Fecal Time Research Articles

Synthesis and quantification of short-chain fatty acid esters of hydroxy fatty acids in rat intestinal contents and fecal samples by LC-MS/MS

Short-chain fatty acid esters of hydroxy fatty acids (SFAHFAs) are a new class of endogenous lipids belonging to the fatty acid esters of the hydroxy fatty acid family. We previously uncovered their chemical structure and discussed their potential biological significance. We anticipate an increased need for SFAHFA measurements as markers of metabolic and inflammatory health. In this study, we synthesized sixty isomeric SFAHFAs by combining 12 hydroxy fatty acids (C16–C24) and five short-chain fatty acids (C2–C6) including a labelled internal standard. SFAHFA enrichment was achieved by solid-phase extraction and established a sensitive method for their quantitation by targeted LC-MS/MS. The method was applied to profile SFAHFAs in intestinal contents and fecal samples collected from rats fed a high-fat diet (HFD). The results demonstrated a significant decrease in SFAHFAs in the intestinal contents of the HFD group compared with the control group. The fecal time course (0–8 weeks) profile of SFAHFAs showed significant downregulation of acetic and propanoic acid esters in just 2 weeks after HFD administration. This study offers the first synthesis and quantitation method for SFAHFAs, demonstrating their potential use in elucidating SFAHFA sources, their role in various diseases, and potential biochemical signalling pathways.

Toxicokinetics of rare earth element oxides administered intravenously to rats

Rare earth elements (REEs) are increasingly used in a wide range of applications. However, their toxicokinetic behaviors in animals and humans are not yet fully documented, hindering health risk assessments. We used a rat experimental model to provide novel data on the toxicokinetics of the insoluble oxide forms of praseodymium (Pr), neodymium (Nd), cerium (Ce) and yttrium (Y) administered intravenously. Detailed blood, urinary and fecal time courses were documented through serial sampling over 21 days in male Sprague-Dawley rats exposed to a mixture of these REE oxides administered at two different doses (0.3 or 1 mg kg−1 bw of each REE oxide commercially sold as bulk μm-sized particles). Tissue REE levels at the time of sacrifice were also measured. Significant effects of the dose on REE time courses in blood and on cumulative urinary and fecal excretion rates were observed for all four REE oxides assessed, as lower cumulative excretion rates were noted at the higher REE dose. In the liver, the main accumulation organ, the fraction of the administered REE dose remaining in the tissue at necropsy was similar at both doses. Toxicokinetic data for the REE oxides were compared to similar data for their chloride salts (also administered intravenously in a mixture, at 0.3 and 1 mg kg−1 bw of each REE chloride) obtained from a previous study. Compared to their chloride counterparts, faster elimination of REE oxides from the blood was observed in the first hours post-dosing. Furthermore, higher mean residence time (MRT) values as well as slower cumulative urinary and fecal excretion were determined for the REE oxides. Also, while liver REE retention was similar for both REE forms, the fractions of the administered REEs recovered in the spleen and lungs were noticeably higher for the REE oxides, at both dose levels. This study highlights the importance of both the dose and form of the administered REEs on their toxicokinetic profiles. Results indicate that chronic exposure and increased doses of REEs may favor bioaccumulation in the body, in particular for insoluble oxide forms of REEs, which are eliminated more slowly from the body.

Genetic Analysis of Cachavirus-Related Parvoviruses Detected in Pet Cats: The First Report From China

In this study, members of the Carnivore chaphamaparvovirus species 1, closely related to a virus previously reported in dog feces named cachavirus was identified for the first time in feces of Chinese cats. Screening tests using rectal swabs from 171 diarrheic and 378 healthy cats collected from Henan, Anhui, and Zhejiang provinces in China revealed two samples from diarrheic cats that were positive for cachavirus, but statistical analysis indicated no association between the presence of the virus and clinical signs (p > 0.05). Subsequently, two partial genome sequences [from nucleotides 479–4123, according to the strains from dogs (cachavirus)] of the two strains from cats (cachavirus-cat1 and -cat2) were amplified. The NS1 and VP1 sites of cachavirus-cat1 and -cat2 shared a high identity of 91.9 and 97.0% with reported cachaviruses, respectively, but lower identity of 74.8 and 73.2% with another carnivore chaphamaparvovirus named fechaviruses detected in cats, respectively, indicated the two strains might origin from dogs. These findings improve our understanding of the diversity and tropism of viruses in Carnivore chaphamaparvovirus species 1 which now include both dogs and now cats viruses.

Whip-LAMP: a novel LAMP assay for the detection of Trichuris muris-derived DNA in stool and urine samples in a murine experimental infection model

BackgroundTrichuris trichiura (human whipworm) infects an estimated 477 million individuals worldwide. In addition to T. trichiura, other Trichuris species can cause an uncommon zoonosis and a number of human cases have been reported. The diagnosis of trichuriasis has relied traditionally on microscopy. Recently, there is an effort to use molecular diagnostic methods, mainly qPCR. LAMP technology could be an alternative for qPCR especially in low-income endemic areas. Trichuris muris, the causative agent of trichuriasis in mice, is of great importance as a model for human trichuriasis. Here, we evaluate the diagnostic utility of a new LAMP assay in an active experimental mouse trichuriasis in parallel with parasitological method by using stool and, for the first time, urine samples.MethodsStool and urine samples were collected from mice infected with eggs of T. muris. The dynamics of infection was determined by counting the number of eggs per gram of faeces. A LAMP based on the 18S rRNA gene from T. muris was designed. Sensitivity and specificity of LAMP was tested and compared with PCR. Stool and urine samples were analysed by both LAMP and PCR techniques.ResultsTrichuris muris eggs were detected for the first time in faeces 35 days post-infection. LAMP resulted specific and no cross-reactions were found when using 18 DNA samples from different parasites. The detection limit of the LAMP assay was 2 pg of T. muris DNA. When testing stool samples by LAMP we obtained positive results on day 35 p.i. and urine samples showed amplification results on day 20 p.i., i.e. 15 days before the onset of T. muris eggs in faeces.ConclusionsTo the best of our knowledge, we report, for the first time, a novel LAMP assay (Whip-LAMP) for sensitive detection of T. muris DNA in both stool and urine samples in a well-established mice experimental infection model. Considering the advantages of urine in molecular diagnosis in comparison to stool samples, should make us consider the possibility of starting the use urine specimens in molecular diagnosis and for field-based studies of human trichuriasis where possible. Further studies with clinical samples are still needed.

After the First Wave of COVID-19: Reflections From Italy.

After the First Wave of COVID-19: Reflections From Italy.

In vitro and in vivo studies of nematophagous fungi Arthrobotrys musiformis and Arthrobotrys robusta against the larvae of the trichostrongylides.

Six isolates of Arthrobotrys musiformis and five isolates of Arthrobotrys robusta were assessed in in vitro test regarding the capacity of prey larvae of the natural mixed trichostrongylides. In 5 isolates of A. robusta, the decrease percentage of infective larvae (L3) of trichostrongylides ranged from 97.71%-99.98% and for the isolates of A. musiformis, 5 isolates ranged from 97.99%-99.95% and only NF015 isolate 60.72%. In the following, the isolate (NPS045) of A. musiformis was selected to assess its excretion time in feces after oral administration of goats. Regarding L3 reduction rate, results demonstrated by NPS045 at each time point after fungal administration were 31.65% (12 h), 51.25% (24 h), 41.07% (48 h), 6.44% (72 h), 0% (96 h) and (120 h) (p<0.05) respectively, when compared to the control group. In the plates of the treated groups, the presence of the isolate (NPS045) was detected in samples at 12, 24 and 48 h after the fungus dose and 72 h later was not done. All native isolates of nematophagous fungi, including 6 isolates of A. musiformis and 5 isolates of A. robusta were assessed in vivo regarding the capacity of supporting the passage through goat gastrointestinal tract. The 3 isolates of A. musiformis could be able to pass through the digestive tract of goats without complete loss of ability of preying larvae of trichostrongylides in feces and their efficacies ranged from 47.60% to 55.93%. The two isolates of A. robusta survived the passage and the percentage reduction of L3 in feces were 41.96% and 66.97%, respectively. The remaining isolates were negative for both the efficacy of L3 reduction and the fungal examination in feces. In this study, the native isolates whose efficacies are good in vivo test have preliminarily demonstrated to be potential for the biological control of small ruminant parasite.

Kinetics of lithium as a lithium chloride dose suitable for conditioned taste aversion in lactating goats and dry sheep.

Lithium chloride (LiCl) is widely used for inducing conditioned taste aversion (CTA) so that livestock will reduce or avoid ingestion of toxic plants and graze groundcover mingled with valuable crops. However, pharmacokinetic studies of LiCl at effective CTA doses are lacking. With this aim, 6 Murciano-Grandina dairy does during late lactation and 6 dry Manchega dairy ewes were orally dosed with 200 and 225 mg LiCl/kg BW, respectively. Does were placed in metabolism cages whereas ewes were group fed in pens. Lithium was measured over 168 (does) and 192 h (ewes) at predefined intervals in plasma, urine, feces, and milk using flame atomic absorption spectroscopy. Plasma Li concentrations reached a maximum at 4 h in does (13.4 ± 1.35 mg Li/L) and 12 h in ewes (17.7 ± 0.8 mg Li/L). The calculated plasma half-lives were 40.3 ± 3.8 and 30.9 ± 2.1 h for does and ewes, respectively. In goats, all Li administered was recovered at 96 h (92 ± 4% in urine, 6.5 ± 1.3% in feces, and 2.8 ± 0.4% in milk); however, the estimated clearance time in feces was 11 and 9 d for does and ewes, respectively. Additionally, maximum Li excretion in doe milk was 15.6 ± 0.5 mg/L, which was approximately half of the calculated effective dose for a 5-kg BW sucking kid. In conclusion, Li kinetics in goats and sheep were similar to cattle and elimination took longer than in monogastric species. The low concentration of Li in feces, urine, and milk, as well as the complete elimination of Li from the body after 1.5 wk allows us to conclude that LiCl is safe and suitable for inducing CTA in ruminants.

Survivability of Highly Pathogenic Avian Influenza H5N1 Virus in Poultry Faeces at Different Temperatures

Highly pathogenic avian Influenza (HPAI) is an important zoonotic disease and is becoming a great threat to poultry industry. India has experienced continual outbreaks of H5N1 HPAI virus since February, 2006 especially in Eastern India. Survivability in poultry faeces is an important determinant in evaluating the persistence of the virus in the poultry sheds and their vicinity. In this paper, survivability of Indian H5N1 HPAI virus in dry and wet poultry faeces at 42, 37, 24 and 4°C, respectively is reported. The effect of different temperatures was determined by linear regression model and defined in terms of linear equation. The virus survived up to 18h at 42°C, 24h at 37°C, 5days at 24°C and 8weeks at 4°C in dry and wet faeces, respectively. The coefficients of determination (R(2)) values for dry and wet faeces revealed that the difference in viral persistence in dry and wet faeces at all temperatures was not very marked. Results of the present study indicated that H5N1 HPAI virus may remain viable for extended periods of time in faeces at low temperatures and may act as a long term source of influenza virus in the environment.

Application ofScutellariae radix,Gardeniae fructus, and Probiotics to PreventSalmonella entericaSerovar Choleraesuis Infection in Swine

Salmonella enterica serovar Choleraesuis, a host-adapted pathogen of swine, usually causes septicemia. Lactic acid bacteria (LAB) strains have been widely studied in recent years for their probiotic properties. In this study, a mouse infection model first screened for potential agents against infection, then a pig infection model evaluated effects of LAB strains and herbal plants against infection. Scutellariae radix (SR) and Gardeniae fructus (GF) showed abilities to reduce bacteria shedding and suppressing serum level of TNF-α induced by infection in swine. Bioactivities of SR and GF were enhanced by combining with LAB strains, which alone could speed up the bacteria elimination time in feces and boost immunity of infected pigs. Baicalein and genipin exhibited stronger cytotoxicity than baicalin and geniposide did, as well as prevent Salmonella from invading macrophages. Our study suggests LAB strains as exhibiting multiple functions: preventing infection, enhancing immunity to prepare host defenses against further infection, and adjusting intestinal microbes' enzymatic activity in order to convert herbal compounds to active compounds. The SR/GF-LAB strain mixture holds potential infection-prevention agents supplied as feed additives.

Survival of Escherichia coli O157:H7 in feces from corn- and barley-fed steers

The survival of Escherichia coli O157:H7 in feces from steers fed corn (CO) or barley (BA) was evaluated at −10, +4 and +22 °C. Fecal pats were inoculated with a four-strain mixture of nalidixic-acid resistant E. coli O157:H7 at two levels: 10 3 CFU g −1 (low, L) and 10 5 CFU g −1 (high, H). At −10 °C, duration of survival of E. coli O157:H7 was reduced ( p < 0.05) in CO-L (35 days) compared to BA-L (49 days), likely due to the effects of fecal volatile fatty acids in combination with a fecal pH of <6.5. At 4 °C, E. coli O157:H7 was detected in BA-H, CO-H, CO-L and BA-L for 77, 77, 56 and 63 days, respectively, with no difference ( p > 0.05) observed in the duration of survival or rate of decline of E. coli O157:H7 among treatments. Survival of E. coli O157:H7 was twice as likely ( p < 0.05) at 22 °C than at 4 °C and −10 °C. While pH and dry matter content increased, and volatile fatty acid concentrations decreased over 84 days at all three temperatures, these changes were most pronounced at 22 °C. Survival of E. coli O157:H7 for extended periods of time in feces from both corn- and barley-fed animals was demonstrated, thus fecal material may serve as a vector for the transmission of the organism. The greater survival of E. coli O157:H7 at 22 °C suggests that temperature may play a role in the seasonality of transmission and prevalence of this bacterium in feedlot cattle. The reported greater prevalence of E. coli O157:H7 in cattle fed barley as compared to those fed corn does not appear to be related to elevated risk of transmission arising from differential survival of the bacterium in feces.

Laboratory study of the chemical composition of aging copepod fecal material

Changes in the chemical composition of aging copepod fecal material were investigated by incubating fresh feces in the dark at 5 and 15 °C and following transformations through time (27 days at 5 °C and 11 days at 15 °C) in terms of carbon (C), nitrogen (N), chlorophyll-type pigments and dissolved free amino acids (DFAA). At both temperatures, feces weight specific C and N decreased during incubation, this decrease being much more important at 15 °C (daily C loss of 2.5 μg·mg feces −1·day −1) than at 5 °C (maximum daily loss = 1.5 μg·mg feces −1·day −1). The C:N atomic ratio ranged between 7 and 8; it showed little temporal variation for both temperature conditions. The relative composition of HPLC-analysed chlorophyll-type pigments was rather constant, as illustrated by the Chl a-based pigment ratios. Six pigments were detected: Chl a, Chl a-type (possibly allomer), Chl c, Chl c-like, pheophorbide a, pheophorbide a-like. Their concentration generally decreased through time. Significant linear decreases were observed for Chl a (rate = −l.15 ng·mg −1 feces·day −1) and pheophorbide a-like at 15 °C. After a rapid decrease within the first 3–5 days of incubation, the total DFAA concentration measured in the filtrates was stable. Most individual amino acids (aspartic and glutamic acids, asparagine, arginine, leucine and ornithine) rapidly became undetectable or were <5–10% of the initial relative molar composition. A few others, considered as waste products (α-amino butyric acid, valine), accumulated with time. This process was temperature-dependent in <1 wk. After that period, the DFAA concentration and composition became stable, whatever the temperature, even when fecal C and pigments continued to decrease. Addition of chloramphenicol at the beginning of the incubation period generally resulted in larger C and pigment content in the fecal material, especially at 15 °C, as well as higher concentrations of DFAA and a more varied composition of amino acids when analysed on the last day of incubation at both temperatures. C or pigment values obtained in the antibiotic-treated samples could not be statistically differentiated from values of the first sampled day of each series, indicating the important role of bacteria in mediating observed transformations. Comparison of the temporal evolution of C and pigments showed differences in the relative rate of transformation of these two elements, particularly at 15 °C, with pigments decreasing faster than C and causing the C: Chl a ratio to increase with time in feces.

The Excretion of Aujeszky’s Disease Virus by Experimentally-infected Pigs

When pigs were infected with Aujeszky’s disease virus they excreted virus into the nose and mouth for periods ranging up to 17 days. Virus could not be detected at any time in faeces or urine.