The fecal microbiota of Holstein cows is heritable and genetically correlated to dairy performances.

The fecal microbiota of ruminants constitutes a diversified community that has been phenotypically associated with a variety of host phenotypes, such as production and health. To gain a better understanding of the complex and interconnected factors that drive the fecal bacterial community, we have aimed to estimate the genetic parameters of the diversity and composition of the fecal microbiota, including heritabilities, genetic correlations among taxa, and genetic correlations between fecal microbiota features and host phenotypes. To achieve this, we analyzed a large population of 1,875 Holstein cows originating from 144 French commercial herds and routinely recorded for production, somatic cell score, and fertility traits. Fecal samples were collected from the animals and subjected to 16S rRNA gene sequencing, with reads classified into Amplicon Sequence Variants (ASVs). The estimated α- and ß-diversity indices (i.e., Observed Richness, Shannon index, Bray-Curtis and Jaccard dissimilarity matrices) and the abundances of ASVs, genera, families and phyla, normalized by centered-log ratio (CLR), were considered as phenotypes. Genetic parameters were calculated using either univariate or bivariate animal models. Heritabilities estimates, ranging from 0.08 to 0.31 for taxa abundances and ß-diversity indices, highlight the influence of the host genetics on the composition of the fecal microbiota. Furthermore, genetic correlations estimated within the microbial community and between microbiota features and host traits reveal the complex networks linking all components of the fecal microbiota together and to their host, thus strengthening the holobiont concept. By estimating the heritabilities of microbiota-associated phenotypes, our study quantifies the impact of the host genetics on the fecal microbiota composition. In addition, genetic correlations between taxonomic groups and between taxa abundances and host performance suggest potential applications for selective breeding to improve host traits or promote a healthier microbiota.

[Oral administration of medications to infants: Implementation of a preliminary risk analysis in a hospital service of pediatrics]. / Administration orale de médicaments au nourrisson : réalisation d'une analyse préliminaire des risques dans un service d'hospitalisation pédiatrique.

PURPOSE: The objective of this study was to reduce the risk of errors when administering oral medications to infants aged 28 days to 2 years. MATERIAL AND METHODS: The method of the preliminary risk analysis (PRA) was implemented by a multidisciplinary group in a hospital service of pediatrics. The study focused on the phase of preparation of drugs by nurses before administration. RESULTS: This analysis revealed 41 scenari, 16 were criticality unacceptable. In particular, their analysis highlighted the impact of the drug dosage form, the lack of scientific information and the human factor on this preparation. Eleven action sheets have been written. DISCUSSION: The risk management requires significant human investment, material resources and organizational solutions: formations, information, i.e. computerized prescribing, dispensing and administering system, centralized drug preparations, automated drugs cabinets or unit drug daily dispensing system. CONCLUSION: Control these risks means to get specific actions at pediatric wards, enhance dispensing system by the hospital pharmacist and the support of the pharmaceutical industry to get commercially available pediatric drugs.

Short communication: Bifidobacterium abundance in the faecal microbiota is strongly associated with milk traits in dairy cattle.

The performance of dairy cows is influenced by the microbial communities hosted within their digestive tract. While the rumen microbiota has long been associated with host phenotypes, the impact of the faecal microbiota remains elusive. In this study, we collected 697 faecal samples from commercial Holstein cows and analysed them with 16S rRNA gene analyses. For each animal, routinely recorded data, i.e., milk yield, fat yield, protein yield, fat content, protein content, and an aggregate production trait (pINEL) based on the French economic dairy index, were available to assess the links between the faecal microbiota and host production. Our findings revealed a strong and significant association between the structure of the bacterial and prokaryote community (ß-diversity) and dairy production. In addition, differential abundance analyses identified 48 genera whose abundances were significantly associated with pINEL, milk, fat and protein yield. Among these genera, the increased abundance of Bifidobacterium, and particularly an amplicon sequence variant with a 16S rRNA V3-V4 gene region identical to B. globosum and B. pseudolongum, was found to be the most important for high-yielding animals. Bifidobacterium seemed to be a potential key member of the bovine faecal microbiota that should be further investigated. Conversely, the p-1088-a5 gut group genus was found more abundant in low-productive cows. In conclusion, this study demonstrates significant associations between the faecal microbiota and the performance of dairy cows at the whole lactation scale. A better understanding of the physiology of the gut microbiota could help to improve dairy cow production.

Characterization of bovine vaginal microbiota using 16S rRNA sequencing: associations with host fertility, longevity, health, and production.

Due to their potential impact on the host's phenotype, organ-specific microbiotas are receiving increasing attention in several animal species, including cattle. Specifically, the vaginal microbiota of ruminants is attracting growing interest, due to its predicted critical role on cows' reproductive functions in livestock contexts. Notably, fertility disorders represent a leading cause for culling, and additional research would help to fill relevant knowledge gaps. In the present study, we aimed to characterize the vaginal microbiota of a large cohort of 1171 female dairy cattle from 19 commercial herds in Northern France. Vaginal samples were collected using a swab and the composition of the microbiota was determined through 16S rRNA sequencing targeting the V3-V4 hypervariable regions. Initial analyses allowed us to define the core bacterial vaginal microbiota, comprising all the taxa observed in more than 90% of the animals. Consequently, four phyla, 16 families, 14 genera and a single amplicon sequence variant (ASV) met the criteria, suggesting a high diversity of bacterial vaginal microbiota within the studied population. This variability was partially attributed to various environmental factors such as the herd, sampling season, parity, and lactation stage. Next, we identified numerous significant associations between the diversity and composition of the vaginal microbiota and several traits related to host's production and reproduction performance, as well as reproductive tract health. Specifically, 169 genera were associated with at least one trait, with 69% of them significantly associated with multiple traits. Among these, the abundances of Negativibacillus and Ruminobacter were positively correlated with the cows' performances (i.e., longevity, production performances). Other genera showed mixed relationships with the phenotypes, such as Leptotrichia being overabundant in cows with improved fertility records and reproductive tract health, but also in cows with lower production levels. Overall, the numerous associations underscored the complex interactions between the vaginal microbiota and its host. Given the large number of samples collected from commercial farms and the diversity of the phenotypes considered, this study marks an initial step towards a better understanding of the intimate relationship between the vaginal microbiota and the dairy cow's phenotypes.

A prospective observational study to evaluate the magnitude of temperature changes in children undergoing elective MRI under general anesthesia.

CONTEXT: Induction of general anesthesia and mandatory low-ambient temperature in the magnetic resonance imaging (MRI) suite renders the pediatric patient prone to fall in core temperature. Previously done studies have shown mixed results with core temperature showing both rise and fall. AIMS: The aim of this study is to evaluate which effect, hypothermia or hyperthermia, predominates in children anesthetized for MRI. Is the change in temperature the same across age groups and for different MRI scanners?. SETTINGS AND DESIGN: Prospective, observational study in a tertiary care teaching hospital. SUBJECTS AND METHODS: Two hundred and fifty children of age between 1 month and 16 years scheduled for MRI under propofol-based total intravenous anesthesia (TIVA) were recruited. A baseline core temperature (pre-scan) was recorded with the pediatric nasopharyngeal temperature probe after induction of anesthesia and also after the scan in the recovery room. RESULTS: The study shows that there is a significant fall in temperature of 1.022°C (CI = 0.964, 1.081) following MRI (P < 0.001) but the difference across different age groups and type of MRI scanner used are not significant. There is a significant correlation between duration in the MRI room and a decrease in temperature (P value = 0.003). Using simple linear regression analysis, it is found that if there is a 1-min increase in the duration of MRI, there is a decrease of 0.006°C in temperature. CONCLUSION: Vigilant temperature preservation strategies have to be maintained during the time the anesthetized child is present in the MRI suite. MRI compatible active warming devices are warranted especially in high turnover centers.

A review of methods for nematode identification.

Nematodes are non-segmented roundworms found in soil, aquatic environment, plants, or animals. Either useful or pathogenic, they greatly influence environmental equilibrium, human and animal health, as well as plant production. Knowledge on their taxonomy and biology are key issues to answer the different challenges associated to these organisms. Nowadays, most of the nematode taxonomy remains unknown or unclear. Several approaches are available for parasite identification, from the traditional morphology-based techniques to the sophisticated high-throughput sequencing technologies. All these techniques have advantages or drawbacks depending on the sample origin and the number of nematodes to be processed. This review proposes an overview of all newly available methods available to identify known and/or unknown nematodes with a specific focus on emerging high-throughput molecular techniques.

Assessment of quantitative models for plasmid ColE1 copy number control.

Two quantitative models of plasmid ColE1 copy number control are compared with respect to mathematical logic of derivation and application to experimental observations. Explanatory background material and clarifications are supplied for selected aspects of each model. Contrasting features are emphasized and experiments are suggested to distinguish between predictions of the models.

Analysis of establishment phase replication of the plasmid ColE1.

The replication regulatory mechanisms by which the small, multicopy plasmid ColE1 maintains a constant steady-state copy number have been extensively characterized by a combination of in vivo genetics and in vitro biochemistry. We have extended the analysis of replication control into the "establishment" phase of replication, when ColE1-directed replicons replicate more than once per cell generation and the intracellular concentrations of plasmid-encoded replication regulatory elements are changing. To study establishment phase replication, in which plasmid-directed replicons amplify from an initially low concentration to the characteristic, steady-state concentration, bacteriophage-plasmid hybrids, termed phasmids, were constructed. Phasmids were shown to exhibit stability, segregation, and incompatibility properties similar to those of the parent plasmid. Establishment phase replication was analyzed by measuring the number of phasmids per cell as a function of time after infection. We observed a linear increase in phasmid concentration until the steady-state concentration characteristic of the ColE1 plasmid component of the hybrid was reached. The number of cell doublings required for the phasmid concentration to reach steady-state was inversely related to cell growth rate. The observed amplification kinetics imply that the frequency of replication initiation per phasmid continually decreases until steady-state is reached. Kinetics of establishment phase amplification were sensitive to rate of expression of RNA II. A phasmid containing an up mutation in the RNA II promoter amplified at a 15-fold faster rate than the wild-type phasmid. Concentration of the ColE1 replication negative regulator (RNA I) was proportional to phasmid concentration throughout the amplification phase. These results suggest that the same elements that regulate steady-state replication also control establishment phase replication.

Phage display mutagenesis of the chimeric dual cytokine receptor agonist myelopoietin.

Myelopoietins comprise a class of chimeric cytokine receptor agonists consisting of an hIL-3 (human interleukin-3) receptor agonist and an hG-CSF (human granulocyte colony-stimulating factor) receptor agonist linked head-to-tail at their respective carboxy and amino termini. The combination of an early acting cytokine (hIL-3) with a late acting one (hG-CSF) allows efficient hematopoeitic reconstruction following myeloablative insult, and drives differentiation of non-myelocytic lineages (ie thrombocytic lineages) that are inaccessible using hG-CSF alone, in both preclinical models and clinical settings. A myelopoietin species was displayed and mutagenized on filamentous bacteriophage: both component agonists of myelopoietin were presented in biologically functional conformations as each recognized its corresponding receptor. Five amino acid positions in a short region of the hG-CSF receptor agonist module of myelopoietin that had been identified as important for proliferative activity were mutagenized. Display was used because it allows very 'deep' mutagenesis at selected residues: >10(5) substitution variants were affinity-screened using the hG-CSF receptor and 130 new, active variants of myelopoietin were identified and characterized. None of the selected variants were significantly more active than the parental myelopoietin species in a hG-CSF-dependent cell proliferation assay, though many were as active. Many of these relatively high-activity variants contained parental amino acids at several positions, suggesting the parental sequence may already be optimal at these positions for the assays used, and potentially accounting for the failure to identify enhanced bioactivity variants. Analysis of substitutions of high-activity variants complements and extends previous alanine scanning, and other genetic and biochemical data for hG-CSF variants.

CD80 and CD86 costimulatory molecules on circulating T cells of HIV infected individuals.

The expression of CD80 and CD86 costimulatory molecules, typical for antigen presenting cells (APC), was measured on circulating T cells of 20 HIV-infected individuals and of 11 HIV-negative healthy controls. The CD80 and CD86 molecules were present on both circulating T subsets of HIV-infected individuals (mean of CD80 expression within CD4+ T cells [CD80/CD4]: 5.0%; and CD86/CD4: 2.6%; CD80/CD8 4.1% and CD86/CD8: 2.7%) and were associated with HLA-DR expression. Some CD80 and CD86 expression was also found in normal controls, and only the expression of CD86 was significantly (P < 0.05) increased on CD4 + and CD8 + T cells of HIV-infected individuals. The expression of CD28 was decreased on T cells of HIV-infected individuals and was negatively correlated to the expression of HLA-DR and CD86 (mean CD28 within CD3+T cells: HIV+ 29.5%, HIV - 67.6%; correlation coefficient, - 0.75 and - 0.71, respectively). The more the disease proceeds, the less CD28 and the more DR and CD86 are found on circulating T cells. This suggests that during HIV infection T cells themselves develop an antigen presenting phenotype by upregulating expression of HLA-DR, CD86 and CD80 molecules.

Improved identification of Staphylococcus aureus using a new agglutination test. Results of an international study.

A new reagent for the identification of Staphylococcus aureus, SLIDEX STAPH-KIT, operates on the principle of a latex and red blood cell combination agglutination: red blood cells are coated with fibrinogen for the detection of clumping factor, and latex particles are sensitized with anti-S. aureus serotype 18 monoclonal antibody for the detection of protein A and antigen 18. French strains belonging to serotype 18 are methicillin-resistant. The performance of this reagent was compared with STAPHYSLIDE and STAPHAUREX in Europe (France, Germany, Italy), in the United States and in Japan using 548 methicillin-resistant S. aureus strains, 392 methicillin-sensitive S. aureus strains, and 441 non-aureus staphylococci. The specificity of the three reagents was equivalent (98.8% for SLIDEX STAPH-KIT, 99.1% for STAPHYSLIDE, 98.1% for STAPHAUREX). SLIDEX STAPH-KIT (97.3%) was more sensitive than STAPHYSLIDE (93.5%) and STAPHAUREX (89.7%) for all S. aureus strains due to a higher rate of identified methicillin-resistant S. aureus strains.

Increased urine histamine and contrast media reactions.

Urine samples were collected from 200 subjects undergoing intravenous pyelography. The urine histamine levels were compared with those of 132 normal control subjects, 11 subjects with systemic mastocytosis, six subjects with idiopathic anaphylaxis, and ten subjects experiencing mild anaphylactic reactions during allergy immunotherapy. Compared with normal controls, as a group, all subjects receiving intravenous contrast media had increased urine histamine (P less than 0.05 by Student's t-test) while those subjects experiencing adverse reactions had considerably larger increases. The urine histamine levels in the subjects experiencing systemic reactions were in the same range as those observed in patients having mild anaphylactic reactions to immunotherapy and somewhat lower than those found in idiopathic anaphylaxis or systemic mastocytosis. These data suggest that some histamine release accompanies infusions of contrast media in all subjects and that larger amounts of histamine release are associated with adverse reactions.

Neurogenic erectile dysfunction: the course of nicotinamide adenine dinucleotide phosphate diaphorase-positive nerve fibers on the surface of the prostate.

OBJECTIVES: Nitric oxide (NO) plays an important role as a neurotransmitter in the peripheral neural regulation of erection. A histochemical stain able to localize fibers releasing NO in combination with an in vivo study were used to evaluate the course and physiologic significance of the nerve fibers innervating the cavernous smooth muscle. METHODS: Morphologic studies in 6 rats and 6 human cadavers were performed, tracing the course of branches of the cavernous nerve branches using a nicotinamide adenine dinucleotide phosphate diaphorase staining technique. Electrostimulations in rats were performed before and after transection of the anterolateral part of the prostate capsule. RESULTS: Multiple nerve fibers were documented running on the lateral and ventral surfaces of the prostate distinct from the classically described dorsolateral neurovascular bundle. Transection of these fibers resulted in a loss of electrically induced intracavernous pressure (59.4 +/- 5.6 cm H2O versus 27.0 +/- 4.6 cm H2O). CONCLUSIONS: These preliminary morphologic and physiologic studies support a significant role for these nerve fibers in erection.

7-Nitroindazole: a selective inhibitor of penile erection: an in vivo study in a rat animal model.

OBJECTIVES: The neuronal isoform of nitric oxide synthase (nNOS) has been localized in neurons innervating the penis and is believed to be an important mediator of erection. Using the selective inhibitor 7-nitroindazole (7-NI), we evaluated the possible role of nNOS in penile erection using a rat animal model. METHODS: Eighteen Sprague-Dawley rats were divided into three study groups. A sham group (n = 6) received the vehicle arachis oil, a low-dose group received 5 mg/kg (n = 6), and a high-dose group received 50 mg/kg (n = 6) of 7-NI prior to measurement of blood pressure and cavernous nerve stimulation-induced rise in intracavernous pressure. RESULTS: A dose-dependent inhibition of erection by 7-NI was seen. Control animals had an intracavernous pressure rise of 55.5 +/- 4.0 cm H2O, whereas the low-dose group had 26.5 +/- 2.8 cm H2O and the high-dose group had 6.2 +/- 2.1 cm H2O. A partial recovery of erection was seen in the low- and high-dose groups after 3 hours. Blood pressure was unaffected by 7-NI administration. CONCLUSIONS: 7-NI induced a reversible, dose-dependent inhibition of erection without affecting blood pressure. This in vivo study provides further evidence of the role played by nNOS in erection.

Evaluation of a no-needle penile injector: a preliminary study evaluating tissue penetration and its hemodynamic consequences in the rat.

OBJECTIVES: Intracavernous needle injection is an effective delivery method for pharmacotherapy of erectile dysfunction. Needle phobia, pain, and concern about local tissue injury have stimulated the search for new, less invasive means of inducing penile erection. In this preliminary communication, we evaluate a jet injector as an alternative to needle injection for intracavernous delivery of vasoactive drugs. METHODS: Jet injection was evaluated in three groups of rats receiving either India ink, saline, or papaverine into the penis. The ability of the jet injection to penetrate through the tunica albuginea and deliver liquid to the corpora cavernosa smooth muscle was assessed by the degree of staining within the corpus cavernosum (ink group), histologic change (saline group), and rise in intracavernous pressure (papaverine group). Erectile capacity following cavernous nerve electric stimulation was compared before and 1 hour after injection of saline or papaverine. RESULTS: Ink traversed the skin and tunica albuginea with extensive deposition noted within the cavernous spaces. Varying degree of subcutaneous hemorrhage were seen with saline jet injection; however, the corpus cavernous smooth muscles showed no evidence of injury. Jet injection of papaverine 3250 micrograms significantly increased cavernous pressure (39.4 +/- 4.6 cm H2O) compared with saline injection (2.8 +/- 1.3 cm H2O). CONCLUSIONS: We conclude that acute jet injection is an effective method for intracavernous delivery of drugs. Long-term effects should be evaluated prior to clinical use.

New insights into the role of endothelin-1 in radiation-associated impotence.

The objectives of this work were to: (1) Determine if prostate and penile tissue levels of endothelin-1 (ET-1) are increased in a rat following pelvic irradiation. (2) Determine if an ETa receptor antagonist (BQ-123) potentiates erectile function in these irradiated animals. Rats were divided into three study groups: control, 1000 cGy and 2000 cGy. The experimental groups received a single dose of radiation to the pelvic region. A time course was established to measure the effects of irradiation on prostate and penile tissue levels of endothelin-1 (ET-1)-like immunoreactivity. The effect of intracavernous injection of BQ-123 (25 microg/30 microl) was evaluated by measuring intracavernous pressure (ICP) following cavernous nerve electrical field stimulation. In the 2000 cGy group, a significant rise in ET-1-like immunoreactivity tissue levels was observed at 20 days. A significant decrease in ICP was recorded in the 1000 and 2000 cGy irradiated rats compared to the control group. Only the 2000 cGy group had a significant improvement in erectile function following BQ-123 administration. A significant improvement was observed 20 min post-administration, lasted 90 min, and was back to pre-administered levels at 120 min. The conclusion made was that radiation-induced impotence in irradiated rats is associated with an increased production of ET-1. Preliminary results are suggestive that ETa receptor antagonist may be of use to reverse such radiation-induced impotence in these irradiated animals.

Phage presentation and affinity selection of a deletion mutant of human interleukin-3.

A deletion derivative of the cytokine human interleukin-3 (hIL-3(15-125), comprising amino acids 15-125 of the native protein) was produced as a fusion to the filamentous phage surface protein pIII. The cytokine was detected in association with phage particles by protein immunoblotting. Compared to an equivalent quantity of soluble-cytokine, phage-presented hIL-3(15-125) exhibited reduced biological activity in a hIL-3-dependent cell proliferation assay. The reduction in activity was attributable to presence of phage particles in the assay, rather than directly owing to physical incorporation of the cytokine into the phage particle. Owing to the position of the amber codon in the phagemid vector, the phagemid-produced free hIL-3(15-125) species (designated hIL-3(15-125) epsilon) had 20 amino acids appended to its C-terminus; hIL-3(15-125) epsilon did not exhibit reduced bioactivity. hIL-3(15-125)-presenting phage were affinity-selected with either a hIL-3-reactive polyclonal antibody or with cells expressing the heterodimeric hIL-3 receptor. These data are consistent with the use of phage-display technology for the affinity selection of hIL-3 variants with modified biological properties.

Monitoring of four DNA extraction methods upstream of high-throughput sequencing of Anisakidae nematodes.

Different methods were evaluated to extract DNA from pooled nematodes belonging to Anisakis, Contracaecum, Pseudoterranova and Hysterothylacium genera isolated from edible fish. Pooled DNA extraction is the first and compulsory step to allow the identification of a large number of samples through high-throughput DNA sequencing with drastic time and cost reductions.