Sero-prevalence of syphilis and associated risk factors among pregnant women attending antenatal care at an urban-poor health centre in Kampala, Uganda: a cross-sectional study.

Introduction: syphilis and its outcomes remain a healthcare system burden with adverse consequences such as stillbirths, neonatal deaths and spontaneous abortions among others. The situation might have worsened because the COVID-19 pandemic has caused a major attention drift from other diseases. Additionally, much as testing for syphilis is a routine practice among pregnant mothers, its proportion is not known in urban health care setting. A study to determine the prevalence of syphilis among pregnant mothers in an urban poor setting is warranted. Methods: a cross-sectional study was conducted among pregnant women who attended antenatal care at Kawaala Health Centre IV in Kampala Capital City between December 2019 to March 2020. Informed consent was sought from study participants prior to data collection using structured questionnaires. Whole blood was collected and tested using SD Bioline HIV/syphilis duo rapid test kit (SD Standard Diagnostics, INC, Korea). Data analysis was done using STATA 14.2. Results: one thousand one hundred and sixty-nine pregnant women participated in the study, with a mean age of 25 years. About 27% of them had completed only primary-level education. Approximately 6% of the participants were HIV seropositive. The prevalence of syphilis was 5.9% (69/1169). HIV positivity (aOR: 4.13, 95%CI: 2.05-8.34), elevated blood pressure (aOR: 2.84, 95%CI: 1.42-5.69), and status of previous pregnancy (aOR: 0.21, 95%CI: 0.05-0.89) were significant predictors of the risk of syphilis among pregnant women in this setting. Conclusion: the prevalence of syphilis among pregnant women in urban poor settings is not low and so must not be underestimated. The potential drivers of syphilis among pregnant women are HIV, elevated blood pressure, and status of previous pregnancy. There should be increased awareness about routine syphilis testing among pregnant mothers attending antenatal care.

Acaricide resistance status of livestock ticks from East and West Africa and in vivo efficacy of acaricides to control them.

Through a collaborative effort across six Sub-Saharan African countries, using recognized international assessment techniques, 23 stocks of three tick species (Rhipicephalus microplus, Rhipicephalus appendiculatus and Amblyomma variegatum) of economic importance for rural small holder farming communities from East and West Africa were collected from cattle, and evaluated in in vitro larval packet tests (LPT). The results demonstrated medium to high resistance to chlorfenvinphos and amitraz across species. Rhipicephalus microplus demonstrated high level alpha-cypermethrin and cypermethrin resistance. Stocks of A. variegatum (West Africa) and R. appendiculatus (Uganda) demonstrated medium level ivermectin resistance. The four least susceptible stocks (East and West African R. microplus, A. variegatum and R. appendiculatus) were taken into in vivo controlled cattle studies where fipronil was found effective against West and East African R. microplus isolates although persistent efficacy failed to reach 90%. Cymiazole and cypermethrin, and ivermectin based acaricides were partially effective against R. microplus without persistent efficacy. Flumethrin spray-on killed A. variegatum within 72 h for up to 10 days posttreatment, however product application was directly to tick attachment sites, which may be impractical under field conditions. A flumethrin pour-on formulation on goats provided persistent efficacy against A. variegatum for up to one-month. Therapeutic control was achieved against R. appendiculatus through weekly spraying cattle with flumethrin, amitraz or combined cymiazole and cypermethrin. A fipronil pour-on product offered four-week residual control against R. appendiculatus (with slow onset of action). Few studies have assessed and directly compared acaricidal activity in vitro and in vivo. There was some discordance between efficacy indicated by LPT and in vivo results. This observation calls for more research into accurate and affordable assessment methods for acaricide resistance. No single active or product was effective against all three tick species, emphasising the need for the development of alternative integrated tick management solutions.

Seroprevalence of contagious bovine pleuropneumonia (CBPP) in cattle from Karamoja region, North-eastern Uganda.

BACKGROUND: Contagious bovine pleuropneumonia [CBPP] is a transboundary animal disease of cattle caused by Mycoplasma mycoides subsp. mycoides [Mmm]. CBPP causes severe economic losses to livestock producers in sub-Saharan Africa mainly due to high mortality, morbidity, reduction in productivity as well as livestock trade restrictions. This study aimed at determining seroprevalence of Mmm in cattle from Karamoja region, north-eastern Uganda; data that are required to design and implement risk based CBPP control program. METHODS: We randomly collected blood samples from 2,300 cattle spread across Karamoja region. Serum was extracted and screened for antibodies against Mycoplasma mycoides subsp. mycoides [Mmm] using the competitive enzyme linked immunosorbent assay [cELISA]. RESULTS: A quarter [25.4%; 95% CI: 23.7-27.3] of the screened cattle [n = 2,300] were sero-positive for Mmm. Amudat and Kaabong districts recorded the lowest [12.3%] and highest [30.7%] Mmm seroprevalence respectively. Increasing age, overnight stay in cattle kraals and location [certain districts, villages, herds and sub counties] of the cattle herds, the factors that promote animal commingling, were the most significant risk factors of seroconversion with Mmm. CONCLUSION: Results from this study indicated a higher seroprevalence of Mmm in Karamoja region cattle herds. This could be due to the increased frequency of CBPP outbreaks in recent years. To be effective, CBPP vaccination programs should target high risk herds along the international borders and other hotspot areas [e.g., parishes or sub counties] where cattle commingling is high.

Odour mapping and air quality analysis of a wastewater treatment plant at a seaside tourist area.

Although wastewater treatment plants (WWTPs) play a fundamental role in protecting the aquatic environment as they prevent organic matter, nutrients and other pollutants from reaching the natural ecosystems, near residential areas they can generate unpleasant smells and noise. The plant studied in the present work is in a seaside tourist area in the Valencian Community, Spain. The main aim was to detect any possible perceptible H2S concentrations from the WWTP by experimental measurement campaigns (including sensor readings and olfactometry measurements by two experts) plus mathematical modelling. After a thorough data analysis of the essential variables involved, such as wind speed, wind direction and H2S concentrations (the main odorant) and comparing their temporal patterns, it was found that the probability of affecting the residential area was highest from June to August before noon and in the late evening. The hourly H2S concentration, influent flow rate and temperature showed a positive correlation, the strongest (R2 = 0.89) being the relationship between the H2S concentration and influent flow rate. These two variables followed a similar daily pattern and indicated that H2S was emitted when influent wastewater was being pumped into the biological reactor. The H2S median concentration at the source of the emission was below 1393.865 µg/m3 (1 ppm), although concentrations 10 times higher were occasionally recorded. The observed H2S peak-to-mean ratio (1 min to 1 h of integration times) ranged from 1.15 to 16.03. This ratio and its attenuation with distance from the source depended on the atmospheric stability. Both H2S concentrations and variability were considerably reduced after submerging the inlet. The AERMOD modelling framework and applying the peak-to-mean ratio were used to map the peak H2S concentration and determine the best conditions to eliminate the unpleasant odour.

Mid- to late Holocene sea-level rise recorded in Hells Bells 234U/238U ratio and geochemical composition.

Hells Bells are underwater secondary carbonates discovered in sinkholes (cenotes) southeast of Cancun on the north-eastern Yucatán peninsula, Mexico. These authigenic calcite precipitates, reaching a length of up to 4 m, most likely grow in the pelagic redoxcline. Here we report on detailed 230Th/U-dating and in-depth geochemical and stable isotope analyses of specimens from cenotes El Zapote, Maravilla and Tortugas. Hells Bells developed since at least ~ 8000 years ago, with active growth until present day. Initial (234U/238U) activity ratios (δ234U0) in Hells Bells calcite decreas from 55 to 15‰ as sea level converges toward its present state. The temporal evolution of the geochemistry and isotope composition of Hells Bells calcites thus appears to be closely linked to sea-level rise and reflects changing hydrological conditions (desalinization) of the aquifer. We suggest that decelerated leaching of excess 234U from the previously unsaturated bedrock traces Holocene relative sea-level rise. Considering this proxy, the resulting mean sea-level reconstruction contains half as much scatter, i.e. improves by a factor of two, when compared to previously published work for the period between 8 and 4 ky BP.

Age-associated alterations of brain mitochondria energetics.

The study aimed to explore the role of age-associated elevated cytosolic Ca2+ in changes of brain mitochondria energetic processes. Two groups of rats, young adults (4 months) and advanced old (24 months), were evaluated for potential alterations of mitochondrial parameters, the oxidative phosphorylation (OxPhos), membrane potential, calcium retention capacity, activity of glutamate/aspartate carrier (aralar), and ROS formation. We demonstrated that the brain mitochondria of older animals have a lower resistance to Ca2+ stress with resulting consequences. The suppressed complex I OxPhos and decreased membrane potential were accompanied by reduction of the Ca2+ threshold required for induction of mPTP. The Ca2+ binding sites of mitochondrial aralar mediated a lower activity of old brain mitochondria. The altered interaction between aralar and mPTP may underlie mitochondrial dysregulation leading to energetic depression during aging. At the advanced stages of aging, the declined metabolism is accompanied by the diminished oxidative background.

Perceptions and practices of farmers of indigenous poultry towards Salmonella infections in North-Central Nigeria.

Background: Salmonella infections continue to be of global concern to poultry health, productivity, and public health. About 44% of the poultry in Nigeria are indigenous and kept in close interaction with farmers who are mostly rural dwellers and have limited access to veterinary and extension services. Aim: The perceptions and practices of farmers of indigenous poultry toward Salmonella infections were assessed to obtain and document baseline data that can be used to create awareness among farmers about these infections and their attendant public health implications. Methods: A cross-sectional approach using a multistage sampling method was used in this survey. A total of 419 farmers keeping indigenous poultry were interviewed using a pre-tested electronic questionnaire in three randomly selected states within North-Central Nigeria. Data were analyzed using descriptive and regression analysis. Results: Out of the 419 respondents, 138 (32.9%), 141 (33.7%), and 140 (33.4%) were from Benue, Kwara, and Plateau States, respectively. Of the 419, 55.4% were females, 40.8% were above 40 years, and 35.8% have over 10 years of farming experience. The majority of the poultry are not housed (58.5%) and farmers predominantly rear chickens (51.8%). Also, 49.9% of the birds were 1-6 months with 41.5% of the flock sizes being 11-20. Respondents had a poor level of perception toward Salmonella infection as the majority did not know that Salmonella affects poultry (89.3%) and that Salmonella infections are zoonotic (94.5%). Significant (p = 0.000) associations existed between categorized perception score and age, educational status, family size, and farming experience of farmers. There were significant (p = 0.000) associations of categorized practice scores with gender, age, education status, family size, and farming experience of farmers. Conclusion: This study has revealed the poor perception of farmers on Salmonella infections and has highlighted their practices. There is a need to raise awareness about these infections to improve indigenous poultry health and productivity as well as public health.

Ecological and Anthropogenic Spatial Gradients Shape Patterns of Dispersal of Foot-and-Mouth Disease Virus in Uganda.

Using georeferenced phylogenetic trees, phylogeography allows researchers to elucidate interactions between environmental heterogeneities and patterns of infectious disease spread. Concordant with the increasing availability of pathogen genetic sequence data, there is a growing need for tools to test epidemiological hypotheses in this field. In this study, we apply tools traditionally used in ecology to elucidate the epidemiology of foot-and-mouth disease virus (FMDV) in Uganda. We analyze FMDV serotype O genetic sequences and their corresponding spatiotemporal metadata from a cross-sectional study of cattle. We apply step selection function (SSF) models, typically used to study wildlife habitat selection, to viral phylogenies to show that FMDV is more likely to be found in areas of low rainfall. Next, we use a novel approach, a resource gradient function (RGF) model, to elucidate characteristics of viral source and sink areas. An RGF model applied to our data reveals that areas of high cattle density and areas near livestock markets may serve as sources of FMDV dissemination in Uganda, and areas of low rainfall serve as viral sinks that experience frequent reintroductions. Our results may help to inform risk-based FMDV control strategies in Uganda. More broadly, these tools advance the phylogenetic toolkit, as they may help to uncover patterns of spread of other organisms for which genetic sequences and corresponding spatiotemporal metadata exist.

The impact of seasonal and event-based infiltration on transition metals (Cu, Ni, Co) in tropical cave drip water.

RATIONALE: The first-row transition metals Cu, Ni, and Co show a strong binding affinity to natural organic matter. Compared to dissolved elements and stable water isotopes, they may be transported rapidly through the soil and host rock into caves in response to infiltration events. This study aims to assess the potential of transition metal ratios as indicators for infiltration changes in response to the seasonal and/or event-based rainfall variation. METHODS: We developed a protocol to analyze Cu, Ni, and Co in the cave drip water using collision cell ICP-QMS without extensive sample pretreatment. The high Ca matrix leads to significant isobaric interferences on all isotope masses. Our method includes a correction of these matrix effects and yields results with comparable accuracy and reproducibility to other published methods. We applied this protocol to drip water samples from Larga Cave (Puerto Rico) covering at least two full annual cycles between 2014 and 2019 on a bimonthly scale. RESULTS: The analysis of external reference materials yielded a reproducibility between 4.7% and 9.2% (relative standard deviation), validating the accuracy of the matrix correction method. The limit of detection is <0.04 ppb for Cu, <0.02 ppb for Ni, and <0.008 ppb for Co. The analysis of drip water samples from Larga Cave reveals pronounced changes of several orders of magnitude in all Element (El) to Ca, Cu/Ni, and Cu/Co ratios in response to seasonal infiltration changes. In addition, we observe a partly even stronger response after major tropical storms and heavy precipitation events of the period of record, for example, tropical storm "Bertha" (2014) and the category 5 hurricanes "Irma" and "Maria" (both 2017). CONCLUSIONS: Transition metal ratios can be accurately measured in cave drip waters with high Ca matrix. At our tropical site, these are promising tracers of infiltration changes in response to changes in the amount of rainfall, providing the first step toward tropical cyclone reconstruction using trace elements in speleothems.

Measurement and sampling error in mixed-methods research for the control of Peste des Petits Ruminants in the Karamoja subregion of Northeastern Uganda: A cautionary tale.

A team of interdisciplinary researchers undertook a mixed methods, participatory epidemiology (PE) based study as part of a pilot project for localized control of Peste des petits ruminants (PPR), a small ruminant disease of high socio-economic impact. Mixed methods research combines qualitative and quantitative methods, allowing iterative comparison of results to arrive at a more comprehensive and informed outcome. In this study, the use of PE and a household survey (HHS) resulted in contradictory results. However, the mixed methods approach also facilitated the detection and the explanation of bias in the HHS, which may have gone undetected and unexplored had only one method been used. Results show that logistical constraints leading to a failure to apply key aspects of the sampling strategy led to problematic gender/ethnic composition of the HHS sample population. Additionally, while PE findings on local disease and terminology were integrated during HHS development and training, there is apparent measurement error related to enumerator bias in HHS results, possibly due to insufficient respondent understanding or a lack of analytic clarity. The extensive nature of the PE, surveillance methodologies used in the initial site assessment, and formative research for the HHS allowed for a critical analysis and interpretation of HHS results as well as reflection on the research process. The findings of this paper underscore the (1) flexibility and utility of participatory methods, (2) the importance of mixed methods research in designing health interventions, and (3) the necessity of tight integration of study design with team planning for implementation of research in environments such as Karamoja, Uganda. If all three are to be achieved not only researchers but funders must provide these space and structure beginning in the study design phase. These findings are relevant in many places, but have particular importance for international, interdisciplinary teams working from various on-and-off-site locations with traditional or indigenous knowledge systems.

Solar forcing of early Holocene droughts on the Yucatán peninsula.

A speleothem record from the north-eastern Yucatán peninsula (Mexico) provides new insights into the tropical hydro-climate of the Americas between 11,040 and 9520 a BP on up to sub-decadal scale. Despite the complex atmospheric reorganization during the end of the last deglaciation, the dominant internal leading modes of precipitation variability during the late Holocene were also active during the time of record. While multi-decadal variations were not persistent, Mesoamerican precipitation was dominated by changes on the decadal- and centennial scale, which may be attributed to ENSO activity driven by solar forcing. Freshwater fluxes from the remnant Laurentide ice sheet into the Gulf of Mexico and the North Atlantic have additionally modulated the regional evaporation/precipitation balance. In particular, this study underlines the importance of solar activity on tropical and subtropical climate variability through forcing of the tropical Pacific, providing a plausible scenario for observed recurrent droughts on the decadal scale throughout the Holocene.

Phylogeographic analysis of foot-and-mouth disease virus serotype O dispersal and associated drivers in East Africa.

The continued endemicity of foot and mouth disease virus (FMDV) in East Africa has significant implications for livestock production and poverty reduction, yet its complex epidemiology in endemic settings remains poorly understood. Identifying FMDV dispersal routes and drivers of transmission is key to improved control strategies. Environmental heterogeneity and anthropogenic drivers (e.g., demand for animal products) can impact viral spread by influencing host movements. Here, we utilized FMDV serotype O VP1 genetic sequences and corresponding spatiotemporal data in order to (i) infer the recent dispersal history, and (II) investigate the impact of external factors (cattle density, human population density, proximity to livestock markets, and drought) on dispersal velocity, location, and direction of FMDV serotype O in East Africa. We identified statistical evidence of long-distance transmission events, and we found that FMDV serotype O tends to remain circulating in areas of high cattle density, high human population density, and in close proximity to livestock markets. The latter two findings highlight the influence of anthropogenic factors on FMDV serotype O spread in this region. These findings contribute to the understanding of FMDV epidemiology in East Africa and can help guide improved control measures.

Invasive cattle ticks in East Africa: morphological and molecular confirmation of the presence of Rhipicephalus microplus in south-eastern Uganda.

BACKGROUND: Rhipicephalus microplus, an invasive tick species of Asian origin and the main vector of Babesia species, is considered one of the most widespread ectoparasites of livestock. The tick has spread from its native habitats on translocated livestock to large parts of the tropical world, where it has replaced some of the local populations of Rhipicephalus decoloratus ticks. Although the tick was reported in Uganda 70 years ago, it has not been found in any subsequent surveys. This study was carried out to update the national tick species distribution on livestock in Uganda as a basis for tick and tick-borne disease control, with particular reference to R. microplus. METHODS: The study was carried out in Kadungulu, Serere district, south-eastern Uganda, which is dominated by small scale livestock producers. All the ticks collected from 240 cattle from six villages were identified microscopically. Five R. microplus specimens were further processed for phylogenetic analysis and species confirmation. RESULTS: The predominant tick species found on cattle was Rhipicephalus appendiculatus (86.9 %; n = 16,509). Other species found were Amblyomma variegatum (7.2 %; n = 1377), Rhipicephalus evertsi (2.3 %; n = 434) and R. microplus (3.6 %; n = 687). Phylogenetic analysis of the 12S rRNA, 16S rRNA and ITS2 gene sequences of R. microplus confirmed the morphological identification. CONCLUSIONS: It is concluded that R. microplus has replaced R. decoloratus in the sampled villages in Kadungulu sub-county, since the latter was not any longer found in this area. There is currently no livestock movement policy in force in Uganda, which could possibly limit the further spread of R. microplus ticks. Future surveys, but also retrospective surveys of museum specimens, will reveal the extent of distribution of R. microplus in Uganda and also for how long this tick has been present on livestock without being noticed.

Rabies in Uganda: rabies knowledge, attitude and practice and molecular characterization of circulating virus strains.

BACKGROUND: Rabies is a deadly preventable viral disease that affects all warm-blooded animals and widespread in many regions including Africa. The disease remains of major public health importance in Uganda. The purpose of this study was to establish Knowledge, Attitude, Practice (KAP) of Rabies in Moyo and Ntoroko districts and to characterize Rabies virus (RABV) strains from seven districts of Uganda with consistent prevalence of rabies. METHODS: KAP survey data were collected based on animal biting history by interviewing the head of the veterinary departments, the medical centers and selected households from the study sites. Data were obtained from 84 households in Ntoroko and Moyo districts. Thirty-five (35) brain samples were collected from bovine, dogs, goats, foxes, jackals ad sheep between 2011 and 2013. Samples were tested using fluorescent antibody test (FAT), One step RT-PCR (following RNA extraction) and partial RABV N gene was sequenced by Sanger method before phylogenetic and phylogeographic analyses of sequences. RESULTS: Scarcity of post-exposure prophylaxis services in the health centers was noted. Poor attitude of wound washing and deficiency of knowledge on how to handle wounds related to dog bites and the significance among household participants lacked. There is a high risk of rabies infection due to a limited dog's vaccination. Dog biting episodes in humans were of 75.00 and 62.50% in Moyo and Ntoroko districts respectively. Twenty-seven (27) samples tested positive for rabies by FAT and PCR. Ugandan sequences were closely related (97% nucleotide id) to the rabies virus sequences from Tanzania, Rwanda, Burundi, Nigeria, Central African Republic and Sudan with both the "Africa 1A" and "Africa 1B" RABV clades represented. A putative new clade 1D was also detected. CONCLUSIONS: Rabies remains a public health hazard in Uganda. There is urgent need to establish advocacy programs in both schools and communities to curtail the spread of rabies. Increasing the knowledge regarding wound washing, post-exposure prophylaxis and dogs vaccination would enhance prevention of rabies. A strong collaboration between medical and veterinary sectors under a one health platform is required to ensure sufficient preventative services to the communities.

Author Correction: 420,000 year assessment of fault leakage rates shows geological carbon storage is secure.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Molecular detection and phylogenetic analysis of lumpy skin disease virus from outbreaks in Uganda 2017-2018.

BACKGROUND: Lumpy skin disease (LSD) is an infectious viral disease of cattle caused by a Capripoxvirus. LSD has substantial economic implications, with infection resulting in permanent damage to the skin of affected animals which lowers their commercial value. In Uganda, LSD is endemic and cases of the disease are frequently reported to government authorities. This study was undertaken to molecularly characterize lumpy skin disease virus (LSDV) strains that have been circulating in Uganda between 2017 and 2018. Secondly, the study aimed to determine the phylogenetic relatedness of Ugandan LSDV sequences with published sequences, available in GenBank. RESULTS: A total of 7 blood samples and 16 skin nodule biopsies were screened for LSDV using PCR to confirm presence of LSDV nucleic acids. PCR positive samples were then characterised by amplifying the GPCR gene. These amplified genes were sequenced and phylogenetic trees were constructed. Out of the 23 samples analysed, 15 were positive for LSDV by PCR (65.2%). The LSDV GPCR sequences analysed contained the unique signatures of LSDV (A11, T12, T34, S99, and P199) which further confirmed their identity. Sequence comparison with vaccine strains revealed a 12 bp deletion unique to Ugandan outbreak strains. Phylogenetic analysis indicated that the LSDV sequences from this study clustered closely with sequences from neighboring East African countries and with LSDV strains from recent outbreaks in Europe. It was noted that the sequence diversity amongst LSDV strains from Africa was higher than diversity from Eurasia. CONCLUSION: The LSDV strains circulating in Uganda were closely related with sequences from neighboring African countries and from Eurasia. Comparison of the GPCR gene showed that outbreak strains differed from vaccine strains. This information is necessary to understand LSDV molecular epidemiology and to contribute knowledge towards the development of control strategies by the Government of Uganda.

New evidence for an early settlement of the Yucatán Peninsula, Mexico: The Chan Hol 3 woman and her meaning for the Peopling of the Americas.

Human presence on the Yucatán Peninsula reaches back to the Late Pleistocene. Osteological evidence comes from submerged caves and sinkholes (cenotes) near Tulum in the Mexican state of Quintana Roo. Here we report on a new skeleton discovered by us in the Chan Hol underwater cave, dating to a minimum age of 9.9±0.1 ky BP based on 230Th/U-dating of flowstone overlying and encrusting human phalanges. This is the third Paleoindian human skeleton with mesocephalic cranial characteristics documented by us in the cave, of which a male individual named Chan Hol 2 described recently is one of the oldest human skeletons found on the American continent. The new discovery emphasizes the importance of the Chan Hol cave and other systems in the Tulum area for understanding the early peopling of the Americas. The new individual, here named Chan Hol 3, is a woman of about 30 years of age with three cranial traumas. There is also evidence for a possible trepanomal bacterial disease that caused severe alteration of the posterior parietal and occipital bones of the cranium. This is the first time that the presence of such disease is reported in a Paleoindian skeleton in the Americas. All ten early skeletons found so far in the submerged caves from the Yucatán Peninsula have mesocephalic cranial morphology, different to the dolicocephalic morphology for Paleoindians from Central Mexico with equivalent dates. This supports the presence of two morphologically different Paleoindian populations for Mexico, coexisting in different geographical areas during the Late Pleistocene-Early Holocene.

Constraining the formation of authigenic carbonates in a seepage-affected cold-water coral mound by lipid biomarkers.

Cold-water coral (CWC) mounds are build-ups comprised of coral-dominated intervals alternating with a mixed carbonate-siliciclastic matrix. At some locations, CWC mounds are influenced by methane seepage, but the impact of methane on CWC mounds is poorly understood. To constrain the potential impact of methane on CWC mound growth, lipid biomarker investigations were combined with mineralogical and petrographic analyses to investigate the anaerobic oxidation of methane (AOM) and authigenic carbonate formation in sediment from a seep-affected CWC mound in the Gulf of Cadiz. The occurrence of AOM was confirmed by characteristic lipids found within a semi-lithified zone (SLZ) consisting of authigenic aragonite, high-magnesium calcite and calcium-excess dolomite. The formation of high-Mg calcite is attributed to AOM, acting as a lithifying agent. Aragonite is only a minor phase. Ca-excess dolomite in the SLZ and upper parts may be formed by organoclastic sulphate reduction, favouring precipitation by increased alkalinity. The AOM biomarkers in the SLZ include isoprenoid-based archaeal membrane lipids, such as abundant glycerol dibiphytanyl glycerol tetraethers (GDGTs) dominated by GDGT-2. The δ13 C values of GDGT-2, measured as ether-cleaved monocyclic biphytanes, are as low as -100‰ versus V-PDB. Further, bacterial dialkyl glycerol diethers with two anteiso-C15 alkyl chains and δ13 C values of -81‰ are interpreted as biomarkers of sulphate-reducing bacteria. The lipid biomarker signatures and mineralogical patterns suggest that anaerobic methane-oxidizing archaea of the ANME-1 group thrived in the subsurface at times of slow and diffusive methane seepage. Petrographic analyses revealed that the SLZ was exhumed at some point (e.g. signs of bioerosion of the semi-lithified sediment), providing a hard substrate for CWC larval settlement. In addition, this work reveals that AOM-induced semi-lithification likely played a role in mound stabilization. Lipid biomarker analysis proves to be a powerful tool to disentangle early diagenetic processes induced by microbial metabolisms.