Animal Gut Research Articles

Widespread stop-codon recoding in bacteriophages may regulate translation of lytic genes.

Bacteriophages (phages) are obligate parasites that use host bacterial translation machinery to produce viral proteins. However, some phages have alternative genetic codes with reassigned stop codons that are predicted to be incompatible with bacterial translation systems. We analysed 9,422 phage genomes and found that stop-codon recoding has evolved in diverse clades of phages that infect bacteria present in both human and animal gut microbiota. Recoded stop codons are particularly over-represented in phage structural and lysis genes. We propose that recoded stop codons might function to prevent premature production of late-stage proteins. Stop-codon recoding has evolved several times in closely related lineages, which suggests that adaptive recoding can occur over very short evolutionary timescales.

Sustainability of the rice-crayfish co-culture aquaculture model: microbiome profiles based on multi-kingdom analyses

While the rice-crayfish culture (RCFP) model, an important aquaculture model in Asia, is generally considered a sustainable model, its sustainability in terms of microbial community profiles has not been evaluated. In this study, multi-kingdom analyses of microbiome profiles (i.e., bacteria, archaea, viruses, and eukaryotes) were performed using environmental (i.e., water and sediment) and animal gut (i.e., crayfish and crab gut) microbial samples from the RCFP and other aquaculture models, including the crab-crayfish co-culture, crayfish culture, and crab culture models, to evaluate the sustainability of the RCFP systematically. Results showed that RCFP samples are enriched with a distinct set of microbes, including Shewanella, Ferroplasma, Leishmania, and Siphoviridae, when compared with other aquaculture models. Additionally, most microbes in the RCFP samples, especially microbes from different kingdoms, were densely and positively connected, which indicates their robustness against environmental stress. Whereas microbes in different aquaculture models demonstrated moderate levels of horizontal gene transfer (HGT) across kingdoms, the RCFP showed relatively lower frequencies of HGT events, especially those involving antibiotic resistance genes. Finally, environmental factors, including pH, oxidation–reduction potential, temperature, and total nitrogen, contributed profoundly to shaping the microbial communities in these aquaculture models. Interestingly, compared with other models, the microbial communities of the RCFP model were less influenced by these environmental factors, which suggests that microbes in the latter have stronger ability to resist environmental stress. The findings collectively reflect the unique multi-kingdom microbial patterns of the RCFP model and suggest that this model is a sustainable model from the perspective of microbiome profiles.

Characterization and Identification of a New Daidzein Reductase Involved in (S)-Equol Biosynthesis in Clostridium sp. ZJ6.

(S)-equol (EQ) is an isoflavone with high estrogen-like activity in the human body, and is only produced by some gut bacteria in vivo. It plays an important role in maintaining individual health, however, the dearth of resources associated with (S)-EQ-producing bacteria has seriously restricted the production and application of (S)-EQ. We report here a new functional gene KEC48-07020 (K-07020) that was identified from a chick (S)-EQ-producing bacterium (Clostridium sp. ZJ6, ZJ6). We found that recombinant protein of K-07020 possessed similar function to daidzein reductase (DZNR), which can convert daidzein (DZN) into R/S-dihydrodaidzein (R/S-DHD). Interestingly, K-07020 can reversely convert (R/S)-DHD (DHD oxidase) into DZN even without cofactors under aerobic conditions. Additionally, high concentrations of (S)-EQ can directly promote DHD oxidase but inhibit DZNR activity. Molecular docking and site-directed mutagenesis revealed that the amino acid > Arg75 was the active site of DHD oxidase. Subsequently, an engineered E. coli strain based on K-07020 was constructed and showed higher yield of (S)-EQ than the engineered bacteria from our previous work. Metagenomics analysis and PCR detection surprisingly revealed that K-07020 and related bacteria may be prevalent in the gut of humans and animals. Overall, a new DZNR from ZJ6 was found and identified in this study, and its bidirectional enzyme activities and wide distribution in the gut of humans and animals provide alternative strategies for revealing the individual regulatory mechanisms of (S)-EQ-producing bacteria.

Gastrointestinal Microbiota and Their Manipulation for Improved Growth and Performance in Chickens.

The gut of warm-blooded animals is colonized by microbes possibly constituting at least 100 times more genetic material of microbial cells than that of the somatic cells of the host. These microbes have a profound effect on several physiological functions ranging from energy metabolism to the immune response of the host, particularly those associated with the gut immune system. The gut of a newly hatched chick is typically sterile but is rapidly colonized by microbes in the environment, undergoing cycles of development. Several factors such as diet, region of the gastrointestinal tract, housing, environment, and genetics can influence the microbial composition of an individual bird and can confer a distinctive microbiome signature to the individual bird. The microbial composition can be modified by the supplementation of probiotics, prebiotics, or synbiotics. Supplementing these additives can prevent dysbiosis caused by stress factors such as infection, heat stress, and toxins that cause dysbiosis. The mechanism of action and beneficial effects of probiotics vary depending on the strains used. However, it is difficult to establish a relationship between the gut microbiome and host health and productivity due to high variability between flocks due to environmental, nutritional, and host factors. This review compiles information on the gut microbiota, dysbiosis, and additives such as probiotics, postbiotics, prebiotics, and synbiotics, which are capable of modifying gut microbiota and elaborates on the interaction of these additives with chicken gut commensals, immune system, and their consequent effects on health and productivity. Factors to be considered and the unexplored potential of genetic engineering of poultry probiotics in addressing public health concerns and zoonosis associated with the poultry industry are discussed.

Methanogenic archaea in subsurface coal seams are biogeographically distinct: an analysis of metagenomically-derived mcrA sequences.

The production of methane as an end-product of organic matter degradation in the absence of other terminal electron acceptors is common, and has often been studied in environments such as animal guts, soils and wetlands due to its potency as a greenhouse gas. To date, however, the study of the biogeographic distribution of methanogens across coal seam environments has been minimal. Here, we show that coal seams are host to a diverse range of methanogens, which are distinctive to each geological basin. Based on comparisons to close relatives from other methanogenic environments, the dominant methanogenic pathway in these basins is hydrogenotrophic, with acetoclastic being a second major pathway in the Surat Basin. Finally, mcrA and 16S rRNA gene primer biases were predominantly seen to affect the detection of Methanocellales, Methanomicrobiales and Methanosarcinales taxa in this study. Subsurface coal methanogenic community distributions and pathways presented here provide insights into important metabolites and bacterial partners for in situ coal biodegradation.

Suitability of Hermetia illucens larvae meal and fat in broiler diets: effects on animal performance, apparent ileal digestibility, gut histology, and microbial metabolites

BackgroundThe possibility of partially replacing soybean meal (SBM) with Hermetia illucens (HI) defatted larvae meal in broiler nutrition has frequently been suggested. For sustainability reasons, however, the larvae fat produced during defatting should also be used and could be particularly beneficial regarding gut health due to its fatty acid composition. To evaluate the suitability of HI larvae as protein and fat source, a 2 × 3 factorial arrangement with two types of protein, i.e. SBM (S) or SBM and 15% of its crude protein replaced by HI larvae meal (L), and three levels of fat sources, namely 0 (0 L), 50% (50 L) or 100% HI larvae fat (100 L) at the expense of soybean oil was applied.ResultsIn the starter phase, an interaction showed higher body weight (BW), average daily gain (ADG) and improved feed conversion ratio (FCR) if 50% or 100% HI larvae fat was fed with HI larvae meal. Moreover, BW, ADG and FCR improved when feeding HI larvae meal as protein source. Additionally, we observed an increased average daily feed intake in the grower, finisher, and overall phase in the L groups and an improved FCR in 0 L compared to 50 L groups during the overall period. Regarding apparent ileal digestibility, HI larvae meal feeding increased dry matter, organic matter, and fat digestibility. Feeding HI larvae meal as protein source decreased the concentrations of agmatine, spermidine, spermine and ammonia in the caecal digesta, whereas fat source affected agmatine with higher concentrations in 50 L compared to 0 L in the colonic digesta. In contrast, caecal ethanolamine concentrations increased in HI larvae meal groups compared to SBM. Caecal butyric acid concentrations decreased with HI larvae meal feeding. An interaction was found for the jejunal villus area, being higher in L + 100 L compared to S + 100 L. Furthermore, L groups had greater villus width.ConclusionsA partial replacement of SBM with HI larvae meal and soybean oil with HI larvae fat in broiler diets without impairing animal performance or gut health seems possible. Feeding HI larvae meal affected broiler performance positively in the starter phase and improved apparent ileal digestibility.

Source Tracking based on Microbial Communities: Sample Collection and ddPCR/qPCR

Fecal contamination serves as a major public health threat, especially in drinking or recreational waters. This is particularly concerning because it promotes the exposure of communities to harmful pathogenic bacteria. Identifying the source of contamination serves as an important context when mitigating the effects of the pollution or preventing it from initially entering the water. Microbial source tracking (MST) is employed to achieve the quantification of dominant sources of fecal contamination. Specifically in this study, a library‐independent method was utilized in the sample level detection of host‐associated 16s rRNA markers. Among the most prevalent bacteria in human and animal gut microbiomes is the phylum Bacteroidetes,which prove to be extremely useful in MST assays. TaqMan primers and probes targeting host‐independent (universal) Bacteroidetes(BacUni), human‐host specific (HF183), and pig host specific (Pig2Bac) were utilized. Dye‐based SYBR assays targeting avian bacteria Helicobacter (GFD) were also investigated. Solid fecal samples were acquired from a local petting zoo and surrounding beach sites. Water samples from various lakes, streams, and wastewater treatment plants were also obtained. Samples were filtered using membrane filtration technology and DNA was isolated using DNEasy PowerSoilkit. Upon DNA isolation, the concentration of DNA was measured using NanoDrop and the samples were subjected to qPCR or ddPCR assays. Results indicate effective primer/probe selectivity. Cross‐reactivity between samples and primers demonstrated that the targets were not exclusive to the presumed host and could overlap. Future research includes testing primers on samples from a wider range of animals and water sources. Additionally, a comparison of qPCR results to those from ddPCR and quantifying any differences within or between the two.

New challenges in the study of the evolution of wild animals and their gut microbiome.

In this viewpoint, by reviewing the recent findings on wild animals and their gut microbiomes, we found some potential new insights and challenges in the study of the evolution of wild animals and their gut microbiome. We suggested that wild animal gut microbiomes may come from microbiomes in the animals' living habitats along with animals' special behavior, and that the study of long‐term changes in gut microbiomes should consider both habitat and special behaviors. Also, host behavior would facilitate the gut microbiome transmission between individuals. We suggested that research should integrate the evolutionary history and physiological systems of wild animals to understand the evolution of animals and their gut microbiomes. Finally, we proposed the Noncultured‐Cultured‐Fermentation‐Model Animal pipeline to determine the function (diet digestion, physiology, and behavior) of these target strains in the wild animal gut.

Occurrence of Extended-spectrum Beta-lactamases and the Antibiogram Pattern of Escherichia coli Isolates from Pus and Urine Samples

Escherichia coli (E. coli) is a significant species and a common dweller in the guts of humans and animals causing urinary tract infections, wound infection, otitis media, bloodstream infections, and other complications in humans. Worldwide resistance to antimicrobials is a serious public health problem. β-lactamases production by E. coli is responsible for resistance to frequently used beta-lactam antibiotics. 1) To determine antibiotic susceptibility pattern of Escherichia coli isolated from pus and urine samples 2) To determine ESBL producing Escherichia coli and its antibiotic susceptibility pattern. Retrospective study of E. coli isolates from urine and pus samples was performed by collecting data from bacteriology registers. Gender, age details of patients, number of E. coli isolates, and their antimicrobial susceptibility profiles were collected from the records. Out of 747 samples 102 E. coli were isolated and among them 60 (59%) were ESBL producers. Male preponderance was seen i.e. 59 (57.84%) and majority 68 (66.66%) were isolated from the age group of <18 years. E. coli showed maximum sensitivity to imipenem 88 (86.27%), followed by piperacillin-tazobactam 84 (82.35%), aminoglycoside group 76 (74.5%) and maximum resistance was seen to penicillin groups 92 (90.19%), followed by cephalosporins 71 (69.6%). Urinary isolates showed maximum sensitivity to nitrofurantoin (93.67%). ESBL producers showed resistance to ciprofloxacin 47 (78%). This study helps in the periodic formulation of regional antimicrobial policies and also giving continuous information to the clinicians with respect to the sensitivity pattern along with ESBL production which can help to avert further drug resistance.

Recent developments in antimicrobial growth promoters in chicken health: Opportunities and challenges

With a continuously increasing human population is an increasing global demand for food. People in countries with a higher socioeconomic status tend to switch their preferences from grains to meat and high-value foods. Their preference for chicken as a source of protein has grown by 70% over the last three decades. Many studies have shown the role of feed in regulating the animal gut microbiome and its impact on host health. The microbiome absorbs nutrients, digests foods, induces a mucosal immune response, maintains homeostasis, and regulates bioactive metabolites. These metabolic activities are influenced by the microbiota and diet. An imbalance in microbiota affects host physiology and progressively causes disorders and diseases. With the use of antibiotics, a shift from dysbiosis with a higher density of pathogens to homeostasis can occur. However, the progressive use of higher doses of antibiotics proved harmful and resulted in the emergence of multidrug-resistant microbes. As a result, the use of antibiotics as feed additives has been banned. Researchers, regulatory authorities, and managers in the poultry industry have assessed the challenges associated with these restrictions. Research has sought to identify alternatives to antibiotic growth promoters for poultry that do not have any adverse effects. Modulating the host intestinal microbiome by regulating dietary factors is much easier than manipulating host genetics. Research efforts have led to the identification of feed additives, including bacteriocins, immunostimulants, organic acids, phytogenics, prebiotics, probiotics, phytoncides, and bacteriophages. In contrast to focusing on one or more of these alternative bioadditives, an improved feed conversion ratio with enhanced poultry products is possible by employing a combination of feed additives. This article may be helpful in future research towards developing a sustainable poultry industry through the use of the proposed alternatives.

Effects of Metal-Polycation Pillaring and Exchangeable Cations on Aflatoxin Adsorption by Smectite

AbstractNatural smectites bind aflatoxins from water effectively, but the complex chemical environment in the guts of mammals and other animals can limit binding of aflatoxins. Many efforts have been made to enhance the adsorption capacity and affinity of smectites for aflatoxins in the presence of biological compounds. The main objective of the present study was to modify smectite structures by pillaring and cation exchange to enhance aflatoxin B1 adsorption capacity and selectivity. Smectite was pillared with Al and Al-Fe polycations or saturated with Ca, Mg, Zn, or Li. Structural changes in smectites with or without heat treatment were determined using X-ray diffraction and Fourier-transform infrared spectroscopy. Equilibrium aflatoxin B1 adsorption to the smectites was measured in aqueous solution and in simulated gastric fluid. Pillaring with the polycations expanded smectites in the z-direction to 18.6 Å and the expansion was stable after heating at 500°C. Changes in the Al–OH–Al infrared bands in the stretching region supported the formation of pillared clays. Migration of Mg, Zn, and Li into the octahedral sites of the smectite was observed as Mg and Zn saturation yielded a d spacing of 15 Å at 200°C which collapsed to 9.6 Å at 400°C. The 14.6 Å peak of the Li-saturated smectite collapsed to 9.6 Å at 200°C while the 15 Å Ca-saturated smectite peak was stable up to 400°C. The unheated Al- and AlFe-pillared smectites adsorbed significantly more aflatoxin B1 from an aqueous suspension than did unpillared clay. In both water and simulated gastric fluid, heat treatment decreased aflatoxin B1 adsorption to pillared smectites, but heat treatment increased aflatoxin B1 adsorption to unpillared smectites. Without heat treatment, smectites saturated with divalent cations (Ca, Mg, Zn) adsorbed more aflatoxin B1 from an aqueous suspension than the smectite saturated with a monovalent cation (Li). Ca-saturated smectite showed the greatest aflatoxin B1 adsorption, 114 g kg–1, from aqueous suspension after 400°C heat treatment. The Zn-, Mg-, and Li-saturated smectites showed maximum aflatoxin adsorption of 107, 93, and 90 g kg–1, respectively, after 200°C heat treatment. From simulated gastric fluid with pepsin, the 200°C heated, Zn-saturated smectite had maximum aflatoxin B1 adsorption of 68 g kg–1. Pillared smectites effectively adsorbed aflatoxin B1 from aqueous suspension, but Ca- and Zn-saturated smectites after heat treatment might improve the selectivity of smectites for aflatoxin B1 over pepsin and enhance the efficacy of smectite as a feed additive.

Changes of antibiotic resistance genes and gut microbiota after the ingestion of goat milk

Antibiotic resistance genes, as newly emerging contaminants, have become a serious challenge to public health through the food chain. The gut of humans and animals is an important reservoir for the development and dissemination of antibiotic resistance genes because of the great abundance and diversity of intestinal microbiota. In the present study, we evaluated the influence of goat milk on the diversity and abundance of antibiotic resistance genes and gut microbial communities, especially pathogenic bacteria. Male mice were used, 12 for each of the 2 groups: a control group that received sterile distilled water and a treated group that received goat milk, and gut microbiota and antibiotic resistance genes were compared in these groups using metagenomic analysis. The results revealed that ingestion of goat milk decreased the diversity and abundance of antibiotic resistance genes in the mice gut. The relative abundance of fluoroquinolone, peptide, macrolide, and β-lactam resistance genes in the total microbial genes significantly decreased after the intervention. Goat milk intake also significantly reduced the abundance of pathogenic bacteria, such as Clostridium bolteae, Clostridium symbiosum, Helicobacter cinaedi, and Helicobacter bilis. Therefore, goat milk intake might decrease the transfer potential of antibiotic resistance gene to pathogenic bacteria in the gut. In addition, bacteria with multiple resistance mechanisms accounted for approximately 4.5% of total microbial communities in the control group, whereas it was not detectable in the goat milk group, indicating the total inhibition by goat milk intake. This study highlights the influence of goat milk on antibiotic resistome and microbial communities in the gut, and provides a new insight into the function of goat milk for further study.

Non-Photosynthetic Melainabacteria (Cyanobacteria) in Human Gut: Characteristics and Association with Health.

Gut microorganisms are comprised of thousands of species and play an important role in the host’s metabolism, overall health status, and risk of disease. Recently, the discovery of non-photosynthetic cyanobacteria (class “Melainabacteria”) in the human and animal gut triggered a broad interest in studying cyanobacteria’s evolution, physiology, and ecological relevance of the Melainabacteria members. In the present paper, we review the general characteristics of Melainabacteria, their phylogeny, distribution, and ecology. The potential link between these microorganisms and human health is also discussed based on available human-microbiome studies. Their abundance tends to increase in patients with selected neurodegenerative, gastrointestinal, hepatic, metabolic, and respiratory diseases. However, the available evidence is correlative and requires further longitudinal studies. Although the research on Melainabacteria in the human gut is still in its infancy, elucidation of their role appears important in better understanding microbiome–human health interactions. Further studies aiming to identify particular gut cyanobacteria species, culture them in vitro, and characterize them on the molecular, biochemical, and physiological levels are encouraged.

Microbiomes of Hadal Fishes across Trench Habitats Contain Similar Taxa and Known Piezophiles.

ABSTRACTHadal snailfishes are the deepest-living fishes in the ocean, inhabiting trenches from depths of ∼6,000 to 8,000 m. While the microbial communities in trench environments have begun to be characterized, the microbes associated with hadal megafauna remain relatively unknown. Here, we describe the gut microbiomes of two hadal snailfishes, Pseudoliparis swirei (Mariana Trench) and Notoliparis kermadecensis (Kermadec Trench), using 16S rRNA gene amplicon sequencing. We contextualize these microbiomes with comparisons to the abyssal macrourid Coryphaenoides yaquinae and the continental shelf-dwelling snailfish Careproctus melanurus. The microbial communities of the hadal snailfishes were distinct from their shallower counterparts and were dominated by the same sequences related to the Mycoplasmataceae and Desulfovibrionaceae. These shared taxa indicate that symbiont lineages have remained similar to the ancestral symbiont since their geographic separation or that they are dispersed between geographically distant trenches and subsequently colonize specific hosts. The abyssal and hadal fishes contained sequences related to known, cultured piezophiles, microbes that grow optimally under high hydrostatic pressure, including Psychromonas, Moritella, and Shewanella. These taxa are adept at colonizing nutrient-rich environments present in the deep ocean, such as on particles and in the guts of hosts, and we hypothesize they could make a dietary contribution to deep-sea fishes by degrading chitin and producing fatty acids. We characterize the gut microbiota within some of the deepest fishes to provide new insight into the diversity and distribution of host-associated microbial taxa and the potential of these animals, and the microbes they harbor, for understanding adaptation to deep-sea habitats.IMPORTANCE Hadal trenches, characterized by high hydrostatic pressures and low temperatures, are one of the most extreme environments on our planet. By examining the microbiome of abyssal and hadal fishes, we provide insight into the diversity and distribution of host-associated life at great depth. Our findings show that there are similar microbial populations in fishes geographically separated by thousands of miles, reflecting strong selection for specific microbial lineages. Only a few psychropiezophilic taxa, which do not reflect the diversity of microbial life at great depth, have been successfully isolated in the laboratory. Our examination of deep-sea fish microbiomes shows that typical high-pressure culturing methodologies, which have largely remained unchanged since the pioneering work of Claude ZoBell in the 1950s, may simulate the chemical environment found in animal guts and helps explain why the same deep-sea genera are consistently isolated.

Acids produced by lactobacilli inhibit the growth of commensal Lachnospiraceae and S24-7 bacteria

ABSTRACT The Lactobacillaceae are an intensively studied family of bacteria widely used in fermented food and probiotics, and many are native to the gut and vaginal microbiota of humans and other animals. Various studies have shown that specific Lactobacillaceae species produce metabolites that can inhibit the colonization of fungal and bacterial pathogens, but less is known about how Lactobacillaceae affect individual bacterial species in the endogenous animal microbiota. Here, we show that numerous Lactobacillaceae species inhibit the growth of the Lachnospiraceae family and the S24-7 group, two dominant clades of bacteria within the gut. We demonstrate that inhibitory activity is a property common to homofermentative Lactobacillaceae species, but not to species that use heterofermentative metabolism. We observe that homofermentative Lactobacillaceae species robustly acidify their environment, and that acidification alone is sufficient to inhibit growth of Lachnospiraceae and S24-7 growth, but not related species from the Clostridiales or Bacteroidales orders. This study represents one of the first in-depth explorations of the dynamic between Lactobacillaceae species and commensal intestinal bacteria, and contributes valuable insight toward deconvoluting their interactions within the gut microbial ecosystem.

Harnessing the potentialities of probiotics, prebiotics, synbiotics, paraprobiotics, and postbiotics for shrimp farming

AbstractThe shrimp farming industry is receiving increasing attention globally. However, extending the frontiers of the industry has been a challenge as high stocking density often increases susceptibility to diseases and compromises shrimp quality. There has been a huge adoption of bacterial‐derived additives in recent decades, particularly in Asia. Accumulating evidence demonstrates the modulatory effect of these functional additives on the animal gut microbiota composition, thus making them potential alternatives to antibiotics in controlling infectious diseases, such as acute hepatopancreatic necrosis disease (AHPND). These functional additives could be introduced in various forms, such as probiotics, synbiotics, paraprobiotics, and postbiotics. This article aims to critically review previous studies that investigated the effects of these bacterial derivatives, particularly on shrimp species. The surge of research in recent years indirectly attested to the immense potentialities of these functional additives. Generally, the introduction of bacterial derivatives not only helped in disease control but also demonstrated encouraging outcomes, including better survival rate, immune enhancement, and growth promotion effects. It is interesting to note that the effectiveness of probiotics extended beyond the viability of the bacteria, thus leading to concepts such as paraprobiotics and postbiotics. Although this is a relatively new research area with limited information, this review aims to consolidate available data and provide a comprehensive overview of the state of the art of this research arena. The challenges in each research domain were also discussed towards the end of this review to provide insights into potential future research niches.

Tetanus Presented as Acute Abdomen

Tetanus is commonly seen in the emergency department with a puncture wound. This is an unusual presentation of tetanus without an open wound. A 59 year old male with a past history of hypertension, hepatitis C, prior injection drug use, and homeless presented to the emergency department with altered mental status and hyperthermia with fever of 105 F on a hot July afternoon. Patient was found in his car sleeping. Initially, the patient declined medical care, but was later found unresponsive with a methadone bottle next to him. Given Narcan on field with improvement of mental status. Patient developed acute ridged abdominal pain. A CT scan raised concern for acute abdomen, with portal venous gas, and intestinal pneumatosis. Surgery consulted and was concerned for ischemic colon with perforation. Vancomycin, ceftriaxone and metronidazole were given. Emergency exploratory laparotomy was performed with a small serosal injury on colon repaired with suture. Blood culture came back positive for Clostridium tetani and Mobiluncus curtisii. Infectious disease consulted. Felt tetanus cannot be ruled out. Patient was given tetanus immune globulin (TIG), metronidazole and later tetanus vaccine. Patient has fully recovered, tolerates oral diets, and has been discharged to a nursing facility without any neurological deficit. Tetanus is a neurological disorder caused by the tetanus toxins produced by clostridium tetani, which is a gram positive obligate anaerobes commonly found in soil. It is relevant to note that this bacterium can also be found inside the gut of animals and humans. The disease is well characterized by ancient Greeks, and common symptoms include trismus (“lockjaw”), opisthotonos, and a board-like rigid abdomen. Incubation is usually 7-21 days and this disease is usually prevented by vaccination. This case is unusual because there is no open wound and PE is limited due to patient’s overdose on opiates (methadone). Possible cause for this particular case may be that since there was no open wound, the blood could have been seeded by the perforated bowel because they may also be found in the gut. The patient was given metronidazole as part of Tx for acute abdomen, which happens to also be a treatment for tetanus. Diseases can be presented atypically. This case illustrates the importance of differential diagnosis when considering treatments in any pathology.

Antibacterial and Antibiofilm Activity of Selected Medicinal Plant Leaf Extracts Against Pathogens Implicated in Poultry Diseases.

Antimicrobial resistant poultry pathogens are responsible for treatment failure and economic losses, and can also be a source of resistant zoonotic infections representing a risk to human health. In 2006 the European Union banned the use of antibiotics as growth promoters in farm animals and other regions are likely to follow suit. Alternative products and strategies are sought to help maintain animal gut health to reduce the prevalence of pathogens in the food chain. The minimum inhibitory concentration (MIC) of organic and aqueous leaf extracts of Alchornea laxiflora, Ficus exasperata, Morinda lucida, Jatropha gossypiifolia, Ocimum gratissimum, and Acalypha wilkesiana were tested against bacterial poultry pathogens including Staphylococcus aureus, Enterococcus faecalis, Salmonella spp., Escherichia coli, Campylobacter spp., and fungal species (Aspergillus fumigatus, Aspergillus flavus, and Candida albicans) using a 2-fold serial microdilution method. Activity of extracts against biofilms of the pathogens was done using a modified crystal violet staining in vitro assay. The safety of extracts was determined against Vero and Caco-2 cells using a tetrazolium-based in vitro assay. Acetone and cold water extracts of M. lucida had the best activity against three bacteria (MIC = 0.05–0.07 mg/ml) and two fungal (MIC = 0.03–0.15 mg/ml) organisms, respectively. The E. coli isolate and A. flavus were the most susceptible bacteria and fungi, respectively. Caco-2 cells generally displayed higher selectivity index (SI) values compared to Vero cells and average SI values against Vero and Caco-2 cells for both bacteria and fungi ranged from 0.01 to 4.48 and 0.005 to 16.41, respectively. All plant extracts had good anti-biofilm activity (>50%) against at least one organism. The disruption of established biofilm growth by the plant samples proved to be more difficult to achieve than efficacy against planktonic forms of bacteria. This study shows that some of the plant species are potential candidates as alternative feed additives in poultry production. In the future, a poultry feed trial to evaluate their in vivo efficacy as herbal feed additives will be conducted.

Extraintestinal Pathogenic Escherichia coli: Beta-Lactam Antibiotic and Heavy Metal Resistance.

Multiple-antibiotic-resistant (MAR) extra-intestinal pathogenic Escherichia coli (ExPEC) represents one of the most frequent causes of human nosocomial and community-acquired infections, whose eradication is of major concern for clinicians. ExPECs may inhabit indefinitely as commensal the gut of humans and other animals; from the intestine, they may move to colonize other tissues, where they are responsible for a number of diseases, including recurrent and uncomplicated UTIs, sepsis and neonatal meningitis. In the pre-antibiotic era, heavy metals were largely used as chemotherapeutics and/or as antimicrobials in human and animal healthcare. As with antibiotics, the global incidence of heavy metal tolerance in commensal, as well as in ExPEC, has increased following the ban in several countries of antibiotics as promoters of animal growth. Furthermore, it is believed that extensive bacterial exposure to heavy metals present in soil and water might have favored the increase in heavy-metal-tolerant microorganisms. The isolation of ExPEC strains with combined resistance to both antibiotics and heavy metals has become quite common and, remarkably, it has been recently shown that heavy metal resistance genes may co-select antibiotic-resistance genes. Despite their clinical relevance, the mechanisms underlining the development and spread of heavy metal tolerance have not been fully elucidated. The aim of this review is to present data regarding the development and spread of resistance to first-line antibiotics, such as beta-lactams, as well as tolerance to heavy metals in ExPEC strains.

Antimicrobial resistance of commensal Enterococcus faecalis and Enterococcus faecium from food-producing animals in Russia.

Background and Aim:Although Enterococcus faecalis and Enterococcus faecium are common members of human and animal gut microbiota, their resistance to different antimicrobials makes them important pathogens. Multidrug-resistant enterococci often contaminate foods of animal origin at slaughterhouses. The World Health Organization and the World Organization for Animal Health recommend including animal-derived enterococci in antimicrobial resistance (AMR) monitoring programs. This study aimed to fill a literature gap by determining the current AMR prevalence of E. faecalis and E. faecium from different food-producing animals in Russia.Materials and Methods:Samples of biomaterial were taken from chickens (n=187), cattle (n=155), pigs (n=49), turkeys (n=34), sheep (n=31), and ducks (n=31) raised at 28 farms in 15 regions of Russia. Isolates of E. faecalis (n=277) and of E. faecium (n=210) (487 isolates in total; 1 isolate per sample) were tested for resistance to 12 antimicrobials from 11 classes using the broth microdilution method. Three criteria were used for the interpretation of minimum inhibitory concentration: Epidemiological cutoff values (ECOFFs) from the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Clinical and Laboratory Standards Institute (CLSI) clinical breakpoints. The AMR cloud online platform was used for data processing and statistical analysis.Results:A difference of >10% was found between E. faecalis and E. faecium resistance to several antimicrobials (erythromycin, gentamycin, tetracycline, chloramphenicol, ciprofloxacin, and streptomycin). In total, resistance to most antimicrobials for enterococci isolates of both species taken from turkeys, chicken, and pigs was higher than cattle, sheep, and ducks. The highest levels were found for turkeys and the lowest for ducks. Among antimicrobials, resistance to bacitracin and virginiamycin was 88-100% in nearly all cases. High levels of clinical resistance were found for both bacteria species: Rifampicin (44-84%) from all animals, tetracycline (45-100%) from poultry and pigs, and erythromycin (60-100%), ciprofloxacin (23-100%), and trimethoprim-sulfamethoxazole (33-53%) from chickens, turkeys, and pigs. No vancomycin-resistant isolates were found. Most isolates were simultaneously resistant to one–three classes of antimicrobials, and they were rarely resistant to more than three antimicrobials or sensitive to all classes.Conclusion:Differences in resistance between enterococci from different farm animals indicate that antimicrobial application is among the crucial factors determining the level of resistance. Conversely, resistance to rifampicin, erythromycin, tetracycline, and ciprofloxacin found in enterococci from farm animals in our study was notably also found in enterococci from wild animals and birds. Our results may be partly explained by the intrinsic resistance of E. faecium and E. faecalis to some antimicrobials, such as trimethoprim/sulfamethoxazole and bacitracin.