Dynamics of antibody response and bacterial shedding of Mycoplasma hyorhinis and M. hyosynoviae in oral fluids from experimentally inoculated pigs.

Mycoplasma hyorhinis (Mhr) and M. hyosynoviae (Mhs) are commensal organisms of the upper respiratory tract and tonsils but may also cause arthritis in pigs. In this study, 8-week-old cesarean-derived colostrum-deprived (CDCD) pigs (n = 30; 3 groups, 10 pigs per group, 2 pigs per pen) were inoculated with Mhr, Mhs, or mock-inoculated with culture medium and then pen-based oral fluids were collected at different time points over the 56 days of the experimental study. Oral fluids tested by Mhr and Mhs quantitative real-time PCRs revealed Mhr DNA between day post inoculation (DPI) 5-52 and Mhs DNA between DPI 5-15. Oral fluids were likewise tested for antibody using isotype-specific (IgG, IgA, IgM) indirect ELISAs based on a recombinant chimeric polypeptide of variable lipoproteins (A-G) for Mhr and Tween 20-extracted surface proteins for Mhs. Mhr IgA was detected at DPI 7 and, relative to the control group, significant (p < 0.05) antibody responses were detected in the Mhr group between DPI 12-15 for IgM and DPI 36-56 for both IgA and IgG. In the Mhs group, IgM was detected at DPI 10 and significant (p < 0.05) IgG and IgA responses were detected at DPI 32-56 and DPI 44-56, respectively. This study demonstrated that oral fluid could serve as an effective and convenient antemortem sample for monitoring Mhr and Mhs in swine populations.

Bacterial shedding and serologic responses following an outbreak of Salmonella Typhi in an endemic cohort.

BACKGROUND: Salmonella enterica serovar Typhi (Salmonella Typhi) is the cause of typhoid fever. Salmonella Typhi may be transmitted through shedding in the stool, which can continue after recovery from acute illness. Shedding is detected by culturing stool, which is challenging to co-ordinate at scale. We hypothesised that sero-surveillance would direct us to those shedding Salmonella Typhi in stool following a typhoid outbreak. METHODS: In 2016 a typhoid outbreak affected one in four residents of a Nursing School in Malosa, Malawi. The Department of Health asked for assistance to identify nursing students that might spread the outbreak to other health facilities. We measured IgG antibody titres against Vi capsular polysaccharide (anti-Vi IgG) and IgM / IgG antibodies against H:d flagellin (anti-H:d) three and six months after the outbreak. We selected participants in the highest and lowest deciles for anti-Vi IgG titre (measured at visit one) and obtained stool for Salmonella culture and PCR. All participants reported whether they had experienced fever persisting for three days or more during the outbreak (in keeping with the WHO definitions of 'suspected typhoid'). We tested for salmonellae in the Nursing School environment. RESULTS: We obtained 320 paired serum samples from 407 residents. We cultured stool from 25 residents with high anti-Vi IgG titres and 24 residents with low titres. We did not recover Salmonella Typhi from stool; four stool samples yielded non-typhoidal salmonellae; one sample produced a positive PCR amplification for a Salmonella Typhi target. Median anti-Vi and anti-H:d IgG titres fell among participants who reported persistent fever. There was a smaller fall in anti-H:d IgG titres among participants who did not report persistent fever. Non-typhoidal salmonellae were identified in water sampled at source and from a kitchen tap. CONCLUSION: High titres of anti-Vi IgG did not identify culture-confirmed shedding of Salmonella Typhi. There was a clear serologic signal of recent typhoid exposure in the cohort, represented by waning IgG antibody titres over time. The presence of non-typhoidal salmonellae in drinking water indicates sub-optimal sanitation. Developing methods to detect and treat shedding remains an important priority to complement typhoid conjugate vaccination in efforts to achieve typhoid elimination.

Neisseria gonorrhoeae Coinfection during Chlamydia muridarum Genital Latency Does Not Modulate Murine Vaginal Bacterial Shedding.

Chlamydia trachomatis and Neisseria gonorrhoeae are the most frequently reported agents of bacterial sexually transmitted disease worldwide. Nonetheless, C. trachomatis/N. gonorrhoeae coinfection remains understudied. C. trachomatis/N. gonorrhoeae coinfections are more common than expected by chance, suggesting C. trachomatis/N. gonorrhoeae interaction, and N. gonorrhoeae infection may reactivate genital chlamydial shedding in women with latent (quiescent) chlamydial infection. We hypothesized that N. gonorrhoeae would reactivate latent genital Chlamydia muridarum infection in mice. Two groups of C. muridarum-infected mice were allowed to transition into genital latency. One group was then vaginally inoculated with N. gonorrhoeae; a third group received N. gonorrhoeae alone. C. muridarum and N. gonorrhoeae vaginal shedding was measured over time in the coinfected and singly infected groups. Viable C. muridarum was absent from vaginal swabs but detected in rectal swabs, confirming C. muridarum genital latency and consistent with the intestinal tract as a C. muridarum reservoir. C. muridarum inclusions were observed in large intestinal, but not genital, tissues during latency. Oviduct dilation was associated with C. muridarum infection, as expected. Contradicting our hypothesis, N. gonorrhoeae coinfection did not reactivate latent C. muridarum vaginal shedding. In addition, latent C. muridarum infection did not modulate recovery of vaginal viable N. gonorrhoeae. Evidence for N. gonorrhoeae-dependent increased C. muridarum infectivity has thus not been demonstrated in murine coinfection, and the ability of C. muridarum coinfection to potentiate N. gonorrhoeae infectivity may depend on actively replicating vaginal C. muridarum. The proportion of mice with increased vaginal neutrophils (PMNs) was higher in N. gonorrhoeae-infected than in C. muridarum-infected mice, as expected, while that of C. muridarum/N. gonorrhoeae-coinfected mice was intermediate to the singly infected groups, suggesting latent C. muridarum murine infection may limit PMN response to subsequent N. gonorrhoeae infection. IMPORTANCE Our work builds upon the limited understanding of C. muridarum/N. gonorrhoeae coinfection. Previously, N. gonorrhoeae infection of mice with acute (actively replicating) vaginal C. muridarum infection was shown to increase recovery of viable vaginal N. gonorrhoeae and vaginal PMNs, with no effect on C. muridarum vaginal shedding (R. A. Vonck et al., Infect Immun 79:1566-1577, 2011). It has also been shown that chlamydial infection of human and murine PMNs prevents normal PMN responses, including the response to N. gonorrhoeae (K. Rajeeve et al., Nat Microbiol 3:824-835, 2018). Our findings show no effect of latent genital C. muridarum infection on the recovery of viable N. gonorrhoeae, in contrast to the previously reported effect of acute C. muridarum infection, and suggesting that acute versus latent C. muridarum infection may have distinct effects on PMN function in mice. Together, these studies to date provide evidence that Chlamydia/N. gonorrhoeae synergistic interactions may depend on the presence of replicating Chlamydia in the genital tract, while chlamydial effects on vaginal PMNs may extend beyond acute infection.

Statistical modeling based on structured surveys of Australian native possum excreta harboring Mycobacterium ulcerans predicts Buruli ulcer occurrence in humans.

Background: Buruli ulcer (BU) is a neglected tropical disease caused by infection of subcutaneous tissue with Mycobacterium ulcerans. BU is commonly reported across rural regions of Central and West Africa but has been increasing dramatically in temperate southeast Australia around the major metropolitan city of Melbourne, with most disease transmission occurring in the summer months. Previous research has shown that Australian native possums are reservoirs of M. ulcerans and that they shed the bacteria in their fecal material (excreta). Field surveys show that locales where possums harbor M. ulcerans overlap with human cases of BU, raising the possibility of using possum excreta surveys to predict the risk of disease occurrence in humans. Methods: We thus established a highly structured 12 month possum excreta surveillance program across an area of 350 km2 in the Mornington Peninsula area 70 km south of Melbourne, Australia. The primary objective of our study was to assess using statistical modeling if M. ulcerans surveillance of possum excreta provided useful information for predicting future human BU case locations. Results: Over two sampling campaigns in summer and winter, we collected 2,282 possum excreta specimens of which 11% were PCR positive for M. ulcerans-specific DNA. Using the spatial scanning statistical tool SaTScan, we observed non-random, co-correlated clustering of both M. ulcerans positive possum excreta and human BU cases. We next trained a statistical model with the Mornington Peninsula excreta survey data to predict the future likelihood of human BU cases occurring in the region. By observing where human BU cases subsequently occurred, we show that the excreta model performance was superior to a null model trained using the previous year's human BU case incidence data (AUC 0.66 vs 0.55). We then used data unseen by the excreta-informed model from a new survey of 661 possum excreta specimens in Geelong, a geographically separate BU endemic area to the southwest of Melbourne, to prospectively predict the location of human BU cases in that region. As for the Mornington Peninsula, the excreta-based BU prediction model outperformed the null model (AUC 0.75 vs 0.50) and pinpointed specific locations in Geelong where interventions could be deployed to interrupt disease spread. Conclusions: This study highlights the One Health nature of BU by confirming a quantitative relationship between possum excreta shedding of M. ulcerans and humans developing BU. The excreta survey-informed modeling we have described will be a powerful tool for the efficient targeting of public health responses to stop BU. Funding: This research was supported by the National Health and Medical Research Council of Australia and the Victorian Government Department of Health (GNT1152807 and GNT1196396).

Effect of intramuscular vaccination against Lawsonia intracellularis on production parameters, diarrhea occurrence, antimicrobial treatment, bacterial shedding, and lean meat percentage in two Danish naturally infected finisher pig herds.

Lawsonia intracellularis (LI) is an economically important enteric pathogen in pigs with a worldwide endemic prevalence. The objective of this study was to evaluate the effect of an intramuscularly administrated LI vaccine (Porcilis®Lawsonia Vet.) in Danish finisher pigs (30-115 kg) measured on key production figures, antimicrobial (AB) treatments, occurrence of diarrhea and LI shedding. The study was a group-randomized block-trial with parallel groups in two herds, Herd 1 and Herd 2, experiencing a natural subclinical-clinical LI infection in early finisher period. Vaccination occurred at weaning, but the study focused on the first eight weeks in the finisher period. Further, slaughterhouse data were included. In total, 52 and 50 finisher pens comprising 2184 and 2254 finisher pigs were included in each of two herds, respectively. LI vaccination significantly reduced feed conversion ratio (FCR) by 0.05 and 0.09 FU/kg (p = 0.007 and p < 0.001) alongside a significantly increased average daily weight gain (ADWG) by 31 and 43 gr/day (p = 0.001 and p < 0.001) in each of the herds, respectively. In the vaccinated group, less variation was found in ADWG compared to the control group (p < 0.001 in both herds) as an expression of a more uniform growth, which was further confirmed by less variation in lean meat percent in the vaccinated group in one herd (p = 0.007). No significant difference between groups were found in mortality and pigs excluded due to welfare reasons. AB flock treatment against diarrhea was significantly reduced in Herd 1 with all pens treated in the control group compared to 30.8 % in the vaccinated group (p < 0.001). In Herd 2, the difference was non-significant with 68.0 % in the control group compared to 50.0 % in the vaccination group (p = 0.252). Low levels of individual treatments against diarrhea were seen in both herds (≤ 5.0 %) but still significantly reduced in vaccinated pigs compared to control pigs (p < 0.050 in both herds). Mean diarrheic blot counts were significantly reduced in vaccinated pens compared to control pens (p < 0.001 in both herds). In vaccinated pigs, shedding of LI was reduced in both prevalence (p < 0.001 in both herds), excretion level in positive samples (p < 0.001 in both herds) and, in one herd, also in duration (p = 0.003) when compared to control pigs. In conclusion, pigs vaccinated with Porcilis®Lawsonia Vet against LI in both of two high-health and high-productive finisher herds had, compared to non-vaccinated pigs, significantly improved key production figures, and reduced AB treatment, occurrence of diarrhea, LI shedding, and growth variation.

A dietary spray-dried plasma feeding programme improves growth performance and reduces faecal bacterial shedding of nursery pigs challenged with enterotoxigenic Escherichia coli K88.

This study aimed to assess the effects of different spray-dried plasma (SDP) feeding programmes to pigs on performance, intestinal histomorphology and faecal bacterial shedding after an Escherichia coli K88 challenge. A total of 96 piglets (5.77 ± 0.01 kg) were weaned at 21 days of age (Day 0) and challenged with 3 ml of 1 × 1010 CFU of E. coli K88 in total 3.0 × 1010 CFU/animal on Days 0, 2 and 4. Pigs were fed nursery diets containing 0.0%, 3.0%, 6.0% or 9.0% SDP from weaning to 35 days of age; 0.0%, 1.5%, 3.0% or 4.5% SDP from 36 to 49 days; and the same control diet (without SDP), for the last 10 days of the experiment (50-59 days of age). Performance was measured from 35 to 59 days of age and faecal bacterial shedding and intestinal histomorphometry were evaluated at Days 28 and 49 of age respectively. From 21 to 35 days of age, there was a linear effect for body weight (BW) and average daily gain (ADG), a trend of linear effect for average daily feed intake (ADFI) and a quadratic effect for feed:gain ratio (FG). From 21 to 49 days, the 9.0:4.5% and 6.0:3.0% SDP feeding programmes improved BW, ADG and FG when compared to the other treatments. At 59 days of age, BW and ADG were increased by the two highest SDP feeding programmes. The 9.0:4.5% SDP feeding programme increased ADFI from 21 to 59 days of age, with 6.0:3.0% being intermediate and the other two treatments being lowest. The CFU counts of E. coli/g of faeces decreased linearly with increasing addition of SDP. These results indicate that an extended inclusion of increased SDP levels in post-weaning diets can improve growth potential and decrease bacterial shedding induced by E. coli K88.

Safety of bivalent live attenuated Salmonella vaccine and its protection against bacterial shedding and tissue invasion in layers challenged with Salmonella.

Nontyphoidal Salmonella infection was one of the predominant foodborne illnesses in humans. The medical burden and antimicrobial resistance of salmonellosis gained importance in public health and requested the poultry industry to seek effective measures to control the disease. The objective of this study was to evaluate the safety and effectiveness of a commercial bivalent live attenuated vaccine (AviPro Salmonella DUO) in specific-pathogen-free (SPF) chickens and field layers. It explored its safety and efficacy against medically important strains, Salmonella Enteritidis (SE) and S. Typhimurium (ST). The results demonstrated that ten vaccine doses in SPF chickens and regular doses in commercial layers showed desirable safety without affecting chicken health. Vaccinated layers demonstrated lower flock mortality and higher egg production performance than the unvaccinated layers during the raising and egg production periods. Additionally, no visceral colonization and egg contaminations were detected. Cloacal shedding of vaccine strains was noted, but the colonization of Salmonella disappeared within four weeks of the last vaccination. Regarding vaccine efficacy, one dose significantly reduced Salmonella cloacal shedding (P = 0.037 for SE and P = 0.027 for ST) and viable cell counts (P = 0.003 for SE and ST) on day 7 post the challenges. Significantly low Salmonella loads of cloacal samples on day 14 after the challenges were also determined in the vaccinated group (P = 0.006 for SE; P = 0.041 for ST). Triple immunizations effectively prevented layers from the cloacal shedding on either day 7 or day 14 post Salmonella challenges. Total viable counts of SE and ST in tissues of vaccinated layers were also reduced on day 14 after the challenges (P = 0.026 for SE; P = 0.002 for ST). To conclude, one dose of vaccine exhibited inhibitory effects on Salmonella shedding and tissue invasions in young layers. Following the regimen of triple vaccinations, Salmonella shedding was completely inhibited, and tissue invasions were significantly reduced. Incorporating this vaccine into a comprehensive Salmonella control program is promising to protect layers from the risks of contaminating the flocks and egg products.

Serologic responses correlate with current but not future bacterial shedding in badgers naturally infected with Mycobacterium bovis.

Bovine tuberculosis is a challenging cattle disease with substantial economic costs in affected countries. Eradication in parts of the United Kingdom and Ireland is hindered by transmission of the causative agent Mycobacterium bovis between cattle and European badgers (Meles meles). Diagnostic tests in badgers are of limited accuracy but may help us understand and predict disease progression. This study aimed to determine the practical ability of a commercially available serologic test, the Dual Path Platform VetTB assay (DPP), to predict mycobacterial shedding (i.e. infectiousness) and disease progression in badgers, and whether test outcomes were associated with re-capture. Clinical samples collected from 2014 to 2019 from a wild, naturally infected population of badgers in southwest England were tested using mycobacterial culture (from sputum, urine, faeces, abscesses and bite wounds), an interferon-gamma release assay and the DPP assay. Data were analysed at both individual badger and social group levels using generalised linear and cumulative-link mixed models, and linear regression. Only the highest DPP readings [optical density relative light unit (RLU) levels] were associated with mycobacterial shedding [odds ratio (OR) for DPP levels > 100 RLU in individual badgers: 79.6, 95%CI: 14.7-848; and for social groups: OR: 7.28, 95%CI: 2.94-21.44; compared with levels < 100 RLU]. For individual badgers, RLU levels at first capture were not associated with disease progression at subsequent captures. Finally, badgers with very high DPP levels (> 1000 RLU) were four times less likely to be recaptured (OR: 0.24, 95%CI: 0.07-0.83) than those without a detectable DPP response, which might indicate enhanced mortality. We conclude that DPP levels of > 100 RLU identify badgers that are likely to be shedding M. bovis. Levels of > 1000 RLU identify badgers that are much less likely to be re-captured. These results provide insights into the potential value of existing tests in intervention strategies for managing M. bovis in badgers.

Evaluation of immune efficacy of Omp2 protein against Lawsonia intracellularis in mice.

Porcine proliferative enteropathy (PPE) is caused by the obligate intracellular bacterium Lawsonia intracellularis. Infection results in an enteric disease characterised by decreased growth performance of pigs, and presents a major economic burden for swine industries worldwide. Since vaccination is an effective technique for controlling PPE, novel effective vaccine platforms are need to be developed. In this study, five proteins of L. intracellularis were screened through animal experiments and the highly immunoprotective Omp2 protein was identified. Then, the immune efficacy of Omp2 was further evaluated based on humoral and cell mediated immune (CMI) responses, faecal bacterial shedding, histopathological lesions, immune barrier function of intestinal mucosa as well as digestive and absorptive capacity following challenge of mice with L. intracellularis. Mice immunised with Omp2 had reduced faecal shedding, fewer histopathological lesions and reduced bacteria colonisation of the ileum. Additionally, Omp2 immunised mice showed stronger serum IgG and IFN-γ levels, up-regulated Occludin and zonula occludens-1 (ZO-1) mRNA levels, as well as increased numbers of intestinal intraepithelial lymphocytes (IELs) and levels of sIgA. On the contrary, the activities of LPS, α-AMS and AKP were significantly increased. Our investigation indicated that immunization with Omp2 reduced the severity of clinical signs and provided efficacious immunoprotection for target animals against L. intracellularis infection in mouse model.

Salmonella Typhi Stool Shedding by Patients With Enteric Fever and Asymptomatic Chronic Carriers in an Endemic Urban Setting.

The burden of Salmonella enterica serotype Typhi (S. Typhi) shedding in stool and its contribution to transmission in endemic settings is unknown. During passive surveillance S. Typhi shedding was seen during convalescence in 332 bacteremic patient with typhoid, although none persisted at 1-year follow-up. Anti-virulence capsule (Vi)-immunoglobulin (Ig) G titers were measured in age-stratified cohort of serosurveillance participants. Systematic stool sampling of 303 participants with high anti-Vi-IgG titers identified 1 asymptomatic carrier with shedding. These findings suggest that ongoing S. Typhi transmission in this setting is more likely to occur from acute convalescent cases, although better approaches are needed to identify true chronic carriers in the community to enable typhoid elimination.

Pertactin contributes to shedding and transmission of Bordetella bronchiseptica.

Whooping cough is resurging in the United States despite high vaccine coverage. The rapid rise of Bordetella pertussis isolates lacking pertactin (PRN), a key vaccine antigen, has led to concerns about vaccine-driven evolution. Previous studies showed that pertactin can mediate binding to mammalian cells in vitro and act as an immunomodulatory factor in resisting neutrophil-mediated clearance. To further investigate the role of PRN in vivo, we examined the functions of pertactin in the context of a more naturally low dose inoculation experimental system using C3H/HeJ mice that is more sensitive to effects on colonization, growth and spread within the respiratory tract, as well as an experimental approach to measure shedding and transmission between hosts. A B. bronchiseptica pertactin deletion mutant was found to behave similarly to its wild-type (WT) parental strain in colonization of the nasal cavity, trachea, and lungs of mice. However, the pertactin-deficient strain was shed from the nares of mice in much lower numbers, resulting in a significantly lower rate of transmission between hosts. Histological examination of respiratory epithelia revealed that pertactin-deficient bacteria induced substantially less inflammation and mucus accumulation than the WT strain and in vitro assays verified the effect of PRN on the induction of TNF-α by murine macrophages. Interestingly, only WT B. bronchiseptica could be recovered from the spleen of infected mice and were further observed to be intracellular among isolated splenocytes, indicating that pertactin contributes to systemic dissemination involving intracellular survival. These results suggest that pertactin can mediate interactions with immune cells and augments inflammation that contributes to bacterial shedding and transmission between hosts. Understanding the relative contributions of various factors to inflammation, mucus production, shedding and transmission will guide novel strategies to interfere with the reemergence of pertussis.

Avaliação histopatológica da influência da obturação do canal radicular no reparo da periodontite apical em animais: uma revisão sistemática / Histopathological evaluation of the influence of root canal obturation in the repair of apical periodontitis in animals: a systematic review

Introdução: A obturação do canal radicular tem sido considerada um fator decisivo no sucesso do tratamento endodôntico. Objetivo: O objetivo da presente revisão sistemática foi avaliar estudos em animais que utilizaram parâmetros histopatológicos para determinar a influência da obturação do canal radicular no processo de reparo da periodontite apical. Métodos: As buscas foram realizadas nas bases de dados PubMed, Lilacs, Scielo, Science Direct e BBO, utilizando palavras-chave indexadas e não indexadas. Foram considerados artigos publicados no período de 2003 a 2019, e aplicados critérios de inclusão e exclusão para seleção dos artigos. Resultados: Cinco artigos atenderam aos critérios de elegibilidade e foram incluídos para análise qualitativa na revisão sistemática. Conclusão: A obturação do canal radicular, no que diz respeito ao seu limite apical, pode influenciar o reparo da periodontite apical, quando há presença de bactérias remanescentes no interior do canal. O uso de cimentos endodônticos com propriedades antimicrobianas não foi efetivo na eliminação de bactérias residuais do canal radicular e na reparação completa da periodontite apical.

Introduction: Root canal fillings are a decisive factor in the success of endodontic treatment. This systematic review evaluated animal studies using histopathological parameters to determine the effect of root canal filling on the healing of apical periodontitis. Methods: A search was conducted in PubMed, Lilacs, Scielo, Science Direct and BBO using indexed and non-indexed keywords. Studies published from 2003 to 2019 were selected according to inclusion and exclusion criteria. Results: Five animal studies met eligibility criteria and were included in the systematic review for a qualitative analysis. Conclusion: The extension of the root canal filling may affect the healing of apical periodontitis when bacteria remain in the canal. Root canal sealers with antimicrobial properties were not effective in eliminating bacteria remaining in the root canal, nor in promoting the complete healing of apical periodontitis (AU).

MicroRNAomes of Cattle Intestinal Tissues Revealed Possible miRNA Regulated Mechanisms Involved in Escherichia coli O157 Fecal Shedding.

Cattle have been suggested as the primary reservoirs of E. coli O157 mainly as a result of colonization of the recto-anal junction (RAJ) and subsequent shedding into the environment. Although a recent study reported different gene expression at RAJ between super-shedders (SS) and non-shedders (NS), the regulatory mechanisms of altered gene expression is unknown. This study aimed to investigate whether bovine non-coding RNAs play a role in regulating the differentially expressed (DE) genes between SS and NS, thus further influencing E. coli O157 shedding behavior in the animals through studying miRNAomes of the whole gastrointestinal tract including duodenum, proximal jejunum, distal jejunum, cecum, spiral colon, descending colon and rectum. The number of miRNAs detected in each intestinal region ranged from 390 ± 13 (duodenum) to 413 ± 49 (descending colon). Comparison between SS and NS revealed the number of differentially expressed (DE) miRNAs ranged from one (in descending colon) to eight (in distal jejunum), and through the whole gut, seven miRNAs were up-regulated and seven were down-regulated in SS. The distal jejunum and rectum were the regions where the most DE miRNAs were identified (eight and seven, respectively). The miRNAs, bta-miR-378b, bta-miR-2284j, and bta-miR-2284d were down-regulated in both distal jejunum and rectum of SS (log2fold-change: -2.7 to -3.8), bta-miR-2887 was down-regulated in the rectum of SS (log2fold-change: -3.2), and bta-miR-211 and bta-miR-29d-3p were up-regulated in the rectum of SS (log2fold-change: 4.5 and 2.2). Functional analysis of these miRNAs indicated their potential regulatory role in host immune functions, including hematological system development and immune cell trafficking. Our findings suggest that altered expression of miRNA in the gut of SS may lead to differential regulation of immune functions involved in E. coli O157 super-shedding in cattle.

Piscirickettsia salmonis shedding and tissue burden, and hematological responses during cohabitation infections in chum Oncorhynchus keta, pink O. gorbuscha and Atlantic salmon Salmo salar.

BACKGROUND: Salmonid rickettsial septicemia is an emergent and geographically widespread disease of marine-farmed salmonids caused by infection with the water-borne bacterium Piscirickettsia salmonis. Very little is known about the route, timing, or magnitude of bacterial shedding from infected fish. METHODOLOGY/PRINCIPAL FINDINGS: A cohabitation challenge model was used to assess shedding from chum Oncorhynchus keta, pink O. gorbuscha and Atlantic salmon Salmo salar. Infections in donor fish were established by intraperitoneal injection of P. salmonis. Naïve recipients were cohabitated with donor fish after which cumulative percent morbidity and mortality (CMM) was monitored, and bacterial burdens in kidney and in tank water were measured by qPCR. All donor fish died with mean days-to-death (MDD) among species ranging from 17.5 to 23.9. Among recipients, CMM ranged from 42.7% to 77.8% and MDD ranged from 49.7 to 56.4. In each trial, two peaks of bacterial DNA concentrations in tank water closely aligned with the MDD values of donor and recipient fish. Bacterial tissue burden and shedding rate, and plasma physiological parameters were obtained from individual donors and recipients. Statistically significant positive correlations between the shedding rate and P. salmonis kidney burden were measured in donor pink and in donor and recipient chum salmon, but not in donor or recipient Atlantic salmon. In Atlantic salmon, there was a negative correlation between kidney bacterial burden and hematocrit, plasma Ca++ and Mg++ values, whereas in infected chum salmon the correlation was positive for Na+ and Cl- and negative for glucose. CONCLUSIONS: A dependency of bacterial shedding on species-specific patterns of pathogenesis was suggested. The coincidence of bacterial shedding with mortality will inform pathogen transmission models.

Horizontal transmission of Piscirickettsia salmonis from the wild sub-Antarctic notothenioid fish Eleginops maclovinus to rainbow trout (Oncorhynchus mykiss) under experimental conditions.

Piscirickettsia salmonis is the aetiological agent of piscirickettsiosis, a bacterial disease that affects farmed salmonids, causing high mortalities and significant economic losses in the Chilean salmon farm industry. Given the Chilean native fish species Patagonian blenny, Eleginops maclovinus, lives in the vicinity of salmon farms, it is relevant to clarify the epidemiological role that this species could play in the transmission and/or dissemination of this pathogen. This study aimed to evaluate the bidirectional transmission of P. salmonis between the Patagonian blenny and Oncorhynchus mykiss (rainbow trout), via a cohabitation challenge model. The results of this study demonstrated the transmission of the bacteria from Patagonian blennies to rainbow trout, considering the specific mortality in cohabitant rainbow trout, reaching 46%: the necropsy of these specimens, evidencing the characteristic pathological lesions of the disease and the positive results of the qPCR analysis for P. salmonis, in the same individuals. In contrast, no mortalities of Patagonian blenny specimens were recorded in the challenged experimental groups. This study is the first report showing the horizontal transmission of P. salmonis from a native non-salmonid species, such as the Patagonian blenny, to a salmonid species, generating the disease and specific mortality in rainbow trout, using a cohabitation challenge.

Type I Interferon Signaling Is a Common Factor Driving Streptococcus pneumoniae and Influenza A Virus Shedding and Transmission.

The dynamics underlying respiratory contagion (the transmission of infectious agents from the airways) are poorly understood. We investigated host factors involved in the transmission of the leading respiratory pathogen Streptococcus pneumoniae Using an infant mouse model, we examined whether S. pneumoniae triggers inflammatory pathways shared by influenza A virus (IAV) to promote nasal secretions and shedding from the upper respiratory tract to facilitate transit to new hosts. Here, we show that amplification of the type I interferon (IFN-I) response is a critical host factor in this process, as shedding and transmission by both IAV and S. pneumoniae were decreased in pups lacking the common IFN-I receptor (Ifnar1-/- mice). Additionally, providing exogenous recombinant IFN-I to S. pneumoniae-infected pups was sufficient to increase bacterial shedding. The expression of IFN-stimulated genes (ISGs) was upregulated in S. pneumoniae-infected wild-type (WT) but not Ifnar1-/- mice, including genes involved in mucin type O-glycan biosynthesis; this correlated with an increase in secretions in S. pneumoniae- and IAV-infected WT compared to Ifnar1-/- pups. S. pneumoniae stimulation of ISGs was largely dependent on its pore-forming toxin, pneumolysin, and coinfection with IAV and S. pneumoniae resulted in synergistic increases in ISG expression. We conclude that the induction of IFN-I signaling appears to be a common factor driving viral and bacterial respiratory contagion.IMPORTANCE Respiratory tract infections are a leading cause of childhood mortality and, globally, Streptococcus pneumoniae is the leading cause of mortality due to pneumonia. Transmission of S. pneumoniae primarily occurs through direct contact with respiratory secretions, although the host and bacterial factors underlying transmission are poorly understood. We examined transmission dynamics of S. pneumoniae in an infant mouse model and here show that S. pneumoniae colonization of the upper respiratory tract stimulates host inflammatory pathways commonly associated with viral infections. Amplification of this response was shown to be a critical host factor driving shedding and transmission of both S. pneumoniae and influenza A virus, with infection stimulating expression of a wide variety of genes, including those involved in the biosynthesis of mucin, a major component of respiratory secretions. Our findings suggest a mechanism facilitating S. pneumoniae contagion that is shared by viral infection.

Prevalence of Coxiella burnetii seropositivity and shedding in farm, pet and feral cats and associated risk factors in farm cats in Quebec, Canada.

Cats represent a potential source of Coxiella burnetii, the aetiological agent of Q fever in humans. The prevalence and risk factors of C. burnetii infection in farm, pet and feral cats were studied in Quebec, Canada, using a cross-sectional study. Serum samples were tested using a specific enzyme-linked immunosorbent assay (ELISA) for the presence of antibodies against C. burnetii, whereas rectal swabs were assayed using real-time quantitative polymerase chain reaction (qPCR) for the molecular detection of the bacteria. Potential risk factors for farm cats were investigated using clinical examinations, questionnaires and results from a concurrent study on C. burnetii farm status. A total of 184 cats were tested: 59 from ruminant farms, 73 pets and 52 feral cats. Among farm cats, 2/59 (3.4%) were ELISA-positive, 3/59 (5.1%) were ELISA-doubtful and 1/59 (1.7%) was qPCR-positive. All pets and feral cats were negative to C. burnetii ELISA and qPCR. Farm cat positivity was associated with a positive C. burnetii status on the ruminant farm (prevalence ratio = 7.6, P = 0.03). Our results suggest that although pet and feral cats do not seem to pose a great C. burnetii risk to public health, more active care should be taken when in contact with cats from ruminant farms.

Tiempo de excreción fecal de Escherichia coliproductora de toxina Shiga en niños con síndrome urémico hemolítico / Duration of fecal shedding of Shiga toxin-producing Escherichia coli among children with hemolytic uremic syndrome

Introducción. Conocer el tiempo de excreción fecal de Escherichia coli productora de toxina Shiga (Shiga toxin-producing Escherichia coli; STEC, por sus siglas en inglés) en pacientes con síndrome urémico hemolítico sería útil para controlar la transmisión de la enfermedad.Objetivos. 1) Analizar las características del tiempo de excreción de STEC. 2) Evaluar la asociación con las variables sexo, edad, necesidad de diálisis, antibióticos y serotipos de STEC.Población y métodos. Estudio prospectivo, observacional, longitudinal y analítico. Período 2013-2019. Se realizaron coprocultivos al ingresar y cada 5-7 días hasta obtener 2 negativos. Se definió tiempo de excreción desde el inicio de la diarrea hasta el primer negativo. Se confirmó STEC por detección de los genes stx1, stx2 y rfbO157 por reacción en cadena de la polimerasa. Se calculó la media (IC 95 %) y percentilos del tiempo de excreción de STEC, y se compararon las variables estudiadas mediante el test de t.Resultados. Se incluyeron 43 pacientes. La media de tiempo de excreción fue 10,2 días (IC 95 %: 8,92-11,59), rango: 3-22 días. El 90 % de los pacientes negativizaron el coprocultivo a los 15 días. No hubo diferencias según sexo (p = 0,419), edad (p = 0,937), necesidad de diálisis (p = 0,917), antibióticos (p = 0,147) ni serotipos (p = 0,231).Conclusión. El 90 % de los pacientes negativizó el coprocultivo a los 15 días del inicio de la diarrea, y todos, al día 22. No se encontró asociación entre el tiempo de excreción y las variables estudiadas.

Introduction. Knowing the duration of fecal shedding of Shiga toxin-producing Escherichia coli(STEC) among patients with hemolytic uremic syndrome would be useful to control disease transmission.Objectives. 1) To analyze the characteristics of STEC shedding duration. 2) To assess the association with sex, age, need of dialysis, antibiotics, and STEC serotypes.Population and methods. Prospective, observational, longitudinal, and analytical study in the 2013-2019 period. Stool cultures were done upon admission and every 5-7 days until 2 negative results were obtained. Shedding duration was defined as the period from diarrhea onset to the first negative result. STEC was confirmed with polymerase chain reaction detection of stx1, stx2, and rfbO157 genes. The mean (95 % CI) and percentile values of the STEC shedding duration were estimated, and the studied outcome measures were compared using the t test.Results. A total of 43 patients were included. The mean duration of shedding was 10.2 days (95 % CI: 8.92-11.59), range: 3-22 days. After 15 days, 90 % of patients had a negative stool culture. There were no differences in terms of sex (p = 0.419), age (p = 0.937), need of dialysis (p = 0.917), antibiotics (p = 0.147) or serotype (p = 0.231).Conclusion. Fifteen days after the onset of diarrhea, 90 % of patients had a negative stool culture, and all patients had one after 22 days. No association was observed between the duration of shedding and studied outcome measures.

Diseminación interhospitalaria de Klebsiella pneumoniae ST512 productora de KPC-3. Detección por MALDI-TOF / Interhospital dissemination of KPC-3 producing-Klebsiella pneumoniae ST512. Detection by MALDI-TOF

Introducción/Objetivo: Describir un brote por Klebsiella pneumoniae (KPN) productora de KPC-3 y determinar la eficacia diagnóstica de MALDI-TOF en su detección. Métodos: Estudio retrospectivo de las KPN-KPC-3 aisladas en 2 hospitales de Ciudad Real. Se buscó el pico a 11,109kDa±15 en el espectro proporcionado por MALDI-TOF para KPN. Resultados: Se aislaron 156 cepas de KPN que portaban el gen blaKPC-3, con un único perfil perteneciente al ST512 (31 cepas estudiadas). Hubo un 25% de infectados. Un 84% tuvieron origen nosocomial o relacionado con la asistencia sanitaria. El 93% tenía alguna enfermedad de base (31% de exitus en el primer mes). La detección del pico mostró una sensibilidad del 90% y una especificidad del 100%. Conclusiones: Detectamos la diseminación clonal de una cepa de KPN ST512 productora de KPC-3 en 3 hospitales de Ciudad Real. Además, evidenciamos la rentabilidad de MALDI-TOF en la detección precoz de KPN-KPC.(AU)

Introduction/Objective: To describe an outbreak of KPC-3-producing Klebsiella pneumoniae (KPN) and determine the diagnostic efficacy of MALDI-TOF in its detection. Methods: Retrospective study of the KPC-3-KPN isolated in 2 hospitals in Ciudad Real. The peak at 11,109kDa±15 was sought in the KPN spectra provided by MALDI-TOF. Results: We isolated 156 KPN strains that carried the blaKPC-3 gene, with a unique profile belonging to ST512 (31 strains studied). There was 25% of infected patients, 84% were nosocomial or related to health care and 93% had some underlying disease (31% of exitus in the first month). The detection of the peak showed 90% sensitivity and 100% specificity. Conclusions: We detected the clonal spread of a KPN ST512 strain producing KPC-3 in 3 hospitals in Ciudad Real. In addition, we show the profitability of MALDI-TOF in the early detection of KPC-KPN.