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Compared to aquatic ecosystems, limited information exists on organochlorine accumulation in terrestrial ecosystems, and this is specifically true for terrestrial carnivores that have received limited attention in terms of studies on pollutant bioaccumulation. The African Leopard, Panthera pardus pardus (Linnaeus, 1758), is a popular focal species for research by ethologists and ecologists, but a noticeable knowledge gap exists with regards to toxicological aspects. To address this gap, the aim of this study was to determine baseline organochlorine pesticide (OCP) concentrations in blood of live wild and captive leopards in South Africa, and to explore the relationship between OCP levels and different conservation management strategies. Peripheral blood samples of seven captive and seven wild leopards, representing regular and melanistic individuals within the captive population, were collected while under sedation. The ΣOCP concentrations in blood serum were detected by means of GC-ECD. Statistical relationships among pesticide content in leopards from three geographical areas, different sexes, age groups and conservation status were examined. Captive leopards from this study had a slightly higher mean ΣOCP concentration (901 pg/ml) than wild leopards (768 pg/ml), and captive females had lower mean levels of ΣOCPs (797 pg/ml) than males (1,058 pg/ml). OCPs accumulated in the following order DDTs (27%) > HCHs (21%) > Heptachlors (15%) > CHLs (15%) > Drins (14%) > HCB (8%). Differences in OCP composition profiles of resampled captive individuals were also found. The sources of OCPs in the leopards can be attributed to the historic and current use of, e.g., DDT and HCHs, for malaria vector control and in insecticides/avicides in South Africa. For the captive leopards in this study the main source is possibly contaminated chicken that formed the major portion of their diet. This is the first report on OCP concentrations from leopards in Africa and highlights the need for this parameter to be considered in terms of the conservation management of healthy populations.

In South Africa, the role of reptilian ticks in the transmission of haemoparasites is lacking, in part, due to limited information on tick diversity and their associated haemoparasites. The aim of this research was to identify tick species parasitizing reptiles and to molecularly screen these ectoparasites for species of the blood apicomplexan genus Hepatozoon. Samples were collected from Ndumo Game Reserve, KwaZulu-Natal, and the Cape Columbine region, Western Cape. Reptiles collected included 2 snakes, 2 monitor lizards of a single species respectively, as well as 17 tortoises of four species. Ticks collected from these were morphologically identified as Amblyomma latum (n = 2) and Amblyomma marmoreum (n = 98), this identification was molecularly confirmed using 16S rRNA and CO1 genes. Screening for Hepatozoon was done by amplifying the 18S rRNA gene. A species of Hepatozoon, Hepatozoon fitzsimonsi, was identified from A. marmoreum ticks, with an overall prevalence of 10%. This Hepatozoon species, has been described parasitizing tortoises from southern Africa, and has been reported from ticks infesting tortoises from Kenya, East Africa. Even though ticks have been suggested to be the likely vector of this Hepatozoon species, with this supported by the findings of Hepatozoon-like developmental stages in ticks collected off of infected tortoises, a recent systematic revision placed this species in a newly erected genus Bartazoon, a genus vectorised by biting insects. The present study thus provides further support for ticks acting as the potential vectors of H. fitzsimonsi.

A need exists to understand how drought and its linked consequences threaten aquatic ecosystems and their associated biota in semi-arid countries, as climate change is predicted to increase the effects of these events. South Africa experienced one of the worst droughts in its history from 2015 to 2017 and, as a result, all but the permanent waterbodies of the lower Phongolo River floodplain (PRF) in northern KwaZulu-Natal dried up. This study examined the resilience of aquatic invertebrates associated with the PRF and what the colonization and succession trends of aquatic invertebrates would be once floodplain pools were to receive water again following a 2-year drought. Water and aquatic invertebrates were collected 11 times from nine field mesocosms over a 5-week period for chemical and biodiversity analysis, respectively. Zooplankton hatched from resting stages and macroinvertebrates colonized the mesocosms from nearby permanent waterbodies within the first week of inundation, suggesting that the drought had not affected the ability of aquatic biota to continue natural colonization and succession. Although the biota appeared unaffected, an extension of such a drought could lead to habitat fragmentation and total desiccation of permanent waterbodies, potentially causing the local extinction of aquatic populations and a loss of biodiversity.

INTRODUCTION Parasites of the genus Gyrodactylus von Nordmann, 1832, are predominantly parasites of freshwater and marine bony fishes. To date, 40 species of the genus have been described from Africa, with only 6 of these reported from South Africa. This represents only a small fraction of the known diversity of almost 500 species known worldwide. METHODS In total, 43 of Straighhfin barb, Enteromius paludinosus were collected from Barberspan Bird Sanctuary, North West Province, South Africa. Monogenean parasites were isolated from the fins and fixed on microscope slides in ammonium picrate glycerine. Morphometric evaluation of opisthaptoral hard parts confirmed the identity of these specimens as a new species. RESULTS The description of Gyrodactylus paludinosus sp. nov. is presented. The new species has well-developed, slightly curved superficial inner roots and well-developed rectangular-like ventral bar membrane. Despite the small differences between the hamuli size, it can be distinguished from its congeners, G. kyogae and G. ivindoensis, the only species known from small African barbs, by the shape of ventral bars and marginal hook sickle propers. CONCLUSION The description of Gyrodactylus paludinosus n. sp. increases the total number of Gyrodactylus spp. known from African cyprinid fish to three. Undoubtedly, more attention should be given to studies on small barbs as potential hosts for new species.

Worldwide the use of pesticides has increased, especially in the industry and agriculture sector even though they contain highly toxic substances. The use of pesticides has various negative effects on the aquatic ecosystem and organisms within these ecosystems. The paper aimed to assess the effects of increased concentrations of malaria vector control insecticides (Dichlorodiphenyltrichloroethane (DDT) and Deltamethrin (DTM)) on the freshwater diatom community structure using a microcosm approach as well as determine whether a mixture (DDT 1:1 Deltamethrin) exposure will have a greater influence on the diatom community when compared to single exposures of these insecticides. Diatoms were exposed to a high and low concentration (based on LC50 data for freshwater Xenopus laevis from the USEPA Ecotox database) of DDT, DTM and a mixture in lentic microcosms over a total period of 28 days. Results indicated that irrespective of exposure concentrations, DDT, DTM and a mixture had negative effects on the diatom community including functionality and vitality as these insecticides induced changes to their chloroplasts. There was an increased percentage dead cells for all exposures compared to the control, with the insecticides having a phototoxic effect on the diatom community. Exposure to the selected insecticides caused a significant decrease in some diatom metrics indicating the negative effects these insecticides have on the diatom metrics. Therefore, diatoms may prove to be useful as bio-indicators in ecotoxicology studies when assessing the effects of any insecticide exposures.

ADP-ribosylation is a pivotal post-translational modification that mediates various important cellular processes producing negatively charged biopolymer, poly (ADP-ribose), the functions of which need further elucidation. Toward this end, the availability of well-defined ADP-ribose (ADPr) oligomers in sufficient quantities is a necessity. In this work, we demonstrate the chemical synthesis of linear ADPr oligomers of defined, increasing length using a modified solid phase synthesis method. An advanced phosphoramidite building block temporarily protected with the base sensitive Fm-group was designed and implemented in the repeating pyrophosphate formation via a P(v)–P(iii) coupling procedure on Tentagel solid support. Linear ADPr oligomers up to a pentamer were successfully synthesized and their affinity for the poly-(ADP-ribose)-binding macrodomain of the human oncogenic helicase and chromatin remodeling enzyme ALC1 was determined. Our data reveal a length-dependent binding manner of the nucleic acid, with larger ADPr oligomers exhibiting higher binding enthalpies for ALC1, illustrating how the activity of this molecular machine is gated by PAR., We report the synthesis of linear ADPr oligomers of defined length up to a pentamer using an improved solid phase method. Binding study with human oncogenic helicase ALC1 shows that ADPr oligomers bind to ALC1 in a length-dependent manner.

The branchial attaching, fish parasitic genus Mothocya Costa, in Hope, 1851 is recorded for the first time from Nigerian brackish waters on Monodactylus sebae (Perciformes: Monodactylidae). Mothocya andonin. sp. is characterised by a trapezoid-shaped cephalon with a subtruncate rostrum; pereonite 1 anterolateral angles only reaching to the posterior margin of the eyes; particularly large and wide coxa 7 extending to pleonite 4; pereopod 7 with a narrow basis; broadly rounded pleotelson; and uropods extending to the pleotelson posterior margin, with subequal rami. Mothocya powellin. sp. is characterised by its small size (7 mm); a posteriorly ovoid body shape; cephalon anterior margin truncate; a short pleon with pleotelson lateral margins converging to a narrowly rounded apex; pleonite lateral margins overlapped by pereonite 7; slender uropods that extend to the posterior margin of the pleotelson with the uropodal exopod almost double the length of the endopod., Graphical abstract Image 1, Highlights • Mothocya andoni n. sp. and Mothocya powelli n. sp. are described. • First records of Mothocya from Nigeria. • Full descriptions and identifying characteristics for the two new species are provided. • Increased knowledge on the understudied eastern South Atlantic.

Thirteen species of sea lice (family Caligidae) are reported from a range of elasmobranch and actinopterygian fishes caught off South Africa or obtained from public aquaria in South Africa. Two new species of Caligus Müller, 1785 are described: C. linearisn. sp. from Pomatomus saltatrix (Linnaeus) and C. tumulusn. sp. from Chrysoblephus cristiceps (Valenciennes). A supplementary description is provided for both sexes of Caligus tetrodontis Barnard, 1948 taken from Amblyrhynchotes honckenii (Bloch) and previous records of this parasite from South African fishes are critically reviewed. It is concluded that Caligus material from Arothron hispidus Linnaeus was previously misidentified as C. tetrodontis and is in urgent need of re-examination. Morphological and molecular observations on Caligus furcisetifer Redkar, Rangnekar & Murti, 1949 indicate that this copepod is phenotypically and genetically identical to Lepeophtheirus natalensis Kensley & Grindley, 1973, and the latter becomes a junior subjective synonym of C. furcisetifer. We include new geographical distribution records for Caligus longipedis Bassett-Smith, 1898, C. rufimaculatus Wilson, 1905 and Lepeophtheirus spinifer Kirtisinghe, 1937, extending into South African waters, as well as both new distribution and host records for Alebion gracilis Wilson, 1905, Caligus dakari van Beneden, 1892 and Lepeophtheirus acutus Heegaard, 1943. The molecular analysis confirmed the monophyly of the genus Caligus. The South African species of Caligus did not cluster together, but the two included South African species of Lepeophtheirus were recovered as sister taxa.

Due to their unusual life cycle that includes parasitic larval and free living adult stages, gnathiid isopods are typically overlooked in biodiversity surveys, even those that focus on parasites. While the Philippines sits within the region of highest marine biodiversity in the world, the coral triangle, no gnathiid species have been identified or described from that region. Here we present the first records of two gnathiid species collected from the Visayas, central Philippines: Gnathia malaysiensis Müller, 1993, previously described from Malaysia, and G. camuripenis Tanaka, 2004, previously described from southern Japan. This paper provides detailed morphological redescriptions, drawings and scanning electron microscope images as well as the first molecular characterisation of both species, Furthermore, a summary of the Central-Indo Pacific Gnathia species is provided.

A fish parasitic isopod recently reported from India as Anilocra leptosoma Bleeker, 1857, was re-examined and morphologically compared to five closely related species: A. capensis Leach, 1818, A. clupei Williams and Bunkley-Williams, 1986, A. leptosoma Bleeker, 1857, A. paulsikkeli Welicky and Smit, 2019 and A. pilchardi Bariche and Trilles, 2006. This species was sequenced and compared to other known Anilocra species based on mitochondrial cytochrome c oxidase subunit I (COI) gene fragments. Both morphological and molecular data corroborate that the Anilocra species parasitising the clupeid fish Tenualosatoli (Valenciennes, 1847) from India should be recognised as a new species, and we describe Anilocra grandmaae n. sp. based on the holotype and paratype females. The key characters of A. grandmaae n. sp. include the body being less than 4.0 times as long as wide; antennula article 3 anterodistal margin expanded, 1.2–1.4 times as wide as long; pleonite 1 concealed by pereonite 7 and lateral margin posteriorly produced; pereopods 1–4 with three nodules on dactylus; endopod of pleopods 3–5 with a proximomedial lobe and folding; and pleotelson ovate, with lateral margins converging smoothly to a caudomedial point., Graphical abstract Image 1, Highlights • Description of a first new Anilocra fish parasitic isopod from India. • First molecular characterisation of a cymothoid isopod from India. • Anilocra grandmaae n. sp. is genetically more closely related to species from the Caribbean than Africa. • First record of a cymothoid isopod parasitising the clupeid fish Tenualosa toil.

Chonopeltis lisikili Van As and Van As, 1996 was original described from the Eastern Caprivi (Namibia) and the Okavango System (Botswana), collected from five Synodontis Cuvier, 1816 (Mochokidae) species. This fish genus is endemic to Africa, with 130 valid species, making it one of the most species-rich and widely distributed mochokid catfish family. During parasitological surveys conducted in the Phongolo River (South Africa), a Chonopeltis Thiele, 1900 species was collected from Synodontis zambezensis Peters, 1852. In total, 21 adult females, four adult males, as well as representatives of the larval developmental stages were found. After morphological comparison, this branchiuran was identified as C. lisikili. This paper provides additional information on body measurements and morphological information of all the life stages of the Phongolo material. Information on Chonopeltis material collected from the Okavango River, Botswana, not yet published is also included as well as the first molecular characterisation of a Chonopeltis species using COI and 18 S rRNA partial sequencing. The endemicity of the host and fish lice necessitates a discussion on the host origin and distribution in more than one river system, particularly referring to host specificity for Chonopeltis species., Graphical abstract Image 1, Highlights • First COI sequence for genus Chonopeltis. • First phylogenetic analysis of Branchiurans using 18 S rRNA. • New host and locality record for Chonopeltis lisikili. • First illustrations of the majority of C. lisikili life stages, in fact for any of the known Chonopeltis species.

The global invasive anguillid gill parasite Pseudodactylogyrus anguillae (Yin and Sproston, 1948) has only recently been documented from eels in South Africa. As there is no known eel trade in South Africa, the source of introduction of this parasite has been debated, and its status as an alien parasite was rendered uncertain. We report on the first infection of Pseudodactylogyrus anguillae from the giant mottled eel Anguilla marmorata from the Phongolo River (South Africa) using classic morphological and molecular methodologies and clarify the introduction status category of this parasite as alien and invasive.

The monogenean genus Heterobothrium (Monogenea: Diclidophoridae) currently consists of 12 accepted species described globally from various pufferfishes (Tetraodontidae). This includes the economically important Heterobothrium okamotoi Ogawa 1991 that causes severe disease in the cultured tiger puffer Takifugu rubripes in Japan. During parasitological surveys targeting near shore marine fishes of South Africa, a new Heterobothrium was discovered on the gills of five evileye pufferfish Amblyrhynchotes honckenii with a prevalence of 100% and mean intensity of 23 (4-72). Specimens were morphologically studied using both light and scanning electron microscopy and molecularly characterized by sequencing the partial ribosomal gene 28S. Heterobothrium victorwepeneri n. sp. differs from its 12 congeners by a combination of morphological characters, such as fourth clamp pair (anteriormost, 180° inverted) as the smallest with some of the sclerites differing in shape from the other pairs, 8 to 9 genital hooks, number of testes 40-50, and absent isthmus. The 28S tree depicted two main branches, one clustering together species of mazocraeidean families while the other clustered together only species of the Diclidophoridae. Heterobothrium victorwepeneri n. sp. is the first species of its genus to be recorded and described from South Africa and from the tetraodontid A. honckenii. This study also provides for the first time 28S sequence for a species of this highly host-specific genus.

Brucethoa bharata gen. et sp. nov. is described from the host fish Glossanodon macrocephalus Bineesh and Endo, 2019 (Argentinidae), at depths of 300–600 metres from the south-west coast of India. T...

Summary Understanding the genetic design principles that determine protein production remains a major challenge. Although the key principles of gene expression were discovered 50 years ago, additional factors are still being uncovered. Both protein-coding and non-coding sequences harbor elements that collectively influence the efficiency of protein production by modulating transcription, mRNA decay, and translation. The influences of many contributing elements are intertwined, which complicates a full understanding of the individual factors. In natural genes, a functional balance between these factors has been obtained in the course of evolution, whereas for genetic-engineering projects, our incomplete understanding still limits optimal design of synthetic genes. However, notable advances have recently been made, supported by high-throughput analysis of synthetic gene libraries as well as by state-of-the-art biomolecular techniques. We discuss here how these advances further strengthen understanding of the gene expression process and how they can be harnessed to optimize protein production.

Significance The Calvin cycle is the most important carbon fixation pathway in the biosphere. However, its carboxylating enzyme Rubisco also accepts oxygen, thus producing 2-phosphoglycolate. Phosphoglycolate salvage pathways were extensively studied in photoautotrophs but remain uncharacterized in chemolithoautotrophs using the Calvin cycle. Here, we study phosphoglycolate salvage in the chemolithoautotrophic model bacterium Cupriavidus necator H16. We demonstrate that this bacterium mainly reassimilates 2-phosphoglycolate via the glycerate pathway. Upon disruption of this pathway, a secondary route, which we term the malate cycle, supports photorespiration by completely oxidizing 2-phosphoglycolate to CO2. While the malate cycle was not previously known to metabolize 2-phosphoglycolate in nature, a bioinformatic analysis suggests that it may support phosphoglycolate salvage in diverse chemoautotrophic bacteria., Carbon fixation via the Calvin cycle is constrained by the side activity of Rubisco with dioxygen, generating 2-phosphoglycolate. The metabolic recycling of phosphoglycolate was extensively studied in photoautotrophic organisms, including plants, algae, and cyanobacteria, where it is referred to as photorespiration. While receiving little attention so far, aerobic chemolithoautotrophic bacteria that operate the Calvin cycle independent of light must also recycle phosphoglycolate. As the term photorespiration is inappropriate for describing phosphoglycolate recycling in these nonphotosynthetic autotrophs, we suggest the more general term “phosphoglycolate salvage.” Here, we study phosphoglycolate salvage in the model chemolithoautotroph Cupriavidus necator H16 (Ralstonia eutropha H16) by characterizing the proxy process of glycolate metabolism, performing comparative transcriptomics of autotrophic growth under low and high CO2 concentrations, and testing autotrophic growth phenotypes of gene deletion strains at ambient CO2. We find that the canonical plant-like C2 cycle does not operate in this bacterium, and instead, the bacterial-like glycerate pathway is the main route for phosphoglycolate salvage. Upon disruption of the glycerate pathway, we find that an oxidative pathway, which we term the malate cycle, supports phosphoglycolate salvage. In this cycle, glyoxylate is condensed with acetyl coenzyme A (acetyl-CoA) to give malate, which undergoes two oxidative decarboxylation steps to regenerate acetyl-CoA. When both pathways are disrupted, autotrophic growth is abolished at ambient CO2. We present bioinformatic data suggesting that the malate cycle may support phosphoglycolate salvage in diverse chemolithoautotrophic bacteria. This study thus demonstrates a so far unknown phosphoglycolate salvage pathway, highlighting important diversity in microbial carbon fixation metabolism.

Fishes of the order Siluriformes, known as catfishes, have a global distribution with more than 3,600 valid species of which 2,087 occur in the Neotropical region. Despite being highly diverse, abundant, and of economic importance as food and ornamental fishes, knowledge about the diversity and distribution of their helminth parasites is fragmentary and scarce. Eight species of catfishes (Pterodoras granulosus, Trachydoras paraguayensis, Pimelodella avanhandavae, Loricariichthys platymetopon, Pterygoplichthys ambrosettii, Rhinelepis aspera, Hemisorubim platyrhynchos, and Sorubim lima) from the Aguapeí River, Upper Paraná River basin, municipality of Castilho, São Paulo State, Brazil, were surveyed for helminth parasites. Collected fishes were weighed, measured, and examined for helminth parasites following standard methodology. Fifty helminth parasite taxa (23 monogeneans, 13 digeneans, 11 nematodes, and three cestodes) were found from a total of 405 fishes screened. The helminth taxon that showed the highest mean intensity of infection and mean abundance was the nematode Rondonia rondoni from P. granulosus, followed by the nematode Parasynodontisia petterae from R. aspera. The ecological analyses were carried out at the component community level and at the infracommunity level. Trachydoras paraguayensis had the richest helminth component community. Pterygoplichthys ambrosettii had the most diverse helminth component community and R. aspera had the lowest. Both hosts are loricariids and have similar diet. However, the high parasite diversity of P. ambrosettii is due to the number of dactylogyrids species found (10), which are directly transmitted, whereas only three dactylogyrid species were found in R. aspera. At infracommunity level, the nematode species R. rondoni and P. petterae dominated the parasite communities. This study presents 38 new host records, contributing considerably to increase the diffuse knowledge of helminth parasites of Neotropical siluriforms., Graphical abstract Image 1, Highlights • Fifty helminth parasite taxa were recovered from 405 catfishes surveyed. • Monogenea was the most diverse group, representing 46% of the helminth taxa found. • Thirty-eight new host records are presented.

Haemogregarine (Apicomplexa: Adeleorina) blood parasites are commonly reported from anuran hosts. Dactylosomatidae (Jakowska and Nigrelli, 1955) is a group of haemogregarines comprising Dactylosoma Labbé, 1894 and Babesiosoma Jakowska and Nigrelli, 1956. Currently Dactylosoma and Babesiosoma contain five re-cognised species each. In the current study, a total of 643 anurans, comprising 38 species, 20 genera, and 13 families were collected from South Africa (n = 618) and Belgium (n = 25), and their blood screened for the presence of dactylosomatid parasites. Three anuran species were found infected namely, Ptychadena anchietae (Bocage, 1868) and Sclerophrys gutturalis (Power, 1927) from South Africa, and Pelophylax lessonae (Camerano, 1882) from Belgium. Based on morphological characteristics, morphometrics and molecular results a new dactylosomatid, Dactylosoma kermiti n. sp. is described form Pty. anchietae and Scl. gutturalis. The species of Dactylosoma isolated from Pel. lessonae could not, based on morphological or molecular analysis, be identified to species level. Phylogenetic analysis shows species of Dactylosoma infecting anurans as a monophyletic group separate from the other haemogregarine groups. Additionally, the mosquitoes Uranotaenia (Pseudoficalbia) ma-shonaensis Theobald, 1901 and U. (Pfc.) montana Ingram and De Meillon, 1927 were observed feeding on Scl. gutturalis in situ and possible dividing stages of this new parasite were observed in the mosquitoes. This study is the first to describe a dactylosomatid parasite based on morphological and molecular data from Africa as well as observe potential stages in possible dipteran vectors. This paper forms part of a VLIR-UOS TEAM funded project (ZEIN21013PR396), co-funded by the National ResearchFoundation of South Africa (NRF project CPRR160429163437, grant105979, NJ Smit, PI). The financial assistance of the NRF towards ECNsupported by the DAAD-NRF doctoral scholarship (Grant UID: 108803), and the VLIR-OUS university scholarship (ID 0620854/Contract000000076310), is acknowledged

In this study we explore the sub-lethal effects of two malaria vector control pesticides, deltamethrin and dichlorodiphenyltrichloroethane (DDT), on Xenopus laevis by incorporating different levels of biological organisation. Pesticide accumulation in frog tissue was measured alongside liver metabolomics and individual swimming behaviour to assess whether changes presented at these different levels, and if such changes could be linked between levels. Results showed evidence of concentration dependent accumulation of DDT and its metabolites, but no measurable accumulation of deltamethrin in adult X. laevis after 96 h of exposure. Both DDT and deltamethrin were shown to cause alterations in the liver metabolome of X. laevis. We also showed that some of these changes can be enhanced in exposure to a mixture of these two pesticides. Initial behavioural responses recorded directly after exposure were seen in the form of decreased activity, less alterations between mobility states, and less time spent at the water surface. This response persisted after 96 h of exposure to a mixture of the two pesticides. This study shows that sub-lethal exposure to pesticides can alter the biochemical homeostasis of frogs with the potential to cascade onto behavioural and ecological levels in mixture exposure scenarios.

Organochlorine pesticides (OCPs) in South Africa have for the most part been banned, except dichlorodiphenyltrichloroethane (DDT) which is still used as malaria vector control. The aim of this study was to determine OCP residues in the aquatic fauna of one of South Africa's most populated areas, Soweto. Risk to human health through OCP exposure via fish consumption was investigated. Clarias gariepinus was chosen as bioindicator because it is an apex predator that is in abundance, but is also a valued food source. Dichlorodiphenyltrichloroethanes (DDTs), hexachlorocyclohexanes (HCHs), and chlordanes (CHLs) were detected in the fish tissue with the DDTs being the most prevalent at all sites. Of the three locations, Fleurhof, Orlando, and Lenasia, the latter location's fish had the highest ΣOCP load, ranging between 81 and 1190 ng/g wm. The DDTs were determined to be from historic use, whereas the CHL levels indicated more recent inputs. Although the possibility of illegal use cannot be excluded completely, the presence of OCPs outside of their allowed areas of use indicate that these compounds not only stay in the aquatic systems long term, but may be of concern in areas previously not considered high risk areas. The OCP residues in C. gariepinus from the study area pose an extremely high risk to human health when consumed, and has a cancer risk as high as 1 in 10. This potential problem should be kept in consideration when developing national health and conservation strategies.

Prokaryotic genes encoding functionally related proteins are often clustered in operons. The compact structure of operons allows for co-transcription of the genes, and for co-translation of the polycistronic messenger RNA to the corresponding proteins. This leads to reduced regulatory complexity and enhanced gene expression efficiency, and as such to an overall metabolic benefit for the protein production process in bacteria and archaea. Interestingly, the genes encoding the subunits of one of the most conserved and ubiquitous protein complexes, the RNA polymerase, are not clustered in a single operon. Rather, its genes are scattered in all known prokaryotic genomes, generally integrated in different ribosomal operons. To analyze the impact of this genetic organization on the fitness of Escherichia coli, we constructed a bacterial artificial chromosome harboring the genes encoding the RNA polymerase complex in a single operon. Subsequent deletion of the native chromosomal genes led to a reduced growth on minimal medium. However, by using adaptive laboratory evolution the growth rate was restored to wild-type level. Hence, we show that a highly conserved genetic organization of core genes in a bacterium can be reorganized by a combination of design, construction and optimization, yielding a well-functioning synthetic genetic architecture.

Schizophrenia is a heritable complex phenotype associated with a background risk involving multiple common genetic variants of small effect and a multitude of environmental exposures. Early twin and family studies using proxy-genetic liability measures suggest gene-environment interaction in the etiology of schizophrenia spectrum disorders, but the molecular evidence is scarce. Here, by analyzing the main and joint associations of polygenic risk score for schizophrenia (PRS-SCZ) and environmental exposures in 1,699 patients with a diagnosis of schizophrenia spectrum disorders and 1,542 unrelated controls with no lifetime history of a diagnosis of those disorders, we provide further evidence for gene-environment interaction in schizophrenia. Evidence was found for additive interaction of molecular genetic risk state for schizophrenia (binary mode of PRS-SCZ above 75% of the control distribution) with the presence of lifetime regular cannabis use and exposure to early-life adversities (sexual abuse, emotional abuse, emotional neglect, and bullying), but not with the presence of hearing impairment, season of birth (winter birth), and exposure to physical abuse or physical neglect in childhood. The sensitivity analyses replacing the a priori PRS-SCZ at 75% with alternative cut-points (50% and 25%) confirmed the additive interaction. Our results suggest that the etiopathogenesis of schizophrenia involves genetic underpinnings that act by making individuals more sensitive to the effects of some environmental exposures. ispartof: WORLD PSYCHIATRY vol:18 issue:2 pages:173-182 ispartof: location:Italy status: published

The branchial-attaching cymothoid genus, Elthusa Schioedte & Meinert, 1884 is a genus with a worldwide distribution of 36 species, including the three species described here. Elthusaraynaudii (Milne Edwards, 1840) is the only species that has been described from southern Africa. All South African material held at the National Museum of Natural History, Paris, France (MNHN) and the Iziko South African Museum, Cape Town (SAMC) identified as, or appearing to belong to, Elthusa was examined. Four species were identified, Elthusaraynaudii and three species that proved to be undescribed. Elthusaxenasp. n. can be distinguished by an evenly rounded pereonite 1 anterior margin, a roughly rectangular pleotelson, and narrowly rounded uropod apices that extend to more than half the length of the pleotelson. Elthusaacutinasasp. n. is identified by the produced and narrowly rounded cephalon anterior margin, acute uropods that are shorter than half the length of the pleotelson, and pereonite 1 anterior margin with medial projection. Elthusarotundasp. n. is characterised by the round body shape, broadly rounded uropod apices, and protrusions on the proximal and lateral margins of the merus and carpus of pereopod 7. A key to the South African Elthusa species is provided, together with a table summarising the hosts and localities of the 33 previously known species of Elthusa.

Simultaneous triggering of Toll-like receptors (TLRs) and NOD-like receptors (NLRs) has previously been shown to synergistically activate monocytes, dendritic cells and macrophages. We applied these properties in a T-cell vaccine setting by conjugating the NOD2-ligand muramyl-dipeptide (MDP) and TLR2-ligand Pam3CSK4 to a synthetic peptide derived from a model antigen. Stimulation of human DCs with the MDP-peptide-Pam3CSK4 conjugate led to a strongly increased secretion of pro-inflammatory and Th1-type cytokines and chemokines. We further show that the conjugated ligands retain their ability to trigger their respective receptors, while even improving NOD2-triggering. Also, activation of murine DCs was enhanced by the dual triggering, ultimately leading to effective induction of vaccine-specific T cells expressing IFNγ, IL-2 and TNFα. Together, these data indicate that the dual MDP-SLP-Pam3CSK4 conjugate constitutes a chemically well-defined vaccine approach that holds promise for the use in the treatment of virus infections and cancer.

The influenza A virus (IAV) interacts with the glycocalyx of host cells through its surface proteins hemagglutinin (HA) and neuraminidase (NA). Quantitative biophysical measurements of these interactions may help to understand these interactions at the molecular level with the long-Term aim to predict influenza infectivity and answer other biological questions. We developed a method, called multivalent affinity profiling (MAP), to measure virus binding profiles on receptor density gradients to determine the threshold receptor density, which is a quantitative measure of virus avidity toward a receptor. Here, we show that imaging of IAVs on receptor density gradients allows the direct visualization and efficient assessment of their superselective binding. We show how the multivalent binding of IAVs can be quantitatively assessed using MAP if the receptor density gradients are prepared around the threshold receptor density without crowding at the higher densities. The threshold receptor density increases strongly with increasing flow rate, showing that the superselective binding of IAV is influenced by shear force. This method of visualization and quantitative assessment of superselective binding allows not only comparative studies of IAV-receptor interactions, but also more fundamental studies of how superselectivity arises and is influenced by experimental conditions.

South African clinids are a major component of the temperate intertidal regions that are also known to participate in life cycles and transmission of several groups of parasites. However, the knowledge of trematode diversity of these fishes is incomplete. In this study, two species of Clinus Cuvier, the super klipfish Clinus superciliosus (Linnaeus) and the bluntnose klipfish Clinus cottoides Valenciennes, were collected from six localities along the South African coast and examined for the presence of trematodes. Metacercariae of Cardiocephaloides Sudarikov, 1959 were found in the eye vitreous humour and brain of C. superciliosus and in the eye vitreous humour of C. cottoides. Detailed analyses integrating morphological and molecular sequence data (28S rDNA, ITS2 rDNA-region, and COI mtDNA) revealed that these belong to two species, Cardiocephaloides physalis (Lutz, 1926) and an unknown species of Cardiocephaloides. This study provides the first report of clinid fishes serving as intermediate hosts for trematodes, reveals that the diversity of Cardiocephaloides in South Africa is higher than previously recorded, and highlights the need for further research to elucidate the life cycles of these trematode species. The broad geographical distribution of Cardiocephaloides spp. was confirmed in the present study based on molecular sequence data. The host-parasite interactions between clinid fishes and metacercariae of Cardiocephaloides are yet to be explored.

Understanding how influenza viruses traverse the mucus and recognize host cells is critical for evaluating their zoonotic potential, and for prevention and treatment of the disease. The surface of the influenza A virus is covered with the receptor-binding protein hemagglutinin and the receptor-cleaving enzyme neuraminidase, which jointly control the interactions between the virus and the host cell. These proteins are organized in closely spaced trimers and tetramers to facilitate multivalent interactions with sialic acid-terminated glycans. This review shows that the individually weak multivalent interactions of influenza viruses allow superselective binding, virus-induced recruitment of receptors, and the formation of dynamic complexes that facilitate molecular walking. Techniques to measure the avidity and receptor specificity of influenza viruses are reviewed, and the pivotal role of multivalent interactions with their emergent properties in crossing the mucus and entering host cells is discussed. A model is proposed for the initiation of cell entry through virus-induced receptor clustering. The multivalent interactions of influenza viruses are maintained in a dynamic regime by a functional balance between binding and cleaving.

The digenetic trematode family Gorgocephalidae comprises just a few species, and the literature devoted to the lineage consists of only a handful of reports. With one exception, all reports have been based on material collected in the Indo-West Pacific, an expansive marine ecoregion stretching from the east coast of Africa to Easter Island, Hawaii and French Polynesia. We collected adult and intramolluscan gorgocephalids from kyphosid fishes and littorinid gastropods from several Australian localities, and from South Africa and French Polynesia. Specimens of Gorgocephalus kyphosi and G. yaaji were collected from, or near, their type-localities, providing new morphological and molecular (COI, ITS2 and 28S) data needed for a revised understanding of species boundaries in the family. Two new species are recognized: Gorgocephalus euryaleae sp. nov. and Gorgocephalus graboides sp. nov. New definitive host records are provided for described species and three new intermediate hosts are identified. These new records are all associated with Kyphosus fishes and littorinid gastropods, reaffirming the restriction of gorgocephalids to these hosts. Most significantly, we provide evidence that G. yaaji is distributed from South Africa to French Polynesia, spanning the breadth of the Indo-West Pacific. Our findings have significant relevance regarding digenean species delineation over geographic range.

The surface of influenza viruses is decorated with two glycoproteins, hemagglutinin (HA) and neuraminidase (NA), that form the characteristic “spikes” and work together in host cell recognition. The receptor specificity of HA and NA is considered a key factor in the adaptation of the virus to different hosts. The receptors for both HA and NA are sialic acid-terminated glycans. NA can cleave the sialic acid from the glycan upon binding. A functional balance exists between the binding and cleaving functions of the two glycoproteins that keeps the virus-receptor interactions dynamic and imparts motility to the virus. A method is presented, called “multivalent affinity profiling”, to quantitatively assess the multivalent binding of influenza viruses on cell-mimicking surfaces through imaging of the virus’ superselectivity on receptor density gradients. The multivalent virus-surface binding has a threshold receptor density above which the binding becomes efficient, which is the hallmark of the multivalent nature of the binding process. The threshold receptor density is stable over time and varies barely with virus concentration, but increases nonlinearly with flow rate due to the shear force exerted on the virus particles. Multivalent affinity profiling was used to compare the binding of an influenza virus to avian and human-type receptors of different lengths. A theoretical model is presented to calculate the avidity, the number of HA-receptor interactions, and their average strength from the threshold receptor density. The influence of NA on the binding of influenza virus as a function of receptor density is studied by varying the concentration of NA inhibitor. With active NA, the virus has a higher avidity due to the contribution of NA to the binding, but the receptor-cleaving activity of NA lowers the receptor density. The virus binding is decreased overall but more strongly at low receptor densities, leading to an enhanced selectivity towards high initial receptor densities. The new insights in the binding of influenza may help to predict properties of emerging influenza strains as well as other viruses, and help the development of broad-spectrum antivirals and targeted anticancer treatments.

Clinus superciliosus was collected for element analysis from six localities along the South African west and south coasts. Concentrations in muscle and liver were determined, considering size and sex. No significant positive correlation between size and concentrations were detected, except for Mn and Sn in liver from Simons Town marina, while no significant differences in sex were detected. The majority of element concentrations were significantly higher in fish from Tsitsikamma in the Garden Route and the small town Chintsa, while some concentrations were significantly higher in muscle at Simons Town marina. Land-use activities had a limited role in element bioaccumulation in klipfish. Element concentrations were influenced by large scale oceanographic processes (currents; upwelling) and localised seasonal geogenic derived run-off. Limited data on element accumulation patterns of intertidal fish species in South Africa, highlights the need for long-term monitoring and further studies on different resident and transient intertidal fish species.

To date, micropollutants from anthropogenic sources cannot be completely removed from effluents of wastewater treatment plants and therefore enter freshwater systems, where they may impose adverse effects on aquatic organisms, for example, on fish. Advanced treatment such as ozonation aims to reduce micropollutants in wastewater effluents and, thus, to mitigate adverse effects on the environment. To investigate the impact and efficiency of ozonation, four different water types were tested: ozonated wastewater (before and after biological treatment), conventionally-treated wastewater, and water from a river (River Ruhr, Germany) upstream of the wastewater treatment plant effluent. Zebrafish (Danio rerio) embryos were used to study lethal and sublethal effects in a modified fish early life-stage test. Mortality occurred during exposure in the water samples from the wastewater treatment plant and the river in the first 24 h post-fertilization, ranging from 12% (conventional wastewater) to 40% (river water). Regarding sublethal endpoints, effects compared to the negative control resulted in significantly higher heart rates (ozonated wastewater), and significantly reduced swimming activity (highly significant in ozonated wastewater and ozone reactor water, significant in only the last time interval in river water). Moreover, the respiration rates were highly increased in both ozonated wastewater samples in comparison to the negative control. Significant differences between the ozonated wastewater samples occurred in the embryonic behavior and heart rates, emphasizing the importance of subsequent biological treatment of the ozonated wastewater. Only the conventionally-treated wastewater sample did not elicit negative responses in zebrafish, indicating that the discharge of conventional wastewater poses no greater risk to embryonic and larval zebrafish than water from the river Ruhr itself. The sublethal endpoints embryonic- and larval behavior, heart rates, and respiration were found to be the most sensitive endpoints in this fish early life-stage test and can add valuable information on the toxicity of environmental samples.

Coculture interactions between lasR loss-of-function and LasR+ Pseudomonas aeruginosa strains may explain the worse outcomes associated with the presence of LasR− strains. More broadly, this report illustrates how interactions within a genotypically diverse population, similar to those that frequently develop in natural settings, can promote unpredictably high virulence factor production., The opportunistic pathogen Pseudomonas aeruginosa damages hosts through the production of diverse secreted products, many of which are regulated by quorum sensing (QS). The lasR gene, which encodes a central QS regulator, is frequently mutated in clinical isolates from chronic infections, and loss of LasR function (LasR−) generally impairs the activity of downstream QS regulators RhlR and PqsR. We found that in cocultures containing LasR+ and LasR− strains, LasR− strains hyperproduce the RhlR/RhlI-regulated antagonistic factors pyocyanin and rhamnolipids in diverse models and media and in different strain backgrounds. Diffusible QS autoinducers produced by the wild type were not required for this effect. Using transcriptomics, genetics, and biochemical approaches, we uncovered a reciprocal interaction between wild-type and lasR mutant pairs wherein the iron-scavenging siderophore pyochelin produced by the lasR mutant induced citrate release and cross-feeding from the wild type. Citrate, a metabolite often secreted in low iron environments, stimulated RhlR signaling and RhlI levels in LasR−but not in LasR+ strains. These studies reveal the potential for complex interactions between recently diverged, genetically distinct isolates within populations from single chronic infections.

The examination of eight spotted skates, Raja straeleni Poll, resulted in the discovery of four new species of Acanthobothrium van Beneden, 1849, namely A. microhabentes sp. n., A. microtenuis sp. n., A. crassus sp. n., and A. dolichocollum sp. n., located off the Western Cape of South Africa. With a total of over 200 valid species of Acanthobothrium recognised worldwide, the use of an integrative approach becomes imperative in the interest of simplifying interspecific comparisons between congeners. In accordance with this, the four new species were incorporated into the category classification system established by Ghoshroy and Caira in 2001, where they were identified as category 2 species, which, at present, includes 47 recognised species of Acanthobothrium. Nevertheless, each of the four new species exhibits postovarian testes, a most intriguing and highly unusual feature among Acanthobothrium, instantly differentiating them from most congeners. This feature has been reported in 12 congeners, which have previously been considered to be restricted to waters of the Indo-Pacific Ocean. Not only do the four new congeners represent the first species of Acanthobothrium reported from southern Africa, but they also represent the first reported species with postovarian testes from the southern Atlantic Ocean. Regarding the legitimacy of the four new species, only two other category 2 species are reported to exhibit this feature, namely A. popi Fyler, Caira et Jensen, 2009, and A. bobconniorum Fyler et Caira, 2010, to which the four congeners were compared to. Acanthobothrium microhabentes sp. n. is the smallest of the congeners and differs from A. popi and A. bobconniorum by having fewer testes and postovarian testes, a shorter body, fewer proglottids, a shorter scolex, and longer cephalic peduncle. Acanthobothrium microtenuis sp. n. differs from A. popi and A. bobconniorum by having fewer testes and postovarian testes, a shorter scolex, longer cephalic peduncle, and the possession of columnar spinitriches on the anterior region of the terminal proglottid. Acanthobothrium crassus sp.n. differs from A. popi and A. bobconniorum by having fewer postovarian testes, a narrower cirrus-sac, larger vitelline follicles, and a longer cephalic peduncle. Acanthobothrium dolichocollum sp. n. is the longest of the four new species and differs from A. popi and A. bobconniorum by having fewer postovarian testes, more postporal testes, a larger body, more proglottids, larger testes and vitelline follicles, and an exceptionally long cephalic peduncle. Apart from differences in overall size, the four new species differ in a combination of measurements for the scolex, vitelline follicles, muscular pad and cephalic peduncle, and the number of proglottids and testes. The four species were recovered from a previously unexplored host and locality, expanding the host associations and geographical distribution of the genus.

Parasitological examination of freshwater fishes of the Phongolo River in north-eastern KwaZulu-Natal, South Africa resulted in the discovery and morphological and molecular characterisation of a new species of Wenyonia Woodland, 1923 (Cestoda: Caryophyllidea). The new species from the plain squeaker, Synodontis zambezensis Peters (Siluriformes: Mochokidae), is morphologically most similar to Wenyonia acuminata Woodland, 1923, a species reported from three species of Synodontis in north-eastern, western and central Africa (Sudan, Nigeria, Democratic Republic of the Congo). Both these species are markedly different from congeners by having a nematoform body and a digitiform scolex. Wenyonia gracilis sp. n. differs from W. acuminata in its general body size, length and width of main body regions (testicular and uterine regions), a posterior extension of the testes into the uterine region, numerous postovarian vitelline follicles filling the entire medulla, eggs c. 1/3 larger in size, and a scolex with an apical introvert but devoid of longitudinal furrows and a well-defined base. Wenyonia gracilis is the seventh species in the genus and the first autochthonous caryophyllidean ever reported and described from southern Africa (south of the Zambezi River).

This study investigated the occurrence and phylogenetic relationship of protozoan parasites and Ehrlichia infecting domestic animals from three municipalities in uMkhanyakude district of KwaZulu-Natal province, South Africa. A total of 208 blood samples collected from clinically healthy cattle, sheep, goats and dogs from uMkhanyakude district were examined by polymerase chain reaction (PCR) assays, using either genus or species-specific primers to determine the occurrence and phylogenetic relationship of various protozoan parasites and Ehrlichia of veterinary importance. A total of 5/109 (4.6%) cattle were PCR-positive for the presence of Toxoplasma gondii , 33/109 (30.3%) for Babesia bovis, 24/109 (22.02%) for Babesia bigemina and 20/109 (18.3%) for Trypanosoma sp., while 3/10 (30%) of sheep were PCR-positive for Theileria ovis and none of the goats were positive for any of the detected pathogens. The co-infection of 4/109 (3.7%) B. bovis and B. bigemina was detected in cattle. Only Ehrlichia canis was detected in dogs with infection rate of 20/48 (41.7%). Sequences of PCR-positive isolates ( B. bovis, B. bigemina, E. canis, T. ovis and T. gondii ) showed that they were closely related to their relevant species from various countries. These findings have expanded our knowledge about the prevalence and phylogenetic similarity between protozoan parasites and Ehrlichia isolates of South African origin. To date, this is the first study in South Africa to detect T. gondii infections from cattle blood using PCR.

Mass Nile crocodile (Crocodylus niloticus) deaths in Africa’s premier conservation area, the Kruger National Park (KNP), prompted numerous studies to determine possible causes of the sudden die-offs. The majority highlighted the involvement of the anthropocene to crocodile mass mortalities. One of the potential causative agents was identified as legacy compounds such as organochlorine pesticides (OCPs) present in the various aquatic ecosystems of the KNP. Thus, this study aimed to quantify OCP levels in wild crocodiles from the aquatic systems associated with the KNP. As part of a larger study, muscle tissue samples were collected from tails of 12 wild crocodiles. Results indicated that eight of the 19 OCPs analysed for were quantifiable. These included DDTs (2130–167968 ng/g lw), chlordanes (BD – 7583 ng/g lw) and cyclodeines (BD – 872 ng/g lw). Measured concentrations indicated spatial- and sex-related differences in accumulation patterns. DDT and its metabolites, paricularly p,p’-DDE accumulated to the highest concentrations of the OCP groups. Levels of the different groups of OCPs were the highest recorded in recent history from any crocodilian tissue. The different measured OCP residues were between 2.5 and 120 times higher than levels reported in literature, depending on the compound. These results are of ecotoxicological significance and have several management implications. In view of individuals being sampled from a conservation area it emphasizes the fact that pollutants know no boundaries and that premier conservation areas such as the world renowned KNP may be some of the most contaminated areas of the world.

Carbon fixation via the Calvin cycle is constrained by the side activity of Rubisco with dioxygen, generating 2-phosphoglycolate. The metabolic recycling of 2-phosphoglycolate, an essential process termed photorespiration, was extensively studied in photoautotrophic organisms, including plants, algae, and cyanobacteria, but remains uncharacterized in chemolithoautotrophic bacteria. Here, we study photorespiration in the model chemolithoautotroph Cupriavidus necator (Ralstonia eutropha) by characterizing the proxy-process of glycolate metabolism, performing comparative transcriptomics of autotrophic growth under low and high CO2 concentrations, and testing autotrophic growth phenotypes of gene deletion strains at ambient CO2. We find that the canonical plant-like C2 cycle does not operate in this bacterium and instead the bacterial-like glycerate pathway is the main photorespiratory pathway. Upon disruption of the glycerate pathway, we find that an oxidative pathway, which we term the malate cycle, supports photorespiration. In this cycle, glyoxylate is condensed with acetyl-CoA to give malate, which undergoes two oxidative decarboxylation steps to regenerate acetyl-CoA. When both pathways are disrupted, autotrophic growth is abolished at ambient CO2. We present bioinformatic data suggesting that the malate cycle may support photorespiration in diverse chemolithoautotrophic bacteria. This study thus demonstrates a so-far unknown photorespiration pathway, highlighting important diversity in microbial carbon fixation metabolism.

Background The African leopard Panthera pardus pardus (L.) is currently listed as a vulnerable species on the IUCN (International Union for the Conservation of Nature) red list of threatened species due to ongoing population declines. This implies that leopard-specific parasites are also vulnerable to extinction. Intracellular apicomplexan haemoparasites from the genus Hepatozoon Miller, 1908 have been widely reported from wild carnivores in Africa, including non-specific reports from leopards. This paper describes two new haemogregarines in captive and wild leopards from South Africa and provides a tabular summary of these species in relation to species of Hepatozoon reported from mammalian carnivores. Methods Blood was collected from nine captive and eight wild leopards at various localities throughout South Africa. Thin blood smears were Giemsa-stained and screened for intraleukocytic haemoparasites. Gamont stages were micrographed and morphometrically compared with existing literature pertaining to infections in felid hosts. Haemogregarine specific primer set 4558F and 2733R was used to target the 18S rRNA gene for molecular analysis. Resulting sequences were compared to each other and with other available representative mammalian carnivore Hepatozoon sequences from GenBank. Results Two species of Hepatozoon were found in captive and wild leopards. Of the 17 leopards screened, eight were infected with one or both morphologically and genetically distinct haemogregarines. When compared with other species of Hepatozoon reported from felids, the two species from this study were morphometrically and molecularly distinct. Species of Hepatozoon from this study were observed to exclusively parasitize a particular type of leukocyte, with Hepatozoon luiperdjie n. sp. infecting neutrophils and Hepatozoon ingwe n. sp. infecting lymphocytes. Phylogenetic analysis showed that these haemogregarines are genetically distinct, with Hepatozoon luiperdjie n. sp. and Hepatozoon ingwe n. sp. falling in well supported separate clades. Conclusions To our knowledge, this is the first morphometric and molecular description of Hepatozoon in captive and wild African leopards in South Africa. This study highlights the value of using both morphometric and molecular characteristics when describing species of Hepatozoon from felid hosts.

The CRISPR-Cas9 nuclease has been repurposed as a tool for gene repression (CRISPRi). This catalytically dead Cas9 (dCas9) variant inhibits transcription by blocking either initiation or elongation by the RNA polymerase complex. Conditional control of dCas9-mediated repression has been achieved with inducible promoters that regulate the expression of the dcas9 gene. However, as dCas9-mediated gene silencing is very efficient, even slightly leaky dcas9 expression leads to significant background levels of repression of the target gene. In this study, we report on the development of optimized control of dCas9-mediated silencing through additional regulation at the translation level. We have introduced the TAG stop codon in the dcas9 gene in order to insert a synthetic amino acid, l-biphenylalanine (BipA), at a permissive site in the dCas9 protein. In the absence of BipA, a nonfunctional, truncated dCas9 is produced, but when BipA is present, the TAG codon is translated resulting in a functional, full-length dCas9 protein. This synthetic, BipA-containing dCas9 variant (dCas9-BipA) could still fully repress gene transcription. Comparison of silencing mediated by dCas9 to dCas9-BipA revealed a 14-fold reduction in background repression by the latter system. The here developed proof-of-principle system thus reduces unwanted background levels of gene silencing, allowing for tight and timed control of target gene expression.

Dichlorodiphenyltrichloroethane (DDT) is an organochlorine insecticide that has been used for indoor residual spraying for the control of mosquito-borne diseases including malaria. However, due to its toxicity and environmental persistence, there are concerns about its potential deleterious effects in humans and wildlife. Therefore, the current study aimed to monitor and estimate the level of DDTs in human communities. The accumulation of DDT and its metabolites was evaluated in house rat (as sentinel) livers collected in an area where DDT was sprayed. DDTs were measured using a gas chromatography / Electron Capture Detector. The results revealed high concentrations of DDTs in the rat livers and the levels of DDTs were similar to findings reported from the same area in 2014.

The opportunistic pathogen Pseudomonas aeruginosa damages hosts through the production of diverse secreted products, many of which are regulated by quorum sensing. The lasR gene, which encodes a central quorum-sensing regulator, is frequently mutated, and loss of LasR function impairs the activity of downstream regulators RhlR and PqsR. We found that in diverse models, the presence of P. aeruginosa wild type causes LasR loss-of-function strains to hyperproduce RhlR/I-regulated antagonistic factors, and autoinducer production by the wild type is not required for this effect. We uncovered a reciprocal interaction between isogenic wild type and lasR mutant pairs wherein the iron-scavenging siderophore pyochelin, specifically produced by the lasR mutant, induces citrate release and cross-feeding from wild type. Citrate stimulates RhlR signaling and RhlI levels in LasR-but not in LasR+ strains, and the interactions occur in diverse media. Co-culture interactions between strains that differ by the function of a single transcription factor may explain worse outcomes associated with mixtures of LasR+ and LasR loss-of-function strains. More broadly, this report illustrates how interactions within a genotypically diverse population, similar to those that frequently develop in natural settings, can promote net virulence factor production.

SUMMARYMitochondrial dysfunction is linked to pathogenesis of Parkinson’s disease (PD). However, individual-mitochondria-based analyses do not show a uniform feature in PD patients. Since mitochondria interact with each other, we hypothesize that PD-related features might exist in topological patterns of mitochondria-mitochondria interaction networks (MINs). Here we showed that MINs form non-classical scale-free supernetworks in colonic ganglia both from healthy controls and PD patients, however, altered topological patterns are observed in PD patients. These patterns highly correlate with PD clinical scores and a machine-learning approach based on the MIN features accurately distinguish between patients and controls with an area-under-curve value of 0.989. The MINs of midbrain dopaminergic neurons (mDANs) derived from several genetic PD patients also display specific changes. CRISPR/CAS9-based genome correction of alpha-synuclein point mutations reverses the changes in MINs of mDANs. Our MIN network analysis opens a new dimension for a deeper characterization of various complex diseases with mitochondrial dysregulation.

The threat to wildlife from chemical exposure exists regardless of the presence of conservation boundaries. An issue exacerbated by the use of environmentally persistent insecticides for vector control and long-range transport of legacy persistent organic pollutants. In this comparative study between two important conservation regions in South Africa, Kruger National Park (KNP) and Ndumo Game Reserve (NGR), we assessed organochlorine pesticide (OCP) accumulation in several anuran species collected from within the conservation regions. The two conservation regions differ in size and subsequent proximity of collection sites to OCP input sources. Detectable concentrations of OCPs were present in ∼ half the frogs analysed from KNP and ∼all frogs from NGR and total OCP loads were similar between regions, where measured in the same species. The OCP profiles in KNP frogs were representative of legacy pesticides likely introduced via long-range transport, whereas NGR profiles showed influence of current use of DDT consistent with close proximity to sources. This indicates amphibians can accumulate OCPs within conservation regions and that the exposure of non-target organisms inside conservation regions to current use pesticides has a strong association with proximity to sources. These results serve to inform conservation management decision making with regard to the non-target organism effects of chemical interventions such as vector control pesticide use in and around conservation regions.

Multiple sclerosis (MS) is an autoimmune disorder manifested via chronic inflammation, demyelination, and neurodegeneration inside the central nervous system. The progressive phase of MS is characterized by neurodegeneration, but unlike classical neurodegenerative diseases, amyloid-like aggregation of self-proteins has not been documented. There is evidence that citrullination protects an immunodominant peptide of human myelin oligodendrocyte glycoprotein (MOG34–56) against destructive processing in Epstein-Barr virus-infected B-lymphocytes (EBV-BLCs) in marmosets and causes exacerbation of ongoing MS-like encephalopathies in mice. Here we collected evidence that citrullination of MOG can also lead to amyloid-like behavior shifting the disease pathogenesis toward neurodegeneration. We observed that an immunodominant MOG peptide, MOG35–55, displays amyloid-like behavior upon site-specific citrullination at positions 41, 46, and/or 52. These amyloid aggregates are shown to be toxic to the EBV-BLCs and to dendritic cells at concentrations favored for antigen presentation, suggesting a role of amyloid-like aggregation in the pathogenesis of progressive MS.

Twenty-one species of acanthocephalans, representative of thirteen genera from ten families of seven orders and three classes, are included in this updated checklist of acanthocephalans in South Africa.Although South Africa appears to have a less diverse acanthocephalan fauna compared to some other countries such as Iran in Asia, or Brazil in South America, this is probably an artefact of fewer parasitological surveys.

Coral reefs harbor the greatest biodiversity per unit area of any ecosystem on earth. While parasites constitute the majority of this biodiversity, they remain poorly studied due to the cryptic nature of many parasites and the lack of appropriate training among coral reef ecologists. Damselfishes (Pomacentridae) are among the most abundant and diverse fishes on coral reefs. In a recent study of blood parasites of Caribbean reef fishes, the first ever apicomplexan blood parasites discovered in damselfishes were reported for members of the genus Stegastes. While these blood parasites were characterized as “Haemohormidium-like”, they appear to be distinct from any other known apicomplexan. In this study, we examined host associations, geographic distributions, and provide further insights on the phylogenetic affiliation of this parasite. A combination of morphological characteristics and 18S rDNA sequences suggest that this parasite may be the same species at multiple sites and occurs from the southern to the northern extreme of the eastern Caribbean, although it appears rare in the north. At present it appears to be limited to members of the genus Stegastes and infects all life history stages. It is most common in benthophagous species that occur in high population densities and appears basal to a major monophyletic clade containing species of coccidia, distinct from the Piroplasmida, the order to which Haemohormidium spp. have been assigned. These findings suggest a possible fecal-oral mode of transmission., Graphical abstract Image 1, Highlights • A new species of Haemorhormidium-like apicomplexan blood parasite has recently been discovered in Caribbean damselfishes. • Morphological and molecular data indicate that it is widespread in the eastern Caribbean. • This parasite is limited to damselfish species of the genus Stegastes and infects juveniles and adults. • It is most common in benthophagous species that occur in high population densities. • This parasite resembles coccidia but 18S rDNA show it to be distinct from known genera, as well as genera of piroplasms.