Identification and composition of the tonsillar and anal enterococcal and streptococcal flora of dogs and cats

Enterococcus faecalis was the most frequently isolated enterococcal species from anal swabs and tonsils of dogs and cats, although in the anal samples from dogs Ent. hirae was found almost as often as Ent. faecalis. Most Ent.faecium strains from dog tonsils differed from those associated with humans and other animals in that they fermented sorbitol. Typical Ent. avium as well as atypical Ent. avium -like strains were seen in dogs, while the related species Ent. raffinosus was associated with cat tonsils. Enterococcus cecorum also occurred mainly in cats. Certain atypical strains, presumptively identified as Ent. cecorum , shared characteristics with Ent. columbae.
The most frequent streptococcal species in tonsils of cats and dogs were Streptococcus suis and Strep. canis. Streptococcus canis and Strep. bovis predominated in anal swabs. The canine Strep. suis differed from the common porcine strains in fermenting mannitol.
Forty-seven of the 288 isolates examined could not be identified or related to known species. The characteristics of two groups of these bacteria, provisionally called 'Ton 31 group' and 'O7 group' are described.     

Identification and composition of the streptococcal and enterococcal flora of tonsils, intestines and faeces of pigs

Streptococcus suis was the most frequent Streptococcus spp. in pig tonsils, followed by the beta-haemolytic porcine 'equisimilis'ecovar of Strep. dysgalactiae. The intestinal streptococcal flora was composed of Strep. bovis, Strep. hyointestinalis and Strep. suis. Many of these intestinal Strep. suis belonged to a beta-glucuronidase-negative biotype which is infrequent in lesions. Nearly half of the strains presumptively identified as Strep. alactolyticus produced acid from lactose. This species was not found in tonsils and intestines but was about equally prevalent as Strep. hyointestinalis in pig faeces and rectal swabs. Other streptococci were rare in this material.
Enterococci were much less frequently identified than streptococci in tonsils and faeces. In intestinal samples Enterococcus faecalis, Ent. faecium, Ent. hirae and Ent. cecorum were most frequently found. In faeces Ent. faecium was the most prevalent enterococcus.
The characteristics of the less well known species Strep. alactolyticus and Strep. hyointestinalis are described in detail, and guidelines for their differentiation from Strep. bovis and Strep. suis given.     

Composition of the enterococcal and streptococcal intestinal flora of poultry

Identification to species level showed that Enterococcus faecalis and Ent. faecium largely dominated the enterococcal and streptococcal gut flora of 1-d-old chicks. Enterococcus faecalis was rare in 3- to 5-week-old broilers. Two species, Ent. faecium and Streptococcus alactolyticus, were isolated from nearly all broilers examined. Enterococcus hirae and Ent. durans were found in the small intestines of this category of poultry. In layers and parent stock of over 12 weeks of age, Ent. cecorum dominated with Strep. alactolyticus ranking next. Other species were isolated irregularly. Enterococcus avium and Ent. gallinarum, originally described from chickens, were rarely found. These species did not appear to belong to the normal intestinal flora of poultry.     

Composition of the enterococcal and streptococcal intestinal flora of poultry

Identification to species level showed that Enterococcus faecalis and Ent. faecium largely dominated the enterococcal and streptococcal gut flora of 1-d-old chicks. Enterococcus faecalis was rare in 3- to 5-week-old broilers. Two species, Ent. faecium and Streptococcus alactolyticus , were isolated from nearly all broilers examined. Enterococcus hirae and Ent. durans were found in the small intestines of this category of poultry.
In layers and parent stock of over 12 weeks of age, Ent. cecorum dominated with Strep. alactolyticus ranking next. Other species were isolated irregularly. Enterococcus avium and Ent. gallinarum , originally described from chickens, were rarely found. These species did not appear to belong to the normal intestinal flora of poultry.     

Enterococcal and streptococcal species isolated from faeces of calves, young cattle and dairy cows.

Faeces from non-ruminating calves were found to contain several species of enterococci: Enterococcus avium, Ent. cecorum, Ent. durans, Ent. faecalis, Ent. faecium and Ent. hirae. Enterococcus faecalis was the most frequent. Few of these animals carried streptococci. Streptococcus bovis largely predominated in ruminating calves, young cattle and dairy cows. Other streptococci as well as enterococci were infrequent in dairy cows, but a variety of other streptococci and enterococci were found in the faeces of young ruminating animals.     

Enterococcal and streptococcal species isolated from faeces of calves, young cattle and dairy cows

Faeces from non-ruminating calves were found to contain several species of enterococci: Enterococcus avium, Ent. cecorum, Ent. durans, Ent. faecalis, Ent. faecium and Ent. hirae. Enterococcus faecalis was the most frequent. Few of these animals carried streptococci.
Streptococcus bovis largely predominated in ruminating calves, young cattle and dairy cows. Other streptococci as well as enterococci were infrequent in dairy cows, but a variety of other streptococci and enterococci were found in the faeces of young ruminating animals.     

The survival of streptococci and enterococci in animal feed and on straw with particular reference to Streptococcus uberis

The survival of Streptococcus uberis, Strep. bovis, Enterococcus faecalis spp. faecalis and Ent. faecium in animal feed and on wheat straw was studied for one month. Streptococcus bovis survived the best in feed and Strep. uberis the worst while on straw Ent. faecium was the most resilient species and there was little to choose between the other three. The recovery of all species, excepting Strep. bovis , on selective and non-selective agar was comparable. The growth of Strep. bovis , on the other hand, was inhibited on both Edward's medium and inulin agar.     

The survival of streptococci and enterococci in animal feed and on straw with particular reference to Streptococcus uberis

The survival of Streptococcus uberis, Strep. bovis, Enterococcus faecalis spp. faecalis and Ent. faecium in animal feed and on wheat straw was studied for one month. Streptococcus bovis survived the best in feed and Strep. uberis the worst while on straw Ent. faecium was the most resilient species and there was little to choose between the other three. The recovery of all species, excepting Strep. bovis, on selective and non-selective agar was comparable. The growth of Strep. bovis, on the other hand, was inhibited on both Edward's medium and inulin agar.     

The distribution of homologous enterococcal plasmid DNA sequences in human faecal isolates.

Hybridization was used to investigate the distribution of enterococcal plasmid sequences among 306 strains of Enterococcus and Streptococcus spp. isolated from faeces of humans of various ages. As DNA probes for the survey three plasmids, whose DNAs did not hybridize each other and designated as pMS13, pTW34 and pHK30, were selected from plasmids borne in Ent. faecalis. pTW34 DNA hybridized only with DNAs from enterococci, with high frequency in Ent. faecalis and low frequency in Ent. faecium. pMS13 DNA hybridized with DNAs of all Enterococcus spp. tested and with Strep. bovis, Strep. equinus and Strep. salivarius. Eighty-five percent of Ent. faecium isolates had sequences homologous to pMS13 but in the other species the values were less than 60%. Some enterococci had DNAs which hybridized with the pHK30 probe. The different distribution of the three DNA sequences indicates the possibility that plasmid DNAs encode advantageous phenotypes for the colonization of bacteria in the lumen of the bowel.     

A new simple medium for the detection of Enterococcus faecalis and Enterococcus faecium by measurement of conductance changes

Streptococcus suis, Streptococcus bovis and the mastitis pathogens Streptococcus dysgalactiae and Streptococcus uberis were the most frequently occurring streptococci in tonsils of cattle. Streptococcus suis dominated in samples from calves between 1 month and 1 year of age, but was much less frequent in calves less than 1 month old. The mastitis pathogen Strep, dysgalactiae was found more often in calves than in older animals. Enterococci were relatively rare, except in the youngest age group. Nearly one third of the strains examined could not be identified to known species.     

The distribution of homologous enterococcal plasmid DNA sequences in human faecal isolates

T. WATANABE, H. KUMATA, M. SASAMOTO AND M. SHIMIZU-KADOTA. 1992. Hybridization was used to investigate the distribution of enterococcal plasmid sequences among 306 strains of Enterococcus and Streptococcus spp. isolated from faeces of humans of various ages. As DNA probes for the survey three plasmids, whose DNAs did not hybridize each other and designated as pMS13, pTW34 and pHK30, were selected from plasmids borne in Ent. faecalis. pTW34 DNA hybridized only with DNAs from enterococci, with high frequency in Ent. faecalis and low frequency in Ent. faecium. pMS13 DNA hybridized with DNAs of all Enterococcus spp. tested and with Strep. bovis, Strep. equinus and Strep. salivarius. Eighty-five percent of Ent. faecium isolates had sequences homologous to pMS13 but in the other species the values were less than 60%. Some enterococci had DNAs which hybridized with the pHK30 probe. The different distribution of the three DNA sequences indicates the possibility that plasmid DNAs encode advantageous phenotypes for the colonization of bacteria in the lumen of the bowel.     

The gram-positive tonsillar and nasal flora of piglets before and after weaning

AIMS: To investigate gram-positive nasal and tonsillar microbial flora of piglets before and after weaning. METHODS AND RESULTS: The nasal and tonsillar gram-positive bacterial flora of 20 non-weaned piglets (2 weeks of age) and 20 weaned piglets (6 weeks of age), obtained from four different piggeries, was quantified by culture and identified by tDNA-PCR. The most widely occurring species from nasal conchae before as well as after weaning in the different piglets investigated were Streptococcus suis and Rothia nasimurium. After weaning a wide variety of Lactobacillus species appeared but in low numbers. In the tonsils, Strep. suis, Strep. dysgalactiae, S. hyicus, S. aureus, Arcanobacterium pyogenes and Actinomyces hyovaginalis were the species isolated from the largest number of pigs before and after weaning. S. aureus and most lactobacilli became more prevalent after weaning. Bacteria not known to be associated with pigs found in the present study included R. nasimurium, Strep. gallolyticus, Pediococcus pentosaceus and some Lactobacillus species. CONCLUSIONS: Over 30 different gram-positive bacterial species may occur in nasal conchae and tonsils of unweaned piglets at 2 weeks of age and of 6-week-old weaned piglets. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated that weaning is associated with changes in prevalence of only a small minority of the highly diversified bacterial flora of the nares and tonsils of pigs.     

Molecular and physiological characterization of dominant bacterial populations in traditional mozzarella cheese processing

The development of the dominant bacterial populations during traditional Mozzarella cheese production was investigated using physiological analyses and molecular techniques for strain typing and taxonomic identification. Analysis of RAPD fingerprints revealed that the dominant bacterial community was composed of 25 different biotypes, and the sequence analysis of 16S rDNA demonstrated that the isolated strains belonged to Leuconostoc mesenteroides subsp. mesenteroides , Leuc. lactis , Streptococcus thermophilus , Strep. bovis , Strep. uberis, Lactococcus lactis subsp. lactis , L. garviae, Carnobacterium divergens , C. piscicola, Aerococcus viridans , Staphylococcus carnosus, Staph. epidermidis , Enterococcus faecalis , Ent. sulphureus and Enterococcus spp. The bacterial populations were characterized for their physiological properties. Two strains, belonging to Strep. thermophilus and L. lactis subsp. lactis , were the most acidifying; the L. lactis subsp. lactis strain was also proteolytic and eight strains were positive to citrate fermentation. Moreover, the molecular techniques allowed the identification of potential pathogens in a non-ripened cheese produced from raw milk.     

THE DISTRIBUTION OF GROUP D STREPTOCOCCI IN CATTLE AND SHEEP

Numbers and types of Lancefield group D streptococci have been determined in samples from the colons of 17 cattle and 9 sheep. Mean total streptococcal counts of 8 × 10⁴/g in cattle and 2 × 10⁶/g in sheep were obtained. Streptococcus bovis was found in every sample and was the predominant species in 15 of the cattle and 6 of the sheep. Other group D streptococci ( Strep. faecalis, Strep. faecium and Strep. durans ) were rare in cattle, but in sheep they formed a significant proportion of the population. Of 60 Strep. faecium, Strep. durans and related strains, 51 fermented raffinose. Many of the strains of Strep. faecium were also atypical in that they fermented sorbitol and appreciably reduced tetrazolium in broth at pH 6.0.
Strep. bovis remained the predominant streptococcus in faeces samples from 4 dairy cows when they were tested again after an interval of 17 and 18 months.     

Identification of enterococci isolated from cow's milk cheese: comparison of the classical methods and the API 20 STREP system (technical note)

A comparison of the results obtained using the classical methods with those of the API 20 Strep system was carried out in identifying 24 enterococci strains isolated from San Simón cow's milk cheese, a traditional Spanish variety. The results of both identification systems coincided exactly in 9 strains (37.5% of the strains studied). In one strain the results obtained using the classical methods did not coincide with those using the API 20 Strep method. 3 strains (12.5%) could not be identified using the API 20 Strep system. However, 11 strains (45%), that remained doubtful between both species E. faecalis and E. faecium on the basis of the classical methods, were identified using the API 20 Strep system. The API 20 Strep system does not include some biochemical tests of importance in identifying of foodborne enterococci and could not identify the atypical strains of Enterococcus. Moreover, this system is adapted to the identification of enterococci of clinical origin and their database does not include some species common in foods. However, it could have an application in combination with the classical methods in order to carry out a reasonably rapid and reliable identification of enterococci related to cheese.     

Distribution and Properties of Serological Types of Streptococcus faecium, Streptococcus durans and Related Strains

S ummary . The distribution of 19 serological types of Streptococcus faecium and related organisms has been studied, using 367 strains isolated mainly from faecal samples. Several types occurred in man as well as in pigs, sheep, cattle or chickens.
Strains of the same serological type showed a diversity of fermentation reactions, so that organisms which could be identified as Streptococcus durans shared a common type antigen with Strep. faecium strains. The evidence given here supports the proposal that Strep. durans should in future be considered as a variant of Strep. faecium .
It has also been shown that, in common with those of Streptococcus faecalis , the type antigens of the Strep. faecium types are cell wall components.     

Population structure and safety aspects of Enterococcus strains isolated from artisanal dry fermented sausages produced in Argentina

Enterococci population from Argentinean artisanal dry fermented sausage was identified and their safety aspects were evaluated. Species-specific PCR was used to distinguish between Enterococcus faecium (56%) and Enterococcus faecalis (17%). Other isolates (27%) were identified as Enterococcus durans , Enterococcus casseliflavus and Enterococcus mundtii by using 16S RNA gene sequence. RAPD analyses showed different biotypes for Ent. faecium and Ent. faecalis species. Low incidence of antibiotic resistance and high virulence traits in Ent. casseliflavus and Ent. faecalis were found; the majority of the Ent. faecium strains were shown to be free of virulence factors. The absence of virulence/resistance traits and the anti-Listeria activity of Ent. faecium isolates may be exploited to enhance natural preservation thereby guaranteeing organoleptic/safety characteristics of artisanal fermented sausages.     

Composition of the enterococcal intestinal flora of chickens in a tropical area

Enterococcus faecalis was the most frequently occurring enterococcal species in caeca and cloacal swabs of chickens in Assam, India. In chickens less than 8 weeks of age Ent. faecium, Ent. durans and Ent. avium were also found, but infrequently. Enterococcus hirae , which is potentially pathogenic in chickens of less than 1 week of age, was found only in chickens of more than 8 weeks. Except for a higher frequency of Ent. faecalis , the composition of this flora was essentially the same as that in European temperate climates.