The COVID-19 pandemic circumstances have varied the pathogens related to acute respiratory infections (ARI), and most specialists have ignored them due to SARS-CoV-2’s similar symptomatology. We identify respiratory pathogens with multiplex PCR in samples with presumptive SARS-CoV-2 but negative RT-qPCR results. We performed a retrospective transversal study employing clinical data and nasopharyngeal swab samples from patients with suspected clinical SARS-CoV-2 infection and a negative PCR result in a private laboratory in Lima, Peru. The samples were analyzed using the FilmArray™ respiratory panel. Of 342 samples, we detected at least one pathogen in 50% of the samples. The main ones were rhinovirus (54.38%), influenza A(H3N2) (22.80%), and respiratory syncytial virus (RSV) (14.04%). The clinical characteristics were sore throat (70.18%), cough (58.48%), nasal congestion (56.43%), and fever (40.06%). Only 41.46% and 48.78% of patients with influenza met the definition of influenza-like illness (ILI) by the World Health Organization (WHO) (characterized by cough and fever) and the Centers for Disease Control and Prevention (CDC) (characterized by fever and cough and sore throat), respectively. A higher prevalence of influenza was associated with ILI by WHO (aPR: 2.331) and ILI by CDC (aPR: 1.892), which was not observed with other respiratory viruses. The clinical characteristic associated with the increased prevalence of rhinovirus was nasal congestion (aPR: 1.84). For patients with ARI and negative PCR results, the leading respiratory pathogens detected were rhinovirus, influenza, and RSV. Less than half of patients with influenza presented ILI, although its presence was specific to the disease.

Artificial insemination is an important assisted reproductive technology that has been applied in several mammalian species. However, successful cryopreservation of semen of South American camelids has been limited, hindering the commercial application of artificial insemination in alpaca species. In this scenario, the addition of antioxidants to semen extenders provides a strategy to improve the freezability of mammalian sperm. Bioactive metabolites from natural extracts of black maca have shown valuable antioxidant properties. Thus, the objective of this study was to evaluate the effect of the addition of atomized black maca in the freezing medium of epididymal spermatozoa of alpacas. Fifteen pairs of epididymis were collected from a local slaughterhouse. Each sample was divided into six groups: (1) fresh, (2) yolk medium (YM), (3) 10 mg/mL maca, (4) 20 mg/mL maca, (5) 30 mg/mL maca, and (6) resveratrol (as an antioxidant control). Sperm cryopreservation was performed through the slow freezing method. Markers associated with functionality, such as motility, viability, and plasma membrane integrity, as well as markers associated with oxidative damage, such as DNA integrity, total ROS production, and mitochondrial function, were analyzed. The results show that the supplementation with black maca (20 mg/mL) improved the sperm motility, viability, plasma membrane integrity, and mitochondrial function evaluated according to an index of formazan deposits. Similarly, the ROS production decreased with maca at 20 mg/mL, although the DNA integrity did not show any differences among the groups. These results suggest that maca at 20 mg/mL has cytoprotective effects during freezing/thawing of epididymal sperm of alpaca species. Further research will be focused on assessing the effects of maca supplementation on semen extenders by using biomolecular markers (proAKAP4) associated with fertility.

The objective of this study was to evaluate the effect of the addition of atomized black Maca in the cryopreservation medium of epididymal spermatozoa of alpaca species. Fifteen pares of testes and epididymis from alpacas of reproductive age were collected from a local (Huancavelica) slaughterhouse and transported in NaCl 0,9% at 10°C to the laboratory. Each sample of epididymal sperm was divided into six groups: 1) Fresh, 2) Yolk medium (YM), 3) YM+10mg/ml Maca, 4) YM+20mg/ml Maca, 5) YM+30mg/ml Maca, and 6) YM+ Resveratrol (antioxidant positive control). Sperm cryopreservation was performed through the slow-freezing method. Markers associated with functionality such as motility, vitality and plasma membrane integrity. Cellular markers associated with oxidative damage, such as DNA fragmentation, total ROS production and lipid peroxidation, were analyzed before and after freezing. The results showed that the supplementation with black Maca at 20mg/ml improved the sperm motility, viability and integrity of the membrane. Similarly, total ROS production decreased with Maca at 10 mg/ml (25 ± 2.23%) and 20mg/ml (20 ± 2.52%) (P<0.05). However, DNA fragmentation and lipid peroxidation were similar in all groups. These results suggest that Maca at 20mg/ml has a protective effect during the freezing/thawing process of epididymal sperm of alpaca species.

Cryopreservation of alpaca (Vicugna pacos) epididymal sperm is an important strategy for preserving valuable genetic resources; however, the freeze–thaw process induces oxidative stress and cellular damage that compromise post-thaw sperm quality. This study aimed to evaluate the effects of resveratrol supplementation during cryopreservation on the post-thaw quality of alpaca epididymal sperm. Two hundred ten cauda epididymides were collected, and only sperm samples with progressive motility ≥50% were considered. Sixty epididymal sperm samples met this criterion and were allocated to cryopreservation experiments. Ten samples were used in a preliminary experiment to select the resveratrol concentration for the main experiment, whereas the remaining 50 samples were used in the main cryopreservation experiment. Spermatozoa were evaluated before cryopreservation and after thawing in a TES–Tris egg yolk extender with or without 50 μM Resveratrol. Post-thaw sperm quality was improved with Resveratrol supplementation (P < 0.05), as evidenced by preserved mitochondrial physiology and sperm membrane integrity, reduced oxidative stress, early apoptosis, and DNA fragmentation, and enhanced homologous sperm binding to the zona pellucida. However, oxidoreductase activity did not differ between the cryopreserved groups. In conclusion, supplementing the freezing extender with 50 μM resveratrol improved the post-thaw quality of alpaca epididymal sperm by preserving sperm physiological integrity and reducing oxidative stress and cryopreservation-induced cellular damage.