Salmonella enterica isolates from faeces of domestic reptiles and a study of their antimicrobial in vitro sensitivity.

From October 2001 to February 2002, the faecal samples of 305 reptiles (165 saurians, 99 ophidians and 41 chelonians) were bacteriologically examined to detect Salmonella enterica. S. enterica was isolated from 73 (23.93%) faecal samples including 44 (60.27%) samples collected from saurians, 15 (20.55%) from chelonians and 14 (19.18%) from ophidians; considering the number of samples taken for each reptile group, S. enterica was isolated from the 36.58% of chelonians, 26.66% of saurians and 14.14% of ophidians. The isolates were distributed among 38 serotypes. Sixty-nine (94.52%) isolates were resistant to erythromycin. About one-third of the isolates was resistant to sulfisoxazole (35.61%), gentamycin (32.88%), amoxycillin (31.51%) and ampicillin (27.40%). All but one of the isolates were sensitive to chloramphenicol. A high percentages of isolates were sensitive to enrofloxacin (84.93%), nitrofurantoin (80.82%), trimethoprim (76.71%) and tetracycline (75.34%).

Epidemiology of leptospirosis: observations on serological data obtained by a "diagnostic laboratory for leptospirosis" from 1995 to 2001.

Serological data on leptospira infection were reported and discussed. From 1995 to 2001, the blood serum samples of 9885 domestic and wild animals and humans, living in Northern and Central Italy, were examined by the macroagglutination test (MAT) employing bratislava, ballum, canicola, grippotyphosa, icterohaemorrhagiae, pomona, hardjo and tarassovi serovars as antigens. Considering sera with > or = 1:400 antibody titers as positive, 674 (6.81%) animals scored positive. Sheep, horses, pigs and dogs gave the highest number of positive responses, particularly against the serovar bratislava and, for dogs, against icterohaemorrhagiae. The percentages of seropositivity observed in the most important animal species were: 12.13% in ovine (132 positive among 1088 tested animals), 11.40% in horses (107 positive animals among 938), 9.46% in swine (123 positive animals among 1299), 6.36% in dogs (278 positive animals among 4369), 2.39% in wild boars (11 positive animals among 459), 1.39% in deer (2 positive animals among 143), 0.48% in cattle (3 positive animals among 626). Among 250 human sera examined, 14 (5.60%) scored positive.

Serological diagnosis of brucellosis caused by Brucella canis.

Blood serum samples from 2,328 dogs were tested to detect antibodies against Brucella canis with the agar gel immunodiffusion (AGID) and 2-mercaptoethanol slide agglutination test (ME-SAT) using Brucella ovis as the antigen. All blood serum samples were also evaluated for antibodies against Brucella abortus and Brucella melitensis using the Rose Bengal test. Twentyfive (1.07%) of the sera evaluated were considered positive with AGID test. Only 4 (16%) of these blood serum samples were positive when evaluated with ME-SAT. The 25 AGID positive samples and 25 AGID negative serum samples were also examined by: the complement fixation test (CFT) using B. ovis hot saline extract (HSE) as the antigen, indirect enzyme linked immunosorbent assay (ELISA) and immunoblotting (IB) using B. canis and B. ovis HSE antigens. Two positive canine sera from culture positive dogs and the serum of an experimentally RM6/66 B. canis-infected rabbit were employed as positive controls and one serum from a known uninfected dog as a negative control. ELISA with B. canis antigen gave 9 (18%) positive results (6 AGID-positive and 3 AGID-negative sera). ELISA performed with B. ovis antigen detected 15 (30%) positive samples (10 AGID-positive, 5 AGID-negative and 8 B. canis ELISA positive sera). IB analysis of known positive controls sera employing B. canis antigen detected bands with molecular weights of 94-80, 64-50, 35, 32-30, 28, 23, 20-18, 15-12 kDa. The same sera tested with B. ovis antigen revealed bands of 35, 32-30, 25, 23, 20-18, 15-12 kDa. No bands were observed with the negative control serum and the 50 canine tested sera.

Cat scratch disease. Survey on the presence of Bartonella henselae among cats of Tuscany.

To verify the presence of Bartonella henselae-infection in cats living in Tuscany (central Italy) serological and bacteriological surveys were carried out. The blood serum samples of 427 cats, 254 living in private houses and gardens and 173 in public or private catteries, were tested for anti-B. henselae antibodies by indirect immunofluorescence assay (IFA). Among these samples, 35 were examined by IFA to detect antibodies against Bartonella quintana. Bacteriological examinations were performed on the blood samples, collected in EDTA (ethylene diaminetetraacetic acid), of 18 cats (10 seropositive to B. henselae and 8 negative). From each of the same 18 specimens DNA was extracted and used as template in polymerase chain reaction (PCR). The primers p24E and p12B were employed in the PCR assay to amplify a 296 bp fragment of the Bartonella 16S rRNA gene. IFA detected 98 (22.95%) B. henselae-positive serum samples (40-40.82% from cats living in houses and gardens and 58-59.18% from cats of catteries) at different antibody titers (70 at 1:64 titer, 4 at 1:128, 22 at 1:256, 2 at 1:512). Among the 35 sera tested to detect antibodies against B. quintana, 9 (25.71%) resulted positive at 1:64 titer; all these samples showed higher antibody titers to B. henselae. Out of the 26 negative sera, 20 were negative to B. henselae too and 6 had antibodies against B. henselae at 1:64. Hemocultures gave negative results. PCR scored positive with DNA of 4 B. henselae-seropositive cats, two of which belonged to two children with cat scratch disease (CSD).

An immunoblotting technique for the serodiagnosis of brucellosis by Brucella ovis.

An immunoblotting (IB) technique was developed for the serodiagnosis of brucellosis caused by Brucella ovis. Immunoblotting was performed, using a B. ovis HS (hot saline extract) antigen, on 44 blood serum samples which came from rams belonging to known brucella-free flocks, 114 samples originating from ten experimentally B. ovis infected rams and 100 from rams of naturally B. ovis infected flocks. No bands were noted on any of the 44 serum samples which originated from known negative flocks. Sera from naturally and experimentally infected rams identified antibodies to antigenic components with molecular masses of 67, 63, 58, 55, 38, 35, 32, 30, 28, 25, 23, 21, 20-18 (proteins) and 15-12 (RLPS) kDa.

Evaluation of tests employed in serological diagnosis of brucellosis caused by Brucella ovis.

A survey was carried out to verify the sensitivity and specificity of various tests (complement fixation test--CF; agar gel immunodiffusion--AGID; indirect enzyme linked immunosorbent assay--ELISA; immunoblotting--IB) employed in the serological diagnosis of brucellosis caused by Brucella ovis. The tests were executed on 44 blood serum samples of rams coming from B. ovis-free flocks, 75 of B. ovis experimentally infected rams and 1139 from rams living in flocks where B. ovis had been previously isolated. All tests were performed using B. ovis hot saline extract (HS) as antigen. Sensitivity results were 97.4% for IB, 98.68% for CF, 100% for AGID and ELISA; specificity was 100% for all methods. Concordance values were 89.62% (CF-AGID), 78.77% (CF-ELISA), 77.74% (AGID-ELISA), 65.45% (IB-CF), 62.93% (IB-ELISA), 67.24% (IB-AGID). IB identified antibodies to antigenic components with molecular weight of 67, 63, 58, 55, 38, 35, 32, 30, 28, 25, 23, 21, 20-18 kDa (proteins) and 15-12 kDa (rough lipopolysaccharide).

Epididymitis by Brucella ovis: experimental infection in virgin ram lambs.

Ten sexually immature rams were experimentally infected with Brucella ovis, to verify the antibody kinetics and its localization in urinary and genital tracts. All animals became positive to the complement fixation test from the 2nd post infection (p.i.) week and reached the maximum titre (1:256) on the 4th p.i. week. Bacteriemia was demonstrated on 3rd, 4th and 5th p.i. weeks. Two animals, respectively slaughtered 11 and 13 weeks after the infection, showed macroscopic and microscopic genital lesions and the etiologic agent was cultured from their urine and genital organs.

Leptospira interrogans in the genital tract of sheep. Research on ewes and rams experimentally infected with serovar hardjo (hardjobovis).

To verify if Leptospira hardjo can colonize the male and female genital organs of sheep, 9 animals (6 non pregnant ewes and 3 mature rams) were infected with a strain of L. hardjobovis recently recovered from the kidneys of a seropositive ewe. Postinfection controls (bacteriologic, serologic, immunohistochemistry and electron microscopy) failed to disclose the presence of leptospires in the uterus and oviducts, testicles, epididymis, prostate and bulbourethral glands of animals used for the experiment and slaughtered from 37 to 242 postinfection days. All animals showed a renal localization of L. hardjobovis lasting for the entire period of the study (over 8 months). These results emphasize the important role of sheep as maintenance hosts of the serovar.

Leptospira interrogans serovar hardjo in the kidneys and genital tracts of naturally infected sheep.

A bacteriological study was carried out to identify possible renal and/or genital carriers of Leptospira interrogans serovar hardjo. L. hardjo was found at slaughter in the kidneys of three seropositive ewes, but not in uterus or salpinges of these animals.

Experimental infection of dogs with Borrelia burgdorferi.

Four beagle dogs were inoculated subcutaneously with the BITs1 Italian strain of Borrelia burgdorferi. Only one dog became infected and B burgdorferi was isolated from its blood and urine three and four weeks after infection. B burgdorferi antibodies were detected by immunofluorescence from four to 11 weeks after infection. An uninoculated dog kept in the same run as the infected dog, developed a positive serological response, but none of the five dogs showed clinical signs.