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  1. Alfadel F, Yimer N, Hiew MWH
    Trop Anim Health Prod, 2023 Feb 10;55(2):76.
    PMID: 36764981 DOI: 10.1007/s11250-023-03490-x
    Bull breeding soundness evaluation (BBSE) is the most common procedure used to predict bull potential fertility. However, the use of traditional methods for semen evaluation can affect its reliability. The inclusion of additional advanced test in BBSE may increase its accuracy. This study aimed to investigate the correlation between the degree of sperm protamination and BBSE main parameters of scrotal circumference (SC), progressive motility (PM), morphologically normal sperm (NS), and different categories of morphological defects. In addition, to determine the correlation between the three methods used for protamine assessment, five Brangus bulls were subjected to the BBSE. Semen samples were collected via electro-ejaculation and evaluated using traditional methods. Three different methods were used to determine the degree of sperm protamination: aniline blue (AB) staining, chromomycin A3 staining with fluorescent microscope (CMA3-FLM), and CMA3 with flow cytometry (CMA3-FCM). Sperm protamine deficiency assessed using the three methods exhibited significant differences among bulls according to their classification by BBSE, and showed significant negative correlation with semen quality parameters of NS and PM. A significant positive correlation was found between AB positivity and morphological abnormalities. The three methods used for protamine assessment also revealed significant positive correlations. Among the three tests, AB staining was the cheapest and easiest test that offers an objective assessment method for sperm protamination. Hence, it can be concluded that the assessment of protamination using AB staining test might serve as an additional valuable parameter or a replacement whenever detail sperm motility and morphology analyses in conducting BBSE to predict bull fertility are not possible.
  2. Constable PD, Megahed AA, Hiew MWH
    J Dairy Sci, 2019 Dec;102(12):11370-11383.
    PMID: 31548071 DOI: 10.3168/jds.2019-16805
    Urine pH (UpH) and net acid excretion (NAE) are used to monitor the degree of systemic acidification and predict the magnitude of resultant hypercalciuria when feeding an acidogenic ration to control periparturient hypocalcemia in dairy cattle. The objectives of this study were to evaluate the diagnostic performance of urine dipstick and pH paper for measuring UpH, and to characterize the UpH-NAE relationship and the association of urine Ca concentration ([Ca]) with UpH and NAE. Urine samples (n = 1,116) were collected daily from 106 periparturient Holstein-Friesian cows fed an acidogenic ration during late gestation. Net acid excretion was measured by titration, and UpH was measured by a glass-electrode pH meter (reference method), Multistix-SG urine dipsticks (Siemens Medical Solutions Inc., Ann Arbor, MI), and Hydrion pH paper (Micro Essential Laboratory Inc., Brooklyn, NY). Diagnostic performance was evaluated using Spearman correlation coefficient (rs), Bland-Altman plots, and logistic regression. Urine pH measured by urine dipstick (rs = 0.94) and pH paper (rs = 0.96) were strongly associated with UpH. Method-comparison studies indicated that the urine dipstick measured an average of 0.28 pH units higher, and pH paper 0.10 pH units lower, than UpH. Urine [Ca] was more strongly associated with UpH (rs = -0.65) than NAE (rs = 0.52). Goals for controlling periparturient hypocalcemia under the study conditions were UpH <6.22 and <6.11, based on achieving urine [Ca] ≥5 mmol/L and estimated urinary Ca excretion ≥4 g/d, respectively. Urine pH was as accurate at predicting urine [Ca] as NAE when UpH >6.11. We conclude that pH paper is an accurate, practical, and low-cost cow-side test for measuring UpH and provides a clinically useful estimate of urine [Ca].
  3. Megahed AA, Hiew MWH, Ragland D, Constable PD
    J Dairy Sci, 2019 Jun;102(6):5550-5565.
    PMID: 30954258 DOI: 10.3168/jds.2018-15063
    High-producing dairy cows experience a state of negative energy balance in the periparturient period that is partially addressed by increasing the rate of fat and protein mobilization. Previous studies have focused on the rate of fat mobilization, and consequently the rate of protein mobilization has not been well characterized. The objective of this study was therefore to determine the change in indicators of muscle mass during early lactation using ultrasonographic measurement of muscle thickness and changes in plasma creatinine concentration. The maximum thickness of the gluteus medius and longissimus dorsi muscles of 106 Holstein cows (34 primiparous, 72 multiparous) was determined ultrasonographically on d -3, 0, 3, 7, 14, 21, and 28 relative to the day of parturition. Plasma creatinine concentration was measured periodically during the same period. Mixed models analysis and Passing-Bablok regression were used to analyze the data. Gluteus medius thickness, longissimus dorsi loin thickness (LDLT), and longissimus dorsi thoracic thickness (LDTT) were decreased at 28 d postpartum compared with d 3 antepartum. Plasma creatinine concentration was weakly associated with gluteus medius thickness, LDLT, and LDTT (Spearman's rho = 0.31, 0.39, and 0.32, respectively). Plasma creatinine concentration in primiparous and multiparous cows at 28 d postpartum decreased by 0.24 and 0.30 mg/dL, respectively, compared with values 3 d antepartum. We concluded that ultrasonographic measurement of LDLT and LDTT and change in plasma creatinine concentration may provide practical methods for monitoring the rate of protein mobilization in periparturient dairy cows. Ultrasonographic examination of LDLT and LDTT therefore complements ultrasonographic measurement of backfat thickness and may be useful in the evaluation of energy reserve mobilization in periparturient dairy cows.
  4. Alamaary MS, Haron AW, Hiew MWH, Ali M
    Vet Med Sci, 2020 11;6(4):666-672.
    PMID: 32602662 DOI: 10.1002/vms3.315
    Present study aimed to investigate the effect of adding antioxidants, cysteine and ascorbic acid on the levels of glutamic oxaloacetic transaminase (GOT), glutamic-pyruvate (GPT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and γ-glutamyl transpeptidase (GGT) enzymes of post-thawed stallion sperm. Ten ejaculates were collected each from four healthy stallions and cryopreserved using HF-20 freezing extender containing either 0 mg/ml cysteine or ascorbic acid, 0.5 mg/ml cysteine and 0.5 mg/ml ascorbic acid. All samples in freezing extender containing cysteine or ascorbic acid or none of them were assessed for sperm motility, viability, plasma membrane integrity, morphology and enzymes concentration. The ALP, LDH and GGT were significantly higher in 0-group compared with cysteine and ascorbic acid groups. The sperm motility of frozen-thawed semen with 0-group was significantly better compared with cysteine and ascorbic acid groups. The variation on viability, sperm membrane integrity and morphology were insignificant between all treated groups. Therefore, these enzymes were reduced when using antioxidants in the freezing extender. Results of the present study suggest that concentration of ALP, LDH and GGT enzymes could be used as parameters for prediction of frozen-thawed stallion semen.
  5. Megahed AA, Hiew MWH, Townsend JR, Constable PD
    Vet Clin Pathol, 2017 Jun;46(2):314-325.
    PMID: 28485837 DOI: 10.1111/vcp.12493
    BACKGROUND: The Precision Xtra electrochemical meter is widely used to measure blood β-hydroxybutyrate concentration (BHBb) in dairy cattle. The meter uses an algorithm optimized for human blood that assumes the HCT in cattle is the same as in people, and that intra-erythrocyte β-hydroxybutyrate (BHBe) and plasma β-hydroxybutyrate (BHBp) concentration are equivalent.

    OBJECTIVES: The first objective was to characterize the analytic performance of the meter for measuring BHBb and BHBp in dairy cattle. The second objective was to characterize the influence of HCT and sample temperature on BHBp concentration measured by the meter.

    METHODS: Blood and plasma samples were obtained from 106 periparturient Holstein cattle and 15 lactating Holstein cows with experimentally induced electrolyte and acid-base imbalances. Meter performance was evaluated using Deming regression and Bland-Altman plots. Multivariable linear regression was used to determine the effect of HCT and sample temperature on BHBb and BHBp concentration, respectively.

    RESULTS: The meter was linear up to BHB = 3.0 mmol/L as measured by the reference method, equivalent to meter values for BHBb > 4.5 mmol/L and BHBp > 5.2 mmol/L. An increase in HCT resulted in higher BHBb concentration. This result was partially explained by BHBe being much lower than BHBp. Changes in sample temperature caused a linear change in measured BHBp whenever BHBp > 3.0 mmol/L.

    CONCLUSIONS: Meter accuracy was markedly dependent on the BHBe-to-BHBp ratio and consequently the HCT. Therefore, the algorithm used by the meter should be revised when applied to bovine blood for improved accuracy.

  6. Hiew MWH, Megahed AA, Horstman LA, Constable PD
    J Dairy Sci, 2020 Jun;103(6):5575-5590.
    PMID: 32307156 DOI: 10.3168/jds.2019-17800
    An accurate, practical, and low-cost method for predicting parturition is urgently needed in the dairy industry. The objective of this study was to evaluate changes in plasma progesterone concentration ([prog]) and glucose concentration in whole blood ([gluc]b) and plasma ([gluc]p) as predictors of parturition within 6, 12, and 24 h in primiparous and multiparous Holstein cows. Blood samples were obtained daily at approximately 0900 h from 34 primiparous and 72 multiparous Holstein cows in late gestation and the time of calving recorded to the nearest hour. Plasma [prog] was measured using an ELISA, and [gluc]b and [gluc]p using a low-cost point-of-care glucose meter. The optimal cut-point for predicting parturition was determined using binomial logistic regression with general estimating equations, because the data set consisted of repeated measures for each cow. Diagnostic test performance was evaluated by comparing the area under the receiver operating characteristic curve (AUC) and calculating the sensitivity, specificity, and κ at the optimal cut-point for predicting parturition. Plasma [prog] was the most accurate predictor of parturition within 24 h (AUC = 0.96) and 12 h (AUC = 0.93), whereas [gluc]b was the most accurate predictor of parturition within 6 h (primiparous, AUC = 0.96; multiparous, AUC = 0.86). We conclude that a decrease in plasma [prog] is currently the most accurate test for predicting calving within 24 h. Measurement of [gluc]b is a promising new test for the cow-side prediction of parturition in dairy cows due to its accuracy, practicality, and low cost.
  7. Megahed AA, Hiew MWH, Constable PD
    J. Vet. Intern. Med., 2018 Mar;32(2):846-852.
    PMID: 29424465 DOI: 10.1111/jvim.15049
    BACKGROUND: Plasma fructosamine concentration ([FRA]) is a widely used long term hyperglycemic biomarker in humans and dogs, but its clinical usefulness as a hypoglycemic biomarker in dairy cattle is uncertain.

    OBJECTIVES: To evaluate the relationship between plasma [FRA] and glucose concentration ([gluc]) as well as indices of energy balance during early lactation in dairy cattle, and to characterize the influence of plasma total protein concentration ([TP]) and albumin concentration ([albumin]) on [FRA].

    ANIMALS: Convenience sample comprising 103 periparturient Holstein-Friesian cattle.

    METHODS: Plasma [gluc], [TP], [albumin], and other clinicopathologic indices of energy status were determined periodically from Day 4 postpartum. Body condition score (BCS) was assessed, and backfat thickness (BFT) and longissimus dorsi muscle thickness (LDT) were measured ultrasonographically. Plasma [FRA] was measured at approximately 28 days postpartum. Associations between plasma [FRA] and study variables were evaluated using Spearman's rho and stepwise forward linear regression. Statistical significance was declared at P 

  8. Alamaary MS, Haron AW, Ali M, Hiew MWH, Adamu L, Peter ID
    Vet World, 2019 Jan;12(1):34-40.
    PMID: 30936651 DOI: 10.14202/vetworld.2019.34-40
    Aim: Different types of extenders have a variety of components which show the tolerance effect on sperm protection during freezing procedures. In the present study, we have examined the impact of the extenders HF-20 and Tris, which were locally manufactured, and they are competing with commercial extenders INRA Freeze® (IMV Technologies, France) and EquiPlus Freeze® (Minitube, Germany) on the quality of horses frozen semen.

    Materials and Methods: A total of 15 ejaculates from three healthy stallions were collected and cryopreserved in the same environment. Each semen sample collected was divided into four equal parts and processed. All samples were analyzed before and after freezing for motility, viability, plasma membrane integrity, and morphology. Furthermore, twenty mares were inseminated using post-thawed semen.

    Results: There were no differences observed among all extenders in all the parameters before freezing. Sperm cryopreserved using HF-20 showed better motility, viability, and plasma membrane integrity than Tris extender. The Tris extender showed the most inferior quality of post-thawed semen between all the extenders. HF-20, INRA Freeze®, and EquiPlus Freeze® extenders revealed the same capacity of semen preservation in vitro and in vivo.

    Conclusion: HF-20 extender has the same quality as INRA Freeze® and EquiPlus Freeze® that can be considered as one of the best extenders for the semen cryopreservation in horses. In contrast, Tris extender needs some degree of improvement.

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