Background: A valuable tool for both research and industry, in vitro fertilization (IVF) has applications range from gamete selection and preservation of traits to cloning. Although IVF has achieved worldwide use, with approximately 339,685 bovine embryos transferred in 2010 alone, there are still continuing difficulties with efficiency. It is rare to have more than 40% of fertilized in vitro cattle oocytes reach blastocyst stage by day 8 of culture, and pregnancy rates are reported as less than 45% for in vitro produced embryos. To investigate potential influences in-vitro fertilization (IVF) has on embryonic development, this study compares in vivo- and in vitro-derived bovine blastocysts at a similar stage and quality grade (expanded, excellent quality) to determine the degree of transcriptomic variation beyond morphology using RNA-Seq.Results: A total of 26,906,451 and 38,184,547 fragments were sequenced for in vitro and in vivo embryo pools, respectively. We detected expression for a total of 17,634 genes, with 793 genes showing differential expression between the two embryo populations with false discovery rate (FDR) < 0.05. There were also 395 novel transcribed units found, of which 45 were differentially expressed (FDR < 0.05). In addition, 4,800 genes showed evidence of alternative splicing, with 873 genes displaying differential alternative splicing between the two pools (FDR < 0.05). Using GO enrichment analysis, multiple biological pathways were found to be significantly enriched for differentially expressed genes (FDR < 0.01), including cholesterol and sterol synthesis, system development, and cell differentiation.Conclusions: Thus, our results support that IVF may influence at the transcriptomic level and that morphology is limited in full characterization of bovine preimplantation embryos.

This study aimed to evaluate the impact of Basella rubra fruit extract (BR-FE) on cryopreserved ram sperm's motility, velocity, and membrane integrity. Thirty ejaculates collected from 3 fertile rams (10 from each) were diluted with semen dilution extender (SDE) in a ratio (1:2) and centrifuged to remove 50% supernatant. The remaining sample was mixed with semen cryopreservation extender (SCE) in 1:4 ratio. Then 1.2 mL of SCE diluted sample was divided in four aliquots (0.3 mL each) that were further extended with [(1) control group (0.7 mL of SCE), (2) BR-FE-0.6% group (0.7 mL of SCE supplemented with 0.6% BR-FE), (3) BR-FE-0.8% group (0.7 mL of SCE supplemented with 0.8% BR-FE), and (4) BR-FE-1.6% group (0.7 mL SCE supplemented with 1.6% BR-FE)]. All extended samples were cooled gradually from 25°C to 4°C in half an hour. The 0.1 mL sample from all aliquots was analyzed for precryopreservation sperm parameters and the remaining sample was loaded in 0.5 mL plastic semen straws, cooled gradually to −20°C, and then dipped in liquid nitrogen. After 24 hours of cryopreservation, the straws were thawed for postcryopreservation sperm evaluations. The results (analysis of variance based) showed significantly enhanced percentage of post-thaw sperm membrane integrity, progressive motility, and velocity in BR-FE-0.6% group at both pre- and postcryopreservation stages as compared with all other groups. However, analysis of covariance revealed concentration-dependent cryoprotective effect of BR-FE with maximum percentage of sperm membrane integrity in the 1.6% group. According to these results, BR-FE supplementation adds enormous sperm protective potential to ram sperm cryopreservation medium.

The aim of the present study was the validation of the already reported Bos taurus SNPs in the Sahiwal breed. A total of nine SNPs of the casein gene were studied. Out of nine, seven Bos taurus SNPs of casein protein genes were found to be significantly associated with milk productivity traits. The genomic DNA was extracted from the mammary alveolar endothelial cells of a flock of 80 purebred Sahiwal lactating dams available at Khizrabad Farm near Sargodha. New allele-specific primers were designed from the NCBI annotated sequence database of Bos taurus to obtain 100 nt-long PCR products. Each dam was tested separately for all the SNPs investigated. Animals with genotype GG for the SNPs rs43703010, rs10500451, and 110323127, respectively, exhibited high milk yield. Similarly, animals with genotype AA for the SNPs rs11079521, rs43703016, and rs43703017 showed high milk yield consistently. For the SNP rs43703015, animals with genotype CC showed high milk productivity. These above-mentioned SNPs have previously been reported to significantly up-regulate casein protein contents in Bos taurus. Our results indicated SNPs that significantly affect the milk protein contents may also significantly increase per capita milk yield. These finding suggest that the above-mentioned reported SNPs can also be used as genetic markers of milk productivity in Sahiwal cattle.

Prescription of antimalarial drugs in the absence of malarial disease is a common practice in countries where malaria is endemic. However, unwarranted use of such drugs can cause side effects in some people and is a financial drain on local economies. In this study, we surveyed the prevalence of malaria parasites in humans, and the prevalence of the malaria transmitting mosquito vectors in the study area. We also investigated the use of antimalarial drugs in the local people. We focused on randomly selected rural areas of eastern Pakistan where no malaria cases had been reported since May 2004.

Background: Carbon tetrachloride (CCl4) is extensively used in various industries and induces oxidative stress in humans. Unfortunately, it is one of the neglected potent male reproductive toxicants. Objective: The present research reports the testicular histopathology of CCl4 and ameliorations by four medicinal fruit pulp extracts (FPEs) in mice. Materials and Methods: Sixty male albino mice were divided into six groups (10/group) as per the following: 1. Vehicle control (Vc); 2. CCl4 (C); 3-6. CCl4+Basella alba (CBa), CCl4+Solanum nigrum (CSn), CCl4+Ficus carica (CFc) and CCl4+Grewia asiatica (CGa). Except for the control group, CCl4 (0.1 mL of 0.2 mL kg−1) was given to the animals in corn oil. The four plant extracts (0.1 mL each) were respectively given to the relevant FPE group animals for the next five consecutive days, while the animals in the Vc and CCl4 groups received water instead of FPE. Results: The CCl4 exposure led to various histometric and histological alterations (loss of interstitial tissue and various dislodged tailless spermatids with enlarged heads) that were recovered in all except Solanum nigrum FPE mice post-treatment. The micrometric data of testicular sections also indicated significant decline in the number of spermatogonia, while the cross-sectional area of the sperm heads remained significantly higher in the CSn and C groups. Conclusion: Except for Solanum nigrum, the three FPEs, especially Ficus carica, showed rehabilitative properties against CCl4 exposure-related modifications in testicular histopathologies. Key words: Basella alba, Grewia asiatica, Solanum nigrum, Ficus carica, testis.

Male infertility has become a global health issue nowadays. Insecticides damage the male reproductive system but can be rescued via the use of medicinal fruit extracts (MFEs). A number of experimental studies have shown that MFEs ameliorate testicular histopathologies induced by environmental (toxin) exposure in albino mice. MFEs show good capacity for testicular rehabilitation because of the presence of phytochemicals, especially phytosterols and natural antioxidants. Additionally, we have recently reported better survival rates and fertilizability of cryopreserved spermatozoa on the fortification of semen extenders with MFEs. On the basis of these findings, it is concluded that chemicals and insecticides are potential disruptors of spermatogenic activity and testicular microarchitecture in vivo, whereas the MFEs have shown excellent rehabilitative potentials in these contexts. Moreover, MFEs have also been shown to contribute to the post -hawed semen quality parameters.