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Study question Does supplementation of human sperm media with L-Proline improve sperm function and chromatin quality during in Vitro Incubation? Summary answer The inclusion of L-proline as an antioxidant in human sperm media improves sperm function and chromatin integrity probably by modulating oxidative stress What is known already Infertility affects millions of people of reproductive age globally. The quality of gametes is one of the most critical aspects that could affect the success rate of infertility therapy. The storage of spermatozoa at the incubator to achieve capacitation could rise sperm DNA fragmentation as a result of prolonged incubation. Although mild and low levels of Reactive Oxygen Species (ROS) could boost fertilization capacity by improving hyperactivation, and capacitation, oxidative stress is one of the major contributors to low sperm quality. L-Proline plays a pivotal role in many aspects of cell metabolism and physiology, including osmotic protection and oxidative stress Study design, size, duration The study was conducted on thirty-five healthy men attending the Motazedi Hospital, Kermanshah of the Division of Andrology, from December 2020 to June 2021. Each semen sample was equally divided into 4 groups: one group as control and three as treatment groups. The control group contained only sperm media (Hams, F10 medium supplemented with 5 mg/ml HSA). In treatment groups, different concentrations of Proline (1, 2, and 4 mmol/L) were also added. Participants/materials, setting, methods 35 normozoospermic men with healthy lifestyles were included. Sperm parameters including motility, viability, and morphology were evaluated. The levels of ROS, lipid peroxidation (LPO), and total antioxidant capacity (TAC) were determined in sperm media. Sperm chromatin integrity was analyzed by Aniline blue (AB), Toluidine blue (TB), and Chromomycin A3 (CMA3) staining tests. DNA fragmentation was measured by the Sperm chromatin dispersion (SCD) test. All assessments were conducted after 1, 4, and 24 hours of incubation. Main results and the role of chance The inclusion of Proline (2 mmol/L) resulted in noticeably improved maintenance of motility and viability after 24 hr of incubation compared to the control group (P < 0.05). Also, 2 mmol/L of Proline was able to significantly maintain normal morphology compared to other groups (P < 0.01). However, the level of ROS production non-significantly diminished in the L-proline groups compared to the control group (P > 0.05). In terms of lipid peroxidation, the presence of 2 mmol/L of Proline kept the MDA production of spermatozoa at lower levels after 4 hr of incubation compared with the control group (P < 0.05). The supplementation of sperm media with 2 mmol/L of Proline led to improved maintenance of TAC level, compared to any other group (P < 0.01). Additionally, the results of AB and TB tests indicated that chromatin quality after supplementation with 2 mmol/L L-proline significantly improved compared to the control group (P < 0.05). In terms of CMA3 assessment, however, there was a minor improvement between the 2 mmol/L Proline group and the control group. Moreover, 2 mmol/L Proline significantly reduced the level of fragmented DNA by comparison to the control group after 24 hr of incubation (P < 0.05). Limitations, reasons for caution Our team was unable to carry out additional studies to verify the impact of L-proline on mitochondrial activity, membrane potential, and apoptosis and to validate the efficiency and safety of L-proline using additional parameters such as the in vitro embryonic development and live birth rate. Wider implications of the findings While further studies are required to uncover the mechanisms underlying the antioxidant properties of Proline, these findings extend our knowledge of human male fertility and pave the way to improving the human sperm culture and in vitro procedure routinely used in infertility clinics for sperm incubation. Trial registration number IR.KUMS.REC.1399.625

Otitis externa is a multifactorial disease that has 5 to 20 percent prevalence in small animal medicine which can be diagnosed according to the history and general dermatological and otoscopic examinations. This study aimed to compare the agents that cause Otitis externa in dogs and cats, taking into account the breeds, gender, and age differences of these animals. The samples were taken from 63 dogs and cats that were suspected of having external ear infections and referred to veterinary clinics from north of Iran, including Mazandaran, Guilan, and Golestan provinces, within 24 months (from April 2019 to April 2021). After obtaining the consent, sampling was performed from the pinna and the end of the vertical ear canal. The prevalence of infection observed in dogs was: 28.57% for Malassezia, 28.57% for gram-positive bacteria, 11.91% gram-negative bacteria, 16.67% and 14.28% for Otobius megnini and Sarcoptes scabiei respectively, while in cats, the prevalence of Malassezia infections was 9.52%, 23.80% gram-positive bacteria, 19.04% gram-negative bacteria, 19.04% Otobius megnini, and 33.34% Notoedres cati. There was no statistically significant difference between the studied dogs and cats with the variables of age, gender, and breed. Identifying the agents causing external ear infections and paying attention to factors such as etiologies, geographical effects, region climate, disease-causing agents, breed differences, gender, and age in Iran is crucial and can improve the rate of success in management and treatment of external ear infections. Therefore, it seems that due to the limited number of these studies in Iran, additional studies in this field are necessary. Among dogs and cats in the north of Iran, this study represents the first assessment of external otitis.

Skin is an organ of living beings afflicted with various disorders such as atopic disease. Atopic disease results from an excess of immunoglobulin E produced due to excessive exposure to non-harmful environmental allergens that affect the skin of various organs, including the ear, leading to external otitis. During this case-control study, 50 dogs referred to clinics in Mazandaran province in Iran with symptoms of atopic otitis were the case group, and 50 dogs with healthy ears were considered the control group. The diagnosis was derived from the clinical findings of the case group, which included itchy skin, redness in the pubic and abdominal area, and ear swelling. The results indicated that 66% of dogs with atopic otitis were in the chronic stage of the disease. Furthermore, there is 4.63 times greater odds of wax secretions in cases of atopic otitis. Additionally, in the case of atopic otitis, the odds of odorous secretions is 5.1 times greater than that of odorless secretions. Lastly, most dogs with this condition had significantly dark ear secretions. Atopic otitis, therefore, is characterized by waxy, odorous and dark secretions in the ears of animals. In general, identifying the causative agent for this disease in dogs is extremely important and can prevent the application of costly and blind treatments. We conducted the first epidemiological study in Iran of external otitis associated with atopy in dogs. These results support a better understanding of the disease.