Phage typization of Staphylococcus aureus strains from staphylococcic mastitis of dairy cows]

Of 62 strains of Staphylococcus aureus, isolated from milk samples of dairy-cows with staphylococcic mastitis from five agricultural enterprises, 68 p. c. could be typified as bacteriophages. In 42 typified strains, altogether 17 different phage pictures were found, in which case, in the different agricultural enterprises, the number of phage picture fluctuated between 1 and 8. Phage picture 42 D was found and dominated in all the agricultural enterprises. Of the typified strains, 76% could be ranked only into one phage group and 24% in the "M" group (= miscellaneous). Of 32 typified strains, 56.3% belonged to phage group IV, 25% to group III, 12.5% to group I, and 6.2% to group II.     

Characteristics of Staphylococcus aureus isolated from lesions of horses.

Seventy-six Staphylococcus aureus strains isolated from various lesions of horses were characterized. All of the 76 strains were identified as biotypes B (38.2%) and C (61.8%). Of 55 strains tested, 42 (76.4%) were differentiated into 7 coagulase types. Coagulase types V and VII were predominant in the metritis strains. Coagulase type II was found most frequently in the strains from phlegmon, dermatitis, sinusitis, empyema sinus, and nasal catarrh. Forty-two (55.3%) of the 76 strains were differentiated into 24 phage patterns. Twenty (58.8%) of 34 typable strains from metritis were lysed by the human group I phage 52, and group II phages 3A, 3C, 55 and 71. Forty-five (59.2%) of the 76 strains were resistant to 1 or more of 6 antibiotics. Strains resistant to penicillin G, irrespective of source, were most frequent (95.6%). Forty (93.0%) of 43 strains resistant to penicillin G alone or in combination with other antibiotics produced beta-lactamase. Only 8 (10.5%) of the 76 strains produced enterotoxins A (n = 2), B (n = 1) or C (n = 5), and they all were isolated from metritis. Only 1 strain isolated from phlegmon and 2 from metritis produced exfoliative toxin (ET) and toxic shock syndrome toxin-1 (TSST-1), respectively. The latter 2 strains also produced enterotoxin C. The results of the present study showed the first evidence of the presence of both ET- and TSST-1-producing S. aureus isolated from horses.     

Epidemiology of Salmonella derby strains isolated from swine, pork and pork products

A total of 35 Salmonella derby strains, isolated from 6 types of samples of porcine origin from 9 different places in Hungary were examined for their characteristics. Thirty-two strains (91%) were of phage type 25, 2 (6%) of phage type 15 and 1 (3%) of phage type 58. Colicin production was observed in 3 (9%) strains. Five strains (14%) were found to be resistant to tetracycline (Tc). The strains harboured plasmids of 2.2, 2.4, 3.4, 4.2 and 72 Md. The 72 Md plasmid appears to be characteristic of S. derby and possibly encodes Tc resistance. The 72 Md plasmid belonged partly to incompatibility (Inc) group I1, while the other plasmid of the same size belonged to Inc. group B. The findings suggest that healthy salmonella carrier pigs carried the infection from the farm to the abattoir. Slaughtering of infected pigs may have led to contamination of the carcasses and, thereafter, that of the pork and pork products.     

Phage typing of Staphylococcus aureus strains isolated in Sweden from bovine milk.

Both the human and the bovine international sets of phages were used for typing of 372 bovine Staphylococcus aureus (Sa) strains, whereas the bovine set alone was used for typing of a further 1183 strains. In addition, 338 of the strains were tested for antibiotic sensitivity. Out of 372 Sa strains 85.5% could be typed with the human and 89.8% with the bovine phage set. Of all the 1555 Sa strains used 92.4% were lysed by the bovine phage set. Several phage types can be present in one and the same herd and some of them can predominate. Resistance to most of the tested antibiotics was very low. The incidence of resistance to penicillin and ampicillin was 10.0% and 4.4% respectively.     

Isolation and characteristics of bacteriophages from staphylococci of chicken origin.

Lysogenicity in chicken coagulase-positive staphylococci was tested by incubating the strains in the presence of mitomycin C. Of 88 strains tested, 84 (95.5%) were proved to be phage carriers and 81 were susceptible to any of the phages. The lysogenic strains were detected with almost equal frequency from both of typeable and untypeable strains by the international phages. Sixteen phages (CH phages) were isolated from chicken lysogenic strains, and their usefulness for the typing of chicken staphylococci was evaluated. Of 122 strains examined, 101 (82.8%) were found to be typeable with the CH phages at a routine test dilution (RTD). About 82% of strains untypeable by the phages of the international series were lysed by one or more of the CH phages. The phages seemed to be highly specific to chicken staphylococci, because they lysed only a few strains of animal origin other than chicken. Thus, the 16 phages newly established were found to have significant advantages in typing chicken strains.     

Association between phage types and antimicrobial resistance among bovine Staphylococcus aureus from 10 countries

This study was conducted to investigate the diversity of phage types and associations between penicillin resistance and phage types among 815 Staphylococcus aureus isolates from bovine mastitis in nine European countries and USA. All isolates were examined for susceptibility to antimicrobial agents and characterised by phage typing. Penicillin resistance was found among strains from all countries with an average occurrence of 32.4% (2-71.4%). A total of 76% of isolates were identifiable by phage typing and 144 different phage types were observed. The most predominant types were phage type 29 (11% of the 815 isolates), phage type 52 (5%), and phage type 80 (5%). Phage type 95 and 29/52/52A/80 were both distributed within seven countries. In the countries with the highest occurrence of penicillin resistance a reduced diversity of phage types and phage groups was observed. Phage group III was significantly associated with penicillin resistance in contrast to phage group I (P=0.0023) and phage complex-80 (P=0.0066). This study confirms that a large number of phage types of S. aureus cause bovine mastitis, but that some types predominate. In addition, these findings could indicate that the use of penicillin in the bovine environment has selected for specific types of S. aureus in countries with a high frequency of resistance.     

Plasmid profile and phage type of Salmonella typhimurium strains encountered in different regions of India

A total of 190 Salmonella typhimurium strains encountered in different parts of India were characterized on the basis of plasmid profile, phage type and antimicrobial resistance pattern. Recent trends in the epidemiology of R-plasmids were also studied.

The majority of S. typhimurium strains (90.5%) were untypable by phage typing. Only 18 strains (9.5%) were phage typable. The phage untypable strains isolated from northern (57) central (65), and southern (50) regions of India could be subgrouped into 24, 12 and 16 different plasmid profiles respectively. Heterogeneity was the prominent feature although most of the plasmid profiles were related among strains isolated from particular place. A great diversity among small plasmids (2.7–8.3 kb) made subgrouping of majority strains (71%) with R-pattern ApCmKmSmSuTcTp possible. Conjugation studies and plasmid profile analysis of transconjugants revealed all the strains to harbour non conjugative non-auto transmissible plasmids with the exception of 7.2 and 2.7 kb plasmids which were not mobilizable.     

Lytic activities, protein profiles and morphologic characteristics of new bacteriophages isolated from canine and human Staphylococcus aureus strains.

The lytic activity, protein profile and morphology of five newly isolated phages from canine Staphylococcus aureus strains and one from a human S. aureus strain were compared with those of selected phages in the international phage sets (IPS). Five canine phages lysed 57 (76.0%) of 75 canine isolates of Staphylococcus aureus from Nigeria at routine test dilution (RTD) while 34 (IPS) phages typed only 31 (41.3%) strains at RTD or/and 100-RTD. The new human phage lysed 11 (14.7%) of 75 strains isolated from human diarrhoea. The new phages were readily propagated, specific in activity and stable during storage at 4 degrees C. Prominent proteins detected by SDS-PAGE indicated similarities between some of the phages but one canine phage was distinctly different, as was its morphology which was an isometric head with a short tail compared to oval heads and long tails which characterized others. IPS phages in the same serologic group had similar protein profiles but no correlation was observed with lytic groups. The use of protein profile and electron micrographs allowed classification of the phages into serogroups. It is concluded that the newly isolated canine phages could be very useful in typing Nigerian canine strains of S. aureus.     

Laboratory-based surveillance of salmonellosis of humans in Germany--safety of Salmonella typhimurium and Salmonella enteritidis live vaccines]

The Paul Ehrlich Institute, Langen, in Germany has been licensed different live vaccines of S. Typhimurium and S. Enteritidis for use in the veterinary medicine since I the 90s. The Robert Koch Institute has established a lab-based surveillance system for these live vaccine strains for an evaluation of recent public health safety. Since 2000 all strains of S. Typhimurium and S. Enteritidis from humans were investigated in respect to their phage types and other vaccine markers. 3676 S. Typhimurium strains and 4489 S. Enteritidis strains mainly from Salmonellose patients were investigated after phage typing according to their auxotrophic or antibiotic resistance markers. The live vaccine strains of Zoosaloral, TAD Salmonella vacT or TAD Salmonella vacE and Salmovac SE could not be found from infections in humans.     

Staphylococcus aureus isolated from poultry in Australia. I. Phage typing and cultural characteristics

The phage typing and cultural characteristics of 574 strains of S. aureus of poultry origin in Australia were examined. With the avian phage set of Shimizu (1979) it was possible to type 74.2% of strains. A number of significant variations in the phage typing patterns of Australian strains compared to those reported from Japan and Europe were observed. A lower proportion of Australian strains were of avian phage group I and a higher proportion of group III. A high proportion of strains were of mixed lytic groups. No locally isolated phages were able to increase significantly the percentage of typeable strains, although four local phages appeared to be of greater value for phage typing poultry strains of S. aureus than some other phages of the avian phage set. The international (human) phage set was of limited value in typing Australian strains of poultry origin although four strains were identified which were indistinguishable from strains of human origin. Using cultural characteristics of the strains in conjunction with phage typing, the Australian strains of S. aureus were assigned to one of three major groups and nine subgroups. A list of typing phages considered to be valuable for use on Australian poultry strains of S. aureus is given.     

The dual role of wild phages for horizontal gene transfer among Salmonella strains

Salmonella bacteriophages seem to mediate horizontal transfer of virulence functions among Salmonella strains in two different ways: by general transduction and also by lysogenic conversion. The majority of wild phages isolated from Salmonella strains belong to the P22 like phages and were able to transduce. Our data show that the lysogenic conversion is generally accompanied by changes in the susceptibility to the typing phages used for epidemiological purposes. Similar phage type conversions to S. Typhimurium DT104 could be detected upon lysogenization with two other S. Typhimurium strains. For some S. Typhimurium strains the typical phage pattern is actually associated with alterations of virulence characteristics. For example, all tested wild type isolates of phage types DT49 and DT204 were found to be SopE phi-lysogens. The Anderson typing phages interfere with the prophages and/or cryptic phages and so the complex genetic short-term evolution can be demonstrated in the lab. This is one reason for the successful application of phage typing in Salmonella epidemiology since the 50s.     

Characteristics of Staphylococcus aureus strains from pathological processes in swine

120 Staphylococcus aureus strains from different and sporadic pathological process in swines have been investigated on biochemical properties and phage patterns. 98 strains (82%) exhibit coagulation of human plasma, no coagulation of bovine plasma, crystal-violet-type C, hemolysin-type beta, no formation of fibrinolysine and sensitivity for phages of group I, III and M. Only in few strains we observed reactions with phages of group II. 26 from 66 strains were resistant to antibiotics, and among them 24 showed resistance to oxytetracacline.     

Experimental horizontal transmission of Salmonella enteritidis strains (phage types 4, 8, and 13a) in chicks

The recent isolation of phage type 4 Salmonella enteritidis strains from poultry and humans in the United States has generated considerable concern because this phage type is predominant in both animals and humans in many other nations. Understanding whether the presence of these strains in poultry flocks poses an elevated threat to public health is a critical issue for developing effective disease control programs. The present study evaluated whether S. enteritidis strains of various phage types found in poultry in the United States (phage types 4, 8, and 13a) differed in their potential for horizontal transmission from experimentally infected chicks to uninoculated chicks housed in the same isolator units. Five days after two seeder chicks in each group of 12 were inoculated with oral doses of approximately 10(3) S. enteritidis cells at 8 days of age, ceca and livers were sampled from seeder chicks and from their contact-exposed penmates. On the basis of the detection of S. enteritidis in cecal samples, phage type 4 strains were transmitted horizontally at a significantly lower frequency than were strains of other phage types. Nevertheless, two of three phage type 4 strains evaluated were very highly invasive.     

The biochemical differentiation between Salmonella and Citrobacter

A number of bacterial isolates which could not be identified as either Salmonella or Citrobacter by conventional biochemical tests and could not be typed as Salmonella with available antisera, were further examined biochemically and by lysis with phage Felix 0.1. Glycerol-positive salmonellae and lysine-positive citrobacters were encountered, which could be confused with the other genus, but when the reactions of such strains were examined in the other tests, accurate identifications could be done. Of the tests examined, glycerol fermentation, the beta-galactosidase test, lysine decarboxylation, sorbose fermentation, galacturonate fermentation and lysis by the phage could be used in the differentiation. These tests in combination, rather than 1 or 2 single tests gave reliable and conclusive differentiation.     

Epidemiology and characterization of animal Salmonella enterica subspecies Enterica serotype typhimurium DT104 in Hungary

Reports on the internationally emerging significance of multiresistant zoonotic Salmonella in animals and man prompted studies to estimate the significance of multiresistant Salmonella enterica subspecies enterica serotype Typhimurium (S. Typhimurium) phage type DT104 of animal origin in Hungary. A collection of 231 strains (primarily of goose, turkey, poultry and porcine origin from the years 1997-1998) was tested for resistance against 7 selected antibiotics (ampicillin, chloramphenicol, enrofloxacin, nalidixic acid, streptomycin, tetracycline and sulphamethoxazole). Strains with resistance against 3 or more were defined as multiresistant. All strains were phage typed using Felix-Callow's S. Typhimurium phage typing system, and 91 of them (suspect DT104) were also typed according to Anderson's definitive typing (DT) system. In this study, 14% of animal strains from 1997-1998 was classified as DT104, for which turkey, pig and duck seemed to be the main carriers, and the multiresistant non-DT104 strains represented a further 6% of this collection. The prevalence of DT104 was highest among strains of turkey origin (50%), followed by strains of pig (29%), chicken (25%), duck (19%), and goose (3%) origin. The other DT104 related phage types (DT12 and U302) were only detected in the case of 4 strains (2 of porcine, and one each of turkey and of goose origin). The DT104 corresponded to the Felix-Callow types 2/3 or 2c/3 in each case, except in the case of 3 turkey strains where they corresponded to type 35/3. Nalidixic acid resistance was detected in all multiresistant turkey strains and in some of other animal origin but none of these strains were resistant to enrofloxacin. A retrospective analysis (based on the above relationship) indicated that S. Typhimurium strains corresponding to DT104 could be present and increase in the Hungarian farm animal population from about 2% to 20% between 1985 and 1990, in a manner similar to the emergence of human DT104, as reported elsewhere (Pászti et al., 2000). The 91 suspect DT104 strains were also tested for plasmid profile and for spvC gene indicating the presence of the large serotype specific plasmid (Ssp). No characteristic plasmid profile could be attributed to S. Typhimurium DT104. The serovar-specific large plasmid was detected by PCR for spvC in 100% of DT104 strains and in 77% of the non-DT104 strains. The virulence of two DT104 strains was tested in orally infected day-old chicks and compared with virulence of 4 non-DT104 strains. Higher colonizing virulence of DT104 strains could be established as compared to the other strains.     

A Comparative Study of Staphylococcus aureus Strains Isolated from Bovine Subclinical Mastitis During 1952–1956 and 1992

Fiftytwo strains of S. aureus isolated from cases of bovine subclinical mastitis in 52 different dairy herds in Denmark, in the periods 1952 to 1956 and 1992, were compared with regard to their phage- and EcoRI ribo-types. Furthermore, susceptibility to penicillin and production of fibrinolysin were used as additional phenotypic markers. Fortynine strains (94%) could be separated into 12 phage types. Ribotyping assigned the 52 strains to 21 different types. Both methods showed that 57% of the 1950’s strains and between 38–45% of the 1992 strains belonged to 3 dominating types. The remaining strains were placed by ribotyping in 8 types occurring among the 1952–1956 strains and 10 types occurring among the 1992 strains. In 87% of the strains the results of the 2 typing methods were in accordance. However, 7 strains gave different results by the 2 methods including 2 strains with major differences. Penicillin resistance only occurred in a single genotype from the 1950’s compared to 6 different genotypes among the 1992 strains.     

Serological, enterotoxin-producing and biochemical properties of Escherichia coli isolated from piglets with neonatal diarrhea in Norway

Altogether 235 strains of Escherichia coli isolated from jejunal content of piglets with neonatal diarrhea were examined for serological, enterotoxin-producing and certain biochemical properties. Of 198 strains examined, 84 (42 %) belonged to O-group 149, while 14 (7 %) strains belonged to each of the O-group s 8 and 64. Seventy strains (30%) could not be grouped with the sera used. The remaining strains were distributed among the following O-groups with only a few strains in each group: 2,6,9,32,45,98 and 141. Eighty out of 84 E. coli strains of O-group 149 possessed the K88 antigen and produced heat labile enterotoxin (LT). Besides, LT production was demonstrated in 3 out of 14 strains of E. coli 08. K88 antigen was demonstrated in only 1 strain not belonging to O-group 149. Among strains of E. coli 064 12 out of 14 were K99 positive. This antigen was not demonstrated in E. coli strains of other O-groups. A close relationship was demonstrated between strains of E. coli 0149 possessing the K88 antigen and the ability to ferment both raffinose and adonitol. This ability was only detected in 2 other strains of E. coli not belonging to O-group 149.     

Nationwide surveillance of salmonella in the faeces of pigs in Japan

The prevalence of faecal carriage of salmonella in 5393 pigs reared on 218 pig farms located in 31 of 47 prefectures in Japan over the period July 2003 to June 2005 was investigated. We isolated 172 strains belonging to 20 serovars and one untypable Salmonella enterica from 169 pig faecal samples (3.1%) collected from 48 farms (22.0%). The most prevalent type of S. enterica was untypable O4,12:d:- which lacks phase 2 flagellar antigen, representing 29.1% (50/172) of all isolates. Of 26 S. enterica serovar Typhimurium isolates, 16 strains appeared to be definitive phage type 104 (DT104) by polymerase chain reaction.     

Genomic lineage of Salmonella enterica serovar Dublin

Thirty five strains of the host adapted Salmonella serotype Dublin (S. Dublin) have been characterized by IS200 patterns, ribotyping, pulsed-field gel electrophoresis (PFGE), restriction fragment polymorphism after hybridization with five randomly cloned DNA-framents of S. enteridis (RFLP), and plasmid profiling in order to divide the strains into ‘genomic lines’. For comparison, 20 other strains of 9 different group-D serotypes were included. The IS200 patterns were identical in all strains of S. Dublin examined. These patterns were different from those observed in other group-D Salmonella with the exception of one strain S. Enteritidis phage type 11 and a strain of S. Rostock. The insertion element IS200 was not detected in strains of S. Dar-es-Salam, S. (II) 9,12:z -, and S. Panama. RFLP, based on probing with five random cloned chromosomal fragments gave the same pattern in all strains except for one isolate from the UK. This strain was also found to have an unique PFGE pattern and ribotype. Among the remaining strains, three different PFGE patterns and 7 different ribotypes were observed. Based on all four typing methods, 8 different ‘genomic lines’ of S. Dublin were identified. The same grouping could be obtained from the use of ribotyping alone, but PFGE and RFLP were found to provide valuable information on possible relationships between ribotypes. Seven different plasmid profiles and a group of strains without plasmids were observed. In several cases, the same plasmid profile was shown to be present in more than one ‘genomic line’.