Long-Term Treatment with Alcaligenes faecalis A12C Improves Host Resistance to Pathogens in Septic Rats: Possible Contribution of Curdlan-Like Immune Trainer.

Curdlan, a ß-1,3/1,6-glucan found in Alcaligenes faecalis (A. faecalis) wall, activates innate and humoral immunity. The aim of this study is to evaluate whether pretreated rats with A. faecalis A12C could prevent sepsis disturbances and identify the immunomodulatory mechanisms involved. Experiments occurred in two stages: a survival study with 16 rats randomly divided into septic (SC) (n = 8) and septic pretreated (SA) (n = 8) groups and 45 rats divided into four groups: healthy (AGUSAN) (n = 9), septic (AGUIC) (n = 13), septic pretreated (AGUIA) (n = 14), and healthy pretreated (AGUSTO) (n = 9). Sepsis was induced by cecal ligation and puncture after 30 days of A. faecalis A12C pretreatment or without. SA group had a higher survival rate of 58% vs. 16% for SC group (P < 0.05). Overall, AGUIA showed better status than AGUIC (P < 0.01). Higher monocytosis was found in AGUIA and AGUSTO vs. AGUIC and AGUSAN, respectively (P < 0.05). A gradual increase in curdlan fecal concentration was observed in AGUIA during pretreatment. Fecal concentrations of Escherichia coli significantly decreased in AGUIA and AGUSTO. Bacterial load in urine, peritoneal lavage fluid (PLF), and bronchoalveolar lavage fluid (BALF) decreased (P < 0.05) in AGUIA vs. AGUIC. Finally, lower inflammation was observed in serum, BALF, and PLF, with reduced IL-6, IL-10, IL-1ß, and TNF-α, along with less damage in lungs and peritoneum in AGUIA vs. AGUIC. These findings suggest the connection between curdlan-produced by A. faecalis A12C-with the immune system and the reduction in severity of experimental sepsis.

Probiotic Properties of Alcaligenes faecalis Isolated from Argyrosomus regius in Experimental Peritonitis (Rat Model).

A strain of Alcaligenes faecalis A12C (A. faecalis A12C) isolated from Argyrosomus regius is a probiotic in fish. Previous experiments showed that A. faecalis A12C had inhibitory effects on the growth of multidrug-resistant bacteria. We aimed to confirm whether A. faecalis A12C is safe and has adequate intestinal colonization in experimental rats, and evaluate its efficacy in an animal model of peritonitis. We used 30 male rats, randomly divided into 6 groups (n = 5): three groups (HA7, HA15, HA30) received A. faecalis A12C in drinking water (6 × 108 CFU/mL) for 7 days, and three control groups received drinking water only. All groups were evaluated at 7, 15, and 30 days. Survival after A. faecalis A12C administration was 100% in all groups. Mild eosinophilia (1.5%, p < 0.01) and increased aspartate aminotransferase (86 IU/L, p < 0.05) were observed in HA7, followed by progressive normalization. No histological signs of organ injury were found. We observed significant E. coli decline in faeces, parallel to an increase in A. faecalis A12C at 7 days. E. coli had a tendency to recover initial values, while A. faecalis A12C disappeared from the intestinal microbiota at 30 days. To evaluate its efficacy against peritonitis, we studied two additional groups of animals: IA group pretreated with A. faecalis A12C before E. coli intra-abdominal inoculation, and IC group inoculated with no A. faecalis A12C. We found an increase in C-reactive protein, alanine aminotransferase, urea, and eosinophils in IC animals when compared with IA. Peritonitis was more evident in IC than in IA animals. Our findings suggest that A. faecalis A12C altered clinically relevant parameters in sepsis and was associated with a lesser spread of infection.

Multicenter Stratified Study From Spain About Acceptance of Living Liver Donation Among Veterinarian Students.

Veterinarians often hold decision-making positions in the public health care system, and therefore can influence public opinion about organ donation. The objective is to analyze the attitude of Spanish veterinarian students toward living liver donation (LLD) and to establish which factors have an influence on this attitude. METHODS: A sociological, interdisciplinary, multicenter, and observational study was carried out in the veterinarian students enrolled in Spain (n = 9000) (university academic year 2010-2011). A sample of 2854 students was stratified by geographic area and academic year. A validated questionnaire (PCID-DVH RIOS) was self-administered and completed anonymously. The questionnaire was applied to each academic year at compulsory sessions at randomly selected veterinary schools. Statistical analysis included t test, χ2 test, and logistic regression analysis. RESULTS: Completion rate was 94% (n = 2683); 89% (n = 2345) were in favor of related LLD, and 40% (n = 1053) supported unrelated LLD. The following variables were associated with a more favorable attitude: (1) age (P < .001), (2) sex (P < .001), (3) academic year (P < .001), (4) believing in the possibility of needing a transplant oneself in the future (P < .001), (5) attitude toward deceased donation (P < .001), (6) attitude toward living kidney donation (P < .001), (7) acceptance of a donated liver segment from a family member if one were needed (P < .001), (8) having discussed the subject with one's family (P = .009) and friends (P < .001), (9) a partner's opinion about the subject (P = .002), and (10) fear of the possible mutilation of the body after donation (P < .001). CONCLUSION: Spanish veterinary students have a favorable attitude toward LLD.

Veterinary Students' Attitude Toward Living Kidney Donation: A Multicenter Stratified Study From Spain.

Living kidney related donations (LKRD) should be promoted because of the current deficit of organs for transplantation. The objective of the study was to analyze the attitude of Spanish veterinary students toward LKRD, because they may influence public opinion in the future, and to determine the factors that condition it. METHODS: A sociological, interdisciplinary, multicenter, and observational study was carried out in the veterinary students enrolled in Spain (n = 9000) in the university academic year 2010-2011. A sample of 2815 students was stratified by geographical area and academic year. The students' attitude toward LKRD was assessed using a psychosocial validated questionnaire (PCID-DVR Rios), was self-administered, and was completed anonymously. Veterinary schools were randomly selected. The questionnaire was applied to each academic year at compulsory sessions. Statistical analysis was preformed using t test, χ2 test, and logistic regression analysis. RESULTS: The completion rate was 95% (n = 2683); 93% (n = 2504) were in favor of LKRD and 36% (n = 945) supported unrelated living kidney donation. The following variables were associated with having a more favorable attitude: (1) sex (P < .001), (2) being a student of southern universities (P = .03), (3) attitude toward deceased donation (P < .001), (4) having a father (P < .001) or a mother (P < .001) in favor of organ donation, and (5) having discussed the subject with friends (P = .03) or family (P = .02). However, only 60% would accept a kidney from a relative. CONCLUSION: Spanish veterinary students have a favorable attitude toward LKRD. However, only 60% would accept a kidney from a relative.

Attitude Toward Organ Donation and Transplantation in Veterinary Students: A Multicenter and Stratified Study in Spain.

BACKGROUND: Veterinarians often hold decision-making positions in the public health care system and can therefore influence public opinion about organ donation and transplantation (ODT). The aim of this work was to analyze the attitude of Spanish veterinary students toward ODT, because they may influence public opinion in the future, and to determine the factors that condition it. METHODS: This was a sociologic, interdisciplinary, multicenter, observational study in Spain. The study population was students studying for a veterinary degree in Spain (n = 9000), and a sample of 2815 students (confidence of 99% and precision of ±1%) was stratified by geographic area and academic year. A validated questionnaire of attitude toward ODT (PCID-DTO-Ríos) was self-administered and anonymous. RESULTS: Of the 2815 selected students (2790 plus the 0.9% per type of sample), 2650 completed the questionnaire (response rate, 94.14%): 83% (n = 2207) of the respondents were in favor of donation and 17% against. The following main variables were related to a favorable attitude: being female (odds ratio [OR], 0.752; P = .034); knowing a donor (OR, 1.834; P = .003); having discussed the matter with one's family (OR, 1.587; P = .002); having spoken about the subject in social circles with friends (OR, 1.633; P < .001), and being in favor of donating a deceased family member's organs (OR, 2.403; P < .001). CONCLUSIONS: Seventeen percent of Spanish veterinary students were not in favor of ODT. It is important to know the factors that determine their attitude, because this will make it possible to optimize the resources invested in campaigns to promote ODT and to take more specific action.

Antimicrobial susceptibility of Enterococcus strains isolated from poultry faeces.

We have investigated the resistance of Enterococcus isolated from poultry faeces to antibiotics commonly used as therapy of enterococcal infections. Identification was made by the method of Facklam and Collins. Minimal inhibitory concentrations of penicillin, ampicillin, vancomycin and teicoplanin were determined and high level aminoglycoside resistance was investigated. Genes codifying high level aminoglycoside resistance (HLAR) were determined by PCR. Fifty five Enterococcus strains were isolated (63.6% E. faecalis, 12.7% E. mundtii, 9.1% E. faecium, 7.3% E. casseliflavus, 3.7% E. durans and 3.6% E. hirae). None of the strains were resistant to VAN, TEC, P or AM. HLAR was found in 34.5% of strains for SM, 27.3% for KM and 7.3% for GM. The gene for the bifunctional enzyme was found only in one strain, that showed HLAR to GM and KM. Fourteen strains harboured the gene aph(3')-III, being 11 resistant to KM and STR, and three resistant to GM, KM and STR. The remaining six strains showed HLAR to STR, but were negative for the three genes tested by PCR. The gene ant(4'4") was not detected in any of the strains. No unexpected vancomycin resistance was detected. The resistance rates among poultry strains were lower than those found among human strains isolated from hospital patients in recent Canary studies.

[Lung transplantation in rats: a viable experimental model]. / Trasplante pulmonar en ratas. Un modelo viable de estudio experimental.

OBJECTIVES: To incorporate a new fast, safe, and reversible anesthetic procedure into the experimental model of lung transplantation (LT) using a cuff technique originally described by Mizuta. MATERIAL AND METHOD: Eighty-eight Sprague-Dawley rats were used in the experimental model. Thirty left LTs were performed, using 60 rats. The donor heart-lung block was excised by median sternotomy with dissection of the left lung and cuffs (intravenous catheters cut into 3-mm sections) were put in place. The left lung was implanted in the recipient by lateral thoracotomy using the cuffs for anastomoses. The duration of surgery and postoperative complications were recorded. Also noted were signs of ischemia-reperfusion injury, and acute rejection of the transplanted lung. RESULTS: We discarded lungs excised from 8 animals when developing the experimental model. Transplants could not be completed in 10 rats due to technical problems, despite satisfactory excision. Of the rats who received a transplant, 4 died in the first 24 hours and 26 survived to 48 hours. They were then killed and examined. The state of the anastomoses was good and signs of ischemia-reperfusion injury, as well as acute rejection were observed in the parenchyma of the transplanted lung. CONCLUSIONS: LT with cuffs in rats is a valid, reliable, reproducible, and cheap model for studying ischemia-reperfusion injury and rejection in LT. The surgical technique is complex, requiring experienced surgeons and a long learning process. Modification of the technique to more closely resemble the surgical procedure in humans is possible, thus facilitating interpretation and allowing more reliable extrapolation to humans.

Group 1 beta-lactamases of Aeromonas caviae and their resistance to beta-lactam antibiotics.

The contribution of beta-lactamase production to beta-lactam antibiotic resistance was examined in an Aeromonas caviae mutant strain, selected in vitro by cefotaxime and derived from a wild-type strain isolated in our laboratory from crude sewage. Both strains produced beta-lactamase. The mutant strain (AC7m) produced beta-lactamase constitutively, in contrast to the parental strain (AC7), which was inducible by cefoxitin. AC7m was regarded as a mutant from AC7, which over-expressed beta-lactamase. The mutant strain showed a remarkable reduction in sensitivity to most of the beta-lactam antibiotics tested, such as (i) aminopenicillins and their combinations with clavulanic acid and sulbactam, (ii) carboxypenicillins, (iii) ureidopenicillins, and (iv) cephalosporins. This strain remained susceptible to ceftazidime, imipenem, and aztreonam. Isoelectric focusing of sonic extracts revealed that both strains AC7 and AC7m shared a common major beta-lactamase band at pI 6.5. The plasmid DNA assays showed that the beta-lactamases expressed by each A. caviae strain were chromosomally encoded. Based on substrate and inhibitor profiles determined in sonic extracts for AC7 and AC7m, the enzymes displayed on isoelectric focusing at pI 6.5 were assigned to chromosomal Group 1 beta-lactamases. Imipenem would therefore be the appropriate choice for therapy of infections caused by A. caviae beta-lactamase over-expressing mutants.

Identification and antimicrobial susceptibility of coagulase positive staphylococci isolated from healthy dogs and dogs suffering from otitis externa.

A total of 67 strains of coagulase positive staphylococci isolated from healthy dogs and dogs suffering from otitis externa were studied. Twenty-two isolates were from healthy dogs (five from hound dogs and 17 from companion dogs) and 45 from dogs suffering otitis externa (14 from hound dogs and 31 from companion dogs). Presumptive identification was attempted using the following tests: production of acetoin, anaerobic utilization of mannitol, acid production from mannitol, presence of beta-galactosidase, and growth on P agar supplemented with different concentrations of acriflavine. Susceptibility of staphylococci to 16 antibiotics was determined. Most effective antibiotics were imipenem, amoxycillin/clavulanic acid, ciprofloxacin, tobramycin, gentamicin and marbofloxacin. Penicillin, ampicillin and polymyxin B showed the lowest activity. There were no significant differences in antimicrobial susceptibility among isolates from healthy dogs and dogs suffering from otitis externa.

Identification and antibiotic resistance of faecal enterococci isolated from water samples.

We have isolated 47 strains of presumptive faecal streptococci from different water samples. Identification was made by the method of Facklam et al. (1989). Antibiotic resistance was studied on Mueller-Hinton Agar. Twelve antibiotics were tested. High-level aminoglycoside resistance (HLAR) and resistance to glycopeptides were studied. Biochemical identification of presumptive faecal streptococci isolates gave the following results: 19 Enterococcus faecalis, 12 E. faecium, 8 E. hirae, 4 E. durans and 4 E. mundtii. E. mundtii is not included among faecal enterococci. None of the strains were resistant to glycopeptides (vancomycin and teicoplanin). Three strains of Enterococci showed HLAR. Two of them were isolated from coastal bathing waters and the other from wastewater. This suggests that water could contribute to spread of HLAR enterococci and it should be a matter of concern for public health authorities.

Antibacterial susceptibility patterns of Pseudomonas strains isolated from chronic canine otitis externa.

Development of antibiotic resistance in bacteria is a problem of great concern. It is important to establish the convenience of antimicrobial susceptibility tests in animal infections. The aim of this study was to test the susceptibility to antibiotics of Pseudomonas strains isolated from chronic canine otitis externa. We tested 23 strains of Pseudomonas: 19 Ps. aeruginosa, three Ps. fluorescens and one Pseudomonas spp. The most effective antibiotics were tobramycin (100% susceptible), marbofloxacin (91.3%) and ceftazidime (91.3%). Ticarcillin and gentamicin, commonly used for the treatment of otitis externa also showed good results (susceptibility of strains was 86 and 65.2% respectively). Lower susceptibility was found using enrofloxacin (52.1%) probably due to its indiscriminate use. We emphasize the need for a rational policy of antibiotic prescribing in order to prevent the selection of resistant strains.