Highly abundant core taxa in the blow within and across captive bottlenose dolphins provide evidence for a temporally stable airway microbiota.

BACKGROUND: The analysis of blow microbiota has been proposed as a biomarker for respiratory health analysis in cetaceans. Yet, we lack crucial knowledge on the long-term stability of the blow microbiota and its potential changes during disease. Research in humans and mice have provided evidence that respiratory disease is accompanied by a shift in microbial communities of the airways. We investigate here the stability of the community composition of the blow microbiota for 13 captive bottlenose dolphins over eight months including both sick and healthy individuals. We used barcoded tag sequencing of the bacterial 16S rRNA gene. Four of the dolphins experienced distinct medical conditions and received systemic antimicrobial treatment during the study. RESULTS: We showed that each dolphin harboured a unique community of zero-radius operational taxonomic units (zOTUs) that was present throughout the entire sampling period ('intra-core'). Although for most dolphins there was significant variation over time, overall the intra-core accounted for an average of 73% of relative abundance of the blow microbiota. In addition, the dolphins shared between 8 and 66 zOTUs on any of the sampling occasions ('inter-core'), accounting for a relative abundance between 17 and 41% of any dolphin's airway microbiota. The majority of the intra-core and all of the inter-core zOTUs in this study are commonly found in captive and free-ranging dolphins and have previously been reported from several different body sites. While we did not find a clear effect of microbial treatment on blow microbiota, age and sex of the dolphins did have such an effect. CONCLUSIONS: The airways of dolphins were colonized by an individual intra-core 'signature' that varied in abundance relative to more temporary bacteria. We speculate that the intra-core bacteria interact with the immune response of the respiratory tract and support its function. This study provides the first evidence of individual-specific airway microbiota in cetaceans that is stable over eight months.

Digestive physiology of captive capybara (Hydrochoerus hydrochaeris).

The capybara (Hydrochoerus hydrochaeris), the largest living rodent, probably has a "mucus-trap" colonic separation mechanism. To test this hypothesis, we measured the mean retention time of a solute marker (MRTSolute ), 2 mm (MRT2 mm ), 10 mm (MRT10 mm ), and 20 mm (MRT20 mm ) particle markers and nutrient digestibility in adult captive capybaras (27-52 kg body mass (BM), 2-11 yr). In addition, total gut fill and the selectivity factor (MRTSolute /MRT2 mm ) were calculated, and mean faecal particle size and metabolic fecal nitrogen of captive capybaras were compared to those of free-ranging specimens. Finally, we also measured methane production in one animal. The MRT2 mm (29.2 ± 8.2 hr) was different (p < 0.01) from MRTSolute (37.0 ± 13.1 hr), MRT10 mm (36.5 ± 8.2 hr), and MRT20 mm (35.1 ± 9.6 hr). The selectivity factor (1.26 ± 0.30) was in the range considered typical for a "mucus-trap" colonic separation mechanism. The estimated total gut fill was 1.50 ± 0.37% and 1.73 ± 0.25% of BM calculated from the results of the 2-mm and 10-mm particle markers, respectively. The CH4 emission was 13.7 L/day. Captive capybaras had greater mean fecal particle size (0.44 ± 0.06 vs. 0.29 ± 0.05 mm, p < 0.001) and metabolic fecal nitrogen (65.5 ± 3.91 vs. 46.8 ± 10.5% of fecal nitrogen, p < 0.001) than free-ranging capybaras. Organic matter digestibility decreased less steeply with increasing dietary crude fiber content in capybaras as compared to published data from rabbits or guinea pigs. Accordingly, the digestive physiology of the capybara is characterized by a comparatively high fiber digestibility, with a "mucus-trap" colonic separation mechanism, allowing capybaras to thrive on forage-only diets.

Methane production by two non-ruminant foregut-fermenting herbivores: The collared peccary (Pecari tajacu) and the pygmy hippopotamus (Hexaprotodon liberiensis).

Methane (CH4) production varies between herbivore species, but reasons for this variation remain to be elucidated. Here, we report open-circuit chamber respiration measurements of CH4 production in four specimens each of two non-ruminant mammalian herbivores with a complex forestomach but largely differing in body size, the collared peccary (Pecari tajacu, mean body mass 17kg) and the pygmy hippopotamus (Hexaprotodon liberiensis, 229kg) fed lucerne-based diets. In addition, food intake, digestibility and mean retention times were measured in the same experiments. CH4 production averaged 8 and 72L/d, 18 and 19L/kg dry matter intake, and 4.0 and 4.2% of gross energy intake for the two species, respectively. When compared with previously reported data on CH4 production in other non-ruminant and ruminant foregut-fermenting as well as hindgut-fermenting species, it is evident that neither the question whether a species is a foregut fermenter or not, or whether it ruminates or not, is of the relevance previously suggested to explain variation in CH4 production between species. Rather, differences in CH4 production between species on similar diets appear related to species-specific differences in food intake and digesta retention kinetics.

Decreasing methane yield with increasing food intake keeps daily methane emissions constant in two foregut fermenting marsupials, the western grey kangaroo and red kangaroo.

Fundamental differences in methane (CH4) production between macropods (kangaroos) and ruminants have been suggested and linked to differences in the composition of the forestomach microbiome. Using six western grey kangaroos (Macropus fuliginosus) and four red kangaroos (Macropus rufus), we measured daily absolute CH4 production in vivo as well as CH4 yield (CH4 per unit of intake of dry matter, gross energy or digestible fibre) by open-circuit respirometry. Two food intake levels were tested using a chopped lucerne hay (alfalfa) diet. Body mass-specific absolute CH4 production resembled values previously reported in wallabies and non-ruminant herbivores such as horses, and did not differ with food intake level, although there was no concomitant proportionate decrease in fibre digestibility with higher food intake. In contrast, CH4 yield decreased with increasing intake, and was intermediate between values reported for ruminants and non-ruminant herbivores. These results correspond to those in ruminants and other non-ruminant species where increased intake (and hence a shorter digesta retention in the gut) leads to a lower CH4 yield. We hypothesize that rather than harbouring a fundamentally different microbiome in their foregut, the microbiome of macropods is in a particular metabolic state more tuned towards growth (i.e. biomass production) rather than CH4 production. This is due to the short digesta retention time in macropods and the known distinct 'digesta washing' in the gut of macropods, where fluids move faster than particles and hence most likely wash out microbes from the forestomach. Although our data suggest that kangaroos only produce about 27% of the body mass-specific volume of CH4 of ruminants, it remains to be modelled with species-specific growth rates and production conditions whether or not significantly lower CH4 amounts are emitted per kg of meat in kangaroo than in beef or mutton production.

Digestive physiology of the plains viscacha (Lagostomus maximus): A large herbivorous hystricomorph rodent.

Plains viscachas (Lagostomus maximus) are large South American, fossorial rodents susceptible to diabetic cataracts. Various aspects of their digestive physiology were studied in three different experiments with nine male and seven female adult animals and six different diets (total n of feeding trials = 35). Viscachas achieved mean retention times of 23­31 hr, which is of a magnitude also recorded in horses; these did not differ for solute or small particle (<2 mm) markers. Secondary marker excretion peaks indicated coprophagy, and were rarer on high-protein as compared to grass hay-only diets. Mean resting metabolic rate was, at 229 kJ/kg0.75/day, lower than expected for a mammal of this size. Digestible energy requirement for maintenance was 445 kJ/kg0.75/day. At 1.6­2.7 L/day, viscachas produced more methane than expected for a hindgut fermenter of their size. On diets that included concentrate feeds, viscachas excreted glucose in their urine, corroborating reports on the susceptibility of this species for diabetes when kept on energy-dense food. Viscachas had a similar apparent digestibility of protein, lipids, and macrominerals as other rodents, rabbits, or domestic horses. This suggests that whether or not a species practices coprophagy does not have a major influence on these measures. Viscachas resemble other hindgut fermenters in their high apparent calcium digestibility. With respect to a digestibility-reducing effect of dietary fiber, viscachas differed from rabbits and guinea pigs but were similar to horses, suggesting that small body size needs not necessarily be linked to lower digestive efficiency on high-fiber diets.

A research synthesis of humans, animals, and environmental compartments exposed to PFAS: A systematic evidence map and bibliometric analysis of secondary literature.

BACKGROUND: Per- and polyfluoroalkyl substances (PFAS) are a class of widely used anthropogenic chemicals. Concerns regarding their persistence and potential adverse effects have led to multiple secondary research publications. Here, we aim to assess the resulting evidence base in the systematic secondary literature by examining research gaps, evaluating the quality of reviews, and exploring interdisciplinary connections. METHODS: This study employed a systematic evidence-mapping approach to assess the secondary literature on the biological, environmental, and medical aspects of exposure to 35 fluorinated compounds. The inclusion criteria encompassed systematic reviews published in peer-reviewed journals, pre-prints, and theses. Comprehensive searches across electronic databases and grey literature identified relevant reviews. Data extraction and synthesis involved mapping literature content and narrative descriptions. We employed a modified version of the AMSTAR2 checklist to evaluate the methodological rigour of the reviews. A bibliometric data analysis uncovered patterns and trends in the academic literature. A research protocol for this study was previously pre-registered (osf.io/2tpn8) and published (Vendl et al., Environment International 158 (2022) 106973). The database is freely accessible through the interactive and user-friendly web application of this systematic evidence map at https://hi-this-is-lorenzo.shinyapps.io/PFAS_SEM_Shiny_App/. RESULTS: Our map includes a total of 175 systematic reviews. Over the years, there has been a steady increase in the annual number of publications, with a notable surge in 2021. Most reviews focused on human exposure, whereas environmental and animal-related reviews were fewer and often lacked a rigorous systematic approach to literature search and screening. Review outcomes were predominantly associated with human health, particularly with reproductive and children's developmental health. Animal reviews primarily focused on studies conducted in controlled laboratory settings, and wildlife reviews were characterised by an over-representation of birds and fish species. Recent reviews increasingly incorporated quantitative synthesis methodologies. The methodological strengths of the reviews included detailed descriptions of study selection processes and disclosure of potential conflicts of interest. However, weaknesses were observed in the critical lack of detail in reporting methods. A bibliometric analysis revealed that the most productive authors collaborate within their own country, leading to limited and clustered international collaborations. CONCLUSIONS: In this overview of the available systematic secondary literature, we map literature content, assess reviews' methodological quality, highlight data gaps, and draw research network clusters. We aim to facilitate literature reviews, guide future research initiatives, and enhance opportunities for cross-country collaboration. Furthermore, we discuss how this systematic evidence map and its publicly available database benefit scientists, regulatory agencies, and other stakeholders by providing access to current systematic secondary literature on PFAS exposure.

Thermal processing reduces PFAS concentrations in blue food - A systematic review and meta-analysis.

Per- and polyfluoroalkyl substances (PFAS) are ubiquitous in the environment and often ingested with food. PFAS exposure in people can have detrimental health consequences. Therefore, reducing PFAS burdens in food items is of great importance to public health. Here, we investigated whether cooking reduces PFAS concentrations in animal-derived food products by synthesizing experimental studies. Further, we examined the moderating effects of the following five variables: cooking time, liquid/animal tissue ratio, cooking temperature, carbon chain length of PFAS and the cooking category (oil-based, water-based & no-liquid cooking). In our systematic review searches, we obtained 512 effect sizes (relative differences in PFAS concentration between raw and cooked samples) from 10 relevant studies. These studies exclusively explored changes in PFAS concentrations in cooked seafood and freshwater fish. Our multilevel-meta-analysis has revealed that, on average, cooking reduced PFAS concentrations by 29%, although heterogeneity among effect sizes was very high (I2 = 94.65%). Our five moderators cumulatively explained 49% of the observed heterogeneity. Specifically, an increase in cooking time and liquid/animal tissue ratio, as well as shorter carbon chain length of PFAS (when cooked with oil) were associated with significant reductions in PFAS concentrations. The effects of different ways of cooking depended on the other moderators, while the effect of cooking temperature itself was not significant. Overall, cooking can reduce PFAS concentrations in blue food (seafood and freshwater fish). However, it is important to note that complete PFAS elimination requires unrealistically long cooking times and large liquid/animal tissue ratios. Currently, literature on the impact of cooking of terrestrial animal produce on PFAS concentrations is lacking, which limits the inference and generalisation of our meta-analysis. However, our work represents the first step towards developing guidelines to reduce PFAS in food via cooking exclusively with common kitchen items and techniques.

Respiratory microbiota of humpback whales may be reduced in diversity and richness the longer they fast.

Humpback whales endure several months of fasting while undertaking one of the longest annual migrations of any mammal, which depletes the whales' energy stores and likely compromises their physiological state. Airway microbiota are linked to respiratory health in mammals. To illuminate the dynamics of airway microbiota in a physiologically challenged mammal, we investigated the bacterial communities in the blow of East Australian humpback whales at two stages of their migration: at the beginning (n = 20) and several months into their migration (n = 20), using barcoded tag sequencing of the bacterial 16S rRNA gene. We show that early in the fasting the whale blow samples had a higher diversity and richness combined with a larger number of core taxa and a different bacterial composition than later in the fasting. This study provides some evidence that the rich blow microbiota at the beginning of their fasting might reflect the whales' uncompromised physiology and that changes in the microbiota occur during the whales' migration.

Does sociality drive diversity and composition of airway microbiota in cetaceans?

The number of social contacts of mammals is positively correlated with the diversity of their gut microbes. There is some evidence that sociality also affects microbes in the respiratory tract. We tested whether the airway microbiota of cetacean species differ depending on the whales' level of sociality. We sampled the blow of blue (Balaenoptera musculus), grey (Eschrichtius robustus), humpback (Megaptera novaeangliae) and long-finned pilot whales (PWs) (Globicephala melas) and analysed the blow microbiota by barcode tag sequencing targeting the V4 region of the bacterial 16S rRNA gene. Humpback whales (HWs) show higher levels of sociality than blue (BW) and grey (GW), while PWs are the most gregarious among the four species. The blow samples of the HWs showed the highest richness and diversity. HWs were also the only species with a species-specific clustering of their microbial community composition and a relatively large number of core taxa. Therefore, we conclude that it cannot be sociality alone shaping the diversity and composition of airway microbiota. We suggest the whale species' lung volume and size of the plume of exhaled air as an additional factor impacting the transmission potential of blow microbiota from one individual whale to another.