Genetic diversity of Clostridium perfringens type A isolates from animals, food poisoning outbreaks and sludge

BACKGROUND: Clostridium perfringens, a serious pathogen, causes enteric diseases in domestic animals and food poisoning in humans. The epidemiological relationship between C. perfringens isolates from the same source has previously been investigated chiefly by pulsed-field gel electrophoresis (PFGE). In this study the genetic diversity of C. perfringens isolated from various animals, from food poisoning outbreaks and from sludge was investigated. RESULTS: We used PFGE to examine the genetic diversity of 95 C. perfringens type A isolates from eight different sources. The isolates were also examined for the presence of the beta2 toxin gene (cpb2) and the enterotoxin gene (cpe). The cpb2 gene from the 28 cpb2-positive isolates was also partially sequenced (519 bp, corresponding to positions 188 to 706 in the consensus cpb2 sequence). The results of PFGE revealed a wide genetic diversity among the C. perfringens type A isolates. The genetic relatedness of the isolates ranged from 58 to 100% and 56 distinct PFGE types were identified. Almost all clusters with similar patterns comprised isolates with a known epidemiological correlation. Most of the isolates from pig, horse and sheep carried the cpb2 gene. All isolates originating from food poisoning outbreaks carried the cpe gene and three of these also carried cpb2. Two evolutionary different populations were identified by sequence analysis of the partially sequenced cpb2 genes from our study and cpb2 sequences previously deposited in GenBank. CONCLUSION: As revealed by PFGE, there was a wide genetic diversity among C. perfringens isolates from different sources. Epidemiologically related isolates showed a high genetic similarity, as expected, while isolates with no obvious epidemiological relationship expressed a lesser degree of genetic similarity. The wide diversity revealed by PFGE was not reflected in the 16S rRNA sequences, which had a considerable degree of sequence similarity. Sequence comparison of the partially sequenced cpb2 gene revealed two genetically different populations. This is to our knowledge the first study in which the genetic diversity of C. perfringens isolates both from different animals species, from food poisoning outbreaks and from sludge has been investigated

Evaluation of LHP® (1% hydrogen peroxide) cream versus petrolatum and untreated controls in open wounds in healthy horses: a randomized, blinded control study

<p>Abstract</p> <p>Background</p> <p>Treatment and protection of wounds in horses can be challenging; protecting bandages may be difficult to apply on the proximal extremities and the body. Unprotected wounds carry an increased risk of bacterial contamination and subsequent infection which can lead to delayed wound healing. Topical treatment with antimicrobials is one possibility to prevent bacterial colonization or infection, but the frequent use of antimicrobials ultimately leads to development of bacterial resistance which is an increasing concern in both human and veterinary medicine.</p> <p>Methods</p> <p>Standardized wounds were created in 10 Standardbred mares. Three wounds were made in each horse. Two wounds were randomly treated with LHP^®or petrolatum and the third wound served as untreated control. All wounds were assessed daily until complete epithelization. Protocol data were recorded on day 2, 6, 11, 16, 21 and 28. Data included clinical scores for inflammation and healing, photoplanimetry for calculating wound areas and swab cytology to assess bacterial colonization and inflammation. Bacterial cultures were obtained on day 2, 6 and 16.</p> <p>Results</p> <p>Mean time to complete healing for LHP^®treated wounds was 32 days (95%CI = 26.9-37.7). Mean time to complete healing for petrolatum and untreated control wounds were 41.6 days (95%CI = 36.2-47.0) and 44.0 days (95%CI = 38.6-49.4) respectively. Wound healing occurred significantly faster in LHP^®wounds compared to both petrolatum (p = 0.0004) and untreated controls (p < 0.0001). There was no significant difference in time for healing between petrolatum and untreated controls. Total scores for bacteria and neutrophils were significantly (p < 0.0001) lower for LHP^®treated wounds compared to petrolatum from day 16 and onwards. <it>Staphylococcus aureus </it>and <it>Streptococcus zooepidemicus </it>were only found in cultures from petrolatum treated wounds and untreated controls.</p> <p>Conclusions</p> <p>Treatment with LHP^®reduced bacterial colonization and was associated with earlier complete wound healing. LHP^®cream appears to be safe and effective for topical wound treatment or wound protection.</p

Management of Animal Botulism Outbreaks: From Clinical Suspicion to Practical Countermeasures to Prevent or Minimize Outbreaks

Botulism is a severe neuroparalytic disease that affects humans, all warm-blooded animals, and some fishes. The disease is caused by exposure to toxins produced by Clostridium botulinum and other botulinum toxin–producing clostridia. Botulism in animals represents a severe environmental and economic concern because of its high mortality rate. Moreover, meat or other products from affected animals entering the food chain may result in a public health problem. To this end, early diagnosis is crucial to define and apply appropriate veterinary public health measures. Clinical diagnosis is based on clinical findings eliminating other causes of neuromuscular disorders and on the absence of internal lesions observed during postmortem examination. Since clinical signs alone are often insufficient to make a definitive diagnosis, laboratory confirmation is required. Botulinum antitoxin administration and supportive therapies are used to treat sick animals. Once the diagnosis has been made, euthanasia is frequently advisable. Vaccine administration is subject to health authorities' permission, and it is restricted to a small number of animal species. Several measures can be adopted to prevent or minimize outbreaks. In this article we outline all phases of management of animal botulism outbreaks occurring in wet wild birds, poultry, cattle, horses, and fur farm animals

Clostridium difficile in horses

In recent years, several animal hospitals in Sweden have reported an increased frequency of acute and often fatal colitis in mature horses. The most common risk group was horses hospitalized and treated for various non-gastrointestinal diseases. After a few days of hospitalization some horses developed acute diarrhea. Another risk group was mares when their foals were treated orally with erythromycin in combination with rifampicin for Rhodococcus equi pneumonia. Some mares developed diarrhea suddenly often after 3-4 days treatment ofthe foals at an animal hospital.In human medicine, the bacterium Clostridium difficile is since many years a wellknown nosocomial pathogen in antibiotic associated diarrhea. This thesis, based on five scientific publications, describes the association of C. difficile colonisation with the occurrence of diarrhea, antibiotic treatment and the age ofthe horses. The occurrence and survival ofthe bacterium in the environment and its antimicrobial susceptibility were also studied. Furthermore, the role ofthe antibiotic erythromycin in induction of acute colitis in horses was investigated.C. difficile is associated with acute colitis in mature horses following treatment with antibiotics, as about 40% of the horses proved positive by culture and 28% in the cytotoxin B test of faeces. No other pathogen was detected in horses affected by antibiotic-associated diarrhea.C. difficile, and/or its cytotoxin, is also associated with acute colitis in mares when their foals are being treated with erythromycin and rifampicin for R. equi pneumonia. The colitis can have resulted from an accidental ingestion of erythromycin by the mares. In an experimental study it was also demonstrated in horses that erythromycin can induce severe colitis associated with proliferation ofC. difficile.A new interesting finding was that in healthy foals younger than 14 days, C. difficile was isolated from every third foal whereas all older foals except one proved negative. Many asymptomatic carriers were also found among non-diarrheic foals treated with antibiotics.Antimicrobial susceptibility testing showed that isolates were susceptible to metronidazole (MIC In conclusion, the work described in this thesis is a contribution to increased knowledge of C. difficile as an etiological factor in antibiotic-associated diarrhea in horses. Preventive measures to avoid accidental ingestion of erythromycin by mares from the treatment oftheir foals are recommended

Faecal bacterial composition in horses with and without free faecal liquid: a case control study

Free faecal liquid (FFL) is a condition in horses which manifests as differential defecation of solid and liquid phases of faeces. The etiology of FFL is currently unknown, but deviances in the hindgut microbiota has been suggested to be of importance. The present study aimed to compare the faecal bacterial composition of farm-matched horses with (case, n=50) and without (control, n=50) FFL. Samples were collected at three different occasions. The V3 and V4 regions of the 16S rRNA gene were amplified and sequenced using Illumina sequencing. Also, samples were cultivated for detection of Clostridioides difficile and Clostridium perfringens. Analysis revealed similar faecal bacterial composition between case and control horses, but an effect of sampling period (p=0.0001). Within sampling periods, 14 genera were present in higher or lower proportions in case compared to control horses in at least one sampling period. Compared to controls, case horses had higher relative abundance of Alloprevotella (adjusted p<0.04) and lower relative abundance of Bacillus spp. (adjusted p<0.03) in at least two sampling periods. All horses tested negative for C. difficile and C. perfringens by culture of faeces. Further studies are required to establish the clinical relevance of specific bacterial taxa in FFL

GIFeGSH : A New Genomic Island Might Explain the Differences in Brucella Virulence

An imported dog was confirmed to be positive with canine brucellosis in Sweden in 2010. The whole genome of Brucella canis SVA10 was subjected to phage analysis (WGS-PA) and was assigned to the Asian B. canis cluster. Further analysis indicated that the genome of B. canis SVA10 is smaller compared to genomes of the same species. A 35,781 bp genomic island (GI) was found to be absent in strain SVA10 which was detected by read mapping the paired reads to the genome of B. canisATCC 23,365T. The lacking genes of genomic island GIFeGSH are mainly coding for iron uptake enzymes and parts of the glutathione pathway. A screening of all available whole genome sequences of Brucella strains confirmed that GIFeGSH is also missing in four more strains of B. canis but present in several strains of B. abortus, B. melitensis, B. suis, B. ovis, B. microti, B. pinnipedialis, and B. ceti. Parts of the GI were present, but scattered in two other B. canis strains. The aim of this study was to find differences in the genomes of Brucella which might explain former described differences in virulence. The analysis was extended to all available Brucella genomes after the detection of a genomic island in strain SVA10

Differential defecation of solid and liquid phases in horses—A descriptive survey

Free faecal liquid (FFL) is a condition in horses where faeces are voided in one solid and one liquid phase. The liquid phase contaminates the tail, hindlegs and area around the anus of the horse, resulting in management problems and potentially contributing to impaired equine welfare. The underlying causes are not known, but anecdotal suggestions include feeding wrapped forages or other feed- or management-related factors. Individual horse factors may also be associated with the presence of FFL. This study, therefore, aimed to characterize horses showing FFL particularly when fed wrapped forages, and to map the management and feeding strategies of these horses. Data were retrieved by a web-based survey, including 339 horses with FFL. A large variety of different breeds, ages, disciplines, coat colours, housing systems and feeding strategies were represented among the horses in the study, meaning that any type of horse could be affected. Respondents were asked to indicate if their horse had diminished signs of FFL with different changes in forage feeding. Fifty-eight percent (n = 197) of the horse owners reported diminished signs of FFL in their horses when changing from wrapped forages to hay; 46 (n = 156) of the horse owners reported diminished signs of FFL in their horses when changing from wrapped forages to pasture; 17% (n = 58) reported diminished signs of FFL when changing from any type of forage batch to any other forage. This indicated that feeding strategy may be of importance, but cannot solely explain the presence of FFL. The results also showed that the horses in this study had a comparably high incidence of previous colic (23%, n = 78) compared to published data from other horse populations. In conclusion, the results showed that FFL may affect a large variety of horse types and that further studies should include detailed data on individual horse factors including gastrointestinal diseases as well as feeding strategies, in order to increase the chance of finding causes of FFL.publishedVersio

Differential defecation of solid and liquid phases in horses—A descriptive survey

Free faecal liquid (FFL) is a condition in horses where faeces are voided in one solid and one liquid phase. The liquid phase contaminates the tail, hindlegs and area around the anus of the horse, resulting in management problems and potentially contributing to impaired equine welfare. The underlying causes are not known, but anecdotal suggestions include feeding wrapped forages or other feed- or management-related factors. Individual horse factors may also be associated with the presence of FFL. This study, therefore, aimed to characterize horses showing FFL particularly when fed wrapped forages, and to map the management and feeding strategies of these horses. Data were retrieved by a web-based survey, including 339 horses with FFL. A large variety of different breeds, ages, disciplines, coat colours, housing systems and feeding strategies were represented among the horses in the study, meaning that any type of horse could be affected. Respondents were asked to indicate if their horse had diminished signs of FFL with different changes in forage feeding. Fifty-eight percent (n = 197) of the horse owners reported diminished signs of FFL in their horses when changing from wrapped forages to hay; 46 (n = 156) of the horse owners reported diminished signs of FFL in their horses when changing from wrapped forages to pasture; 17% (n = 58) reported diminished signs of FFL when changing from any type of forage batch to any other forage. This indicated that feeding strategy may be of importance, but cannot solely explain the presence of FFL. The results also showed that the horses in this study had a comparably high incidence of previous colic (23%, n = 78) compared to published data from other horse populations. In conclusion, the results showed that FFL may affect a large variety of horse types and that further studies should include detailed data on individual horse factors including gastrointestinal diseases as well as feeding strategies, in order to increase the chance of finding causes of FFL.publishedVersio

Genetic diversity of <it>Clostridium perfringens </it>type A isolates from animals, food poisoning outbreaks and sludge

Abstract Background Clostridium perfringens, a serious pathogen, causes enteric diseases in domestic animals and food poisoning in humans. The epidemiological relationship between C. perfringens isolates from the same source has previously been investigated chiefly by pulsed-field gel electrophoresis (PFGE). In this study the genetic diversity of C. perfringens isolated from various animals, from food poisoning outbreaks and from sludge was investigated. Results We used PFGE to examine the genetic diversity of 95 C. perfringens type A isolates from eight different sources. The isolates were also examined for the presence of the beta2 toxin gene (cpb2) and the enterotoxin gene (cpe). The cpb2 gene from the 28 cpb2-positive isolates was also partially sequenced (519 bp, corresponding to positions 188 to 706 in the consensus cpb2 sequence). The results of PFGE revealed a wide genetic diversity among the C. perfringens type A isolates. The genetic relatedness of the isolates ranged from 58 to 100% and 56 distinct PFGE types were identified. Almost all clusters with similar patterns comprised isolates with a known epidemiological correlation. Most of the isolates from pig, horse and sheep carried the cpb2 gene. All isolates originating from food poisoning outbreaks carried the cpe gene and three of these also carried cpb2. Two evolutionary different populations were identified by sequence analysis of the partially sequenced cpb2 genes from our study and cpb2 sequences previously deposited in GenBank. Conclusion As revealed by PFGE, there was a wide genetic diversity among C. perfringens isolates from different sources. Epidemiologically related isolates showed a high genetic similarity, as expected, while isolates with no obvious epidemiological relationship expressed a lesser degree of genetic similarity. The wide diversity revealed by PFGE was not reflected in the 16S rRNA sequences, which had a considerable degree of sequence similarity. Sequence comparison of the partially sequenced cpb2 gene revealed two genetically different populations. This is to our knowledge the first study in which the genetic diversity of C. perfringens isolates both from different animals species, from food poisoning outbreaks and from sludge has been investigated.</p

Differential defecation of solid and liquid phases in horses—A descriptive survey

Free faecal liquid (FFL) is a condition in horses where faeces are voided in one solid and one liquid phase. The liquid phase contaminates the tail, hindlegs and area around the anus of the horse, resulting in management problems and potentially contributing to impaired equine welfare. The underlying causes are not known, but anecdotal suggestions include feeding wrapped forages or other feed- or management-related factors. Individual horse factors may also be associated with the presence of FFL. This study, therefore, aimed to characterize horses showing FFL particularly when fed wrapped forages, and to map the management and feeding strategies of these horses. Data were retrieved by a web-based survey, including 339 horses with FFL. A large variety of different breeds, ages, disciplines, coat colours, housing systems and feeding strategies were represented among the horses in the study, meaning that any type of horse could be affected. Respondents were asked to indicate if their horse had diminished signs of FFL with different changes in forage feeding. Fifty-eight percent (n = 197) of the horse owners reported diminished signs of FFL in their horses when changing from wrapped forages to hay; 46 (n = 156) of the horse owners reported diminished signs of FFL in their horses when changing from wrapped forages to pasture; 17% (n = 58) reported diminished signs of FFL when changing from any type of forage batch to any other forage. This indicated that feeding strategy may be of importance, but cannot solely explain the presence of FFL. The results also showed that the horses in this study had a comparably high incidence of previous colic (23%, n = 78) compared to published data from other horse populations. In conclusion, the results showed that FFL may affect a large variety of horse types and that further studies should include detailed data on individual horse factors including gastrointestinal diseases as well as feeding strategies, in order to increase the chance of finding causes of FFL