Exploring Beneficial/Virulence Properties of Two Dairy-Related Strains of Streptococcus infantarius subsp. infantarius

The genus Streptococcus includes various species, remarkably different in their behavior, applications, virulence, and safety. Taxonomically Streptococcus infantarius subsp. infantarius belonging to the Streptococcus bovis group, which includes several pathogen species, however, has been found as predominant species in some African dairy products that are widely consumed and considered to be safe. Streptococcus infantarius subsp. infantarius’ safety may be questioned due to the association of this species with clinical cases. In this study, isolates from dairy origin were selected based on their bacteriocinogenic potential and differentiated by their RAPD-PCR profiles. Two strains were identified by 16S rRNA sequencing as St. infantarius subsp. infantarius and investigated regarding their potential beneficial properties and factors related to virulence and safety. A series of in vitro tests included properties related to survival in the gastrointestinal tract and beneficial intestinal activities. Production of bacteriocin/s, detection of related genes, and partial characterization of expressed antimicrobial protein were evaluated. Genes related to folate biosynthesis were detected in both studied strains. Evaluation of physiological tests related to strains virulence, adhesion, and resistance to antibiotics and detections of virulence and biogenic amines production-related genes were also investigated. Taking in consideration all the aspects of the specific nature of St. infantarius subsp. infantarius K1–4 and K5–1 (beneficial properties and virulence characteristics), both strains cannot be considered safe for human or other animals application, even though they have been isolated from dairy products. This study is highlighting the importance of evaluation for presence of potential virulence factors in newly characterized strains in order to be confident in their safety.

     

Focus: Preventive Medicine: 2020 Evaluation of Portable Vision Screening Instruments

Amblyopia is the most common cause of preventable visual impairment in children and occurs as a result of unilateral or bilateral impairment in best-corrected visual acuity. Early diagnosis and proper treatment are crucial to prevent poor visual outcomes in adulthood. Advances in technology have provided more objective diagnostic tools, which can now be used by a wide range of healthcare providers. Here, we highlight tools that have gained popularity in the past two decades and compare clinically relevant parameters to guide primary care providers seeking to incorporate instrumental vision screening in pediatric patient care.     

Stejneger's beaked whale strandings in Alaska, 1995–2020

Presented here is the first comprehensive and updated compilation of history, distribution, and findings of Stejneger's beaked whales (Mesoplodon stejnegeri) in Alaska. Stejneger's beaked whales are a poorly understood, elusive, deep-diving cetacean species found in the North Pacific Ocean. Since Stejneger's beaked whale strandings data in Alaska through 1994 were last published, 35 additional strandings have been documented. Twenty-seven animals stranded in the Aleutian Islands, seven stranded in Southcentral Alaska, and one animal stranded on St. Lawrence Island. Twenty-two carcasses were necropsied, but only four were fresh. Seventeen of the 22 died during mass stranding events and cause of death could not be definitively determined. Barotrauma was suspected in three cases and infectious disease possibly complicated by barotrauma occurred in two cases. We documented an expansion of strandings into the northern Bering Sea, characterized a sex bias, examined stomach contents that included macroplastic, and identified parasites not previously associated with Stejneger's beaked whales. Also included are data on the largest known mass stranding of Stejneger's beaked whales, which occurred on Adak Island in 2018. The history, distribution, and findings presented here are central to further our understanding of this species.     

Genetic evidence for seasonal consumption of monkfish (Lophius spp.) and salmonids (Salmo spp.) by gray seals

Recoveries of gray seal (Halichoerus grypus) populations across their eastern Atlantic distribution have led to a steady increase in seal-fishery interactions. Fishers have estimated depredation of salmonids (Salmo spp.) and monkfish (Lophius spp.) as high as 40% and 59% respectively in Ireland. However, empirical evidence for the consumption of these species has been extremely limited due to diagnostic hard part remains not being found in scats or stomach samples. We applied species-specific primers and tested for the presence of monkfish and salmonids in gray seal diet genetically using quantitative polymerase chain reaction (qPCR) on scats. Monkfish occurred in 29.7% of sampled scats, while salmonids occurred in 12.7%. Seasonal and regional variability in occurrence were noted for both species, likely related to the migratory behavior of the prey species and proximity of seal haul-outs to aquaculture sites. Traditional hard part analysis of scats, including scats that tested positive for monkfish and salmonid DNA, failed to find any evidence of either species. This study provides important empirical evidence for the consumption of these species in Ireland that can inform management.     

Efficacy of RyR2 inhibitor EL20 in induced pluripotent stem cell-derived cardiomyocytes from a patient with catecholaminergic polymorphic ventricular tachycardia

Catecholaminergic polymorphic ventricular tachycardia (CPVT) is an inherited cardiac arrhythmia syndrome that often leads to sudden cardiac death. The most common form of CPVT is caused by autosomal-dominant variants in the cardiac ryanodine receptor type-2 (RYR2) gene. Mutations in RYR2 promote calcium (Ca²⁺) leak from the sarcoplasmic reticulum (SR), triggering lethal arrhythmias. Recently, it was demonstrated that tetracaine derivative EL20 specifically inhibits mutant RyR2, normalizes Ca²⁺ handling and suppresses arrhythmias in a CPVT mouse model. The objective of this study was to determine whether EL20 normalizes SR Ca²⁺ handling and arrhythmic events in induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) from a CPVT patient. Blood samples from a child carrying RyR2 variant RyR2 variant Arg-176-Glu (R176Q) and a mutation-negative relative were reprogrammed into iPSCs using a Sendai virus system. iPSC-CMs were derived using the Stemdiff^TM kit. Confocal Ca²⁺ imaging was used to quantify RyR2 activity in the absence and presence of EL20. iPSC-CMs harbouring the R176Q variant demonstrated spontaneous SR Ca²⁺ release events, whereas administration of EL20 diminished these abnormal events at low nanomolar concentrations (IC₅₀ = 82 nM). Importantly, treatment with EL20 did not have any adverse effects on systolic Ca²⁺ handling in control iPSC-CMs. Our results show for the first time that tetracaine derivative EL20 normalized SR Ca²⁺ handling and suppresses arrhythmogenic activity in iPSC-CMs derived from a CPVT patient. Hence, this study confirms that this RyR2-inhibitor represents a promising therapeutic candidate for treatment of CPVT.     

The association of Amyloid P-Component (AP) With The Amyloid Fibril: An updated Method for Amyloid Fibril Protein Isolation

Abstract

An amyloid fibril isolation procedure is proposed which uses citrate as well as saline washes to dissociate the calcium dependent linkage of amyloid P-component (AP) from the amyloid fibril. In two amyloid rich tissues, the amount of AP was quantitated in each saline and citrate wash and totalled 13.8% and 20.8% of the amyloid fibrils isolated. The amount of AP removed from these and 22 additional amyloid rich tissues was greater than had previously been recognized.

AP protein was present in tissue only when amyloid fibrils were present. It could not be found in normal non-amyloidotic tissue, nor could it be found in tissue sediment after the fibrils were removed.     

Developing a taxonomic identification system of Phytophthora species based on microsatellites

BackgroundPhytophthora is the most important genus of the Oomycete plant pathogens. Nowadays, there are 117 described species in this genus, most of them being primary invaders of plant tissues. The different species are causal agents of diseases in a wide range of crops and plants in natural environments. In order to develop control strategies against Phytophthoraspecies, it is important to know the biology, ecology and evolutionary processes of these important pathogens.AimsThe aim of this study was to propose and validate a low cost identification system for Phytophthora species based on a set of polymorphic microsatellite (SSRs) markers.MethodsThirty-three isolates representing Phytophthora infestans, Phytophthora andina, Phytophthora sojae, Phytophthora cryptogea, Phytophthora nicotianae, Phytophthora capsici and Phytophthora cinnamomi species were obtained, and 13 SSRs were selected as potentially transferable markers between these species. Amplification conditions, including annealing temperatures, were standardized for several markers.ResultsA subset of these markers amplified in all species, showing species-specific alleles.ConclusionsThe adaptability and impact of the identification system in Colombia, an Andean agricultural country where different Phytophthora species co-exist in the same or in several hosts grown together, are discussed.