Detection and genotyping of Chlamydia species responsible for reproductive disorders in Algerian small ruminants.

Chlamydiosis in small ruminants is a zoonotic disease mainly related to Chlamydia abortus. This bacterium is responsible for abortions and reproductive disorders in sheep and goats. Stillbirth and infertility, leading to important economic losses, are also associated with this pathology. In Algeria, abortion cases are frequently reported by veterinarians but, except for brucellosis which is a notifiable disease in this country, abortive diseases are in general poorly studied. In order to detect and genotype Chlamydia species in small ruminants in different areas of Algeria, a study was conducted on samples collected from females (164 blood samples and 199 vaginal swabs) between October 2011 and March 2013. Serum samples were tested with a C. abortus-specific indirect ELISA test. Fourteen samples (8.5 %), from six farms (6/20, 30 %) were tested positive. Vaginal swabs were analysed with a real-time PCR targeting all Chlamydiaceae spp. Thirty samples (15 %) were diagnosed positive in 16 farms (16/25, 64 %). Positive samples were all re-tested with a C. abortus- and a C. pecorum-specific real-time PCR. Finally, 13/30 (43.3 %) and 6/30 (20 %) were identified as C. abortus and C. pecorum, respectively. Enough concentrated C. abortus samples were genotyped by multi-loci variable number of tandem repeat (VNTR) analysis (MLVA), and all were related to the genotype [2] group which mainly includes French C. abortus isolates. C. pecorum-positive samples were genotyped by multi-locus sequence typing (MLST). Interestingly, two of them were successfully genotyped and showed identical MLST sequences to VB2, AB10, E58 and SBE, a group which includes C. pecorum isolates considered as highly pathogenic. These findings suggest a possible role of C. abortus and C. pecorum strains in the aetiology of abortion in Algerian small ruminants.

Multilocus variable-number tandem-repeat analysis scheme for chlamydia felis genotyping: comparison with multilocus sequence typing.

Chlamydia felis is an important ocular pathogen in cats worldwide. A multilocus variable-number tandem-repeat analysis (MLVA) system for the detection of tandem repeats across the whole genome of C. felis strain Fe/C-56 was developed. Nine selected genetic loci were tested by MLVA in 17 C. felis isolates, including the C. felis Baker vaccine strain, and 122 clinical samples from different geographic origins. Analysis of the results identified 25 distinct C. felis MLVA patterns. In parallel, a recently described multilocus sequence typing scheme for the typing of Chlamydia was applied to 13 clinical samples with 12 different C. felis MLVA patterns. Rare sequence differences were observed. Thus, the newly developed MLVA system provides a highly sensitive high-resolution test for the differentiation of C. felis isolates from different origins that is suitable for molecular epidemiological studies.

Public interest and awareness regarding general health, sleep quality and mental wellbeing during the early COVID-19 pandemic period: An exploration using Google trends.

The internet network continues to be a major source of health-related information. Our research provides insights into the online health-seeking behaviors of the general population, and evaluates the potential relationship between the COVID-19 pandemic and public interest and awareness of general sleep health, mental health and wellbeing. Google Trends' weekly relative search volumes (RSVs) were examined during 2020 for searches specifically related to COVID-19 symptoms, and for searches related to general health, sleep and wellbeing, in the United Kingdom, the United States of America, France, Italy and Japan. To obtain insight into the association between the initiation of public restrictions and online search trends, we assessed a six-week period; the 'early pandemic period' (EPP) (01 March 2020 - 11 April 2020). To provide a meaningful pre-pandemic comparison, a similar period during 2019 (03 March - 13 April 2019) was compared for RSV and median difference analysis. The EPP was associated with increased online searches related to COVID-19 symptoms, as compared with those related to more general sleep health, mental health and wellbeing. The latter search terms frequently showed a decrease or minimal change in RSV during the EPP compared with the equivalent period in 2019. This finding illustrates the potential link between the COVID-19 pandemic and online search behavior and corroborates existing findings regarding internet searches during this period. Proactive communication by healthcare professionals during future pandemics and as an ongoing measure could help prevent public neglect of general health and wellbeing symptoms, and encourage reporting and early intervention.

Effect of a Combination of Magnesium, B Vitamins, Rhodiola, and Green Tea (L-Theanine) on Chronically Stressed Healthy Individuals-A Randomized, Placebo-Controlled Study.

The effect of a combination of magnesium, vitamins B6, B9, B12, rhodiola and green tea/L-theanine (Mg-Teadiola) on stress was evaluated in chronically stressed, otherwise healthy individuals. Effects on stress-related quality-of-life parameters (sleep and perception of pain) were also explored. Adults with stress for ≥1 month, scoring ≥14 points on the Depression Anxiety Stress Scale (DASS)-42 questionnaire, were randomized (1:1) to receive oral Mg-Teadiola (n = 49) or a placebo (n = 51), for 28 days, with a follow-up assessment on Day 56 (NCT04391452). The primary endpoint was the change in the DASS-42 stress score from baseline to Day 28 with Mg-Teadiola versus placebo. The DASS-42 stress scores significantly decreased from baseline to Day 28 with Mg-Teadiola versus placebo (effect size, 0.29; 95% CI [0.01, 0.57]; p = 0.04). Similar reductions were observed on Day 14 (p = 0.006) and Day 56 (p = 0.02). A significant reduction in sensitivity to cold pain (p = 0.01) and a trend for lower sensitivity to warm pain was observed (p = 0.06) on Day 28. Improvements in daytime dysfunction due to sleepiness (Pittsburgh Sleep Quality Index-7 component score) were reported on Day 28, and were significant on Day 56 (p < 0.001). Mg-Teadiola is effective in managing stress in otherwise healthy individuals. Its beneficial effects on sleep and pain perception need further investigation.

A genetic variant of Burkholderia mallei detected in Kuwait: Consequences for the PCR diagnosis of glanders.

Glanders is a contagious zoonotic disease caused by Burkholderia mallei. Following the detection of glanders positive horses using the OIE complement fixation test, the tissues of two horses were analysed by PCR. While PCR systems targeting the Burkholderia pseudomallei complex gave positive signals, the species-specific PCR systems targeting B. mallei (fliP-IS407A) and B. pseudomallei (orf11)-the OIE recommended targets-resulted in negative signals. However, the presence of B. mallei in these tissues was confirmed with a recently described B. mallei-specific real-time PCR system and genotyping with MLST- and SNP-based methods, performed on the most positive tissue, identified a genotype closely related to B. mallei strains recently isolated in the Middle East. This study leads to recommendations regarding the use of PCR systems for the molecular diagnosis of glanders, especially in regions where the circulating B. mallei strains have not yet been fully genetically characterized.

Chlamydial infections in duck farms associated with human cases of psittacosis in France.

Five severe cases of psittacosis in individuals associated with duck farms were notified in France between January and March 2006. Diagnostic examination included serology and/or molecular detection by PCR from respiratory samples. As a consequence, we investigated all duck flocks (n=11) that were housed in the three farms where human infections occurred. While serology by complement fixation test was negative for all samples, cloacal and/or tracheal chlamydial excretion was detected by PCR in all three units. Notably, one duck flock was tested strongly positive in 2 of the 3 affected farms, and Chlamydophila (C.) psittaci strains were isolated from cloacal and/or tracheal swab samples from both farms. Human samples and duck isolates exhibited the same PCR-RFLP restriction pattern, which appeared to be an intermediate between genotypes A and B. Analysis of ompA gene sequences and comparison to those of the type strains showed that the isolates could not be strictly assigned to any of the generally accepted genotypes of C. psittaci. Further analysis by MLVA of the PCR-positive human samples revealed two distinct patterns, which were related to previously isolated C. psittaci duck strains.

Effects of heart rate variability biofeedback training in athletes exposed to stress of university examinations.

INTRODUCTION: Heart rate variability biofeedback (HRV-BFB) training, a method whereby one controls an unusually low breathing rate to reach cardiac coherence, has been shown to reduce anxiety and improve cardiac autonomic markers in diseased people, but much less is known about HRV-BFB benefits in healthy people. Here we investigated potential benefits in young competitors experiencing stress during university examinations as well as persistence of benefits after HRV-BFB training cessation. METHODS: A group of sports students (n = 12) practiced 5-min HRV-BFB training twice a day for 5-weeks using URGOfeel® (URGOTECH) and was compared to a control group (n = 6). University examinations occurred immediately after HRV-BFB training (Exam1), then 12-weeks later (Exam2). Anxiety markers and cardiac autonomic markers were assessed at baseline, Exam1 and Exam2. Principal Component Analyses (PCA) that combined all these markers were computed at Exam1 and Exam2 to emphasize covariations. RESULTS: At Exam 1, immediately after HRV-BFB training cessation, the experimental group demonstrated greater autonomic markers but similar states of anxiety when compared to the Control group. Twelve weeks later at Exam2, autonomic markers were greater and anxiety scores were lesser among the experimental group. PCA highlighted covariations only within cardiac autonomic markers at Exam1. Rather, variations in cardiac markers were associated with anxiety markers at Exam2. CONCLUSION: Short sessions of HRV-BFB training for a brief period of 5 weeks bring substantial benefits to autonomic markers and anxiety levels in young competitors. Here beneficial effects persisted for 12 weeks. Dissociated profiles of anxiety and cardiac autonomic adaptations shed new light on the role of the amygdala in heart-brain interactions after cardiac coherence training.

Randomized clinical trial: efficacy of a food supplement, TRANSITECH, on healthy individuals with mild intermittent constipation.

BACKGROUND: Constipation is a common disorder in the general population and can be observed in healthy individuals. A natural product leading to an increase in bowel movements and decrease in colonic transit time (CTT), without bloating, could be useful for the patient's care. OBJECTIVES: To investigate the effects of TRANSITECH, a food supplement composed of plants and lactic ferments, on bowel movements, CTT and bloating. METHODS: A total of 100 healthy participants, presenting two to five stools per week, were selected and followed over a 6-day baseline period. They were randomly assigned to receive daily two tablets of TRANSITECH or placebo during 10 days. They were then followed up over 28 days after intervention. Participants daily recorded in a home questionnaire the characteristics of stools (frequency and consistency), and the importance of bloating during the preintervention period (from D-6 to D0), the intervention period (from D0 to D10) and the postintervention period (from D10 to D38). Their CTTs were also evaluated by following the propagation of radiopaque markers at D0 and D10. RESULTS: At D10, the food supplement group showed, compared with the placebo group, higher daily stool emission (0.95±0.50, 0.70±0.20, P<0.001), softer stool consistency (2.5±0.6 vs. 3.0±0.8, P<0.001) and lower CTT (33.8±28.2 vs. 56.4±36.2 h, P=0.01). The active group also showed a sustained increase in daily stool emissions observed at D38 compared with D0 (P=0.03). CONCLUSION: TRANSITECH is an efficient natural solution for the treatment of constipation. It increases the number of bowel movements, decreases the oroanal and segmental CTT, is well tolerated, and presents sustained effects after treatment completion.

A longitudinal study of the Mycobacterium avium subspecies paratuberculosis infection status in young goats and their mothers.

The dynamics between Mycobacterium avium subspecies paratuberculosis (MAP) infection and the immune response of goats naturally exposed to MAP were studied in a herd where the clinical expression of paratuberculosis had been observed. Four generations of goats were observed over a 33-month period: mothers of three different generations (G1, G2, G3) and their daughters, generation 4 (G4). A MAP infection status was defined according to the combined results of an IFN-γ assay, antibody response, faecal culture and post-mortem examination. Goats were defined as non-infected (NI), infected and non-shedder (INS), infected and shedder (IS) or atypical (A). Twenty-nine percent of goats were NI, 66% were infected and either shedding (14%) or not shedding (52%) MAP, and 5% were atypical. IFN-γ responses were detected first, followed by faecal shedding and antibody responses. The results showed that in goats naturally exposed to MAP, IFN-γ responses were regularly detected earlier in non-shedders than in young infected shedder goats and were stronger in shedder than in non-shedder goats. They were also higher in the mother goats than in their daughters. Goats shedding MAP or with positive antibody response at the beginning of their pregnancy are more likely to have an infected daughter positive to an IFN-γ assay by the age of 15 months.

Detection of atypical Chlamydiaceae in roe deer (Capreolus capreolus).

Investigations on fecal samples, vaginal swabs and sera from roe deer (Capreolus capreolus) in south-western France led to the detection of a non-classified Chlamydiaceae strain. A total of 85 vaginal swabs were sampled from roe deer that had been captured in 2012 (n=42) and 2013 (n=43). Using a Chlamydiaceae family-specific real-time PCR, only one vaginal swab out of the 42 samples done in 2012 tested positive and was subsequently identified as Chlamydia (C.) psittaci. In contrast, 6/43 vaginal swab samples were positive in 2013. Four of these positive samples came from a single group of roe deer, captured in the Fabas plain. Fecal samples from this group of 9 females were subsequently analyzed, with 6 of them testing positive with the Chlamydiaceae-specific PCR. All positive samples collected in 2013 were negative when re-tested with C. abortus-, C. pecorum- and C. suis-specific real-time PCR assays. Sera from this group of 9 females were analyzed with two immunoassays (recomLine and ELISA). Whereas intense positive reactions with C. pneumoniae antigens were observed for all sera when tested with the recomLine test, none was positive with the C. abortus specific ELISA test. Comparative sequence analysis of the 16S, 23S rRNA and ompA gene sequences from 3 animals, as well as the MLST analysis from 2 animals, showed that this roe deer group likely harbored the same bacterium related to members of the family Chlamydiaceae. Notably, the roe deer strain formed a separate entity different from the currently recognized chlamydial species, with C. trachomatis, C. suis and C. muridarum appearing as its closest relatives.

Agreement between indirect calorimetry and traditional tests of lactose malabsorption.

BACKGROUND: Lactose malabsorption occurs frequently and the variable consequent intolerance may seriously impair quality of life. No reliable and convenient test method is in routine clinical practice. A recent animal study showed that the respiratory quotient changed significantly after ingestion of sucrose and lactose in naturally lactase-deficient rats. AIMS: This exploratory study evaluated the relevance of monitoring the respiratory quotient after lactose ingestion to detect malabsorption. METHODS: Healthy volunteers were identified and classified lactose absorbers and malabsorbers by a lactose tolerance test (25 g). After an overnight fast, a second lactose challenge was performed to monitor hydrogen excretion and respiratory quotient kinetics over 4h. Participants also completed questionnaires to score and localise their gastrointestinal symptoms. RESULTS: 20 subjects were enrolled (10 per group, 60% males, mean age 34 ± 4 years). Respiratory quotient kinetics were different between absorbers and malabsorbers during the first 100 min after lactose ingestion (p<0.01) and during the initial 30-50 min period. Respiratory quotient was significantly, positively correlated to peak glycaemia (R=0.74) and negatively correlated to hydrogen excretion (R=-0.51) and symptoms score (R=-0.46). CONCLUSIONS: Indirect calorimetry could improve the reliability of lactose malabsorption diagnosis. Studies on larger populations are needed to confirm the validity of this test and propose a simplified measurement.

Genotyping of Chlamydophila abortus strains by multilocus VNTR analysis.

Chlamydophila (C.) abortus is the causative agent of ovine enzootic abortion with zoonotic potential whose epidemiology has been held back because of the obligate intracellular habitat of the bacterium. In the present study, we report on a molecular typing method termed multiple loci variable number of tandem repeats (VNTR) Analysis (MLVA) for exploring the diversity of C. abortus. An initial analysis performed with 34 selected genetic loci on 34 ruminant strains including the variant Greek strains LLG and POS resulted in the identification of five polymorphic loci, confirming the widely held notion that C. abortus is a very homogeneous species. Analysis of additional 111 samples with the selected five loci resulted in the classification of all strains into six genotypes with distinct molecular patterns termed genotypes [1] through [6]. Interestingly, the classification of the isolates in the six genotypes was partly related to their geographical origin. Direct examination of clinical samples proved the MLVA to be suitable for direct typing. Analysis of the genomic sequences in six C. abortus prototypes of amplicons generated with each of the five selected VNTR primers revealed that variation between genotypes was caused by the presence or absence of coding tandem repeats in three loci. Amplification of Chlamydophila psittaci reference strains with the five selected VNTR primers and of the six C. abortus prototype strains with the eight VNTR primers established for the typing of C. psittaci [Laroucau, K., Thierry, S., Vorimore, F., Blanco, K., Kaleta, E., Hoop, R., Magnino, S., Vanrompay, D., Sachse, K., Myers, G.S., Bavoil, P.M., Vergnaud, G., Pourcel, C., 2008. High resolution typing of Chlamydophila psittaci by multilocus VNTR analysis (MLVA). Infect. Genet. Evol. 8(2), 171-181] showed that both MLVA typing systems were species-specific when all respective VNTR primer sets were used. In conclusion, the newly developed MLVA system provides a highly sensitive, high-resolution and easy-to-perform tool for the differentiation of C. abortus isolates of different origin, which is suitable for molecular epidemiological studies.