Influence of the coincubation of post-thawed bull semen with elevated ß-hydroxybutyrate concentrations on sperm characteristics.

In this study, the relationships between post-thaw bull sperm characteristics and hyperketonemic conditions after coincubation with cow plasma or media were determined to investigate if such a condition could affect bull sperm characteristics. Two experiments were conducted. In experiment 1, blood samples were collected from 31 cows to prepare plasma. Cows were independently categorized into two groups according to plasma ß-hydroxybutyrate (BHB) concentrations (above or below 1.2 mM). Thawed bull semen was diluted and incubated with diluted plasma; motility parameters were evaluated using Computer Assisted Semen Analysis (CASA). In experiment 2, a pooled sample of thawed semen was diluted and divided into three aliquots: without BHB (control) and treated with either 1.2 mM (1.2) or 3 mM (3) BHB. In addition to motility, flow cytometric analyses were carried out. In experiment 1, the overall motility decreased significantly in plasma containing high (≥1.2 mM) BHB compared to plasma containing low (<1.2 mM) BHB. In experiment 2, the overall motility tended to be lower in BHB (3 mM)-supplemented samples. The supplementation of 3 mM BHB increased the proportion of live superoxide-positive sperm and sperm with high mitochondrial potential, while the DNA fragmentation index decreased.

The Effect of Vitamin E and Poly Ethylene Glycol (PEG) Association on Chilled Rabbit Sperm: Impact on Sperm Motility and Oxidative Stress Status.

BACKGROUND: Vitamin E is the major lipid-soluble antioxidant, however, its potent effect is limited by its poor solubility. OBJECTIVE: The present study aimed to investigate whether polyethylene glycol 6000 (PEG) can enhance Vitamin E solubility and help protect sperm motility and against oxidative status. MATERIALS AND METHODS: Sperm groups consisted of the control aliquot diluted with Tris buffer and aliquots treated with Tris buffer containing polyethylene glycol (PEG), vitamin E (Vit E) or vitamin E- PEG complex (PEG/Vit E). Sperm motility was measured using a Computer Aided Semen Analysis at 0, 1, 3, 6 hours of cooling at 4°C. The oxidative stress status was measured at 4 hours using ABTS radical scavenging capacity. RESULTS: Sperm motility and oxidative status were significantly protected when using PEG and Vit E individually; however the most potent effects were observed in PEG/Vit E treatment. CONCLUSION: The present study demonstrated that treating rabbit semen with vitamin E complexed to PEG 6000 (PEG/Vit E) is effective in protecting sperm cells during chilling at 4°C.

Association of pregnancy per artificial insemination with gonadotropin-releasing hormone and human chorionic gonadotropin administered during the luteal phase after artificial insemination in dairy cows: A meta-analysis.

One strategy for improving fertility in cattle is administration of GnRH or human chorionic gonadotropin (hCG) during the luteal phase, which increases progesterone (P4) secretion and delays luteolysis. To provide an overview of how GnRH or hCG treatment between 4 and 15 d after artificial insemination (AI) improves pregnancy per AI (P/AI) in cows, a meta-analysis was performed on 107 different trials from 52 publications. Data from 18,082 treated cows and 18,385 untreated controls were meta-analyzed. The meta-analysis explained the relative risk for P/AI with GnRH or hCG treatment under various circumstances. The results did not show any difference in P/AI between cows treated with hCG and cows treated with GnRH. Compared with no treatment, treatment with GnRH or hCG improved the chances of P/AI in cows with very poor (<30%) and poor (30.1 to 45%) fertility, whereas treatment did not benefit cows with very good fertility (>60.1%). Moreover, treatment with GnRH and hCG improved the chances of P/AI in primiparous cows. The improvement was much better in primiparous cows with very low fertility. Treatment with buserelin at a dose above 10 µg and with hCG at a dose above 2,500 IU was associated with increased chances of P/AI compared with lower doses. Treatment with GnRH 10 d after AI was also associated with increased chances of P/AI compared with earlier treatment. The present meta-analysis showed that the use of GnRH and hCG after AI should be focused on cows expected to have low or moderate fertility. Day and dose of treatment have to be considered as well.

Comparative efficacy of two-dimensional mode and color Doppler sonography in predicting gender of the equine fetus.

BACKGROUND AND AIM: Ultrasonographic fetal sexing is of utmost economic importance for horse breeders. Relatively, a few studies have been conducted to determine fetal sex in mare using transrectal Doppler ultrasound. This study aimed to compare two sexing techniques, two-dimensional (2D) mode and color Doppler ultrasonography. MATERIALS AND METHODS: The study was conducted on 39 mares under field conditions. Examinations were performed using the ultrasonic model device (Medison SonoAce Pico, South Korea), equipped with real-time 3-7 MHz convex multifrequency transducer. Fetal sex diagnosis was carried out in two periods of pregnancy, early period (57-80 days of gestation) and late period (80-150 days of gestation). RESULTS: No difference (p=0.4) was observed between the efficiency of the 2D mode and Doppler ultrasound in sex determination with the respective frequency of 74% and 85%. The best time to sex the early fetus was between 57 and 70 days of gestation and between 90 and 120 days for the older fetus. CONCLUSION: The accuracy of sex determination can be increased using Doppler mode compared to 2D mode, especially in the early period between 57 and 70 days, in male more than female sex.

Prevalence and factors associated with a higher or lower risk of exposure to Coxiella burnetii, Chlamydia abortus and Toxoplasma gondii in dairy cows that have aborted in Algeria. / Prévalences et facteurs associés à un risque augmenté ou diminué d'exposition à Coxiella burnetii, Chlamydia abortus et Toxoplasma gondii chez la vache laitière ayant avorté en Algérie.

In Algeria, the prevalence of causes of abortion on dairy cattle farms (whether infectious causes or not) has been little studied. The current study involved a serological analysis conducted between October 2014 and June 2016 in northern Algeria using an enzyme-linked immunosorbent assay test on blood samples taken from 368 cows that had aborted on 124 farms. It was complemented by a survey to identify the factors associated with a higher or lower risk of exposure to Coxiella burnetii, Chlamydia abortus and Toxoplasma gondii, using univariate logistic regression and then multivariate logistic regression. The individual serological prevalences obtained were 8.4% (31/368) for C. burnetii and 12.2% (45/368) for C. abortus. For T. gondii, the individual seroprevalence was 13.8% (51/368); the factors associated with a higher risk of individual exposure were the fourth month of gestation (odds ratio [OR] = 22.68; 95% confidence interval [CI]: 1.38-392.97) and the fifth month of gestation (OR = 25.51; 95% CI: 1.47-442.11). All the other factors identified by the multivariate logistic regression were associated with a lower risk of exposure. They are the inspection visits in 2015 (OR = 0.0006; 95% CI: 0.000004-0.12) and in 2016 (OR = 0.0005; 95% CI: 0.000002-0.13) and artificial insemination (OR = 0.15; 95% CI: 0.05-0.44) for C. burnetii ; winter (OR = 0.39; 95% CI: 0.15-1.00), spring (OR = 0.45; 95% CI: 0.20-0.97), and artificial insemination (OR = 0.27; 95% CI: 0.13-0.56) for C. abortus; and the number of gestations (OR = 0.38; 95% CI: 0.16-0.92) for T. gondii. The seroprevalence at herd level was 16.1% (20/124) for C. burnetii and 29.8% (37/124) for both C. abortus and T. gondii. At herd level, the risk factors associated with a higher risk of exposure to C. abortus and T. gondii were the practice of deworming (OR = 3.89; 95% CI: 1.53-9.89) and drilling individual wells as a source of drinking water (OR = 7.50; 95% CI: 2.11-26.69). For C. burnetii, the inspection visit in 2015 (OR = 0.02; 95% CI: 0.0008-0.65) and in 2016 (OR = 0.01; 95% CI: 0.0003-0.36), artificial insemination (OR = 0.21; 95% CI: 0.06-0.69) and rodent eradication (OR = 0.19; 95% CI: 0.06-0.57) were factors that reduced the risk of exposure.

En Algérie, la prévalence des causes d'avortement dans les élevages bovins laitiers (que ces causes soient d'origine infectieuse ou non infectieuse) a été peu étudiée. Cette étude concerne une analyse sérologique conduite d'octobre 2014 à juin 2016 dans le nord de l'Algérie à l'aide d'un test ELISA (méthode immuno-enzymatique) sur des prélèvements sanguins issus de 368 vaches ayant avorté provenant de 124 élevages et complétée par un formulaire d'enquête visant à identifier, par une régression logistique uni puis multivariée, les facteurs associés à un risque augmenté ou diminué d'exposition à Coxiella burnetii, Chlamydia abortus et Toxoplasma gondii. Les prévalences sérologiques individuelles obtenues ont été respectivement de 8,4 % (31/368) pour C. burnetii et de 12,2 % (45/368) pour C. abortus. Pour T. gondii, la séroprévalence individuelle était de 13,8 % (51/368), avec comme facteurs associés à un risque accru d'exposition individuelle, le quatrième mois de gestation (rapport des cotes [odds ratio : OR] = 22,68 ; intervalle de confiance [IC] à 95 % : 1,38-392,97) et le cinquième mois de gestation (OR = 25,51 ; IC à 95 % : 1,47-442,11). Les autres facteurs identifiés par la régression logistique multivariée étaient tous des facteurs associés à un risque diminué d'exposition. Ils concernaient l'année de visite en 2015 (OR = 0,0006 ; IC à 95 % : 0,000004-0,12) et en 2016 (OR = 0,0005 ; IC à 95 % : 0,000002-0,13) et l'insémination artificielle (OR = 0,15 ; IC à 95 % : 0,05-0,44) pour C. burnetii ; l'hiver (OR = 0,39 ; IC à 95 % : 0,15-1,00), le printemps (OR = 0,45 ; IC à 95 % : 0,20-0,97) et l'insémination artificielle (OR = 0,27 ; IC à 95 % : 0,13-0,56) pour C. abortus ; et le nombre de gestations (OR = 0,38 ; IC à 95 % : 0,16-0,92) pour T. gondii. La séroprévalence obtenue à l'échelle du troupeau a été respectivement de 16,1 % (20/124) pour C. burnetii et de 29,8 % (37/124) pour C. abortus et T. gondii. À l'échelle des troupeaux, les facteurs associés à un risque accru d'exposition à C. abortus et T. gondii concernent respectivement la pratique du déparasitage (OR = 3,89 ; IC à 95 % : 1,53-9,89) et le forage personnel comme source d'abreuvement (OR = 7,50 ; IC À 95 % : 2,11-26,69). Pour C. burnetii, l'année de visite en 2015 (OR = 0,02 ; IC à 95 % : 0,0008-0,65) et en 2016 (OR = 0,01 ; IC à 95 % : 0,0003-0,36), l'insémination artificielle (OR = 0,21 ; IC à 95 % : 0,06-0,69) et l'éradication des rongeurs (OR = 0,19 ; IC à 95 % : 0,06-0,57) sont des facteurs de diminution du risque d'exposition.

En Argelia está poco estudiada la prevalencia de las causas de aborto en las explotaciones bovinas de producción láctea (ya sean causas de tipo infeccioso o no infeccioso). Los autores exponen aquí un análisis serológico realizado entre octubre de 2014 y junio de 2016 en el norte de Argelia. Empleando una técnica de ensayo inmunoenzimático (ELISA), se examinaron muestras sanguíneas de 368 vacas que habían sufrido un aborto procedentes de 124 explotaciones, análisis que se complementó con un formulario de investigación destinado a determinar, por regresión logística de una sola variable y después multifactorial, los factores ligados a un aumento y a un decremento del riesgo de exposición a Coxiella burnetii, Chlamydia abortus y Toxoplasma gondii. Las prevalencias serológicas individuales obtenidas fueron, respectivamente, de un 8,4% (31/368) para C. burnetii y un 12,2% (45/368) para C. abortus. Por lo que respecta a T. gondii, se obtuvo una seroprevalencia individual del 13,8% (51/368). Los factores ligados a un mayor riesgo de exposición individual eran el cuarto mes de gestación (razón de probabilidades [odds ratio: OR] = 22,68; intervalo de confianza [IC] al 95%: 1,38-392,97) y el quinto mes de gestación (OR = 25,51; IC al 95%: 1,47-442,11). Todos los otros factores determinados por regresión logística multifactorial venían asociados a una reducción del riesgo de exposición. Se trataba concretamente de: el hecho de que la visita se hubiera efectuado en 2015 (OR = 0,0006; IC al 95%: 0,000004-0,12) o en 2016 (OR = 0,0005; IC al 95%: 0,000002-0,13) y la inseminación artificial (OR = 0,15; IC al 95%: 0,05-0,44), en el caso de C. burnetii; el hecho de que fuera invierno (OR = 0,39; IC al 95%: 0,15-1,00) o primavera (OR = 0,45; IC al 95%: 0,20-0,97) y la inseminación artificial (OR = 0,27; IC al 95%: 0,13-0,56), en el caso de C. abortus; y el número de gestaciones (OR = 0,38; IC al 95%: 0,16-0,92), en el caso de T. gondii. La seroprevalencia de rebaño obtenida fue respectivamente de un 16,1% (20/124) para C. burnetii y de un 29,8% (37/124) para C. abortus y T. gondii. A la escala de los rebaños, los factores vinculados a un mayor riesgo de exposición a C. abortus y T. gondii eran, respectivamente, la práctica de desparasitaciones (OR = 3,89; IC al 95%: 1,53-9,89) y el uso de un pozo propio como abrevadero (OR = 7,50; IC al 95%: 2,11-26,69). En cuanto a C. burnetii, los factores que reducían el riesgo de exposición del rebaño eran el hecho de que el año de visita fuera 2015 (OR = 0,02; IC al 95%: 0,0008-0,65) o 2016 (OR = 0,01; IC al 95%: 0,0003-0,36), la inseminación artificial (OR = 0,21; IC al 95%: 0,06-0,69) y la erradicación de los roedores (OR = 0,19; IC al 95%: 0,06-0,57).

Effects of female bovine plasma collected at different days of the estrous cycle on epididymal spermatozoa motility.

The aim of this study was to assess the effects of female bovine plasma collected at different days of the reproductive cycle on epididymal spermatozoa motility and to test hypothesis that the subpopulations pattern of motile spermatozoa is affected by this treatment. Blood plasma samples were collected from five Holstein Friesian cows at different stages of the estrous cycle (days 0, 5, 10, 12 and 18), one pregnant cow and one adult bull and were diluted 1:9 (V/V) with normal saline. Female charcoal-treated plasma, Bull plasma and saline were used as controls. Semen samples were obtained from cauda epididymidis through retrograde flushing and diluted in saline to approximately 60 × 106 sperm/ml. The extended semen was diluted 1:2 (V/V) with tested media and motility was evaluated at 15 min and then every hour for 6 h using a computer-assisted semen analysis. Multivariate clustering procedure was applied to identify and quantify specific subpopulations within the semen samples. The statistical analysis clustered all the motile spermatozoa into three separate subpopulations with defined patterns of movement: Subpopulation 1 poorly motile and non-progressive spermatozoa (39.3%), subpopulation 2 including the fastest and the most vigorous spermatozoa (46.4%) and subpopulation 3 represented by slow, non-vigorous but linear spermatozoa (14.3%). Initially, sperm samples supplemented with female, male or female charcoal-treated plasma stimulated equally total motility and spermatozoa belonging to subpopulation 2 regardless of the estrous cycle stage. After 1-h incubation, the motility of these both categories of spermatozoa (total motile and those assigned to subpopulation 2) is enhanced and maintained more in day 12, 18 and pregnant cow plasma than in female plasma from earlier stage of the estrous cycle (day 0, 5 and 10), male plasma and female-charcoal treated plasma. In conclusion, the overall results showed that female plasma stimulated significantly sperm motility, especially at the late stage of the estrous cycle. Additionally, to the diverse compounds contained in blood plasma, progesterone may play a key role in such motility activation.

Effect of equilibration time on the motility and functional integrity of canine spermatozoa frozen in three different extenders.

The present work aimed to assess the effect of equilibration time on post-thaw motility parameters of canine sperm frozen in three extenders: 6% low-density lipoproteins (LDL), 6% liposomes (LIPO), and 40% egg yolk plasma (EYP). A second experiment is aimed at evaluating the functional integrity of canine spermatozoa frozen in the three extenders at the best equilibration time found in the experiment one. In the first experiment, 20 ejaculates harvested from 7 dogs, were frozen in three extenders (LDL, LIPO, and EYP) after four equilibration times (30min, 1h, 3h, and 6h). The semen was evaluated after thawing using an image analyser (HT-IVOS 14.0). The 6h equilibration time gave better results of motility and progressive motility in the three studied extenders. (LDL: 58.9% vs. 42.7%; LIPO: 54.4% vs. 31.9%; EYP: 55.4% vs 40.5% for motility 6 vs. 1h). In the second experiment, 10 ejaculates taken from 6 dogs were frozen under the same conditions as the previous experiment, after 6h equilibration time. The integrity parameters of the spermatozoal membrane (hypo-osmotic swelling test, and SYBR14/propidium Iodide staining), acrosome (FITC-Pisium sativum Aglutinin staining), and DNA (acridine orange staining) were evaluated at three different stages: post-dilution (T0), post-equilibration, and post-thawing. Post-thaw results were as follows: membrane integrity (HOSt: 62;6% vs 58% vs 64.4%; SYBR14/IP: 63.6% vs 57.9% vs 64.8%); acrosome integrity (FITC-PSA: 79.4% vs 74% vs 76.2%) and DNA integrity (Acridine-orange: 98.9% vs 98.5% vs 98.7%) respectively for LDL vs. LIPO vs. EYP. No significant difference existed between the extenders tested; thus 6%LIPO and 40%EYP could be good candidates for replacement of 6%LDL in the protection of canine sperm during the freeze-thaw process without altering motility and integrity parameters.

In-house preparation and evaluation of (125)I-histamine progesterone tracer for radioimmunoassay of progesterone.

The progesterone tracer for the development of in-house radioimmunoassay was prepared in two steps clearly described. First, activation of 11α-hemisuccinate progesterone (progesterone-11 α HS) and labeling of histamine with (125)I, and second, conjugation of activated 11α-HS progesterone with iodinated histamine. The purification of the tracer was carried out by gel filtration on Sephadex G-25. The radiochemical purity of purified the tracer was 95%. The maximum binding using solid phase (coated tubes) reached 41% and the non specific binding didn't exceed 5%. The tracer stored at different temperatures 10°C and -6°C was stable during 12 weeks. The skim milk provided assay better sensitivity (0.22 ng/mL) than serum (0.28 ng/mL).

First epidemiological study on exposure to Neospora caninum in different canine populations in the Algiers District (Algeria).

Neospora caninum is an important cause of abortion in cattle worldwide. Dogs act as final hosts shedding oocysts in the environment. They can also harbour the extraintestinal stage of the parasite and this may be associated with a fairly rare neuromuscular condition. The sera of 781 dogs from the Algiers District were screened by IFAT for the presence of anti-N. caninum antibodies. These dogs were distributed into four populations: local stray dogs, police dogs, dogs from breeding kennels and farm dogs. The overall seroprevalence was 21.90%. Significant differences were observed between the different populations, the highest prevalence being observed in farm (44.44%) and stray dogs (22.55%). Additionally, the highest titres were observed in farm dogs. Among studied epidemiological parameters, breed, dog origin, season and vaccination status were significantly associated with IFAT results. Additionally, a recently described real time PCR was used on the blood of 100 pound dogs and the results were compared with the serological data. A higher proportion of dogs was found to be positive by PCR when compared to the IFAT results. There was only a fairly low agreement between PCR and IFAT results which suggests that these techniques measured different aspects of the host-parasite relationship. This study indicates that the level of exposure of the canine population of Algiers area to N. caninum is very high. This would indicate a potentially high risk for N. caninum induced abortion in cattle in this region and in Algeria.

Uterine collagen during pregnancy in cattle.

Uteri were collected at the slaughter of non-pregnant dairy cattle and cattle at various stages of gestation. The weight of the whole uterus increased about 12-fold during the 9 month gestation period. The greatest increase was in the weight of the pregnant horn. The increase in weight of the uterine horns was accompanied by an increase in collagen content. The collagen concentration did not change. The collagen crosslink pyridinoline was identified in pregnant uterine tissues at a level of about 0.13 residues per mole of collagen; this level was the same at the beginning and end of pregnancy.

Uterine collagen during involution in cattle.

Post-partum involution of the bovine uterus was assessed by clinical examination and monitored biochemically by measuring the total urinary excretion of hydroxyproline and the collagen cross-link, pyridinoline. Uterine tissue contained 0.13 +/- 0.04 residues of pyridinoline per mole of collagen and negligible (less than 0.005 residues/mole) deoxypyridinoline. The pyridinoline/creatinine ratio was found to be a clearer and more reliable indicator of uterine resorption than the hydroxyproline/creatinine ratio. The amount of collagen cross-link excreted above baseline was equivalent to the amount of cross-link estimated to have been accumulated during pregnancy. Maximal excretion of pyridinoline occurred at day 6 and returned to baseline values in 2-3 weeks after parturition. An electron microscope study during the period of peak resorption revealed macrophage-like cells apparently engulfing or containing collagenous material, indicating phagocytic removal of the connective tissue.