The intestinal microbiota of a Risso’s dolphin (Grampus griseus): possible relationships with starvation raised by macro-plastic ingestion / La microbiota intestinal del delfín gris (Grampus griseus): posibles relaciones con la inanición provocada por la ingestión de macroplásticos

Ingesting marine plastics is increasingly common in cetaceans, but little is known about their potential effects. Here, by utilizing 16S rRNA gene sequencing, we profiled the intestinal bacterial communities of a stranded Risso’s dolphin (Grampus griseus) which died because of the ingestion of rubber gloves. In this study, we explored the potential relationships between starvation raised by plastic ingestion with the dolphin gut microbiota. Our results showed significant differences in bacterial diversity and composition among the different anatomical areas along the intestinal tract, which may be related to the intestinal emptying process under starvation. In addition, the intestinal bacterial composition of the Risso’s dolphin showed both similarity and divergence to that of other toothed whales, suggesting potential roles of both host phylogeny and habitat shaping of the cetacean intestinal microbiome. Perhaps, the microbiota is reflecting a potentially disordered intestinal microbial profile caused by the ingestion of macro-plastics which led to starvation. Moreover, two operational taxonomic units (0.17% of the total reads) affiliated with Actinobacillus and Acinetobacter lwoffii were detected along the intestinal tract. These bacterial species may cause infections in immunocompromised dolphins which are malnourished. This preliminary study profiles the intestinal microbiota of a Risso’s dolphin, and provides an additional understanding of the potential relationships between starvation raised by ingesting macro-plastics with cetacean gut microbiota.(AU)

The intestinal microbiota of a Risso's dolphin (Grampus griseus): possible relationships with starvation raised by macro-plastic ingestion.

Ingesting marine plastics is increasingly common in cetaceans, but little is known about their potential effects. Here, by utilizing 16S rRNA gene sequencing, we profiled the intestinal bacterial communities of a stranded Risso's dolphin (Grampus griseus) which died because of the ingestion of rubber gloves. In this study, we explored the potential relationships between starvation raised by plastic ingestion with the dolphin gut microbiota. Our results showed significant differences in bacterial diversity and composition among the different anatomical areas along the intestinal tract, which may be related to the intestinal emptying process under starvation. In addition, the intestinal bacterial composition of the Risso's dolphin showed both similarity and divergence to that of other toothed whales, suggesting potential roles of both host phylogeny and habitat shaping of the cetacean intestinal microbiome. Perhaps, the microbiota is reflecting a potentially disordered intestinal microbial profile caused by the ingestion of macro-plastics which led to starvation. Moreover, two operational taxonomic units (0.17% of the total reads) affiliated with Actinobacillus and Acinetobacter lwoffii were detected along the intestinal tract. These bacterial species may cause infections in immunocompromised dolphins which are malnourished. This preliminary study profiles the intestinal microbiota of a Risso's dolphin, and provides an additional understanding of the potential relationships between starvation raised by ingesting macro-plastics with cetacean gut microbiota.

Robinsoniella peoriensis: an emerging pathogen with few virulence factors / Robinsoniella peoriensis: un patógeno emergente con pocos factores de virulencia

Robinsoniella peoriensis is a Gram-positive bacterium which is anaerobic, spore-forming, and non-motile. It was initially isolated and characterized from feces and swine manure. Strains of this species have since been identified from different mammalian and non-mammalian gastrointestinal tracts. Strains have also been isolated from a variety of human infections, such as bacteremia, bone infections, and skin structures. R. peoriensis has recently been reported as causative for pyometra, which could result in death in the absence of sufficient antimicrobial treatment. However, to the author’s knowledge, there has not been a single virulence factor identified. A major challenge of modern medicine is the failure of conventional procedures to characterize the capability of an emerging pathogen to cause disease. The goal of this study is to initially characterize the pathogenicity of this bacterium using a pathogenomics approach.(AU)

Robinsoniella peoriensis: an emerging pathogen with few virulence factors.

Robinsoniella peoriensis is a Gram-positive bacterium which is anaerobic, spore-forming, and non-motile. It was initially isolated and characterized from feces and swine manure. Strains of this species have since been identified from different mammalian and non-mammalian gastrointestinal tracts. Strains have also been isolated from a variety of human infections, such as bacteremia, bone infections, and skin structures. R. peoriensis has recently been reported as causative for pyometra, which could result in death in the absence of sufficient antimicrobial treatment. However, to the author's knowledge, there has not been a single virulence factor identified. A major challenge of modern medicine is the failure of conventional procedures to characterize the capability of an emerging pathogen to cause disease. The goal of this study is to initially characterize the pathogenicity of this bacterium using a pathogenomics approach.

Draft genome sequence data of Clostridium perfringens FA isolated from the faecal material of the critically endangered African wild dog, Lycaon pictus.

In this present article the draft sequence data for Clostridium perfringens FA, which was isolated from the faecal material of a critically endangered African Wild dog, is reported. The bacterium is widely distributed in the environment and in the normal intestinal flora of humans and animals. The genome of strain C. perfringens FA was assembled into 21 contigs with a total length of 3, 044, 349 bp and a GC content of 28.20%. There are 2742 CDS, 70 tRNAs and 5 rRNAs. Five putative virulence genes were detected. There were no plasmid replicons found. The genome of few environmental isolates has been sequenced. The draft genome of strain FA can be compared to disease causing isolates cultured from humans to aid in a better understanding of the pathogenesis of the bacterium.

Biogeographic patterns of potential pathogenic bacteria in the middle and lower reaches of the Yangtze River as well as its two adjoining lakes, China.

Understanding the distribution patterns and shaping factors of bacterial pathogens in aquatic ecosystems, especially in natural waters, are critical to the control of pathogen transmission. In this study, using 16S rRNA gene amplicon sequencing, we explored the composition and biogeographic dynamics of potential bacterial pathogens in the middle and lower reaches of the Yangtze River, as well as its two vast adjoining lakes (Dongting Lake and Poyang Lake). The pathogen community belonged to 12 potential pathogenic groups, with "intracellular parasites," "animal parasites or symbionts" and "human pathogens all" occupying 97.5% in total. The potential pathogen community covered seven phyla with Proteobacteria (69.8%) and Bacteroidetes (13.5%) the most predominant. In addition, 53 genera were identified with Legionella (15.2%) and Roseomonas (14.2%) the most dominant. The average relative abundance, alpha diversity and microbial composition of the potential bacterial pathogens exhibited significant biogeographical variations among the different sections. An in-depth analysis reflected that environmental variables significantly structured the potential bacterial pathogens, including water physiochemical properties (i.e., chlorophyll-a, total nitrogen and transparency), heavy metals (i.e., As and Ni), climate (i.e., air temperature) and land use type (i.e., waters). Compared to the overall bacterial community which was composed of both pathogenic and non-pathogenic bacteria, the pathogen community exhibited distinct microbial diversity patterns and shaping factors. This signifies the importance of different variables for shaping the pathogen community. This study represents one attempt to explore pathogen diversity patterns and their underlying drivers in the Yangtze River, which provides a foundation for the management of pathogenic bacteria.

Biofilm formation, antimicrobial assay, and toxin-genotypes of Clostridium perfringens type C isolates cultured from a neonatal Yangtze finless porpoise.

This is a culture-dependent study with the objective of pure culturing and characterizing pathogenic bacteria from the blowhole, lung, stomach and fecal samples of a neonatal crucially endangered Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis) that died 27 days after birth. Bacteria were inoculated using a swab onto blood and MacConkey agar plates and representative isolates were identified through 16S rRNA gene sequence analysis. A total of three Clostridium perfringens type C strains from the fecal samples were isolated. Toxin genes, including cpa, cpb and cpb2, were detected by PCR amplification, whereas the etx, iap and cpe genes were not detected. Biofilm formation of the three strains was then examined. Only one strain was capable of biofilm formation. In addition, isolates showed strong resistance against the antibiotics amikacin (3/3), erythromycin (1/3), gentamicin (3/3), streptomycin (3/3), and trimethoprim (3/3), while sensitivity to ampicillin (3/3), bacitracin (3/3), erythromycin (2/3), penicillin G (3/3), and tetracycline (3/3). The results suggested C. perfringens type C could have contributed to the death of this neonatal porpoise.

Analysis of the microbial diversity in the fecal material of the critically endangered African wild dog, Lycaon pictus.

The African wild dog (AWD) (Lycaon pictus) is a critically endangered species. These animals are hypercarnivores, hunting mostly medium-sized antelope. In this study, using bacterial tag-encoded FLX-Titanium amplicon pyrosequencing (bTEFAP®), the microbiota in the fecal material of AWDs living in the Great Plains Zoo & Delbridge Museum of Natural History was investigated. In both samples, the most predominant bacterial phylum was the Firmicutes with members of the genus Blautia spp. being the most dominant bacteria.

Characterization of the Biodiesel Degrading Acinetobacter oleivorans Strain PT8 Isolated from the Fecal Material of a Painted Turtle (Chrysemys picta).

Acinetobacter species are gram-negative, non-fermenting bacteria with coccobacilli morphology. The bacteria are found ubiquitously and have the ability to occupy niches which include environmental sites, animals, and humans. The original purpose of this study was to determine if painted turtles (Chrysemys picta) living in the wild in Western Wisconsin were colonized with carbapenem-resistant bacteria. Fecal samples from ten turtles were examined for carbapenem-resistant bacteria. None of the isolates were found to be carbapenem resistant by antimicrobial susceptibility testing. However, all the isolates were resistant to other ß-lactams and chloramphenicol classes of antimicrobials. One isolate, Acinetobacter oleivorans strain PT8, was selected for additional characterization, including whole-genome sequencing (WGS). Strain PT8 is capable of degrading biodiesel, forming biofilms, and has a putative type 6 gene cluster. Finally, the taxonomic position of the available whole-genome sequences of 25 A. oleivorans genomes from purified isolates was determined.

Searching for putative virulence factors in the genomes of Shewanella indica and Shewanella algae.

Bacterial pathogens are a major threat to both humans and animals worldwide. It is crucial to understand the mechanisms of various disease processes at the molecular level. Shewanella species are widespread in the environment and some are considered as emerging opportunistic human and marine mammal pathogens. In this study, putative virulence factors on the genome of Shewanella indica BW, a bacterium isolated from the Bryde's whale (Balaenoptera edeni), were determined. Additionally, for comparative purposes, putative virulence factors from two other S. indica and ten S. algae strains were also determined using the Pathosystems Resource Integration Center (PATRIC) pipeline. We confirmed the presence of previously reported virulence factors and we are proposing several new candidate virulence factors. Interestingly, the putative virulence factors were very similar between the two species with the exception of microbial collagenase which was present in all S. algae genomes, but absent in all S. indica genomes.

The intestinal microbiome of an Indo-Pacific humpback dolphin (Sousa chinensis) stranded near the Pearl River Estuary, China.

The mammalian intestinal microbiome is critical for host health and disease resistance. However, the cetacean intestinal microbiota remains relatively unexplored. By using high-throughput 16S rRNA gene sequencing, we analyzed intestinal bacterial samples from an Indo-pacific humpback dolphin (Sousa chinensis) stranded near the Pearl River Estuary in China. The samples included 3 anatomical regions (foregut, midgut, and rectum) and 2 anatomical locations (content and mucus). Our analyses revealed that the dolphin intestinal bacteria contained 139 operational taxonomic units (OTUs), dominated at the phyla level by Firmicutes (47.05% in the content; 94.77% in the mucus), followed by Bacteroidetes (23.63% in the content; 1.58% in the mucus) and Gammaproteobacteria (14.82% in the content; 2.05% in the mucus). The intestinal bacteria had a small core community (15 OTUs, accounting for 99.74% of the reads), some of which could be potentially pathogenic to both human and dolphins. As an alternative to sampling the dolphin intestinal bacteria, fecal sampling could be used. Additionally, function potentials such as, xenobiotics biodegradation, beta-lactam resistance, and human disease-related pathways, were detected in the dolphin intestinal bacteria. These findings provide the first baseline knowledge of the intestinal microbiome of the Indo-Pacific humpback dolphin, which may offer new insights into cetacean conservation by using microbial surveillance.

Isolation and characterisation of carbapenem-resistant Xanthomonas citri pv. mangiferaeindicae-like strain gir from the faecal material of giraffes.

The purpose of this study was to determine if giraffes (Giraffa camelopardalis) living in captivity at the Jacksonville Zoo and Gardens, Jacksonville, FL were colonised with carbapenem-resistant bacteria and, if found, to identify underlying genetic mechanisms contributing to a carbapenem-resistant phenotype. Faecal samples from seven giraffes were examined for carbapenem-resistant bacteria. Only one isolate (a Xanthomondaceae) was found to be carbapenem-resistant by antibiotic susceptibility testing. This isolate was selected for additional characterization, including whole genome sequencing (WGS). Based on average nucleotide identity, the bacterium was identified as Xanthomonas citri pv. mangiferaeindicae-like strain gir. Phenotypic carbapenemase tests and PCR for the most common carbapenemase genes produced negative results, suggesting that carbapenem resistance was mediated by another mechanism. Resistance gene profile analysis of WGS results confirmed these results. Among identified resistance genes, a chromosomal class A beta-lactamase with 71% identity to the penP beta-lactamase gene from Xanthomonas citri ssp. citri was identified, which could contribute to a carbapenem-resistant phenotype.

Analysis of the Microbiota in the Fecal Material of Painted Turtles (Chrysemys picta).

We used high-throughput sequencing analysis, which targeted the hypervariable V3-V4 region of the bacterial 16S rRNA gene, to investigate the microbiota in fecal material from ten wild painted turtles (Chrysemys picta) captured in southeastern Wisconsin. The most predominant bacterial phylum detected in all samples was the Firmicutes (relative abundance for all samples 96.4% to 68.3%). The next most predominant phylum was Bacteroidetes (relative abundance for all samples 23.9% to 7.8%) in eight samples. Fusobacteria (relative abundance for all samples 22.2% to 0%) was the second most predominant in the other two samples.

Putative virulence factors of Plesiomonas shigelloides.

Plesiomonas shigelloides is a Gram-negative rod-shaped bacterium which has been isolated from humans, animals and the environment. It has been associated with diarrhoeal disease in humans and various epizootic diseases in animals. In this study P. shigelloides strains were isolated from the faecal material of a captive Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis; YFP) living in semi-natural conditions in China. Plesiomonas shigelloides strain EE2 was subjected to whole genome sequencing. The draft genome was then compared to the genome sequences of ten other P. shigelloides isolates using the Pathosystems Resource Integration Center pipeline. In addition to several virulence factors which have been previously reported, we are proposing new candidate virulence factors such as a repeats-in-toxin protein, lysophospholipase, a twin-arginine translocation system and the type VI secretion effector Phospholipase A1.

The Possible Effects of High Vessel Traffic on the Physiological Parameters of the Critically Endangered Yangtze Finless Porpoise (Neophocaena asiaeorientalis ssp. asiaeorientalis).

Background: Poyang is the largest freshwater lake in China, where the acoustic environment and space for the critically endangered Yangtze finless porpoises (YFPs) has been altered by heavy vessel traffic and dredging activities. The density of vessel and the rate of dredging increases annually, especially in the area with the highest density of YFPs. The heavy vessel traffic can cause an increase in the physical activities and direct physical injuries to the YFPs. Furthermore, noise is a potent stressor to all cetaceans irrespective of age and can compromise all their physiological functions. The objective of this study was to examine the possible effects of heavy vessel traffic and dredging on the biochemistry, hematology, adrenal, thyroid, and reproductive hormones of two different YFP populations. One population was living in Poyang Lake and the second living in the Tian-E-Zhou Oxbow which is a semi-natural resserve. Results: The results showed statistically significantly higher levels of serum cortisol, fT3, fT4, and lowered testosterone in both adult and juvenile YFPs living in Poyang Lake vs. adult YFPs living in the Tian-E-Zhou Oxbow. The serum biochemical parameters (Aspartate Amino Transferase, Alkaline Phosphatase, High Density Lipoprotein cholesterol ratio, Globulin, Uric acid, Glucose, K+, and Amylase) and the hematology parameters (Red Blood Cells, Hematocrit, Mean Corpuscular Volume, White Blood Cells, and Eosinophils) were statistically significantly higher in the adult Poyang Lake YFPs vs. adult Tian-E-Zhou Oxbow YFPs. On the other hand, adult males of the Tian-E-Zhou Oxbow also showed significantly higher levels of the serum biochemical parameters (Total Cholesterol, Light Density Lipoprotein cholesterol, Direct Bilirubin, Albumin, Lactate Dehydrogenase, CO2, and Na+) and the blood parameters (Mean Corpuscular Hemoglobin and Mean Corpuscular Hemoglobin Concentration). In Poyang Lake YFPs, various parameters showed significantly positive (fT4, amylase, neutrophil, Ca+2) or negative (total protein, lymphocyte) correlations with cortisol levels. Conclusions: The hyperactivity of adrenal glands in response to heavy vessel traffic and dredging resulted in significantly elevated cortisol levels in Poyang Lake YFPs. The higher cortisol level could possibly have affected various hormonal, hematological, and biochemical parameters, and ultimately the YFPs physiology.

Microbial communities in different regions of the gastrointestinal tract in East Asian finless porpoises (Neophocaena asiaeorientalis sunameri).

Mammalian gastrointestinal (GI) tract microbial communities are critical for host health. However, the microbiota along the GI tract in cetaceans has not been well characterized compared to other animals. In this study, the bacteria and fungi present in the stomach, foregut, hindgut and feces, of East Asian finless porpoises (Neophocaena asiaeorientalis sunameri, EAFPs) were characterized using high-throughput sequencing analysis. The bacterial and fungal diversity and richness in the stomach, hindgut and fecal samples tended to be higher than those in the foregut. Bacterial taxonomic compositions found in the hindgut and feces were different from those seen in the stomach and foregut. A greater proportion of strict anaerobic bacteria including Clostridia, Fusobacteria, and Ruminococcaceae were found in the hindgut and fecal samples. The fungal communities present in stomach samples differed from those detected in other regions to some extent. Zygomycota and Neocallimastigomycota were more predominant in the stomach. Some potential pathogens, such as Helicobacter spp. and Vibrio spp., were commonly present along the GI tract. Our study confirms that the fecal microbiota can represent the whole GI tract to some extent because of their relatively higher microbial diversity and presence of potential pathogens. Our study provides the first comprehensive characterization of the EAFPs GI microbiota, expanding on the current knowledge about the bacterial diversity in the GI tract of cetaceans. In addition, this is the first study characterizing the fungal diversity of any species of porpoise.

The possible effects of anthropogenic acoustic pollution on marine mammals' reproduction: an emerging threat to animal extinction.

For about 119 species of cetaceans and other aquatic animals, sound is the key source of learning about the environment, navigation, communication, foraging, and avoiding predators. However, in the recent era, the introduction of large quantities of anthropogenic noise into the ocean has significantly altered the ocean's acoustic environment. The anthropogenic noises travel very long distances, blanketing enormous areas. This can affect cetaceans, either by direct killing or compromising hearing, navigation, communication, predation, as well as normal behaviors. It has been suggested that acoustic pollution could possibly negatively affect cetacean reproduction, which is harmful for endangered and threatened species. However, it is still unknown how acoustic pollution can suppress cetacean reproduction. This is the first comprehensive review article, which focuses on the possible consequences affecting the reproduction of marine mammals resulting from acoustic pollution.

Analysis of the Microbial Diversity in the Fecal Material of Giraffes.

Using bacterial and fungal tag-encoded FLX-Titanium amplicon pyrosequencing, the microbiota of the fecal material of seven giraffes living in captivity at the Jacksonville Zoo and Gardens, Jacksonville, FL was investigated. In all samples, the most predominant bacterial phylum was the Firmicutes followed by Bacteroidetes. The most predominant fungi were members of the phylum Ascomycota followed by Neocallimastigomycota in five of seven samples. The reverse was true in the other two samples.

Isolation of culturable aerobic bacteria and evidence of Kerstersia gyiorum from the blowhole of captive Yangtze finless porpoises.

Bacterial respiratory illnesses are problematic in aquatic mammals such as the Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis; YFP), which is now at a critically endangered status. Yet little is known about the bacteria inhabiting the respiratory tract of YFPs. In this study, we preliminarily characterized the culturable aerobic bacteria from blow samples of captive YFPs. The bacterial diversity was assessed through cultivation by direct exhalation onto Columbia blood agar plates and identification of representative isolates through 16S rRNA gene sequence analysis. In total, eleven bacterial species belonging to four phyla Proteobacteria (71 %), Firmicutes (25 %), Bacteroidetes (3 %) and Actinobacteria (1 %) were identified. Most of these isolates were opportunistic pathogens found in respiratory illnesses in humans and animals. We also reported the first case of Kerstersia gyiorum isolated from an animal. This work provides a preliminary assessment of the bacteria present in the respiratory tract of captive YFPs, which will be an important first step in elucidating the roles of normal microbiota in maintaining respiratory health of YFPs. This study also points out the necessity of future long-term monitoring of blowhole microorganisms in the YFPs and making emergency preparedness plans for respiratory tract infections. These measures can aid in assessing the pathogenic risk of the critically endangered YFP populations.

Analysis of the microbial diversity in faecal material of the endangered blue whale, Balaenoptera musculus.

Using bacterial and fungal tag-encoded FLX-Titanium amplicon pyrosequencing, the microbiota of the faecal material of two blue whales living in the wild off the coast of California was investigated. In both samples the most predominant bacterial phylum was the Firmicutes with Clostridium spp. being the most dominant bacteria. The most predominant fungi were members of the phylum Ascomycota with Metschnikowia spp. being the most dominant. In this study, we also preliminarily characterised the culturable anaerobic bacteria from the faecal material, using traditional culture and 16S rRNA gene sequencing approaches. In total, three bacterial species belonging to the phylum Firmicutes were identified.