Search

To assess bacterial translocation through needle punctures and the impact of preparation protocols using equine cadaver skin and different needle sizes and types. Nonprepared, disinfected, clipped and disinfected, and clipped and antiseptically washed skin specimens were sampled sterilely using swabs and biopsies to compare disinfection efficacy. The skin specimens were punctured using 18G hollow needles (18G), 18G needles with stylet (18GM), and 22G hollow needles (22G) and were flushed with sterile saline solution. The saline was collected in blood culture systems or homogenization test tubes and processed for qualitative (ql) or semiquantitative microbiologic evaluation. Disinfection and antiseptic preparation of skin specimens led to a significant reduction of bacteria (P < .05) sampled with swabs and biopsies, whereas clipping did not produce further significant differences. Bacteria were detected in many needle punctures, depending on the preparation protocols and needle types and sizes used. The highest amount of bacteria was detected with 18GM and the lowest with 22G (ql; nonprepared; nonsignificant). Disinfection and antiseptic preparation reduced bacterial translocation significantly in 18G punctures (ql; P < .05), whereas clipping did not result in any further significant difference. Overall, more bacteria were detected after 18G needle punctures when samples were processed qualitatively. Disinfection before injections reduces the number of bacteria translocated into deeper tissue. Larger needles and needles with stylet increase the number of bacteria translocated through the skin.

Among pathogenic Nocardia species in humans and animals, infections caused by Nocardia (N.) veterana have rarely been described and so far, all non-human cases are linked to bovine mastitis in Brazil. The aim of this study was to identify the causative microorganism involved in the death of a three-month-old dog suffering from dyspnea and neurological deficits ante mortem. Pathomorphological investigation revealed (pyo-)granulomatous lesions in various organs. Bacteriological examination was performed and the respective bacteria were subjected to matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS), 16S rDNA sequencing, and antimicrobial susceptibility testing by broth microdilution. Gram-staining and colony morphology suggested the presence of an actinomycete which was identified as N. veterana by MALDI-TOF MS. This identification was confirmed by 16S rDNA sequence analysis. Distemper-associated immunosuppression may have played a role in the pathogenesis of systemic nocardiosis in this dog. Retrospective analysis of the antimicrobial susceptibility status showed that the N. veterana isolate was multiresistant and displayed high minimal inhibitory concentrations to all antimicrobial agents used for the dog's therapy. To the best of our knowledge, this is the first report of a systemic nocardiosis caused by N. veterana in a dog with a concurrent canine distemper virus infection.

Acute hemorrhagic diarrhea syndrome (AHDS), formerly named canine hemorrhagic gastroenteritis, is one of the most common causes of acute hemorrhagic diarrhea in dogs, and is characterized by acute onset of diarrhea, vomiting, and hemoconcentration. To date, histologic examinations have been limited to postmortem specimens of only a few dogs with AHDS. Thus, the aim of our study was to describe in detail the distribution, character, and grade of microscopic lesions, and to investigate the etiology of AHDS. Our study comprised 10 dogs with AHDS and 9 control dogs of various breeds, age, and sex. Endoscopic biopsies of the gastrointestinal tract were taken and examined histologically (H&E, Giemsa), immunohistochemically ( Clostridium spp., parvovirus), and bacteriologically. The main findings were acute necrotizing and neutrophilic enterocolitis (9 of 10) with histologic detection of clostridia-like, gram-positive bacteria on the necrotic mucosal surface (9 of 10). Clostridium perfringens isolated from the duodenum was identified as type A (5 of 5) by multiplex PCR (5 of 5). In addition, each of the 5 genotyped isolates encoded the pore-forming toxin netF. Clostridium spp. (not C. perfringens) were cultured from duodenal biopsies in 2 of 9 control dogs. These findings suggest that the pore-forming netF toxin is responsible for the necrotizing lesions in the intestines of a significant proportion of dogs with AHDS. Given that the stomach was not involved in the process, the term “acute hemorrhagic diarrhea syndrome” seems more appropriate than the frequently used term “hemorrhagic gastroenteritis.”

The aim of this study was to determine the frequency of carriage of methicillin-resistant Staphylococcus pseudintermedius (MRSP) among dogs admitted to a small animal hospital during a 17-month period, to characterize these isolates and to initially screen for possible factors associated with MRSP carriage. Swabs were taken from the nose/pharynx and the perineum as well as from wounds and skin infections (if present) of 814 dogs before entering the small animal hospital. A questionnaire for background information was completed. The staphylococcal species and methicillin resistance were confirmed pheno- and genotypically. The identified MRSP isolates were characterized by SCCmec typing, testing for susceptibility to 25 antimicrobial agents and SmaI-directed pulsed-field gel electrophoresis. A first screening for possible risk factors for MRSP carriage was performed by means of unifactorial contingency tables and CART analysis. Sixty (7.4%) dogs were positive for MRSP. All MRSP isolates harboured a type II-III SCCmec cassette and showed extended resistance to antimicrobial agents. Fifteen different SmaI patterns were observed. The major factors that clustered with MRSP carriage were former hospitalization and antibiotic treatment within the last six months before sampling. This study showed that only a minor part of the sampled dogs carried multi-resistant MRSP isolates. The facts that prior hospitalization and/or antibiotic therapy are potential associated factors for MRSP carriage underline the necessity of a judicious use of antibiotics in small animal medicine.

Based on cryo-SEM, standard and high resolution TEM, glycoconjugate histochemistry, and microbiological differentiation, the present study demonstrates the colonisation of the epithelium of the equine oesophagus with microorganisms. As particularly apparent using cryo-SEM to illustrate natural conditions, the present microbiota were clearly dominated by bacteria, forming a one-layer system, as attached to and embedded in concentrated mannose/mannan substances covering the outer stratum corneal cells. Bacterial numbers ranged from 5600 to 7200 per mm(2) in the central part of the oesophagus, the number of fungi was less than 1% of the amount of bacteria. The compact stratum corneal cells showed numerous short protrusions sometimes as part of desmosomal contacts, but mainly projecting into distinct intercellular spaces, containing a mixture of acid and neutral glycoconjugates. The outermost corneal cells exhibited intact mitochondria and cytoplasmic vesicles, and a number of short cell processes toward the oesophageal lumen; i.e. into the glycoconjugate layers on the surface of the oesophagus. The diverse spectrum of bacteria found indicated a permanent mucosal flora, predominated by facultative and obligate anaerobic species. The genera isolated most frequently and in highest numbers included streptococci, Prevotella spp., Fusobacterium spp. and Actinobacillus equuli. Only two groups of Enterobacteriaceae (Escherichia coli, Pantoea spp.) were regularly found and their abundance was lower than that of the other bacterial groups mentioned above. Yeasts were very rarely identified as the typically present fungi.

In a field study with fattening pigs, effects of feed particle size at the dietary presence of organic acids on Salmonella prevalence were measured. On two farms (f1/f2), each holding ∼800 pigs, diets based on finely ground (control) or coarsely ground ingredients (experiment) were fed as crumbs. On f1 both control and experimental grower and finisher diets contained identical concentrations of formic and propionic acid (0.4% and 0.2% respectively). On f2 only finisher diet of the experimental group contained 1.2% potassium diformate. At the start of the fattening period no statistical differences were measured between Salmonella prevalence in animals fed control and experimental diets on both farms. At slaughter Salmonella prevalence in caecal contents was lower (p < 0.05) on f1 in animals fed the experimental diet. Furthermore, the number of seronegative meat juice samples taken from these animals [optical density (OD)

The aim of this study was to examine whether and to what extent the addition of potassium diformate (pdf) or free organic acids (fpa) to the diet and the grinding intensity might affect the course of infection and the passage of orally applied Salmonella and Escherichia coli in pigs. Experiments were carried out using 80 reared piglets allotted to four groups. Pigs were fed pelleted diets ad libitum (except during a 15 h feed-withholding-period before infection). The control diet contained finely ground cereals (2 mm screen). To two test diets (also finely ground) 1.2% pdf, 0.9% organic acids (75% formic and 25% propionic acid, fpa) respectively were added. The fourth diet (without acids) was based on coarsely ground cereals (6-mm screen). After experimental infection alternately with S. Derby or E. coli, the course of infection was examined (rectal swab technique). Pigs were sacrificed 4-5 h after a further oral application of approximately 10(9)-10(10) CFU S. Derby or E. coli to determine the counts of Salmonella or E. coli in chyme (classical culture methods). Adding pdf or fpa to the diet led to reduced Salmonella shedding and resulted in significantly lower counts of Salmonella and E. coli in the stomach content indicating an improved efficacy of the stomach barrier. In the distal parts of the digestive tract, the effect was less obvious concerning counts of E. coli, whereas counts of Salmonella were reduced markedly as well. The diet based on coarsely ground cereals failed to demonstrate positive effects concerning infection and passage of orally applied bacteria as well, but this diet was also pelleted and showed unintentionally, comparable amounts of fine particles. Results obtained in this study allow the recommendation of using pdf or organic acids as additives when dietary measures against Salmonella or E. coli in pigs are required.

Summary In bronchoalveolar lavage fluid (BALF) of pigs originating from different herds bacteria, cells and the antibacterial peptide PR‐39 were examined to gain information about the lung health status. In a high health nucleus herd 56% and in low health herds 20–100% of the examined pigs were found positive for potentially pathogenic bacteria. Based on these findings, a novel definition for bacterial respiratory tract disease was established using an 8% cut‐off for the relative number of neutrophils in bronchoscopic and a 40% cut‐off in transtracheal BALF in combination with the occurrence of potentially pathogenic microorganisms. The antibacterial peptide PR‐39 was highly correlated to this definition of respiratory disease. An assessment of the bacteriological respiratory health status appears to be possibly based on the determination of PR‐39 concentrations in BALF using different cut‐off values according to the lavage method (2.5 nM for bronchoscopic and 5 nM for transtracheal BALF).

SummaryBackground Transient bacteraemia can occur during tooth extraction in humans, and dogs and can lead to severe infectious sequelae. Several case reports describe distant site infections following equine tooth extraction, but the occurrence of bacteraemia during dental surgery has not been evaluated in the horse. Objectives To determine if transient bacteraemia occurs during tooth extraction in horses, describe isolated organisms and compare these with those found in the diseased teeth. Study design Prospective, observational study. Methods Blood was collected aseptically for blood culture before, during and after oral extraction of incisor, canine or cheek teeth from 20 adult horses undergoing dental extraction that had not received antimicrobial agents for at least 4 weeks prior to surgery. Bacteria found in blood cultures were compared with those found in swab samples obtained from the extracted teeth. Results Eighteen of 20 horses had positive blood cultures at one or more time points. Streptococcus spp., Actinomyces spp., Fusobacterium spp. and Prevotella spp. were most commonly found. Bacterial genera isolated from swab samples of extracted teeth largely corresponded with those identified in blood cultures. Main limitations This study was limited by its use of only conventional bacterial culture, the lack of statistical analysis to explore associations between gingiva score and the occurrence of bacteraemia, and the lack of an age-matched control group of horses not undergoing exodontia. Conclusions Transient bacteraemia of oral origin commonly occurs during dental extraction in horses. As none of the horses developed complications associated with bacteraemia during the observation period after surgery, the significance of this bacteraemia remains uncertain. The Summary is available in Chinese – see Supporting Information.

To prove the hypothesis that the physical form of diet affects the outcome of an artificial infection with Salmonella Enteritidis in broilers, 7-day-old birds were allotted to one of four groups and fed botanically, and nearly also chemically identical diets, differing in grinding and further compaction. In total, two birds from each group (age 14 days) were administered on average 1.06 x 10(8) colony-forming units (CFU) of Salmonella Enteritidis directly into the crop by gavage and immediately put back as "seeder birds" into their respective groups. The salmonella status of each bird was analyzed by cloacal swabs, and at postmortem examination, cecal content and liver tissue samples were taken. Shedding (measured by cloacal swabs) was reduced significantly (P0.05) in groups offered the coarsely ground and pelleted diet and the diet including whole wheat compared with the groups fed the finely ground and pelleted and the coarsely ground and extruded diet. Nevertheless, only broilers fed the diet containing whole wheat showed a significantly (P0.05) lower frequency of Salmonella Enteritidis isolation in the cecal content and liver tissue. This diet was characterized by the highest percentage of particles2 mm. In this study the physical form of diet affected the outcome of an artificial infection with Salmonella Enteritidis significantly.

HAEMORRHAGIC septicaemia (HS), also termed acute pasteurellosis, is an infectious disease characterised by high morbidity and mortality, predominantly affecting juvenile ungulates (cattle, pigs) in Africa, Asia, Middle East and southern Europe. The clinical presentation is characterised by a peracute or acute period of illness consisting of fever, subcutaneous oedema, and fibrinous pleuropneumonia or haemorrhagic enteritis. The causative agent is Pasteurella multocida , a Gram-negative, coccoid rod, non-sporulating, immobile, facultatively anaerobic, facultatively pathogenic bacterium. Five capsule types (A, B, D, E, F) and 16 serotypes exist, but only types B (Asia) and E (Africa) have been associated with this disease. A …

cin, as the MIC90 of cethromycin was 0.12 mg/L (4-fold lower) compared with an MIC90 of telithromycin of 0.5 mg/L (Table 1). Among penicillin-non-susceptible (MIC 4 mg/L) isolates, the MIC90 of ceftaroline was 0.25 mg/L and the MIC90 of ceftobiprole was 1.0 mg/L, whereas that of ceftriaxone was 2 mg/L (Table 1). Similarly, the MIC90 of cethromycin was 0.12 mg/L, which was 4-fold lower than that of telithromycin. Among erythromycin-non-susceptible (MIC 0.5 mg/L) isolates, the MIC90 of cethromycin was 0.12 mg/L compared with an MIC90 of telithromycin of 0.5 mg/L (Table 1). Similarly, the results showed that ceftaroline, ceftobiprole and cethromycin were more active against emerging MDR serotype 19A than currently available drugs. In summary, among b-lactams tested, ceftaroline was the most potent against MDR S. pneumoniae isolates. Similarly, the ketolide cethromycin was more active than telithromycin.

Campylobacter spp., Salmonella enterica, and Yersinia enterocolitica are common causes of foodborne infections in humans with pork as a potential source. Monitoring programs at farm level are, to date, only implemented for S. enterica, while epidemiological knowledge of the other two pathogens is still lacking. This study aimed to assess the pathogen load (in the pigs' environment) in fattening pig herds, their simultaneous occurrence, and the occurrence of Campylobacter spp. and Y. enterocolitica in herds in different Salmonella risk categories. In 50 fattening pig herds in northern Germany, four pooled fecal samples and 10 swab samples from the pigs' direct environment (pen walls, nipple drinkers), indirect environment (hallways, drive boards), and flies and rodent droppings were collected from each herd and submitted for cultural examination. Campylobacter spp. were detected in 38.1% of fecal, 32.7% of direct environment, 5.3% of indirect environment, and 4.6% of flies/pests samples collected, and Y. enterocolitica in 17.1, 8.1, 1.2, and 3.1% and S. enterica in 11.2, 7.7, 4.1, and 1.5%, respectively. For Campylobacter spp., Y. enterocolitica, and S. enterica, 80, 48, and 32% of herds were positive, respectively; 22 herds were positive for both Campylobacter spp. and Y. enterocolitica, 12 for Campylobacter spp. and S. enterica, and 7 for Y. enterocolitica and S. enterica. There was no significant association between the pathogens at herd level. Campylobacter spp. and Y. enterocolitica were found more often in samples from the low Salmonella risk category (odds ratio, 0.51; confidence interval, 0.36 to 0.73, and 0.3, 0.17 to 0.57), and this was also the case for Y. enterocolitica at herd level (odds ratio, 0.08; confidence interval, 0.02 to 0.3). This study provides evidence that the pigs' environment should be accounted for when implementing control measures on farms against Campylobacter spp. and Y. enterocolitica. An extrapolation from the current Salmonella monitoring to the other two pathogens does not seem feasible.

This study was conducted to assess the antimicrobial susceptibility rate of Salmonella and Campylobacter spp. isolated from Northern German fattening pigs. From 540 lymph node samples, 16 Salmonella Typhimurium, 1 Salmonella Brandenburg, 37 Campylobacter coli, and 11 Campylobacter jejuni strains were isolated. Antibiotic susceptibility testing was carried out by the broth dilution method. The 14 tested antibiotics for Salmonella were ampicillin, cefotaxime, ceftazidime, chloramphenicol, ciprofloxacin, colistin, florfenicol, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim. The eight tested antibiotics for Campylobacter spp. were ampicillin, ampicillin-sulbactam (2:1), ciprofloxacin, erythromycin, gentamicin, nalidixic acid, sulfamethoxazole-trimethoprim (1:19), and tetracycline. In total, 93.7% (n = 16) of Salmonella Typhimurium, 75.7% (n = 37) of C. coli, and 54.5% (n = 11) of C. jejuni isolates were resistant to at least one of the tested antibiotics. Multiresistance to three antibiotics was observed in 75% of Salmonella Typhimurium, 16.2% of C. coli, and 0% of C. jejuni isolates. Pansusceptibility was detected in 6.3% of Salmonella Typhimurium, 24.3% of C. coli, and 45.5% of C. jejuni isolates. Multiresistance is defined as resistance to three or more antibiotics, and pansusceptibility is defined as not having resistance to any antibiotic. Regarding drugs of last resort--cefotaxime, ciprofloxacin, and nalidixic acid--resistance was not common among Salmonella (6.3%). The resistance rate of Campylobacter spp. to last-resort drugs--erythromycin, ciprofloxacin, and nalidixic acid--varied between species. The observed trend was not statistically significant. No C. coli isolates and few C. jejuni isolates (9.1%) were resistant to erythromycin. In contrast to C. jejuni, the C. coli isolates were more likely to be resistant to ciprofloxacin (9.1 and 18.9%, respectively) and nalidixic acid (0 and 13.5%, respectively). The same phenomenon was detected for tetracycline (27.3 and 62.2%, respectively), sulfamethoxazole (9.1 and 43.2%, respectively), and ampicillin (9.1 and 21.6%, respectively).

Methicillin-resistant Staphylococcus aureus (MRSA) isolates from pet animals were characterized and compared with human isolates from clonal complexes most prevalent in Central Europe.S. aureus isolates were investigated for their in vitro susceptibility to antimicrobial agents by broth microdilution. Resistance genes and the Panton-Valentine leucocidin gene lukF-lukS were identified by PCR. All isolates were characterized by SmaI macrorestriction analysis and spa typing to assess their genomic relationships. Representative isolates were additionally analysed by multilocus sequence typing and PCR-directed SCCmec typing.All pet isolates carried the resistance genes mecA and erm(C) and proved to be resistant to beta-lactams and MLS(B) antibiotics. In addition, all isolates were resistant to fluoroquinolones. None of the pet isolates carried the Panton-Valentine leucocidin gene lukF-lukS. Macrorestriction analysis revealed that the pet MRSA isolates exhibited four closely related SmaI fragment patterns. Moreover, all isolates revealed spa type t032. Further analysis of representatives of the different PFGE types revealed the presence of multilocus sequence type ST22 in combination with a type IV SCCmec element. Thus, molecular typing results were similar for pet strains and human ST22 reference strains.Based on their strain characteristics, the MRSA isolates from pets investigated in this study closely resembled ST22 MRSA isolates which are widely disseminated in German hospitals. The results of this study indicate that cross-transmission of MRSA between pet animals and humans might have occurred.

Background Actinobacillus pleuropneumoniae (A.pp.) is the causative agent of porcine pleuropneumonia leading to high economic losses in the pig industry. Infrared thermography (IRT) of the thorax might offer a new method to select swine with lung alterations for further diagnostics. In this study 50 german landrace pigs were infected with A.pp. in an established model for respiratory tract disease, while 10 healthy pigs served as control animals. To avoid drift errors during IR measurements absolute skin temperatures and temperature differences between a thoracal and an abdominal region were assessed for its diagnostic validity. Results IRT findings during the course of experimental A.pp.-infection were verified by computed tomography (CT) before and on days 4 and 21 after infection. Significant correlations were found between clinical scores, CT score and lung lesion score. Ambient temperature, body temperature and abdominal surface temperature were factors influencing the skin surface temperature of the thorax. On day 4 but not on day 21 after infection the right thoracal temperature was significantly higher and the difference between a thoracal region in the height of the left 10th vertebra and an abdominal region was significantly lower in infected pigs than in control pigs. At a cut off of 28°C of right thoracal temperature the specificity of the method was 100% (CI 95%: 69-100%) and the sensitivity 66% (CI 95%: 51-79%). At a cut off of 2°C temperature difference between thoracal and abdominal region on the left body site the specificity of the method was 100% (CI 95%: 69-100%) and the sensitivity 32% (CI 95%: 19-47%) with all control pigs detected negative. Orientation for lung biopsy by IRT resulted in 100% specificity and sensitivity (CI 95%: 69-100%) of bacteriological examination of tissue samples during the acute stage of infection. Conclusion IRT might be a valuable tool for the detection of inflammatory lung alterations in pigs, especially during the acute stage of infection and if ambient temperatures are constant during individual measurements. External and internal factors interfere with this method, so that its application in the field might be restricted to a selection of pigs for further diagnostic with adequate specificity.

Background Actinobacillus (A.) pleuropneumoniae is the causative agent of porcine pleuropneumonia and causes significant losses in the pig industry worldwide. Early host immune response is crucial for further progression of the disease. A. pleuropneumoniae is either rapidly eliminated by the immune system or switches to a long-term persistent form. To gain insight into the host-pathogen interaction during the early stages of infection, pigs were inoculated intratracheally with A. pleuropneumoniae serotype 2 and humanely euthanized eight hours after infection. Gene expression studies of inflammatory cytokines and the acute phase proteins haptoglobin, serum amyloid A and C-reactive protein were carried out by RT-qPCR from the lung, liver, tonsils and salivary gland. In addition, the concentration of cytokines and acute phase proteins were measured by quantitative immunoassays in bronchoalveolar lavage fluid, serum and saliva. In parallel to the analyses of host response, the impact of the host on the bacterial pathogen was assessed on a metabolic level. For the latter, Fourier-Transform Infrared (FTIR-) spectroscopy was employed. Results Significant cytokine and acute phase protein gene expression was detected in the lung and the salivary gland however this was not observed in the tonsils. In parallel to the analyses of host response, the impact of the host on the bacterial pathogen was assessed on a metabolic level. For the latter investigations, Fourier-Transform Infrared (FTIR-) spectroscopy was employed. The bacteria isolated from the upper and lower respiratory tract showed distinct IR spectral patterns reflecting the organ-specific acute phase response of the host. Conclusions In summary, this study implies a metabolic adaptation of A. pleuropneumoniae to the porcine upper respiratory tract already during early infection, which might indicate a first step towards the persistence of A. pleuropneumoniae. Not only in lung, but also in the salivary gland an increased inflammatory gene expression was detectable during the acute stage of infection. Electronic supplementary material The online version of this article (doi:10.1186/s12917-017-0979-6) contains supplementary material, which is available to authorized users.