In the cow, cryopreserved semen is inseminated into the uterus, and most of sperm are removed by backflow and phagocytes. Nevertheless, the mechanism responsible for sperm phagocytosis is unclear. Here, we used cultured bovine uterine epithelial cells (BUECs) to investigate the uterine response to sperm and the mechanism that activates polymorphonuclear neutrophils (PMNs). BUEC monolayers were co-cultured with different numbers of washed sperm obtained from cryopreserved semen (10 , 10 , and 10 sperm/ml) for 3 hr. Sperm dose-dependently up-regulated IL8 (Interleukin 8). Sperm at 10 /ml increased mRNA expression of TNFA (Tumor necrosis factor alpha), IL1B (Interleukin 1B), NFKB2 (Nuclear factor kappa B2), and C3 (Complement factor 3), as well as PGES (Prostaglandin E synthase) expression and PGE release. Live sperm, but not dead sperm, attached to BUECs, and dead sperm did not induce an acute inflammatory response. Time-dependent effects were evaluated by co-culture of 10 /ml washed sperm with BUECs for 0, 1, 3, and 6 hr. The number of detached sperm increased gradually toward 6 hr. Maximum mRNA expression of IL8, TNFA, IL1B, and NFKB2 was induced at 3 hr, while C3 continued to increase toward 6 hr. Sperm did not stimulate mRNA expression of anti-inflammatory cytokines TGFB1 (Transforming growth factor beta 1) or IL10 (Interleukin 10). Medium conditioned by sperm co-incubated with BUECs stimulated PMNs phagocytosis of sperm in vitro. Fresh media supplemented with low levels of IL1B, TNFA, and PGE up-regulated sperm phagocytosis by PMNs as well. In conclusion, our findings strongly suggest that the active sperm attach to BUECs and trigger uterine local innate immunity with induction of a pro-inflammatory response that enhances sperm phagocytosis by PMNs.

Graphical abstract

Hyperglycemia, the mark normal for diabetes and associated disorders are the main goals of natural diabetes therapies. In this context, the present research was designed to study the effects of fenugreek sprouts juice (FS), barley sprouts juice (BS), cell-free probiotic extract (cell-free PE), whey protein hydrolysate (WPH) and their mixture on diabetic rats. Free radical scavenging activity, total phenolic contents (TPC) and total flavonoid contents (TFC) of each item mentioned were determined. Diabetes was induced through the injection of male rats with a single intraperitoneal dose (45 mg/kg) of streptozotocin. After the development of diabetes, diabetic rats were orally administered daily with 1ml of with fenugreek sprouts juice, barley sprouts juice, cell-free probiotic extract, whey protein hydrolysate or their mixture until the end of the study period (45 day). Oral administration of fenugreek sprouts juice, barley sprouts juice, cell-free probiotic extract, whey protein hydrolysate and their mixture to diabetic rats significantly reduced fasting blood glucose levels and improved the lipid profile. All the studied items limit the reductions of haemoglobin concentrations and plasma α-amylase activities. Also all the studied items suppressed the elevation of malondialdehyde values and the reduction of catalase activities. Histopathological investigation of pancreas, liver and kidneys of the diabetic rats showed histological alterations. On the other hand, supplementations with the tested materials lead to relieving these injuries. Results revealed that fenugreek sprouts juice, barley sprouts juice, cell-free probiotic extract, whey protein hydrolysate and their mixture had promising effects towards hyperglycemia and associated disorders.

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The effect of Honey bee supplementation into Tris extender on semen quality and bacterial activity of chilled-stored Barky ram' semen was studied. Semen samples were processed in Barky ram's semen extender with or without honey bee. Seven adult Barky rams aged 24-36 month and weighed 43.0± 1.5 kg were used. A total of 210 ejaculates were collected from the seven rams using an artificial vagina, 30 ejaculates each, twice weekly throughout the period of the study (July-October, 2016). After collection, each ejaculate was diluted (1:10) with Tris-based extender at 37˚C immediately after dilution, the specimens were split into 4 aliquots; the first aliquot served as control (C), whereas the other 3 aliquots were supplemented with 1.5 (HB 1.5), 3 (HB 3) and 4.5 (HB 4.5) ml honey bee/100 ml extender. Both control and treated specimens were subjected to chilled preservation at 5˚C for a period of 48 h, during which semen physical characteristics were evaluated, immediately after dilution time and every 24 h thereafter until 48 h storage at 5˚C. Microbial contamination in all specimens was also determined at 0, 24 and 48 h. The results showed that supplementing semen extender with honey bee improved (P<0.05) sperm motility and reduced (P < 0.05) percent of dead and abnormal spermatozoa and acrosomal damage. In conclusion, supplementing semen extender with honey bee for ram semen preservation at 5°C has reduced the deleterious effects due to chilling condition. This would facilitate the application of assisted reproductive technologies; i.e. artificial insemination and in vitro fertilization in sheep by extending the storagability and survival of sheep sperm.

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Dairy products are exposed to contamination by aflatoxin M 1 (AFM 1 ) directly or by cross-contamination with aflatoxin B 1 . Strategies applied to reduce their impacts include probiotics and prebiotics. Zizyphus (Ziziphus spina-christi) fruit contains bioactive compounds that may act as prebiotics. The objective of this research was to discover what function encapsulated probiotics, Zizyphusz fruit, and their combination play in preventing oxidative liver damage and genotoxicity in rats. Probiotics (Streptococcus thermophilus, Lactobacillus acidophilus,, and Bifidobacterium bifidum) and Zizyphusz were encapsulated separately using a soy protein concentrate and maltodextrin as carriers. Encapsulated probiotics and Zizyphusz as well as their combination with camel milk were evaluated in both normal and AFM 1 -exposed rats. Biochemical alterations and histological changes were determined. Also, DNA damage was assessed by the comet assay protocol. At the studied doses, the encapsulated probiotics (2.7 × 10 11 CFU/ml) and Zizyphusz (1 mg) reduced the elevation of liver functions, oxidative marker (malondialdehyde), inflammatory marker (tumor necrosis factor-alphaα), and DNA damage in rats exposed to AFM 1 . Possibly, the encapsulated probiotics or Zizyphusz can effectively protect the liver from oxidative stress and DNA damage caused by AFM 1 .

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