A proteomic approach to identifying spermatozoa proteins in Indonesian native Madura bulls.

Proteins assist sperm mature, transit the female reproductive tract, and recognise sperm oocytes. Indigenous Indonesian bulls, Madura bulls, have not been studied for reproductive proteomics. As local Indonesian beef livestock, Madura cattle assist in achieving food security; hence, their number must be improved. Thus, the identification of molecular proteomics-based bull fertility biomarkers is needed. This study aimed to characterise the sperm fertility function of the superior Madura bull (Bos indicus × Bos Javanicus) spermatozoa proteome. Frozen semen from eight Madura superior bulls (Bos indicus × Bos javanicus) aged 4-8 years was obtained from the artificial insemination centre (AIC) in Singosari and Lembang. Madura superior bulls are those that have passed the bull breeding soundness evaluation. Frozen sperm were thawed and centrifuged at 3000 × g for 30 min. Proteins in sperm were characterised through proteomic analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The resulting gene symbols for each protein were then subjected to bioinformatics tools, including UniProt, DAVID, and STRING databases. Regarding sperm fertility, the analysis revealed that 15 proteins were identified in the sperm of Madura bulls. Amongst the identified proteins, the superior Madura bull sperm contained several motilities, energy-related proteins, and chaperone proteins. A substantial portion of characterised proteins are linked to metabolic pathways and the tricarboxylic acid (TCA) cycle, contributing to sperm energy production. In conclusion, the first in-depth proteome identification of sperm related to sperm quality and bull fertility of a unique indigenous Madura breed of Indonesia was performed using the LC-MS/MS proteomic method. These findings may serve as a reference point for further studies related to the functions of bovine sperm and biomarkers of fertility and sperm quality.

The reproductive success of Simmental bovine after sex-sorting under various incubation and centrifugation protocols.

Background and Aim: To enhance the reproductive potential and increase productivity and population of cows, spermatozoa sex-sorting technology is required. This study aimed to examine the effect of sexing sperm, separated using a bovine serum albumin (BSA) column with varying incubation durations and centrifugation methods, for successful artificial insemination. Materials and Methods: Six Simmental bulls and 30 cows (n = 30) as the recipients were selected for this study at Balai Pembibitaan Hijauan Pakan Ternak Sembawa Indonesia. The study parameters included sperm motility, viability, plasma membrane integrity, and conception rate (CR). The experiment was divided into three protocols to find out differences in some parameters: (1) BSA incubation time effect (P) with P1 (40 min), P2 (50 min), and P3 (60 min); (2) freezing time effect with before freezing and after-thawing treatments; and (3) CR determined by measuring the proportion of pregnant cows following insemination with non-sexed, X-bearing, and Y-bearing sperms without centrifugation (n = 15) (A0, A1, and A2) and with centrifugation (n = 15) (B0, B1, and B2) in the acquired data, which were counted using the Statistical Package for the Social Sciences version 21 program. Analysis of variance was utilized to evaluate all treatments at various levels. Results: The results demonstrated that centrifugation time influenced all sperm quality metrics for sperm containing X and Y (p < 0.05). The non-return rate (NRR) of non-sexed frozen semen, both centrifuged (A0) and not centrifuged (B0), was more significant than frozen semen produced by sexing X and Y spermatozoa. The NRR indicated a value of 80% based on the number of lactating cows. Conclusion: Bovine serum albumin incubation and centrifugation protocols influenced and decreased all sperm quality indicators throughout the sexing procedure and could still be used as a sexing protocol. Furthermore, regarding NRR and service per conception, non-sexual treatment is superior to sexing treatment.

A Molecular Interaction Analysis Reveals the Possible Roles of Graphene Oxide in a Glucose Biosensor.

In this paper, we report the molecular docking study of graphene oxide and glucose oxidase (GOx) enzyme for a potential glucose biosensing application. The large surface area and good electrical properties have made graphene oxide as one of the best candidates for an enzyme immobilizer and transducer in the biosensing system. Our molecular docking results revealed that graphene oxide plays a role as a GOx enzyme immobilizer in the glucose biosensor system since it can spontaneously bind with GOx at specific regions separated from the active sites of glucose and not interfering or blocking the glucose sensing by GOx in an enzyme-assisted biosensor system. The strongest binding affinity of GOx-graphene oxide interaction is -11.6 kCal/mol and dominated by hydrophobic interaction. Other modes of interactions with a lower binding affinity have shown the existence of some hydrogen bonds (H-bonds). A possibility of direct sensing (interaction) model of glucose by graphene oxide (non-enzymatic sensing mechanism) was also studied in this paper, and showed a possible direct glucose sensing by graphene oxide through the H-bond interaction, even though with a much lower binding affinity of -4.2 kCal/mol. It was also found that in a direct glucose sensing mechanism, the sensing interaction can take place anywhere on the graphene oxide surface with almost similar binding affinity.