Use of platelet lysate for in-vitro embryo production and treatment of repeat breeding in cows.

Platelet-rich plasma (PRP) is a biological hemocomponent derived from blood after the complete removal of red blood cells and the partial or complete removal of white blood cells to concentrate platelets in an appropriate volume of plasma. Platelets have important growth factors, cytokines, and active metabolites that improve the endometrial environment and positively affect implantation. This study evaluated the effect of the addition of activated PRP (platelets lysate; PL) on in vitro bovine oocyte maturation and embryonic development and the effect of intrauterine (IU) infusion of autologous PL in repeat breeder (RB) cows. Experiment 1 examined the effects of allogeneic PL, fetal calf serum (FCS), mixed PL + FCS, or platelet-poor plasma (PPP) supplementations to in vitro maturation and development media on in vitro oocyte maturation and embryo development in good- and poor-quality oocytes of Japanese Black cows. Experiment 2 examined the IU infusion of autologous PL, 24 h post-insemination, in 21 RB Holstein-Friesian dairy cows. The cleavage rate of good-quality oocytes was higher in the PL group (85.93 ± 2.50%) than in the PPP group (67.16 ± 3.41%) (P < 0.05), while the cleavage rate of the poor-quality oocytes was higher in the PL alone (76.13 ± 4.04%) and mixed PL + FCS treated (73.59 ± 4.22%) groups than in the PPP group (54.64 ± 2.93%) (P < 0.05). The blastocyst rate of the good-quality oocytes was higher in the PL group (40.97 ± 3.03%) than in the FCS (27.97 ± 3.31%) and PPP (25.33 ± 2.15%) groups (P < 0.05). The blastocyst rate of poor-quality oocytes and the hatching rates of both good and poor-quality oocytes showed no significant differences among all groups. The conception rate in the autologous PL-treated group was 41.67% (5/12), while it was 11.11% (1/9) in the control group. The platelets' count in the pregnant PL-treated cows (n = 5; mean ± SEM, 1.07 ± 0.10 × 109/mL) was higher than in the non-pregnant ones (n = 7; 0.67 ± 0.10 × 109/mL) (P < 0.05). In conclusion, allogeneic PL was effective in stimulating the in vitro oocyte maturation and embryonic development in both good and poor-quality bovine oocytes, and post-insemination IU infusion of autologous PL derived from high platelets' count-PRP would be recommended for the treatment of RB cows.

Exogenous gonadotropin-releasing hormone counteracts the adverse effect of scrotal insulation on testicular functions in bucks.

This study determined the effects of scrotal insulation on testicular functions in bucks and evaluated the impact of exogenous gonadotropin-releasing hormone (GnRH) administration before scrotal insulation on sperm production and testicular vascular dynamics. Twelve bucks were randomly divided into three groups: scrotal-insulated animals without GnRH treatment (INS), scrotal-insulated animals treated previously with GnRH (GnRH + INS), and animals without insulation as controls (CON). Doppler ultrasonography was used to evaluate testicular vascular changes, and semen samples were collected to assess seminal parameters. Testicular samples were collected from slaughtered bucks at the end of the experiment for histological investigations and immunohistochemical analysis for caspase 3 (apoptotic marker), and a vascular endothelial growth factor (VEGF; hypoxic marker) evaluation. Sperm motility drastically decreased (33%) in the INS group on day 8 compared with those in the GnRH + INS and CON groups (58% and 85%, respectively). Testicular blood flow significantly decreased for 3 and 2 weeks in the INS and GnRH + INS groups, respectively. The pulsatility index (PI) reached pretreatment values at 5 and 4 weeks after insulation in the INS and GnRH + INS groups, respectively. The resistance index (RI) values increased in both insulated groups for the first 2 weeks and decreased to control values 4 weeks after insulation. However, the maximum velocity (VP) started to increase reaching pretreatment values by the 5th and 3rd weeks after insulation in the INS and GnRH + INS groups, respectively. Histological investigations showed a marked reduction in lipid inclusions in Sertoli cells in the GnRH + INS group compared with those in the INS group. The distributions of both caspase 3 and VEGF decreased in the GnRH + INS group compared with those in the INS group. This study showed that the administration of a single dose of GnRH delayed the negative effects of scrotal insulation on different seminal traits and revealed the pivotal role of GnRH in compensating testicular insulation in bucks.

Toxoplasma gondii seropositivity and the associated risk factors in sheep and pregnant women in El-Minya Governorate, Egypt.

BACKGROUND AND AIM: The cosmopolite protozoan, Toxoplasma gondii, has a significant economic and medical impact. Cats traditionally play a predominant role in the disease maintenance cycle; however, humans can be infected as a result of milk and meat consumption of Toxoplasma-infected livestock. In addition, infected pregnant women, even symptomless, can pass the disease to their unborn fetus. The limited clinical records and absence of specific national educational programs in countries like Egypt underscore the need for periodic toxoplasmosis disease evaluation. Here, we identified T. gondii seroprevalence among sheep and pregnant women and the associated risk factors in El-Minya Governorate, Egypt. MATERIALS AND METHODS: Using peripheral blood, we detected T. gondii-specific antibodies in 151 sheep and 96 pregnant women sera from El-Minya Governorate using latex agglutination and indirect enzyme-linked immunosorbent assay. The impact of different environmental and behavioral risk factors identified with in-person interviews and serology results on acquiring toxoplasmosis was statistically analyzed. RESULTS: The overall toxoplasmosis seroprevalence was 39.1% and 22.9% in sheep and pregnant women, respectively. Significantly higher seroprevalence was correlated with increasing sheep age and geographical location. Nonetheless, no statistical significance was found based on abortion history and pregnancy status of the examined sheep. Exposure factors important for pregnant women included pregnancy trimester, contact with cats, and the habit of eating undercooked sheep meat, which all had a statistically significant association with Toxoplasma seropositivity. CONCLUSION: The current study confirms increased antibodies against toxoplasmosis in both sheep and pregnant women in El-Minya Governorate and a clear association between women's age, contact with cats, and the habit of eating undercooked sheep meat and seroreactivity to T. gondii. These results strongly suggest the need for a more comprehensive epidemiological study and public health awareness education for toxoplasmosis.

Effect of exogenous estradiol Benzoate on uterine blood flow in postpartum dairy cows.

The objective of this study was to assess the uterine blood flow following estradiol benzoate administration in Holstein-Friesian dairy cows by trans-rectal color Doppler ultrasonography. Six healthy lactating Holstein-Friesian cows were examined daily for 10 days starting at 4 weeks postpartum. All the cows, which were clinically healthy based on vaginal mucus scoring and endometrial cytology, were examined by trans-rectal Doppler ultrasonography to measure pulsatility index (PI), resistance index (RI), time average maximum velocity (TAMAX), blood flow volume (BFV) and diameter in the uterine arteries ipsilateral and contralateral to the previously pregnant uterine horn. On the third day of the experiment, the six cows were administered 10 mg intramuscular injection of estradiol-17ß (E2).Blood samples were collected at the time of daily examination for the assessment ofE2concentrations.The PI and RI values decreased while TAMAX, BFV and diameter of uterine arteries increased in response toE2 administration (P < 0.05).There was a high correlation between both the ipsilateral and contralateral uterine arteries for all variables that were studied(r = 0.860, P < 0.0001, r = 0.922, P < 0.0001, r = 0.651, P < 0.0001, r = 0.879, P < 0.0001, r = 0.861, P < 0.0001 for the PI, RI, TAMAX, BFV and uterine arteries diameter, respectively).In conclusion, the greater blood concentrations ofE2may be responsible for the greater TAMAX, BFV, increased diameters and decreased PI and RI of the uterine arteries during the puerperium in dairy cows.

Sero-diagnosis of brucellosis in sheep and humans in Assiut and El-Minya governorates, Egypt.

Egypt is an endemic area with brucellosis, so wherever herd problem associated abortion is present, brucellosis should be suspected, and its sero-diagnosis is needed. The present study aimed to estimate the seroprevalence of brucellosis in sheep and their contact humans in Assiut and El-Minya Governorates where a history of abortion in sheep was the chief complaint of the farmers; besides; the appearance of signs of undulant fever among some contact humans. Also, to identify the risk factors for brucellosis seropositivity at human and animal level. Serum samples were collected from 189 sheep and 53 in contact humans in Assiut and El-Minya Governorates, Egypt, during 2017. Antibodies against brucellosis were measured in the serum samples using Rose Bengal Plate test (RBPT) and were further confirmed using Serum Tube Agglutination Test (STAT). The overall seroprevalence of brucellosis using RBPT was 15.87% in sheep and that in humans was 9.44% (5 of 53), respectively. STAT confirmed that 40% of the RBPT-positive reactor sheep were infected by Brucella abortus, 16.67% were containing Brucella melitensis and 20% were experiencing a mixed infection of both Br. abortus and Br. melitensis. Additionally, 80% of the RBPT-positive reactor humans were infected with Br. abortus only and 20% were having both species of Brucella. Among different analyzed variables in this study, the age and farming system of the examined sheep were statistically significant. There was no significant effect in relation to gender, abortion history and pregnancy status of the examined animals, however, higher exposure rates were found among males, abortive animals, and recently calved animals. Gender of the tested humans and presence of seropositive sheep in contact differed significantly in acquiring the infection. Nevertheless, other factors, including age, education, and profession were statistically insignificant on getting human brucellosis. These results indicate the high seroprevalence of brucellosis in humans and sheep in the study areas and generally in Egypt, and therefore, control programs should be implemented.

Cross-sectional study for determining the prevalence of Q fever in small ruminants and humans at El Minya Governorate, Egypt.

OBJECTIVE: Q fever is a febrile illness caused by the bacterial pathogen Coxiella burnetii (C. burnetii) and is transmitted to humans from small ruminants via contaminated secreta and excreta of infected animals. This pathogen threatens public health; however, little is known regarding Q fever prevalence in humans and small ruminants. Therefore, we employed a cross-sectional design to determine the Q fever seroprevalence and the associated risk factors in small ruminants and their owners in El Minya Governorate, Egypt between August 2016 and January 2017. RESULTS: The seroprevalence of C. burnetii IgG antibodies was 25.68% (28 of 109), 28.20% (11 of 39) and 25.71% (9 of 35) in sheep, goats, and humans, respectively. None of the studied variables in small ruminants differed significantly between the seropositive and seronegative animals. There was a significantly higher prevalence (P = 0.0435) and increased odds of exposure was also observed among women (odds ratio, OR = 5.43 (95% CI 1.058-27.84) when compared to men; nevertheless, no significant difference was noted between the infection rate in small ruminants and humans. This study clearly points out that Q fever may be emerging in the area which lay the foundation for early prediction and better management of possible future outbreaks.

Effects of intrauterine infusion of povidone-iodine on endometrial cytology and bacteriology in dairy cows with clinical endometritis.

Endometritis is one of the major diseases causing infertility in the cow. Intrauterine infusion of povidone-iodine (PVP-I) is a common treatment. However, the optimal concentration of PVP-I for treating endometritis effectively remains unknown. We tested concentrations of 2.0% or 0.5% PVP-I for treating clinical endometritis in dairy cattle. In Experiment 1, bacteria isolated from the uterus were incubated with either 2.0% or 0.5% PVP-I, and the numbers of bacterial colonies were counted. In Experiment 2, 18 cows with clinical endometritis were treated with either 2.0% or 0.5% PVP-I (n=9 in each group). Cytology samples and bacteria were collected using a cytobrush on weeks 0 (W0), 1 (W1) and 2 (W2) after treatment. Subsequent reproductive performance was compared between the two groups. In Experiment 1, both concentrations had a similar antiseptic outcome. In Experiment 2, the percentage of polymorphonuclear neutrophils (PMN%) in the endometrial epithelium at W2 in the 2.0% group was significantly lower (P<0.05) than in the 0.5% group, although the PMN% decreased significantly from W0 to W2 (P<0.01) in both groups. Decreases in bacterial infection rates from W0 to W2 were similar in both groups. The first service conception rate was higher, numbers of services per conception were fewer, and time to conception was shorter in the 2.0% group than in the 0.5% group. Thus, an intrauterine infusion of 2.0% PVP-I was better than 0.5% in treating clinical endometritis in these dairy cattle.

Quantification of epicardial fat: Which method can predict significant coronary artery disease?

AIM: To compare the predictive value of three methods of epicardial fat (EF) assessment for presence of significant coronary artery disease (CAD) [i.e., epicardial fat volume (EFV), EFV indexed with body surface area (EFV/BSA) and EFV indexed with body mass index (EFV/BMI)]. METHODS: The study was performed on 170 patients (85 women and 85 men) with clinical suspicion of CAD. They aged 26-89 years with a median age of 54 years. The patients were classified into three groups: Group 1: 58 patients with normal coronary arteries; group 2: 48 patients with non-significant CAD and group 3: 64 patients with significant CAD. The three methods for assessment of epicardial fat were retrospectively studied to determine the best method to predict the presence of significant CAD. RESULTS: The three methods for epicardial fat quantification and measurements, i.e., EFV, EFV/BSA and EFV/BMI with post- hoc analysis showed a significant difference between patients with significant coronary artery disease compared to the normal group. Receiver operating characteristic curve analysis showed no significant difference between the three methods of epicardial fat measurements, the area under curve ranging between 0.6 and 0.62. The optimal cut-off was 80.3 cm(3) for EFV, 2.4 cm(3)/m(2) for EFV indexed with BMI and 41.7 cm(3)/(kg/m(2)) for EFV indexed with BSA. For this cut-off the sensitivity ranged between 0.92 and 0.94, while specificity varied from 0.31 to 0.35. CONCLUSION: Any one of the three methods for assessment of epicardial fat can be used to predict significant CAD since all have the same equivalent predictive value.