Prevalence and genetic diversity of azole-resistant Malassezia pachydermatis isolates from canine otitis and dermatitis: a two-year study.

Despite previous reports on the emergence of Malassezia pachydermatis strains with decreased susceptibility to azoles, there is limited information on the actual prevalence and genetic diversity of azole-resistant isolates of this yeast species. We assessed the prevalence of azole resistance in M. pachydermatis isolates from cases of dog otitis or skin disease attended in a veterinary teaching hospital during a 2-year period and analyzed the ERG11 (encoding a lanosterol 14-α demethylase, the primary target of azoles) and whole genome sequence diversity of a group of isolates that displayed reduced azole susceptibility. Susceptibility testing of 89 M. pachydermatis isolates from 54 clinical episodes (1-6 isolates/episode) revealed low minimum inhibitory concentrations (MICs) to most azoles and other antifungals, but 11 isolates from six different episodes (i.e., 12.4% of isolates and 11.1% of episodes) had decreased susceptibility to multiple azoles (fluconazole, itraconazole, ketoconazole, posaconazole, ravuconazole, and/or voriconazole). ERG11 sequencing of these 11 azole-resistant isolates identified eight DNA sequence profiles, most of which contained amino acid substitutions also found in some azole-susceptible isolates. Analysis of whole genome sequencing (WGS) results revealed that the azole-resistant isolates from a same episode of otitis, or even different episodes affecting a same animal, were more genetically related to each other than to isolates from other dogs. In conclusion, our results confirmed the remarkable ERG11 sequence variability in M. pachydermatis isolates of animal origin observed in previous studies and demonstrate the value of WGS for disentangling the epidemiology of this yeast species.

We analyzed the prevalence and diversity of azole-resistant Malassezia pachydermatis isolates in a veterinary hospital. A low prevalence of multi-azole resistance (c.10% of isolates and cases) was found. Whole genome and ERG11 sequencing of resistant isolates revealed remarkable genetic diversity.

Active surveillance of antimicrobial resistance in companion animals: A pilot study in a Spanish Veterinary Teaching Hospital.

The role of small animal veterinary hospitals in the onset and dissemination of antimicrobial-resistant organisms (AMROs) is still not clear, and the implementation of an internal surveillance systems is a cost-effective tool to better understand their impact. The aim of this study was to describe a pilot program of active surveillance in a Spanish Veterinary Teaching Hospital, developed to estimate the detection frequency of AMROs in the commensal flora of patients and in the environment. Surveillance was focused on Methicillin-resistant Staphylococci (MRS), third generation cephalosporins resistant gram-negative bacteria (3GCR-GNB), and carbapenems-resistant gram-negative bacteria (CR-GNB). Oral and perirectal swabs were collected in the same dogs and cats hospitalized > 48 h, at their admission and before their discharge. Out of 50 patients sampled, 24% (12/50) were carriers at admission of at least one of the three investigated AMROs. Twenty-eight percent of patients (14/50) acquired at least one AMRO during the hospital stay. MRS detection frequency at admission was 12% (6/50), while acquisition was 6% (3/50). 3GCR-GNB detection frequency was 14% at admission (7/50) and acquisition 22% (11/50), while CR-GNB detection frequency was 2% at admission (1/50) and acquisition 2% (1/50). Environmental surveillance (98 samples) showed a total detection frequency of 22.4% for MRS (22/98), 2% for 3GCR-GNB and CR-GNB (2/98). Clinical staff' shoe soles showed high detection frequency for MRS (50%). 3GCR Escherichia coli was the most isolated species in patients (n = 17). The results show how active surveillance can be used as a tool to assess the impact of AMROs in veterinary hospitals to subsequently build up tailored control plans based on specific issues.

Analysis of the culturable gut yeast microbiota of dogs with digestive disorders.

Despite the increasing interest in studying the gut mycobiota of dogs, the association between fungal colonization and the development of digestive disorders in this species remains largely understudied. On the other hand, the high prevalence of antifungal-resistant yeasts detected in previous studies in samples from animals represents a major threat to public health. We analyzed the presence of culturable yeasts in 112 rectal swab samples obtained from dogs with digestive disorders attended in a veterinary teaching hospital. Our results revealed that Malassezia pachydermatis was frequently isolated from the studied dog population (33.9% of samples), and that the isolation of this yeast was significantly associated to the age of animals, but not to their sex, disease group, or the presence of vomits and/or diarrhea. In contrast, other yeast species were less prevalent (17.9% of samples in total), and their isolation was not significantly associated to any variable included in the analysis. Additionally, we observed that 97.5% of the studied M. pachydermatis isolates (n = 158, 1-6 per positive episode) displayed a minimum inhibitory concentration (MIC) value >4 µg/ml to nystatin, 31.6% had a MIC ≥32 µg/ml to fluconazole, and 27.2% had a MIC >4 µg/ml to amphotericin B. The antifungal susceptibility profiles of non-Malassezia (n = 43, 1-7 per episode) were more variable and included elevated MIC values for some antifungal-species combinations. These results confirm that the intestine of dogs is a reservoir of opportunistic pathogenic yeasts and suggest that the prevalence of M. pachydermatis colonization depends more on the age of animals than on any specific digestive disorder.

In vitro activity of fidaxomicin and combinations of fidaxomicin with other antibiotics against Clostridium perfringens strains isolated from dogs and cats.

BACKGROUND: Previous studies have demonstrated that fidaxomicin, a macrocyclic lactone antibiotic used to treat recurrent Clostridioides difficile-associated diarrhea, also displays potent in vitro bactericidal activity against Clostridium perfringens strains isolated from humans. However, to date, there is no data on the susceptibility to fidaxomicin of C. perfringens strains of animal origin. On the other hand, although combination therapy has become popular in human and veterinary medicine, limited data are available on the effects of antibiotic combinations on C. perfringens. We studied the in vitro response of 21 C. perfringens strains obtained from dogs and cats to fidaxomicin and combinations of fidaxomicin with six other antibiotics. RESULTS: When tested by an agar dilution method, fidaxomicin minimum inhibitory concentrations (MICs) ranged between 0.004 and 0.032 µg/ml. Moreover, the results of Etest-based combination assays revealed that the incorporation of fidaxomicin into the test medium at a concentration equivalent to half the MIC significantly increased the susceptibility of isolates to metronidazole and erythromycin in 71.4% and 61.9% of the strains, respectively, and the susceptibility to clindamycin, imipenem, levofloxacin, and vancomycin in 42.9-52.4% of the strains. In contrast, » × MIC concentrations of fidaxomicin did not have any effect on levofloxacin and vancomycin MICs and only enhanced the effects of clindamycin, erythromycin, imipenem, and metronidazole in ≤ 23.8% of the tested strains. CONCLUSIONS: The results of this study demonstrate that fidaxomicin is highly effective against C. perfringens strains of canine and feline origin. Although fidaxomicin is currently considered a critically important antimicrobial that has not yet been licensed for veterinary use, we consider that the results reported in this paper provide useful baseline data to track the possible emergence of fidaxomicin resistant strains of C. perfringens in the veterinary setting.

Presence of Aspergillus fumigatus with the TR34/L98H Cyp51A mutation and other azole-resistant aspergilli in the air of a zoological park.

Antifungal-resistant fungi, including Aspergillus fumigatus and other Aspergillus species, pose an urgent threat to human and animal health. Furthermore, the environmental route of azole resistance selection due to the widespread use of azole fungicides in crop protection and other applications is a major public health issue. Although environmental surveillance of fungi is frequently performed in many zoological parks and wildlife rehabilitation centers, the antifungal susceptibility of recovered isolates is only rarely analyzed, which precludes a clear assessment of the threat posed by these fungi to captive animals. In this study, we assessed the presence of airborne azole-resistant Aspergillus spp., including the so-called 'cryptic species' (i.e., species which are phenotypically similar to more well-known aspergilli but clearly constitute different phylogenetic lineages) in a zoological park located in the city of Madrid, Spain. In general, our results revealed a low prevalence A. fumigatus and cryptic aspergilli with decreased susceptibility to azoles. However, we detected an A. fumigatus isolate with the TR34/L98H mutation in the gene encoding the lanosterol 14α-demethylase (Cyp51A), consisting of a tandem repeat of 34 base pairs in the promoter region and a lysine to histidine substitution at codon 98. Notably, this TR34/L98H mutation has been linked to the environmental route of azole resistance selection, thus highlighting the 'One Health' dimension of the emerging problem of antifungal resistance. In this context, continuous environmental surveillance of azole-resistant aspergilli in zoological parks and other similar animal facilities is recommended.

Psychometric evaluation and validation of the Postpartum Specific Anxiety Scale for the Spanish-speaking population: PSAS-ES.

OBJECTIVE: The transition to motherhood is a period of risk for the development of mood disorders. Postpartum anxiety has not been as thoroughly studied as other emotional disorders despite its impact on mothers and their babies. The absence of standardized programmes for early detection and specific tools for its diagnosis means postpartum anxiety is often underestimated or overshadowed. This study aimed to translate and validate the Postpartum Specific Anxiety Scale [PSAS] for the Spanish population and to analyse its reliability as an exploratory tool for specific anxiety in mothers. METHOD: Four stages were followed in this research: translation and back-translation to obtain the Spanish version [PSAS-ES]; preliminary pilot study to explore the comprehensibility and ease of responding the items (n = 53); convergent validity analyses (n = 644); and test-retest reliability (n = 234). RESULTS: The PSAS-ES has shown to have good acceptability, convergent validity and high internal consistency with a Cronbach's α coefficient of 0.93 for the overall scale of PSAS. The four factors had good reliability. The results of test-retest was 0.86, indicating excellent stability over time in the first 16 weeks. CONCLUSION: The psychometric results show that the PSAS-ES is a valid tool to explore and detect anxiety in Spanish mothers between 0 and 16 weeks postpartum.

Susceptibility testing of Prototheca bovis isolates from cases of bovine mastitis using the CLSI reference broth microdilution method and the Sensititre YeastOne colorimetric panel.

A total of 62 Prototheca bovis isolates from cases of bovine mastitis were tested for susceptibility to different antifungal compounds by the Clinical and Laboratory Standards Institute (CLSI) reference microdilution method and a commercial colorimetric microdilution panel (Sensititre YeastOne). All isolates displayed low susceptibility to echinocandins (MICs > 8 µg/ml for anidulafungin, caspofungin, and micafungin), flucytosine (MIC > 64 µg/ml), and the azoles enilconazole and fluconazole (MICs > 4 and > 64 µg/ml, respectively). Moreover, 45.2, 32.3, and 1.6% of isolates had MICs > 4 µg/ml for ketoconazole, terbinafine, and voriconazole, respectively, when tested by the CLSI method. In contrast, all isolates were more susceptible to the polyene compounds amphotericin B and nystatin, and itraconazole, posaconazole, and ravuconazole (MICs ≤ 2 µg/ml, in all cases). Comparison of the results obtained in the CLSI and Sensititre methods showed excellent essential agreement (EA) for azoles (98.4% for itraconazole and posaconazole, and 100% for voriconazole) and moderate EA for amphotericin B (72.6%), when MICs were read after 48 and 24 h of incubation, respectively. In contrast, much lower EA values were obtained in some cases when the MICs for both techniques were determined after 48 h of incubation (e.g., 9.7% for amphotericin B and 69.4% for posaconazole). Therefore, the CLSI broth microdilution method and the Sensititre YeastOne panel can be used indistinctly for susceptibility testing of P. bovis isolates against azoles but not against amphotericin B until further optimization of the test conditions. LAY SUMMARY: The antifungal susceptibility of Prototheca bovis isolates was analyzed. All tested isolates displayed low susceptibility to echinocandins, flucytosine, and some azoles. Excellent agreement of the results of two different test methods was obtained for azoles, but not for the polyene amphotericin B.

The Impact of the COVID-19 Pandemic on Physical and Mental Health in China and Spain: Cross-sectional Study.

BACKGROUND: Differences in physical and mental health impact across continents during the COVID-19 pandemic are unknown. OBJECTIVE: This study compared the levels of impact of COVID-19 on mental health among people from Spain and China and correlated mental health parameters with variables relating to symptoms similar to COVID-19, COVID-19 knowledge, and precautionary measures. METHODS: We collected information on demographic data, physical symptoms, contact history with persons with a confirmed COVID-19 diagnosis, COVID-19 knowledge, and precautionary measures. Participants completed the Impact of Event Scale-Revised (IES-R) and the Depression, Anxiety and Stress Scale-21 Items (DASS-21). To analyze the differences in the mental health parameters, the mean scores between Chinese and Spanish respondents were compared using the independent samples t test. The differences in categorical variables between the two samples were analyzed by the chi-square test. Linear regression was used to calculate the univariate associations between the independent variables and mental health parameters for both groups separately, with adjustments made for age, gender, and education. RESULTS: A total of 1528 participants (Spain: n=687; China: n=841) were recruited. The mean age of the Chinese respondents was 24.73 years (SD 7.60; range 18-65 years), and the mean age of the Spanish respondents was 43.06 years (SD 11.95; range 18-76 years). Spanish participants reported significantly more symptoms similar to COVID-19 infection (eg, fever, sore throat, and breathing difficulties), contact history with COVID-19, higher perceived risk of contracting COVID-19, frequent use of medical services, and less confidence in medical services compared with their Chinese counterparts (P<.001). Spanish participants reported significantly higher DASS-21 stress and depression scores, while Chinese participants reported significantly higher IES-R scores (P<.001). Chinese participants encountered more discrimination from other countries (P<.001). Significantly more Chinese participants reported using face masks than Spanish ones (P<.001). More exposure to health information was associated with adverse mental health in Spain (depression: P=.02; anxiety: P=.02; stress: P=.001). CONCLUSIONS: Our study found that Spanish respondents reported higher levels of stress and depression as well as more symptoms and use of medical services. In preparation for the next pandemic, Spain needs to establish a prompt policy to implement rapid response and enhance medical services to safeguard physical and mental health.

A chain mediation model on COVID-19 symptoms and mental health outcomes in Americans, Asians and Europeans.

The novel Coronavirus-2019 (COVID-19) was declared a pandemic by the World Health Organization (WHO) in March 2020, impacting the lifestyles, economy, physical and mental health of individuals globally. This study aimed to test the model triggered by physical symptoms resembling COVID-19 infection, in which the need for health information and perceived impact of the pandemic mediated the path sequentially, leading to adverse mental health outcomes. A cross-sectional research design with chain mediation model involving 4612 participants from participating 8 countries selected via a respondent-driven sampling strategy was used. Participants completed online questionnaires on physical symptoms, the need for health information, the Impact of Event Scale-Revised (IES-R) questionnaire and Depression, Anxiety and Stress Scale (DASS-21). The results showed that Poland and the Philippines were the two countries with the highest levels of anxiety, depression and stress; conversely, Vietnam had the lowest mean scores in these areas. Chain mediation model showed the need for health information, and the perceived impact of the pandemic were sequential mediators between physical symptoms resembling COVID-19 infection (predictor) and consequent mental health status (outcome). Excessive and contradictory health information might increase the perceived impact of the pandemic. Rapid COVID-19 testing should be implemented to minimize the psychological burden associated with physical symptoms, whilst public mental health interventions could target adverse mental outcomes associated with the pandemic.

A validation of the Postpartum Specific Anxiety Scale 12-item research short-form for use during global crises with five translations.

BACKGROUND: Global crises inevitably increase levels of anxiety in postpartum populations. Effective and efficient measurement is therefore essential. This study aimed to create a 12-item research short form of the 51-item Postpartum Specific Anxiety Scale [PSAS] and validate it for use in rapid response research at a time of global crises [PSAS-RSF-C]. We also present the same 12-items, in five other languages (Italian, French, Chinese, Spanish, Dutch) to increase global accessibility of a psychometric tool to assess maternal mental health. METHODS: Twelve items from the PSAS were selected on the basis of a review of their factor loadings. An on-line sample of UK mothers (N = 710) of infants up to 12 weeks old completed the PSAS-RSF-C during COVID-19 'lockdown'. RESULTS: Principal component analyses on a randomly split sample (n = 344) revealed four factors, identical in nature to the original PSAS, which in combination explained 75% of the total variance. Confirmatory factor analyses (n = 366) demonstrated the four-factor model fit the data well. Reliability of the overall scale and of the underlying factors in both samples proved excellent. CONCLUSIONS: Findings suggest the PSAS-RSF-C may prove useful as a clinical screening tool and is the first postpartum-specific psychometric scale to be validated during the COVID-19 pandemic. This offers psychometrically sound assessment of postpartum anxiety. By increasing the accessibility of the PSAS, we aim to enable researchers the opportunity to measure maternal anxiety, rapidly, at times of global crisis.

Oxidative Stability of Vegetal Oil-Based Lubricants.

Lipids are widely distributed in nature and are one of the most important components of natural foods, synthetic compounds, and emulsions. To date, there is a strong social demand in the industrial sector for the use of sustainable products with a minimal environmental impact. Depending on their origin and composition, lipids can be employed as a plausible alternative as biodegradable lubricants in order to reduce the use of conventional mineral oil lubricants and mitigate their environmental impact. This perspective provides an overview of the advantages and constrains of vegetal oils under different lubrication regimes and the tribochemical reactions that can take place. Also, the different factors and pathways that influence their oxidation, the key role of moisture, and the changes of physical properties under pressure and temperature are reviewed. Special emphasis is devoted to the oxidation instability of fatty acids and vegetal oils and the physical and chemical approaches to improve oxidative and thermal stability are described in detail.

Transition Trajectories: Contexts, Difficulties and Consequences Reported by Young Transgender and Non-Binary Spaniards.

The transition process can have a significant impact on young transgender and non-binary individuals (TNBI), especially regarding their mental health. As such, this study aimed to explore the encountered difficulties and expectations of 225 young adults, between 14 and 25 years old, who identify themselves as TNBI. Four different aspects were analyzed: difficulties related to the transition process; main current difficulties; if the current difficulties are due to the participants' sexual orientation or gender identity; and if participants feel the need of changing something in their lives. Several obstacles either before, during or after the transition process were identified; and these obstacles have serious consequences in TBNI's mental health. Questions regarding legislation, discrimination, transphobia, lack of social support, the pathologization of transgenderism, low self-esteem, anxiety symptomatology, among others, were pointed out by the participants. The narratives collected and analyzed in the present study appear as important inputs to the literature, which can be translated into the basis of further investigations and the improvement of professional practices in the health field.

Detection of Clostridium difficile in the environment in a veterinary teaching hospital.

The present study focused on detecting the presence of Clostridium difficile on veterinary hospital surfaces of large and small animal areas at the Universidad Complutense of Madrid. Isolated C. difficile strains were further characterized and investigated for antimicrobial susceptibility testing. Of n = 23 sampling area, 17% were positive for the presence of C. difficile. The isolates belonged to PCR ribotypes 078, 014, 039, and 154, of which RT 078 and 014 are also frequently found as human pathogens. Two isolates had high level resistance to metronidazole. These results suggest that the veterinary hospital environment constitutes a potential reservoir of zoonotical transferable C. difficile.

In vitro activity of amphotericin B-azole combinations against Malassezia pachydermatis strains.

Combination therapy has become popular in clinical practice, but limited data on the effects of combinations of antifungal agents is still available for most fungal pathogens. We studied the in vitro response of 30 genetically diverse clinical strains of the basidiomycetous lipophilic yeast Malassezia pachydermatis obtained from cases of canine otitis to several amphotericin B (AMB)-azole combinations. Broth microdilution checkerboard tests revealed that AMB antagonized the effects of itraconazole (ITC) and voriconazole (VRC) in 50% and 6.7% of the strains, respectively, but did not interact with fluconazole or posaconazole (fractional inhibitory concentration index (FICI) values were <4 in all cases). Subsequent Etest-based assays performed for a subset of strains did not confirm the antagonism between AMB and ITC or AMB and VRC. In summary, the results of this study suggest that antagonistic combination effects between AMB and azoles might occur when tested against M. pachydermatis. Nevertheless, as observed for other fungi, different in vitro analyses yielded contrasting results, and the response to AMB-azole combinations was compound- and strain-dependant.

Distribution and tracking of Clostridium difficile and Clostridium perfringens in a free-range pig abattoir and processing plant.

The presence and genetic diversity of Clostridium difficile and C. perfringens along the slaughtering process of pigs reared in a free-range system was assessed. A total of 270 samples from trucks, lairage, slaughter line and quartering were analyzed, and recovered isolates were toxinotyped and genotyped. C. difficile and C. perfringens were retrieved from 14.4% and 12.6% of samples, respectively. The highest percentage of positive samples for C. difficile was detected in trucks (80%) whereas C. perfringens was more prevalent in cecal and colonic samples obtained in the slaughter line (85% and 45%, respectively). C. difficile isolates (n = 105) were classified into 17 PCR ribotypes (including 010, 078, and 126) and 95 AFLP genotypes. C. perfringens isolates (n = 85) belonged to toxinotypes A (94.1%) and C (5.9%) and were classified into 80 AFLP genotypes. The same genotypes of C. difficile and C. perfringens were isolated from different pigs and occasionally from environmental samples, suggesting a risk of contaminated meat products.

Recreational sandboxes for children and dogs can be a source of epidemic ribotypes of Clostridium difficile.

Different studies have suggested that the sand of public playgrounds could have a role in the transmission of infections, particularly in children. Furthermore, free access of pets and other animals to the playgrounds might increase such a risk. We studied the presence of Clostridium difficile in 20 pairs of sandboxes for children and dogs located in different playgrounds within the Madrid region (Spain). Clostridium difficile isolation was performed by enrichment and selective culture procedures. The genetic (ribotype and amplified fragment length polymorphism [AFLP]) diversity and antibiotic susceptibility of isolates was also studied. Overall, 52.5% (21/40) of samples were positive for the presence of C. difficile. Eight of the 20 available isolates belonged to the toxigenic ribotypes 014 (n = 5) and 106 (n = 2), both regarded as epidemic, and CD047 (n = 1). The other 12 isolates were non-toxigenic, and belonged to ribotypes 009 (n = 5), 039 (n = 4), and 067, 151 and CD048 (one isolate each). Nevertheless, all isolates (even those of a same ribotype) were classified into different AFLP genotypes indicating non-relatedness. In conclusion, our results revealed the presence of epidemic ribotypes of C. difficile in children's and dog's sandboxes located nearby, which constitutes a major health risk.

Data from a survey of Clostridium perfringens and Clostridium difficile shedding by dogs and cats in the Madrid region (Spain), including phenotypic and genetic characteristics of recovered isolates.

This article contains information related to a recent survey of the prevalence of fecal shedding of Clostridium perfringens and C. difficile by dogs and cats attended in veterinary clinics located in the Madrid region (Spain). Specifically, we provide detailed information about the clinics that participated in the survey, the demographic and clinic characteristics of recruited animals and the genetic and phenotypic characteristics (including antimicrobial susceptibility data), of recovered bacterial isolates.

Prevalence and characteristics of Clostridium perfringens and Clostridium difficile in dogs and cats attended in diverse veterinary clinics from the Madrid region.

Despite extensive research on the epidemiology of pathogenic clostridia in dogs and cats, most published studies focus on a selected animal population and/or a single veterinary medical centre. We assessed the burden of Clostridium perfringens and C. difficile shedding by small animals in 17 veterinary clinics located within the Madrid region (Spain) and differing in size, number and features of animals attended and other relevant characteristics. In addition, we studied the genetic diversity and antibiotic susceptibility of recovered isolates. Selective culture of all fecal specimens collected during a single week from dogs (n = 105) and cats (n = 37) attended in participating clinics yielded C. perfringens/C. difficile from 31%, 4.8% of the dogs, and 20%, 0% of the cats analyzed, respectively, and three dogs yielded both species. Furthermore, 17 animals (15 dogs and two cats) that yielded a positive culture for either species were recruited for a follow-up survey and C. perfringens was again obtained from nine dogs. Considerable differences in prevalence were observed among participating clinics for both clostridial species. C. perfringens isolates (n = 109) belonged to toxinotypes A (97.2%) and E (three isolates from one dog), whereas C. difficile isolates (n = 18) belonged to the toxigenic ribotypes 106 (33.3%) and 154 (16.7%), a 009-like ribotype (33.3%) and an unknown non-toxigenic ribotype (16.7%). Amplified fragment length polymorphism-based fingerprinting classified C. perfringens and C. difficile isolates into 105 and 15 genotypes, respectively, and tested isolates displayed in vitro resistance to benzylpenicillin (2.8%, 88.8%), clindamycin (0%, 16.7%), erythromycin (0.9%, 16.7%), imipenem (1.8%, 100%), levofloxacin (0.9%, 100%), linezolid (5.5%, 0%), metronidazole (4.6%, 0%) and/or tetracycline (7.3%, 0%). All animals from which multiple isolates were retrieved yielded ≥2 different genotypes and/or antimicrobial susceptibility profiles. Future studies should focus on the seasonal and geographical variations of prevalence and diversity patterns of clostridial species in small animals.

Clostridium perfringens Type A Isolates of Animal Origin with Decreased Susceptibility to Metronidazole Show Extensive Genetic Diversity.

Metronidazole (MTZ) is a first-line drug for the treatment of human infections caused by anaerobic bacteria. Although most Clostridium perfringens isolates are susceptible to metronidazole, a number of strains with decreased susceptibility have been obtained in clinical and environmental surveys. However, little is known yet about the toxinotype and genetic diversity of metronidazole-resistant C. perfringens strains. We tested for susceptibility to metronidazole and assessed the toxigenic status and amplified fragment length polymorphism (AFLP) diversity of a collection of 80 C. perfringens isolates of animal origin. In vitro susceptibility testing by the standard agar dilution technique revealed that 22 isolates (27.5% of total) were susceptible to metronidazole, 54 (67.5%) intermediate susceptible, and four (5%) resistant. All isolates belonged to toxinotype A, and two, 41 and 17, of them yielded a positive PCR result for presence of the gene encoding for enterotoxin (cpe), the consensus form of ß2 toxin (cpb2) and the atypical form of ß2 toxin, respectively. AFLP fingerprinting classified isolates into 65 genotypes indicating a high molecular diversity among isolates in this study, and this diversity was reflected among isolates from each susceptibility category. In conclusion, the results of this study revealed extensive genetic diversity among C. perfringens type A isolates regardless of their susceptibility to metronidazole.

Subtyping and antimicrobial susceptibility of Clostridium difficile PCR ribotype 078/126 isolates of human and animal origin.

The Clostridium difficile PCR ribotype complex 078/126 (RT078/126) is often involved in human disease and is also frequently isolated from diverse animal species. The high genetic relatedness between human and animal RT078/126 isolates found in different regions has encouraged discussion about the zoonotic potential of this lineage. We compared for the first time the genetic diversity and antimicrobial susceptibility profiles of human and animal C. difficile RT078/126 isolates from Spain. A collection of 96 isolates (50 of human and 46 of animal origin; 63 and 33 of ribotypes 078 and 126, respectively) was subtyped by an improved amplified fragment length polymorphism (AFLP) fingerprinting method and tested for in vitro antimicrobial susceptibility. A total of 67 genotypes were distinguished, three of which grouped together isolates of human and animal origin. Furthermore, two main groups of isolates that mostly correlated with PCR ribotypes could be distinguished in the AFLP dendrogram. Human origin was significantly associated with resistance to ertapenem, erythromycin and moxifloxacin; resistance to clindamycin and erythromycin was associated with RT126 and AFLP group 1. Twenty-nine isolates (30.2% of total) displayed heteroresistance to metronidazole. Substantial differences were observed in the susceptibility profiles of isolates belonging to a same genotype. Altogether, these results provide a valuable baseline for future studies on the epidemiology of C. difficile RT078/126.