The widespread species Parmotrema crinitum (Ach.) M. Choisy and Parmotrema perlatum (Huds.) M. Choisy are mainly distinguished by their reproductive strategies. While P. crinitum propagates by isidia, P. perlatum produces soredia. In this study, we aim to evaluate the phylogenetic relationship between both species and to critically examine their species boundaries. To this purpose, 46 samples belonging to P. crinitum and P. perlatum were used in our analysis, including 22 for which we studied the morphology and chemistry, before extracting their DNA. We used 35 sequences of the internal transcribed spacer region of nuclear ribosomal DNA (ITS) of Parmotrema perlatum from Europe and Africa (20 of which were newly generated), and 11 of Parmotrema crinitum from Europe, North America and North Africa (two newly generated). Additionally, 28 sequences of several species from Parmotrema were included in the ITS dataset. The ITS data matrix was analyzed using different approaches, such as traditional phylogeny (maximum likelihood and Bayesian analyses), genetic distances, automatic barcode gap discovery (ABGD) and the coalescent-based method poisson tree processes (PTP), in order to test congruence among results. Our results indicate that all samples referred to P. crinitum and P. perlatum nested in a well-supported monophyletic clade, but phylogenetic relationships among them remain unresolved. Delimitations inferred from PTP, ABGD and genetic distance analyses were comparable and suggested that P. crinitum and P. perlatum belong to the same lineage. Interestingly, two samples of P. perlatum separate in a different monophyletic clade, which is supported as a different lineage by all the analyses.

Although protected areas (PAs) play an important role in ecosystem conservation and climate change adaptation, no systematic information is available on PA protection of high-elevation freshwater ecosystems (e.g., lakes and watersheds with glaciers), their biodiversity and their ecosystem services in the tropical Andes. We therefore combined a literature review and map analysis of PAs of International Union for Conservation of Nature (IUCN) and national systems of PAs and freshwater ecosystems. We found that seven national parks were created for water resources protection but were not designed for freshwater conservation (i.e., larger watersheds). High-value biodiversity sites have not been protected, and new local PAs were created due to water resource needs. We quantified 31 Ramsar sites and observed that PAs cover 12% of lakes, 31% of glacial lakes and 12% of the total stream length in the tropical Andes. Additionally, 120 watersheds (average area 631 km2) with glaciers and 40% of the total glacier surface area were covered by PAs. Future research into the role of PAs in ecosystem services provision and more detailed freshwater inventories within and around PAs, especially for those dependent on glacier runoff, will fill key knowledge gaps for freshwater conservation and climate change adaptation in the tropical Andes.

The species of the Parmelia saxatilis complex occurring in the Iberian Peninsula were revised. Eight species are accepted, including a new species found in southern Spain, described as P. rojoi A. Crespo, V. J. Rico & Divakar. The new species, which forms a sister-group relationship with P. saxatilis s. str., is rare in the Iberian Peninsula and is restricted to higher altitudes of northern and central Spain. Parmelia rojoi differs from P. saxatilis by generally narrower isidia and a more fragile thallus. The segregation of the new species is also supported by ITS (rDNA) and Mcm7 (MS456) phylogeny and multispecies coalescent-based approaches, including StarBEAST and BP&P. Furthermore, the divergence of P. rojoi is dated back to the Pleistocene, c. 2.13 Ma. A key to the identification of species from the P. saxatilis complex with their diagnostic features is provided. All species of the complex known from Europe are also found in the Iberian Peninsula. We hypothesize that P. rojoi is a relict species that survived the Pleistocene glaciations in refugia in Spain and has been unable to extend its distributional range in postglacial periods.

This study aims to investigate the influence of work experience variables on the emotional state of worker-caregivers based on stress process model. The emotional state (depression, negative and positive affect and worry-strain), caregiver and care recipient features, caregiving stressors and appraisal, and role strains/work-related variables were assessed in 83 worker-caregivers of elderly dependent relatives. Hierarchical multiple linear regression analyses were performed for each of the four emotional outcome variables. Caregiving overload and positive job experience were the best predictors of depression and positive affect. The predictors of negative affect were reaction to memory and behavior problems, overload and role captivity. The predictors of worry and strain were daily hours of caregiving worries, reaction to memory and behavior problems, overload, role captivity and job-caregiving conflicts. The explained variances for the four models were 58.8%, 40.2%, 62.9% and 78.8%, respectively; the role strain contributions were 8.2%, 13.2%, 7.2% and 6%. The results indicate that the effect of perceived job experiences on caregivers’ emotional status is more relevant than objective job conflicts. In addition, caregivers’ emotional state is primarily related to the subjective indicators of caregiving stressors, with a lower contribution of work-related variables.

A 2,4-D-resistant tall waterhemp population (FS) from Nebraska was evaluated for resistance to other TIR1 auxin receptor herbicides and to herbicides having alternative mechanisms of action using greenhouse bioassays and genetic markers. Atrazine, imazethapyr, lactofen, mesotrione, glufosinate, and glyphosate were applied in a single-dose bioassay, and tissue was collected from marked plants for genetic analysis. The FS population was not injured by atrazine or by imazethapyr. Approximately 50% of the plants survived lactofen and were actively growing 28 d after treatment. The population was susceptible to mesotrione, glufosinate, and glyphosate. Ametryn, chlorimuron-ethyl, 2,4-D, aminocyclopyraclor, aminopyralid, and picloram were applied in dose–response studies. The FS population was sensitive to ametryn, and the Ser-264-Gly substitution in the D1 protein was not detected, suggesting the lack of response to atrazine is not due to a target-site mutation. The FS population exhibited less than 50% injury to chlorimuron-ethyl at application rates 20 times the labeled use rate. The Ser-653-Asn acetolactate synthase (ALS) substitution, which confers resistance to imidazolinone herbicides, was present in the FS population. However, this does not explain the lack of response to the sulfonylurea herbicide, chlorimuron-ethyl. Sequencing of a portion of the PPX2L gene did not show the ΔG210 mutation that confers resistance to protoporphyrinogen oxidase–inhibiting herbicides, suggesting that other factors were responsible for waterhemp survival after lactofen application. The FS population was confirmed to be at least 30-fold resistant to 2,4-D relative to the susceptible populations. In addition, it was at least 3-fold less sensitive to aminopyralid and picloram, two other TIR1 auxin receptor herbicides, than the 2,4-D-susceptible populations were. These data indicated that the FS population contains both target and non–target site mechanisms conferring resistance to herbicides spanning at least three mechanisms of action: TIR1 auxin receptors, ALS inhibitors, and photosystem II inhibitors.

Macro-morphological features traditionally used to segregate genera in Parmeliaceae have been shown to be highly plastic, placing limits on their taxonomic value. Here we aim to elucidate the evolutionary relationships of the genera Relicina and Relicinopsis and reassess the phenotypic features traditionally used to separate these genera. To this end, we gathered ribosomal DNA sequences of ITS, nuLSU and mtSSU and analyzed them in a phylogenetic framework. Relicina was recovered as paraphyletic, with Relicinopsis nested within, and three different clades were identified within Relicina. Alternative hypothesis tests significantly rejected the monophyly of Relicina. Our results indicate that the presence or absence of bulbate cilia is of limited taxonomic value in this clade. Based on differences in conidia, however, we propose to accept Relicinopsis as a subgenus within Relicina as Relicina subgen. Relicinopsis (Elix & Verdon) Kirika, Divakar & Lumbsch. It is proposed that five new combinations of species previously classified in Relicinopsis be placed in Relicina.

This research communication aimed to compare somatic cell count (SCC), the main marker of udder health status, in organic farms not using antibiotics (O, n = 6), organic farms using antibiotics (OA, n = 7) and conventional farms (CA, n = 5) using antibiotic treatments, all of them at pasture. SCC was statistically significantly higher in O (173780) compared to CA (93325) and OA (107152). Milk yield had a significant diluting effect on SCC and differences between groups increased with parities. Stratified analysis of SCC depending on lactation number and % of monthly SCC test with different linear scores (LS) indicated that there is no difference in udder health in the primiparous heifers from the three groups of farms, but it deteriorates in older cows because of chronic infections in O (possibly due to lack of antibiotic use). Our results suggest that the non-use of antibiotics had an effect in udder health leading to higher occurrence of clinical and subclinical mastitis. Therefore, preventive management practices for mastitis control are essential in organic farms.

The results of the first molecular phylogenetic study of Pseudephebe are presented; a three-locus phylogeny. The genus is confirmed as monophyletic within the alectorioid clade of Parmeliaceae. Two major clades were recovered, which can be assigned to the two traditional taxa, P. minuscula and P. pubescens, with modifications of the species delimitation, especially the variable P. minuscula. These species are cryptic and cannot be confidently distinguished morphologically due to phenotypic convergence. Therefore, the use of P. pubescens aggr. is recommended for samples not molecularly analyzed. Contrary to previous studies, specimens of both species might have indistinct pseudocyphellae and also contain lichen substances; norstictic acid was detected in c. 60% of specimens tested. An SSU 1516 Group I intron is usually present in P. minuscula but always absent in P. pubescens. The species-level nomenclature is summarized and sequenced reference specimens (RefSpec) for both Pseudephebe species are selected. Sequences from Bryoria mariensis established that this name was a synonym of P. minuscula.

As part of a comprehensive revision of the Rhizocarpon geographicum species group using molecular and morphological approaches, we examined the name-bearing types of 15 species. We report ambiguities and inconsistencies with the reported features of some type specimens, original descriptions, and circumscriptions employed in keys for the identification of taxa within the complex. The reporting of chemical reactions and some morphological characters was found to be inconsistent and likely to lead to errors of identification. This is unfortunate in view of the widespread use of the complex in glaciology for lichenometry. Issues surrounding the typification of the basionym Lichen geographicus are clarified, and the epitype is illustrated. The complex nomenclatural situation regarding the name Rhizocarpon lecanorinum and its typification is also made clear. The examination of the anatomical and morphological characters presented here needs to be considered together with molecular phylogenetic information to provide a more informed new taxonomy of the group.

In order to confirm and investigate the extent of reported mismatches between chemotypes and molecular sequence data in Bryoria fuscescens s. lat., we examined 15 morphologically similar thalli from each of three Pinus forest sites in the Sistema Central of central Spain. Three thalli were rejected due to infections by Phacopsis huuskonenii (not previously published from Spain). The remaining 42 thalli represented nine ITS rDNA haplotypes and four chemotypes (by TLC): fumarprotocetraric and protocetraric acids; norstictic and connorstictic acids; psoromic acid; and fumarprotocetraric, protocetraric and psoromic acids. The molecular phylogenetic tree was characterized by extremely short branch lengths, often only with a single mutational difference, and a single haplotype could have different chemical products. In some cases, adjacent specimens represented different chemotypes, and three thalli appeared to be mixed individuals. Consistency of both molecular and chemical data within individual specimens was demonstrated by examining four different parts of each thallus, which showed only a difference in the location of psoromic acid in some. This is the first population-level study of this taxon, and so it is premature to propose taxonomic changes at this time. Further populations in different parts of the geographical range of this widespread complex now need to be analyzed, and more sensitive chemical analyses conducted, in order to understand the basis of the variability and determine the appropriate taxonomic treatment.

The phylogenetic position of the genus Pseudoparmelia was addressed using molecular data from five loci (mtSSU, nuLSU, ITS, Mcm7, RPB1), generated from three species and aligned with sequences from 293 samples representing all major clades of Parmeliaceae. Pseudoparmelia species form a well-supported monophyletic group that is the sister group of a clade consisting of the genera Relicina and Relicinopsis. These three genera share a thallus with a pored epicortex, isolichenan as cell wall polysaccharide, and relatively small ascospores. Morphological and chemical characters that distinguish Pseudoparmelia from the closely related Relicina and Relicinopsis are discussed. To further elucidate the relationships of these three genera, we assembled a second dataset including 15 additional samples of Relicina and Relicinopsis using three loci (mtSSU, nuLSU, ITS). All three genera are monophyletic but monophyly of Relicina lacks support and, in the mtSSU single locus tree, the genus is paraphyletic with Relicinopsis nested within. Additional studies including more Relicina species are necessary to test delimitation of the genera Relicina and Relicinopsis.

The importance of the anatomy and fine morphology of reproductive structures for the systematics of the family Parmeliaceae is highlighted by the new genus Notoparmelia, described here for Australasian species of Parmelia. These species were known to form a monophyletic lineage but correlated characters for its delimitation were lacking. A major characteristic used here for the circumscription of this genus is the overlooked apothecial anatomy. The proper exciple is reduced to one layer formed exclusively by large hyphae without any clear branching pattern and embedded in an abundant polysaccharide matrix. This feature differs from the rest of Parmeliaceae which have a stratified three-layered proper exciple composed of a thin hyaline layer, intermediate layer and basal cortex-like structure. The anatomy of proper exciple, together with the thickness of spore walls and other morphological characters such as lobe morphology, pseudocyphellae and rhizine type, allow a precise diagnosis of the new genus. The area of distribution is also useful for characterizing this genus. Sixteen new combinations are proposed.

Several researchers have found that caring for relatives with dementia has negative consequences on the caregiver’s physical and emotional health during the caregiving period. However, less attention has been paid to caregiver´s emotional state after the patient´s death. The aims of this study are 1) to evaluate the emotional state of the Spanish former caregivers after the care recipient death, and 2) to analyze the patient’s death circumstances and their influence in the current emotional state of caregivers. The sample was composed of 50 former caregivers of relatives diagnosed with Alzheimer’s disease. The dependent variables were assessed using the Beck Depression Inventory and the Texas Revised Inventory of Grief. Most of the caregivers did not develop a complicated grief or depressive symptoms. However, caregivers are not uniform in their responses to bereavement. Specifically, grief was higher in spouses than in adult child, even after controlling the effect of gender and age. Furthermore, this study provides first evidence of the positive relationship between saying good-bye to the patient and caregivers’ emotional status.

The organization and the histological characteristics of Leptodactylus chaquensis testis throughout the reproductive cycle were analyzed in the presented study. Gonads of adult males, processed with routine techniques for optical microscopy, revealed that during the reproductive period the seminiferous tubules were characterized by presentation of a large number of cysts, germ cells at the same maturation stage supported by Sertoli cells. All the germ line cells were also present in the postreproductive period and maintained their morphological characteristics. Primary spermatogonia were large-sized cells found isolated or in small groups. The rest of the cells of the germ line formed cysts. Secondary spermatogonia showed morphological characteristics similar to their predecessors, although they were smaller. Primary and secondary spermatocytes showed images of the different stages of the first and second meiotic division respectively. One finding was the presence of intercytoplasmic bridges between the secondary spermatocytes. Primary spermatids were rounded cells with an acrosomal vesicle associated with the nucleus and had cysts that were characterized by large intercellular spaces. Secondary spermatids were elongated cells with a well defined acrosome, which in the spermatozoa had the shape of an arrowhead. Another peculiar characteristic of this species was the fusion of the walls of the seminiferous tubule with the efferent duct that formed a path for spermatozoa during spermiation. The presence in the seminiferous tubules of all stages of the spermatogenic line during the two periods of the cycle studied indicated that Leptodactylus chaquensis had a potentially continuous reproductive cycle.

In amphibians, the components of the jelly coats that surround the oocytes at the time of fertilization and coordinate gamete interaction are secreted by the oviduct. We analysed the histological variations in the mucosa of the oviductal pars convoluta (PC) of Rhinella arenarum during the reproductive cycle and its relationship with secretion. During the preovulatory period, the mucosa reaches a high degree of morphological and functional development, with a large number of epithelial (ESC) and glandular secretory cells (GSC) loaded with contents that are secreted into the oviductal lumen. During the ovulatory period, the secretory cells (SC) of both layers present maximum secretory activity through apocrinia and merocrinia. While the ESC located at the tips of the folds release their content directly in contact with the oocytes, the GSC secrete material from the bottom of the epithelial folds that, by interaction with the secretion of the ESC in the lateral faces, form a product with a certain degree of organization. Secretion is a continuous process with formation of coats of increasing complexity from the intermediate proximal zone (IPZ) to the pars convoluta (pc) itself, and the passage of the oocyte is a requisite for the organization of the jelly coats around the gamete.

During the early postovulatory period, although there is a marked decrease in the number and volume of the SC, the ESC still release material into the oviductal lumen. In the late postovulatory period the morphological characteristics of the PC begin to recovery although there is no evidence of secretion.