmin And 2-Methylisoborneol From Aquaculture Water By Novel, Alginate-Based Carriers: Performance And Metagenomic Analysis. JOURNAL OF WATER PROCESS ENGINEERING 2021, 42.Abstract
Hydrophobic carriers were examined for geosmin and 2-methylisoborneol removal from water derived from an aquaculture system. A combination of adsorption and biodegradation was found to underlie the removal of the off-flavor compounds. Adsorption of these compounds by the carriers was unaffected by the presence of organic matter in the water to be treated. A model based on adsorption/desorption and first-order degradation kinetics provided an accurate prediction for experimentally determined 2-methylisoborneol removal rates. Steady removal of geosmin and 2-methylisoborneol as well as nitrate reduction were observed during long-term operation of the plug-flow reactors with water derived from an aquaculture facility. Metagenomic analysis of the microbial community on the carriers during long-term operation of the reactors revealed a predominance of denitrifying bacteria. It was found that geosmin and 2-methylisoborneol led to statistically significant changes in the abundances of 21 contigs that contained genes involved in terpene degradation. This study shows that at low ambient concentrations of geosmin and 2-methylisoborneol in nitrate and organic-rich water, such as found in aquaculture systems, their biodegradation can be accomplished by terpene-degrading denitrifiers that develop on hydrophobic carriers used for filtration of the contaminated water.
Roth, Z. ; Kressel, Y. Z. ; Lavon, Y. ; Kalo, D. ; Wolfenson, D. . Administration Of Gnrh At Onset Of Estrus, Determined By Automatic Activity Monitoring, To Improve Dairy Cow Fertility During The Summer And Autumn. ANIMALS 2021, 11.Abstract
Simple Summary We used an automatic activity-monitoring system to determine onset of estrus in dairy cows. Within 5 h of onset, we administered a single injection of GnRH analogue to improve fertility during the summer and autumn. The treatment increased pregnancy per insemination during the autumn, but not in the summer. The subgroups for which the treatment specifically tended to improve conception risk during the autumn were: mature (2nd plus parity) cows and cows with uterine disease and ketosis after calving. Detection of estrus onset by activity monitoring and GnRH administration shortly thereafter could be incorporated into a synchronization program, to improve fertility of positively-responding subpopulations of cows. We examined gonadotropin-releasing hormone (GnRH) administration at onset of estrus (OE), determined by automatic activity monitoring (AAM), to improve fertility of dairy cows during the summer and autumn. The study was performed on two dairy farms in Israel. The OE was determined by AAM recorded every 2 h, and a single im dose of GnRH analogue was administered shortly after OE. Pregnancy was determined by transrectal palpation, 40 to 45 d after artificial insemination (AI). Conception risk was analyzed by the GLIMMIX procedure of SAS. Brief visual observation of behavioral estrus indicated that about three-quarters of the events (n = 40) of visually detected OE occurred within 6 h of AAM-detected OE. Accordingly, the GnRH analogue was administered within 5 h of AAM-detected OE, to overlap with the expected endogenous preovulatory LH surge. Overall, pregnancy per AI (P/AI) was monitored over the entire experimental period (summer and autumn) in 233 first, second or third AI (116 and 117 AI for treated and control groups, respectively). Least square means of P/AI for treated (45.8%) and control (39.4%) groups did not differ, but group-by-season interaction tended to differ (p = 0.07), indicating no effect of treatment in the summer and a marked effect of GnRH treatment (n = 58 AI) compared to controls (n = 59 AI) on P/AI in the autumn (56.6% vs. 28.5%, p < 0.03). During the autumn, GnRH-treated mature cows (second or more lactations), and postpartum cows exhibiting metabolic and uterine diseases, tended to have much larger P/AI than their control counterparts (p = 0.07-0.08). No effect of treatment was recorded in the autumn in first parity cows or in uninfected, healthy cows. In conclusion, administration of GnRH within 5 h of AAM-determined OE improved conception risk in cows during the autumn, particularly in those exhibiting uterine or metabolic diseases postpartum and in mature cows. Incorporation of the proposed GnRH treatment shortly after AAM-detected OE into a synchronization program is suggested, to improve fertility of positively responding subpopulations of cows.
Kogan, T. ; Dahan, D. G. ; Laor, R. ; Argov-Argaman, N. ; Zeron, Y. ; Komsky-Elbaz, A. ; Kalo, D. ; Roth, Z. . Association Between Fatty Acid Composition, Cryotolerance And Fertility Competence Of Progressively Motile Bovine Spermatozoa. ANIMALS 2021, 11.Abstract
Simple Summary:& nbsp;Spermatozoa move forward in the female reproductive tract in a linear pattern defined as progressive motility (PM). PM is associated with fertilization competence and used as a parameter to evaluate the spermatozoa's quality. The livestock-breeding industry is based on artificial insemination with cryopreserved spermatozoa, making the relationship between PM and cryosurvival important. This study explores the association between PM and spermatozoa's quality, cryotolerance and fertilization competence. The number of progressively motile spermatozoa within a standard insemination straw positively correlates with conception rate, making progressively motile spermatozoa's survival during cryopreservation a potentially relevant parameter for spermatozoa evaluation.
An association between progressive motility (PM) and spermatozoa fertility competence has been suggested. However, the mechanism that underlies PM is not clear enough. We examined physiological characteristics and fatty acid composition of fresh spermatozoa with high and low PM. Additional analysis of fatty acid composition and structural characteristics was performed on spermatozoa samples with high and low progressively motile spermatozoa's survival (PMSS), i.e., the ratio between the proportion of progressively motile spermatozoa after and before cryopreservation. Finally, a fertility field trial was conducted to examine the association between the number of PM spermatozoa within the insemination straw post thawing and conception rate. Analysis of fresh spermatozoa revealed a higher omega-6 to omega-3 ratio in ejaculates with low PM relative to those with high PM (p < 0.01). The proportion of polyunsaturated fatty acids was higher in low-PMSS fresh samples (p < 0.05) relative to their high-PMSS counterparts. Fresh samples with high-PMSS expressed a higher mitochondrial membrane potential (p < 0.05) and a higher proportion of viable cells that expressed reactive oxygen species (ROS; p < 0.05). Post-thawing evaluation revealed a reduced proportion of progressively motile sperm, with a prominent effect in samples with high PM relative to low PM, defined before freezing (p < 0.01). No differences in spermatozoa mitochondrial membrane potential or ROS level were found post-thawing. A fertility study revealed a positive correlation between the number of progressively motile spermatozoa within a standard insemination straw and conception rate (p < 0.05). Considering these, the bull PMSS is suggested to be taken into account at the time of straw preparation.
Reicher, N. ; Uni, Z. . Intestinal Brush Border Assembly During The Peri-Hatch Period And Its Contribution To Surface Area Expansion. POULTRY SCIENCE 2021, 100.Abstract
Microvilli generate the small intestinal brush border, the main site of nutrient digestion and absorption. Mucosal structuring of the small intestine of chicken during the perihatch period has been widely researched, yet the developmental dynamics of micro-villi during this period have not been fully character-ized. In this study, we examined the structural and molecular characteristics of microvilli assembly and maturation during the perihatch period. Small intes-tines of broiler embryos and chicks were sampled at prehatch ages 17 E and 19 E, at day of hatch (DOH) and at 1, 3, 7, and 10 d posthatch. Morphological evalu-ations and measurements were conducted by scanning electron microscopy (SEM) and light microscopy (LM) (n = 3/timepoint), and expression of microvilli structural genes Plastin 1, Ezrin, and Myo1a was exam-ined by Real-Time qPCR (n = 6/timepoint). Results revealed dissimilar patterns of microvilli and villi devel-opment during the perihatch period. From 19 E to 1 d, microvilli lengths increased 4.3-fold while villi lengths increased 2.8-fold (P < 0.0001). From 3 to 7 d, villi lengths increased by 20% (P < 0.005), while microvilli lengths decreased by 41% (P = 0.001). At 10 d, micro-villi lengths stabilized, while villi continued to elongate by 26% (P < 0.0001). Estimations of the microvilli amplification factor (MAF) and total enterocyte surface area (TESA) revealed similar trends, with peak values of 78.53 and 1961.67 mu m(2), respectively, at 3 d. Microvilli structural gene expression portrayed diverse patterns. Expression of Plastin 1, which bundles and binds actin cores to the terminal web, increased 8.7-fold between 17 E and DOH (P = 0.005), and gradually increased up to 7 d (P = 0.045). Ezrin and Myo1a, both actin core-cell membrane cross-linkers, portrayed dif-ferent expression patterns throughout the perihatch period, as Ezrin expression was relatively stable, while Myo1a expression increased 15.8-fold between 17 E and 10 d (P < 0.0001). We conclude that microvilli assem -bly during the perihatch period is a rapid, coordinated process, which dramatically expands the digestive and absorptive surface area of the small intestine before the completion of villi maturation.