Ant bites on dogs belly

Ant bites on dogs belly

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Ant bites on dogs belly is one of the most important problems occurring in veterinary practice. Various bacteria have been considered as etiological agents of pyogranulomatous fasciitis (PF) and dermatitis (PD) ([@B01],[@B02]). *Acinetobacter* and *Citrobacter* have been mentioned as possible etiologic agents of PD ([@B03],[@B04]). Other investigators reported that some *Pseudomonas* strns have been isolated from cases of PF ([@B05],[@B06]).

*Citrobacter* are gram-negative, non-sporulating, catalase-negative, oxidase-negative, aerobic bacteria found ubiquitously in nature ([@B07],[@B08]). They have the ability to ferment carbohydrates such as glucose and maltose, they can also hydrolyze esters and glycols. *Citrobacter* spp. have been isolated from raw foods and are not sensitive to many antibiotics. *Citrobacter* spp. isolated from food have a great potential to cause diarrhea, and this potential has been confirmed in our laboratory as well as in others ([@B09]).

Although various investigators have tried to isolate bacteria responsible for cases of PD and PF, only *A.baumannii* and *C.freundii* were identified in such cases ([@B10],[@B11]). There are no published data in the literature regarding the isolation and identification of *Citrobacter* species as an etiological agent of PD and PF. The purpose of this study was to identify *Citrobacter* species isolated from cases of PD and PF.

Seventy six dog skin biopsies were collected from different skin areas affected by PD and PF from August 2006 to November 2009. Skin lesions were excised and samples were placed in sterile phosphate-buffered saline (PBS) and transported to the laboratory on ice. Biopsies (~0.5 cm^2^) were used for microbiological examination.

After washing and homogenizing the tissue, one loopful was streaked onto Columbia agar (Oxoid) and incubated overnight at 37ºC. One or two isolated colonies from each positive culture were selected and biochemically tested for Gram stning, catalase, hemolysis on blood agar, and oxidase activity.

All isolates were characterized for biotype and serotype. For the biotype, *Citrobacter* species were subjected to biochemical tests such as growth on trypticase soy agar, growth in 6.5% NaCl, methyl red-voges-proskauer and citrate utilization, and acetate utilization ([@B12]). The serotype was determined by tube agglutination with antiserum using commercial antisera (Denka Seiken, Tokyo, Japan). Colonies with the same biotype and serotype were considered as the same genotype and serotype, respectively ([@B5]).

Antibiotic susceptibility was determined by the disk-diffusion method. Mueller-Hinton agar (Merck, Darmstadt, Germany) was inoculated with a bacterial suspension (0.5 McFarland) and antibiotic disks (Difco, Detroit, MI) (0.12--16 μg of penicillin G per disk) were placed on the agar surface. The antibiotic-sensitive *Citrobacter* strns were used as controls. Plates were incubated at 37ºC for 18 h. The inhibition zone diameters were interpreted as susceptible, intermediate, and resistant according to Clinical and Laboratory Standards Institute ([@B6]).

The susceptibility data were analyzed with the computer software package SPSS for Windows (version 19.0, Chicago, IL, USA). The chi-square test or Fisher exact test was used to determine the differences in proportions of antimicrobial-resistance among the *Citrobacter* spp. isolates. A *P* value <, 0.05 was considered to be statistically significant.

This study was approved by the institutional review board of the College of Medicine, The Catholic University of Korea (SC13LISE0019).



The prevalence of *Citrobacter* spp. in the patients


The overall frequency of *Citrobacter* spp. isolation was 1.24% (14/1,132, *C. freundii* 2, *C. youngae* 1, and *C. braakii* 11). The proportion of *Citrobacter* spp. among the patients with diarrhea was significantly higher than that of the patients with urinary tract infection or bacteremia (4.9% vs. 0.55%, *P* <, 0.001). The proportion of *Citrobacter* spp. among the patients who underwent operation was significantly lower than that of those without operation (0.58% vs. 1.64%, *P* = 0.039).

Microbiological characteristics of *Citrobacter* spp.


Of the 14 *Citrobacter* spp. isolates, 11 were from stools, one from urine, and two from blood. Of the 11 isolates from stools, four were from the patients with diarrhea, and seven from the patients with fever or non-diarrhea. Of the four isolates from the patients with diarrhea, two were from the patients with watery diarrhea, one from the patients with bloody diarrhea, and the other from the patient with both diarrhea and fever. Of the seven isolates from the patients without diarrhea, one from the patient with fever and two from the patients with non-diarrhea were isolated from blood. The source of isolation of *Citrobacter* spp. was not identified in five patients. We identified eight unique STs of *Citrobacter* spp. ([Figure 1](#f1-idr-11-1215){ref-type="fig"}) in this study, four STs were new STs and four STs were new and the ST type of other *Citrobacter* spp. were not identical (two for *C. youngae*, one for *C. braakii*, and one for *C. freundii*).

Antimicrobial susceptibility of *Citrobacter* spp.


The MICs and MBCs of the 14 isolates agnst 11 antimicrobials are shown in [Table 3](#t3-idr-11-1215){ref-type="table"}. According to the results, ciprofloxacin and minocycline were the most potent antimicrobials for all isolates. All the isolates were susceptible to carbapenems, except for one *C. freundii* isolate. The rate of nonsusceptibility to imipenem was 95.2%. All isolates showed MDR (except for one *C. freundii* isolate). The prevalence of resistance to quinolones was 50.0% and the resistance rate to cephalosporin was 28.6%. *Citrobacter* spp. were resistant to ampicillin, erythromycin, and penicillin. In addition, the isolates were resistant to more than three antimicrobials. The rates of resistance to other antimicrobials were variable.

Cluster analysis of *Citrobacter*

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