High-efficient separation of deoxyribonucleic acid from pathogenic bacteria by hedgehog-inspired magnetic nanoparticles microextraction.

Efficient separation of deoxyribonucleic acid (DNA) through magnetic nanoparticles (MN) is a widely used biotechnology. Hedgehog-inspired MNs (HMN) possess a high-surface-area due to the distinct burr-like structure of hedgehog, but there is no report about the usage of HMN for DNA extraction. Herein, to improve the selection of MN and illustrate the performance of HMN for DNA separation, HMN and silica-coated Fe3O4 nanoparticles (Fe3O4@SiO2) were fabricated and compared for the high-efficient separation of pathogenic bacteria of DNA. Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) are typical Gram-negative and Gram-positive bacteria and are selected as model pathogenic bacteria. To enhance the extraction efficiency of two kinds of MNs, various parameters, including pretreatment, lysis, binding and elution conditions, have been optimized in detail. In most separation experiments, the DNA yield of HMN was higher than that of Fe3O4@SiO2. Therefore, a HMN-based magnetic solid-phase microextraction (MSPE) and quantitative real-time PCR (qPCR) were integrated and used to detect pathogenic bacteria in real samples. Interestingly, the HMN-based MSPE combined qPCR strategy exhibited high sensitivity with a limit of detection of 2.0 × 101 CFU mL-1 for E. coli and 4.0 × 101 CFU mL-1 for S. aureus in orange juice, and 2.8 × 102 CFU mL-1 for E. coli and 1.1 × 102 CFU mL-1 for S. aureus in milk, respectively. The performance of the proposed strategy was significantly better than that of commercial kit. This work could prove that the novel HMN could be applicable for the efficient separation of DNA from complex biological samples.

Dermatologic Diseases of Four-Toed Hedgehogs.

Dermatologic diseases are frequently diagnosed in the four-toed hedgehog. Common clinical signs include spine loss, scales and crusts, pruritus, alopecia, and masses. Caparinia tripilis, family Psoroptidae, is the most common mite affecting hedgehogs, and is diagnosed via light microscopy. Dermatophytosis from Trichophyton erinacei infection is a common zoonotic dermatologic disease in pet hedgehogs. Diagnosis is through fungal culture. Neoplasia is another major cause of dermatologic diseases in four-toed hedgehogs. The diagnostic approach typically begins with a fine-needle aspirate but often requires a biopsy for a definitive diagnosis.

Zoonotic pathogens linked with hedgehog diphtheric disease.

Hedgehog diphtheric disease (HDD), an ulcerative skin disease with a high fatality rate, is an emerging threat to European hedgehogs (Erinaceus europaeus). We explored the potential role of a panel of zoonotic pathogens in the presumed multifactorial nature of HDD in 188 hedgehogs from 3 wildlife rescue centres in Belgium. As expected, and with a prevalence of 67% in 57 hedgehogs with skin lesions, characteristic of HDD, the occurrence of Corynebacterium ulcerans was strongly associated with the disease. Remarkably, with a prevalence of 42% in affected animals, infections with Borrelia burgdorferi sensu lato were 3.92 times more likely to be detected in HDD (95% confidence interval: 1.650-9.880; p = .0024). Overall, 40 hedgehogs tested positive for the B. burgdorferi sensu lato complex, including Borrelia afzelii (n = 30), Borrelia bavariensis (n = 7) and Borrelia spielmanii (n = 7). Other widely occurring pathogens included Salmonella (prevalence of 19%, with three pulsed-field gel electrophoresis profiles) and Leptospira sp. (prevalence of 11%, including Leptospira interrogans and Leptospira borgpetersenii), but these were not associated with the occurrence of HDD. These findings show that hedgehogs in Belgium represent a significant reservoir of multiple zoonotic bacteria, of which toxigenic C. ulcerans and B. burgdorferi sensu lato are associated with widespread hedgehog skin pathology and mortality.

Emergence of methicillin resistance predates the clinical use of antibiotics.

The discovery of antibiotics more than 80 years ago has led to considerable improvements in human and animal health. Although antibiotic resistance in environmental bacteria is ancient, resistance in human pathogens is thought to be a modern phenomenon that is driven by the clinical use of antibiotics1. Here we show that particular lineages of methicillin-resistant Staphylococcus aureus-a notorious human pathogen-appeared in European hedgehogs in the pre-antibiotic era. Subsequently, these lineages spread within the local hedgehog populations and between hedgehogs and secondary hosts, including livestock and humans. We also demonstrate that the hedgehog dermatophyte Trichophyton erinacei produces two ß-lactam antibiotics that provide a natural selective environment in which methicillin-resistant S. aureus isolates have an advantage over susceptible isolates. Together, these results suggest that methicillin resistance emerged in the pre-antibiotic era as a co-evolutionary adaptation of S. aureus to the colonization of dermatophyte-infected hedgehogs. The evolution of clinically relevant antibiotic-resistance genes in wild animals and the connectivity of natural, agricultural and human ecosystems demonstrate that the use of a One Health approach is critical for our understanding and management of antibiotic resistance, which is one of the biggest threats to global health, food security and development.

European Hedgehogs (Erinaceus europaeus L.) as a Reservoir of Dermatophytes in Poland.

The European hedgehog (Erinaceus europaeus Linnaeus) frequently colonises areas located close to human life in cities, as these are more suitable nest sites offering an abundance of food and allowing avoidance of predators. However, urbanisation has a significant impact on the epidemiology of infectious diseases, including dermatophytoses, the primary source of which are wild animals. In this study, we determined the spectrum of dermatophytes isolated from the European hedgehog and assessed their susceptibility profile to antifungal drugs. Symptomatic and asymptomatic dermatophyte infections were observed in 7.7% and 8% of the 182 examined free-living hedgehogs, respectively. In the pool of the isolated dermatophyte strains, Trichophyton erinacei was dominant (29.9%), followed by Trichophyton mentagrophytes (17.9%), Trichophyton benhamiae (13.4%), Nannizzia gypsea (11.9%), Microsporum canis (10.4%), Nannizzia nana (7.5%), Paraphyton cookei (6.0%), and Nannizzia fulva (3.0%). Susceptibility tests revealed the highest activity of luliconazole and the lowest of activity fluconazole among the azole drugs applied. Although terbinafine generally exhibited high efficacy, two Trichophyton mentagrophytes isolates showed resistance to this drug (MIC = 2 µg/ml) resulting from missense mutations in the SQLE gene corresponding to the amino acid substitution Leu393Phe. Summarising, our study has also revealed that such wildlife animals as hedgehogs can be a reservoir of pathogenic human dermatophytes, including harmful strains resistant to commonly used antifungal drugs.

Benzylpenicillin-producing Trichophyton erinacei and methicillin resistant Staphylococcus aureus carrying the mecC gene on European hedgehogs - A pilot-study.

BACKGROUND: A high carriage rate of methicillin-resistant Staphylococcus aureus with the mecC gene (mecC-MRSA) has been described among Wild European hedgehogs (Europeaus erineaus). Due to this frequent occurrence, it has been suggested that hedgehogs could be a natural reservoir for mecC-MRSA. However, the reason why hedgehogs carry mecC-MRSA remains unknown, but it has been hypothesized that mecC-MRSA could have evolved on the skin of hedgehogs due to the co-occurrence with antibiotic producing dermatophytes. The aim of this pilot-study was therefore to investigate if hedgehogs in Sweden carry Trichophyton spp. and to provide evidence that these dermatophytes are able to produce penicillin or similar substances. In addition, the study aimed to identify if dermatophytes co-occurred with mecC-MRSA. METHODS: Samples were collected from hedgehogs (Europeaus erineaus) that were euthanized or died of natural causes. All samples were screened for dermatophytes and mecC-MRSA using selective cultivation methods. Suspected isolates were characterized using PCR-based methods, genome sequencing and bioinformatic analyses. Identification of penicillin was performed by ultra-high-performance liquid chromatography-tandem mass spectrometry. RESULTS: In total 23 hedgehogs were investigated, and it was shown that two carried Trichophyton erinacei producing benzyl-penicillin, and that these hedgehogs also carried mecC-MRSA. The study also showed that 60% of the hedgehogs carried mecC-MRSA. CONCLUSION: The pilot-study demonstrated that Trichophyton erinacei, isolated from Swedish hedgehogs, can produce benzylpenicillin and that these benzylpenicillin-producing T. erinacei co-occurred with mecC-MRSA. The study also reconfirmed the high occurrence of mecC-MRSA among hedgehogs.

Red Fluorescence of European Hedgehog (Erinaceus europaeus) Spines Results from Free-Base Porphyrins of Potential Microbial Origin.

Bioluminescence has been recognized as an important means for inter- and intra-species communication. A growing number of reports of red fluorescence occurring in keratinaceous materials have become available. The fluorophore(s) in these cases were shown to be, or suspected to be, free base porphyrins. The red fluorescence found in the downs of bustards was associated with inter-species signaling in mate selection. First reported in 1925, we confirm that spines of the European hedgehog (Erinaceus europaeus) when irradiated with UV (365-395 nm) light display red fluorescence localized in the light-colored sections of their proximal ends. Using reflectance fluorescence spectroscopy, we confirmed that the fluorophores responsible for the emission are free-base porphyrins, as suspected in the original report. Base-induced degradation of the spine matrix and subsequent HPLC, UV-vis, and ESI+ mass spectrometry analysis revealed the presence of a mixture of coproporphyrin III and uroporphyrin III as predominant porphyrins and a minor fraction of protoporphyrin IX. Investigation of the spine microbiome uncovered the abundant presence of bacteria known to secrete and/or interconvert porphyrins and that are not present on the non-fluorescing quills of the North American porcupine (Erethizon dorsatum). Given this circumstantial evidence, we propose the porphyrins could originate from commensal bacteria. Furthermore, we hypothesize that the fluorescence may be incidental and of no biological function for the hedgehog.

Non-tuberculous Mycobacterial Granulomatous Dermatitis in an African Pygmy Hedgehog (Atelerix albiventris).

A 2-year-old female African pygmy hedgehog (Atelerix albiventris) with a cutaneous nodular lesion on the dorsal surface of the right forelimb was presented for clinical examination. Histopathological findings included granulomatous dermatitis with extensive necrosis. Long and slender acid-fast bacilli were seen within the cytoplasm of macrophages and in extracellular spaces. Bacteriological culture of skin revealed acid-fast bacilli and non-tuberculous mycobacterial infection was confirmed by gene sequencing and identity analysis using the BLAST tool. To our knowledge, this is the first report of non-tuberculous granulomatous dermatitis in hedgehogs.

Detection of Intestinal Parasites and Genetic Characterization of Enterocytozoon bieneusi in Hedgehogs from China.

Microsporidia are a group of unicellular and opportunistic intestinal parasites in which Enterocytozoon bieneusi is a frequent species causing microsporidial infections in humans. Many domesticated and wild animals have been shown to be hosts of E. bieneusi and other microsporidia. The role of hedgehogs in the ecology of microsporidia is unclear; therefore, we investigated the prevalence and genetic diversity of E. bieneusi, Cryptosporidium, and Blastocystis spp. in hedgehogs (Erinaceus amurensis) collected from Hubei Province in Central China. PCR amplification of the internal transcribed spacer region of the ribosomal DNA indicated that 9.8% (4/41) hedgehogs were positive to E. bieneusi, but none (0/41) was positive to Cryptosporidium and Blastocystis spp. Phylogenetic analysis showed the strains detected from the hedgehogs belong to four novel genotypes (EA1-EA4), which were most closely related to type IV of group 1c. This study demonstrated that hedgehogs are hosts of E. bieneusi and may play a role in the transmission of E. bieneusi to humans in the process of being caught and slaughtered.

Occurrence and Genotyping of Coxiella burnetii in Hedgehogs in China.

Coxiella burnetii is the causative agent of query fever (Q fever), and distributes broadly in environment. Livestock are identified as main reservoirs, which may infect people through their contaminative urine, feces, milk, and birth products. Wild animals can also be the potential carriers and transmitters of C. burnetii. To understand the geographic distribution and host species of C. burnetii in China, we investigated the prevalence of C. burnetii in hedgehogs (Erinaceus amurensis) in Hubei Province. Hedgehogs were tested for C. burnetii with PCR targeting three genes (com1, rrs, and icd) followed by multispacer sequence typing (MST). We found that 12.2% (5/41) hedgehogs were PCR positive for C. burnetii. MST revealed presence of two novel genotypes and phylogenetic analysis revealed that the strains were similar to a group of isolates from chronic Q fever patients and mammals. This study showed that C. burnetii are highly prevalent in hedgehogs in Hubei Province in central China, suggesting that hedgehogs may play an important role in the ecology and transmission of C. burnetii to humans because it is captured and used as traditional medicine in China.

Salmonellosis in Hedgehogs.

African pygmy hedgehogs are popular pets worldwide. The knowledge and understanding of pet hedgehog common veterinary conditions are increasing as new information and research are published; however, there is still much to learn about this fascinating animal. Salmonella is one of the most common zoonoses worldwide and is naturally isolated from the intestinal tract of many animal species, including hedgehogs. This article discusses the cause, clinical signs, diagnosis, treatment, and prevention of salmonella infection in hedgehogs, primarily focusing on African pygmy hedgehogs, with some reference to European hedgehogs.

Detection of Leptospira interrogans in Hedgehogs from Central China.

Background:Leptospira is the causative agent of leptospirosis, a zoonotic disease of global importance. To have a better understanding on the host species of Leptospira, we investigated the prevalence of Leptospira species in hedgehogs in Central China. Materials and Methods: Hedgehogs were captured in Hubei Province, China in May and October, 2018. Total DNA was extracted from the kidney tissues of hedgehogs for determining the Leptospira species by PCR amplification of the rrs2, secY, and flaB genes with genus-specific primers. Results: PCR amplification indicated that the positive rate of hedgehogs to the rrs2, secY, and flaB genes were 19.5% (8/41), 12.2% (5/41), and 9.8% (4/41), respectively. The homology of the partial sequence of rrs2, secY, and flaB genes were 99.0-100% among the Leptospira strains from hedgehogs. Phylogenetic analysis revealed that Leptospira species detected in this study clustered together with Leptospira interrogans.Conclusions: We detected L. interrogans from hedgehogs in Central China, suggesting hedgehogs are the hosts of L. interrogans.

Detection of Tetracycline Resistance Genes in European Hedgehogs (Erinaceus europaeus) and Crested Porcupines (Hystrix cristata).

Relatively little is known regarding the role of wildlife in the development of antibiotic resistance. Our aim was to assess the presence of the tetracycline resistance genes, tet(A), tet(B), tet(C), tet(D), tet(E), tet(G), tet(K), tet(L), tet(M), tet(O), tet(P), tet(Q), tet(S), and tet(X), in tissue samples of 14 hedgehogs (Erinaceus europaeus) and 15 crested porcupines (Hystrix cristata) using PCR assays. One or more tet genes were found in all but three hedgehogs and one crested porcupine. Of the 14 tetracycline resistance genes investigated, 13 were found in at least one sample; tet(G) was not detected. We confirmed the potential role of wild animals as bioindicators, reservoirs, or vectors of antibiotic-resistant bacteria in the environment.

European hedgehogs (Erinaceus europaeus) as a natural reservoir of methicillin-resistant Staphylococcus aureus carrying mecC in Denmark.

OBJECTIVES: A recent study from Sweden showed that European hedgehogs may constitute a reservoir for methicillin-resistant Staphylococcus aureus (MRSA), but this host-parasite relationship remains to be investigated in other countries. In this study, we therefore sought to: 1) determine the dissemination of MRSA in European hedgehogs throughout Denmark; 2) investigate determinants of MRSA carriage in hedgehogs; 3) determine the potential for zoonotic transmission of MRSA from hedgehogs to humans; and 4) characterise the detected MRSA on both a phenotypic and molecular level. METHODS: Nasal swabs were taken from 188 dead hedgehogs collected by volunteers throughout Denmark to determine the occurrence of MRSA. Additionally, 16 hedgehog rehabilitators were tested for potential zoonotic transmission of MRSA from hedgehogs to humans. The swabs were incubated in tryptic soy broth supplemented with 6.5% NaCl, followed by spread of 10 µl on Brilliance MRSA 2 agar. One presumptive MRSA colony from each plate was subcultured on 5% blood agar. All S. aureus subcultures were verified by a PCR assay detecting mecA, mecC, lukF-PV, scn, and spa, followed by spa typing. RESULTS: A total of 114 (61%) hedgehogs carried mecC-MRSA, whereas none carried mecA-MRSA. The detected mecC-MRSA belonged to two genetic lineages CC130 (spa-types: t528, t843, t1048, t3256, t3570, t6220, t17133) and CC1943 (spa-types: t978, t2345, t3391, t8835, t16868), 52% of which were spa-type t843 (CC130).The detection rate of mecC-MRSA in the hedgehogs was similar regardless of cause of death, sex, region and habitat type. None of the hedgehog rehabilitators carried MRSA. CONCLUSIONS: This nationwide study confirms a high occurrence of mecC-MRSA in hedgehogs, which could serve as a natural reservoir for this specific type of MRSA. Furthermore, our study did not find signs of zoonotic transmission of mecC-MRSA to hedgehog rehabilitators.

Tox-positive Corynebacterium ulcerans in hedgehogs, Germany.

Toxigenic Corynebacterium ulcerans may cause both respiratory and cutaneous diphtheria in humans. As a zoonotic emerging pathogen it has been isolated from a wide variety of animals living in captivity, such as livestock, pet, zoo and research animals and additionally in a large number of different wild animals. Here we report the isolation of tox-positive C. ulcerans in four hedgehogs with cutaneous diphtheria and pneumonia, respectively.

Tinea Corporis Due to Trichophyton erinacei Probably Transmitted from a Hedgehog.

A 26-year-old female homemaker presented with an approximately 2-month history of an erythematous lesion with agminated seropapules in the right popliteal fossa associated with scales and crusts. The lesion was initially treated as contact dermatitis, but there was no improvement. KOH examination revealed filamentous fungi. The fungal culture was positive, and the morphological characteristics were identical to those of Trichophyton mentagrophytes complex. The fungus was identified as T. erinacei based on genetic analysis. This is the second case report of human tinea corporis due to this fungus in Japan.

[Mexican case of tinea incognito and granuloma de Majocchi acquired from a hedgehog]. / Tiña incógnita y granuloma de Majocchi adquirido de un erizo. Un caso mexicano.

Exotic pets, such as the ground hedgehog, are capable of transmitting to the human being different zoonoses, such as salmonellosis, mycobacteria, protozoa such as Cryptosporidium parvum, and dermatophytosis. We present the case report of a male adult patient, who had recently acquired a ground hedgehog, who presented in his hand a ringworm lesion incognito and a Majocchi granuloma. The etiological agent was identified as Trichophyton erinacei by mycological culture and molecular biology. The patient was treated with terbinafine oral, with excellent response.