Isolations of aerobic bacteria from wild desert bighorn sheep (Ovis canadensis nelsoni and O. c. mexicana) in Arizona

Nasal, pharyngeal, cervical and vaginal swab specimens were obtained from 74 desert bighorn sheep for the purpose of investigating the normal aerobic bacterial flora of wild sheep. A total of 281 isolates was obtained and identified by standard microbiologic tests. One hundred seven of these isolates were gram positive and included Bacillus sp. (36%), Staphylococcus epidermidis (8%), S. aureus (4%), Corynebacterium sp. (diphtheroids, 4%), and Streptococcus sp. (48%). gram negative isolates totaled 174 and included Neisseria sp. (18%), Citrobacter sp. (3%), Enterobacter sp. (2%), Escherichia coli (2%), Proteus sp. (2%) and non-fermentative bacilli (NFB) (73%). Of the NFB isolates, Pseudomonas sp. (25%), Acinetobacter sp. (18%), Moraxella sp. (15%) were identified.     

Antibiotic sensitivity pattern; experience at University Hospital, Riyadh, Saudi Arabia

Results of sensitivity testing were discussed based on examination of 5192 isolates of the various bacteria isolated from clinical specimens from King Khalid University Hospital in Riyadh, Saudi Arabia. Streptococcus pyogenes and Streptococcus pneumoniae were sensitive to penicillin and erythromycin. The sensitivity pattern of Staphylococcus aureus was also predictable as they were fairly sensitive to both methicillin (98%) and erythromycin (96%). Neisseria gonorrhoeae (27%) showed a high level of resistance to penicillin. The resistance of Haemophilus influenzae to ampicillin and chloramphenicol was low. Brucella species was sensitive to tetracycline and rifampicin; resistance to streptomycin and cotrimoxazole was minimal being 1% and 6% respectively. The resistance of E. coli, Klebsiella species and Proteus species to second and third generation cephalosporins and amikacin was fairly low ranging from 1.3% to 3%. The gentamicin resistance for these organisms was also within the acceptable range (3%-10%). Gentamicin and amikacin resistance for Pseudomonas aeruginosa was low (2-8%). Salmonella typhi was sensitive to ampicillin, cotrimoxazole, and chloramphenicol. Salmonella enteritidis, Shigella species, and enteropathogenic E. coli were highly resistant to various antibiotics. Campylobacter jejuni was sensitive to gentamicin but 6% of isolates were resistant to erythromycin. Ninety six percent of Gram-negative rods except P. aeruginosa isolated from urine of patients having urinary tract infections were sensitive to amoxycillin-clavulanic acid. In addition, P. aeruginosa showed fairly low resistance to norfloxacin which is given orally to treat cystitis caused by this organism.     

Universal primers for detection of common bacterial pathogens causing prosthetic joint infection

The diagnosis of low grade prosthetic joint infection is difficult and time consuming. Nested-PCR for universal bacterial DNA segments detection of "orthopaedic" bacteria was tested in a laboratory setting. This method is based on amplification of the 16S bacterial ribosomal RNA coding sequences. 11 species of the most frequent bacterial pathogens (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus agalactiae, Enterococcus faecium, Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Serratia marcescens) involved in prosthetic joint infections were studied. All could be detected rapidly and sensitively by this method.     

A Microbiological Study of Pneumonic Calf Lungs

BITSCH, V., N. F. FRIIS and H. V. KROGH: A microbiological study of pneumonic calf lungs. Acta vet. scand. 1976, 17, 32–42.–Fifty pneumonic calf lungs were subjected to microbiologic screening with regard to bacteria, mycoplasmas, and viruses. Of bacteria the species most commonly found were Pasteurella multocida (eight lungs), Pasteurella hemolytica (eight lungs), and Corynebacterium pyogenes (13 lungs). Of special interest was the demonstration of Neisseria spp. in five lungs. Mycoplasma dispar was found in 31 lungs, Mycoplasma bovirhinis in 16 lungs, and Urea-plasma in 26 lungs. Cytopathogenic agents were demonstrated in 14 lungs. Four isolates were found to be bovine respiratory syncytial virus, three were bovine viral diarrhea virus, and two were bovine parainfluenza 3 virus. The remaining five cytopathogenic agents were not identified.     

Rapid extraction of bacterial genomic DNA with guanidium thiocyanate

A method is described for the rapid isolation and purification of bacterial genomic DNA. A total of 215 bacterial strains representing species of Campylobacter, Corynebacterium, Escherichia, Legionella, Neisseria, Staphylococcus and Streptococcus , were lysed with guanidium thiocyanate. DNA was prepared using just three other reagents and one high-speed centrifugation step. The method, which was applicable to both Gram-positive and Gram-negative bacteria, eliminated endogenous nuclease activity and avoided the need for phenol, RNase and protease treatments. The DNA was of high purity, high molecular mass and double-stranded.     

Agar Medium for the Selective Enumeration of Coagulase-positive Staphylococcus from the Rat Alimentary Tract

A selective agar medium (medium J(1)) is proposed for the quantitative enumeration of egg yolk-positive (EYP) and egg yolk-negative (EYN) Staphylococcus pyogenes from the digestive tract and feces of the rat. This medium, buffered at pH 5.0, is composed of acid casein hydrolysate and yeast extract with 7.5% sodium chloride, 1.6% sodium pyruvate, 0.0008% 2,3,5-triphenyltetrazolium chloride (TTC), and 6% egg yolk emulsion. Inoculation is by the pour-plate method and incubation is at 38 C in a water-jacketed incubator for 36 hr. Colonies of S. pyogenes reduce TTC; EYP strains are surrounded by a halo of opacity; and EYN strains may be surrounded by a red halo, but no opacity. Small, white colonies of S. epidermidis may develop, but Micrococcus, and all other groups of Staphylococcus recognized in the rat intestinal flora, are inhibited. Other bacterial genera, notably Bacillus, Corynebacterium, Proteus, and Streptococcus, are also inhibited.     

In Vitro Action of Carbenicillin Against Bacteria Isolated from Clinical Material

More than 500 bacteria isolated from patient material were tested against carbenicillin (disodium alpha-carboxybenzylpenicillin) by diffusion and dilution modalities. The same bacteria, which included Pseudomonas aeruginosa, Escherichia coli, Klebsiella-Aerobacter-Enterobacter group, various species of Proteus, Staphylococcus aureus and epiddermidis, enterococci, pneumococci, Streptococcus pyogenes, etc., were examined for susceptibility to other antibiotics commonly used with special emphasis on ampicillin and cephalothin. The responses of pyocine-typed P. aeruginosa were the most remarkable. The majority of these bacteria displayed susceptibility to carbenicillin by both the dilution and the diffusion techniques. The concentrations of this antibiotic used in the laboratory were of the same order of magnitude as that of the other drugs. The laboratory behavior of the other bacteria, toward this new semisynthetic penicillin derivative approximated their response to ampicillin and cephalothin.     

Effect of chlorhexidine/EDTA/Tris against bacterial isolates from clinical specimens

Chlorhexidine/EDTA/Tris was more active compared with chlorhexidine against the following species of organisms: Acinetobacter species, Citrobacter species, Enterobacter species, Escherichia coli, Proteus mirabilis, Providence species, Pseudomonas species, Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus faecalis, when tested in isosensitest agar. The addition of 20% serum to the medium reduced its effectiveness mainly against Providence species, Proteus species, and Streptococcus faecalis. However, the potential for this solution as a bladder instillation or topical antiseptic should be considered because of its reduced side effects compared with chlorhexidine alone, and its increased general effectiveness against all isolates tested.     

In-vitro antibacterial, antifungal and cytotoxic activities of some coumarins and their metal complexes

A series of new antibacterial and antifungal coumarin-derived compounds and their transition metal complexes [cobalt (II), copper (II), nickel (II) and zinc (II)] have been synthesized, characterized and screened for their in vitro antibacterial activity against Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella typhi, Shigella dysenteriae, Bacillus cereus, Corynebacterium diphtheriae, Staphylococcus aureus and Streptococcus pyogenes bacterial strains and for in vitro antifungal activity against Trichophyton longifusus, Candida albicans, Aspergillus flavus, Microsporum canis, Fusarium solani, Candida glaberata. The results of these studies show the metal complexes to be more antibacterial and antifungal as compared to the uncomplexed coumarins. The brine shrimp bioassay was also carried out to study their in vitro cytotoxic properties.     

Metalloantibiotics: synthesis, characterization and in-vitro antibacterial studies on cobalt (II), copper (II), nickel (II) and zinc (II) complexes with cloxacillin

The synthesis and characterization of cloxacillin (clox) complexes with divalent metal ions [Co (II), Cu (II), Ni (II) and Zn (II)] is described. The nature of bonding of the chelated cloxacillin and the structures of the metal complexes have been elucidated on the basis of their physical and spectroscopic data. In all the complexes, the cloxacillin acts as a uninegatively charged bidentate ligand with coordination involving the carboxylate-O and endocyclic-N of the beta-lactam ring. The new compounds have been screened for in-vitro antibacterial activity against Escherichia coli (a), Klebsiella pneumonae (b), Proteus mirabilis (c), Pseudomonas aeruginosa (d), Salmonella typhi (e), Shigella dysentriae (f), Bacillus cereus (g), Corynebacterium diphtheriae (h), Staphylococcus aureus (j) and Streptococcus pyogenes (k) bacterial strains. The brine shrimp bioassay was also carried out to study their in-vitro cytotoxic properties. All compounds, respectively, showed a promising activity (90%) against five bacterial species at 10 microg/ml concentration and a significant activity (52%) against the same test bacteria at 25 microg/ml concentration.     

Multilocus sequence typing

Multilocus sequence typing (MLST) provides a new approach to molecular epidemiology that can identify and track the global spread of virulent or antibiotic-resistant isolates of bacterial pathogens using the Internet. MLST databases, together with interrogation software, are available for Neisseria meningitidis and Streptococcus pneumoniae and databases for Streptococcus pyogenes and Staphylococcus aureus will be released shortly.     

In-vitro antibacterial and cytotoxic activity of cobalt (ii), copper (ii), nickel (ii) and zinc (ii) complexes of the antibiotic drug cephalothin (Keflin)

Keflin (kefl) interacts with Co(II), Cu(II), Ni(II) and Zn(II) metal ions leading to complexes of the type M(kefl)2Cl2 and M(kefl)Cl2, which have been characterized by physicochemical and spectroscopic methods. Magnetic moment, IR, electronic spectral and elemental analyses data suggest that keflin behaves tridentately forming octahedral or trigonal bipyramidal complexes with the metal ions mentioned above. The new compounds have been screened in-vitro for antibacterial and cytotoxic activity against Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella typhi, Shigella dysentriae, Bacillus cereus, Corynebacterium diphtheriae, Staphylococcus aureus and Streptococcus pyogenes bacterial strains. Compounds, 4 and 8 showed promising activity (90%) against seven, compound 6 showed significant activity (52%) against four and, compounds 1 and 5 showed activity (40%) against three test bacterial strains at concentration of 10 microM.     

Comparative in vitro activity of first, second and third generation cephalosporins

Minimum inhibitory concentrations (MIC) were determined against 662 recent clinical isolates for eight cephalosporins representing first, second and third generation compounds. All four third-generation cephalosporins tested (cefoperaxone, cefotaxime, ceftazidime and moxalactam) were significantly more active against aerobic gram-negative bacteria than the older compounds (cephalothin, cefamandole, cefoxitin, and cefuroxime). Cefotaxime and moxalactam were most active against Enterobacteriaceae with extremely low MIC-values. Ceftazidime was definitely most active against Pseudomonas aeruginosa with more than 90% of strains inhibited at 4 micro g/ml. MIC-values for cefotaxime against Staphylococcus aureus were for all strains 1-2 micro g/ml, slightly higher for cefoperazone, while the effect of ceftazidime and moxalactam was more limited. All third generation cephalosporins demonstrated efficiency against Streptococcus pyogenes, cefotaxime being most active and moxalactam least active, but were essentially ineffective against Streptococcus faecalis. Moxalactam demonstrated higher activity against Bacteroides fragilis than other second and third generation cephalosporins including cefoxitin. Previous studies have demonstrated a very high activity of all third generation cephalosporins against Haemophilus influenzae and Neisseria gonorrhoeae, including beta-lactamase producing strains.     

Thymidine kinase of bacteria: activity of the enzyme in actinomycetes and related organisms

Various micro-organisms were studied for their thymidine kinase (adenosine 5'-triphosphate:thymidine 5'-phosphotransferase, EC 2.7.1.21) (TK) activity. The sonicated cell extract of Escherichia coli K12 had a TK activity of 35-66 pmol thymidine monophosphate formed min-1 (mg protein)-1. The cell extracts of Salmonella typhimurium and Klebsiella pneumoniae showed a markedly higher (5- to 11-fold) TK activity. Somewhat lower but significant TK activity was detected in the cell extracts of Staphylococcus aureus, Streptococcus pyogenes, Bacillus subtilis and Proteus mirabilis. In contrast, weak TK activity, if any, was detected in the cell extracts of Pseudomonas aeruginosa. This was also the case with respect to the cell extracts of various actinomycetes (such as Nocardia and Streptomyces) and related organisms (such as Corynebacterium, Mycobacterium and Rhodococcus).     

Milk chloride level as an indicator of bovine mastitis

The aim of this study was to determine an influence of udder infection on milk chloride level and on milk productivity of cows of black and white race. Bacteriological analysis was performed by bacterial isolation from milk collected in sterile conditions from single lobes of mammary gland. The study was aimed to detect the following bacteria: Streptococcus agalactiae, Streptococcus uberis, Streptococcus agalactiae, Staphylococcus aureus, Staphylococcus epidermidis, Actinomyces pyogenes, Enterobacteriaceae rods, Corynebacterium bovis, and Micrococcus sp. Milk chloride level was determined by burette method in 1250 milk samples collected from entire udder. Milk productivity was determined on the day of bacterial isolation. Statistical analysis of the results of the study on udder infection, milk chloride level, and milk productivity of mammary gland did reveal lack of a simple correlations between those parameters and in the indirect manner indicated an influence of inappropriate maintenance conditions of tested cows on the health condition of their udder. It seems possible that alkalosis and acidosis in cows, taken into consideration in discussion section, could constitute a factor influencing the frequency of mastitis incidence.     

Cleavage of influenza A virus H1 hemagglutinin by swine respiratory bacterial proteases.

Cleavage of influenza A virus hemagglutinin (HA) is required for expression of fusion activity and virus entry into cells. Extracellular proteases are responsible for the proteolytic cleavage activation of avirulent avian and mammalian influenza viruses and contribute to pathogenicity and tissue tropism. The relative contributions of host and microbial proteases to cleavage activation in natural infection remain to be established. We examined 23 respiratory bacterial pathogens and 150 aerobic bacterial isolates cultured from the nasal cavities of pigs for proteolytic activity. No evidence of secreted proteases was found for the bacterial pathogens, including Haemophilus parasuis, Pasteurella multocida, Actinobacillus pleuropneumoniae, Bordetella bronchiseptica, and Streptococcus suis. Proteolytic bacteria were isolated from 7 of 11 swine nasal samples and included Staphylococcus chromogenes, Staphylococcus hyicus, Aeromonas caviae, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Enterococcus sp. Only P. aeruginosa secreted a protease, elastase, that cleaved influenza virus HA. However, compared to trypsin, the site of cleavage by elastase was shifted one amino acid in the carboxy-terminal direction and resulted in inactivation of the virus. Under the conditions of this study, we identified several bacterial isolates from the respiratory tracts of pigs that secrete proteases in vitro. However, none of these proteolytic isolates demonstrated direct cleavage activation of influenza virus HA.     

Surface microbial consortia from Livarot, a French smear-ripened cheese

The surface microflora (902 isolates) of Livarot cheeses from three dairies was investigated during ripening. Yeasts were mainly identified by Fourier transform infrared spectroscopy. Geotrichum candidum was the dominating yeast among 10 species. Bacteria were identified using Biotype 100 strips, dereplicated by repetitive extragenic palindromic PCR (rep-PCR); 156 representative strains were identified by either BOX-PCR or (GTG)(5)-PCR, and when appropriate by 16S rDNA sequencing and SDS-PAGE analysis. Gram-positive bacteria accounted for 65% of the isolates and were mainly assigned to the genera Arthrobacter , Brevibacterium , Corynebacterium , and Staphylococcus . New taxa related to the genera Agrococcus and Leucobacter were found. Yeast and Gram-positive bacteria strains deliberately added as smearing agents were sometimes undetected during ripening. Thirty-two percent of the isolates were Gram-negative bacteria, which showed a high level of diversity and mainly included members of the genera Alcaligenes , Hafnia , Proteus , Pseudomonas , and Psychrobacter . Whatever the milk used (pasteurized or unpasteurized), similar levels of biodiversity were observed in the three dairies, all of which had efficient cleaning procedures and good manufacturing practices. It appears that some of the Gram-negative bacteria identified should now be regarded as potentially useful in some cheese technologies. The assessment of their positive versus negative role should be objectively examined.     

A review of antimicrobial resistance in Canada

Antimicrobial resistance is a global concern. Over the past 10 years, considerable efforts and resources have been expended to detect, monitor, and understand at the basic level the many different facets of emerging and increasing resistance. This review summarizes our current understanding of bacterial antimicrobial resistance issues in Canada with particular emphasis given to the Enterobacteriaceae, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus, Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis, and Streptococcus pyogenes. In addition, future concerns and programs for ongoing surveillance are discussed.     

In vitro antibacterial, antifungal & cytotoxic activity of some isonicotinoylhydrazide Schiff's bases and their cobalt (II), copper (II), nickel (II) and zinc (II) complexes

Isonicotinoylhydrazide Schiff's bases formed by the reaction of substituted and unsubstituted furyl-2-carboxaldehyde and thiophene-2-carboxaldehyde with isoniazid and, their Co (II), Cu (II), Ni (II) and Zn (II) complexes have been synthesized, characterized and screened for their in vitro antibacterial activity against Mycobacterium tuberculosis, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella typhi, Shigella dysenteriae, Bacillus cereus, Corynebacterium diphtheriae, Staphylococcus aureus and Streptococcus pyogenes bacterial strains and for in vitro antifungal activity against Trichophyton longifusus, Candida albicans, Aspergillus flavus, Microsporum canis, Fusarium solani and Candida glabrata. The results of these studies show the metal complexes to be more antibacterial and antifungal against one or more bacterial/fungal strains as compared to the uncomplexed compounds. The brine shrimp bioassay indicated Schiff's bases, L3 and L6 and, their Cu (II) and Ni (II) metal complexes to be cytotoxic against Artemia salina, while all other compounds were inactive (LD50 > 1000).     

The microflora of ulzerous zone mucosa in patients with duodenal ulcer

Bacteriological study of the biopsies taken from gastric and duodenal mucosa of 10 healthy volunteers and 74 patients with duodenal ulcer, was carried out. In the gastroduodenal zone of healthy subjects microorganisms of 6 genera (Streptococcus, Candida, Staphylococcus, Bacillus, Helicobacter and Lactobacillus) were detected. H. pylori was isolated in 20% of cases only in biopsy specimens taken from the antral section of the stomach of healthy as monoculture or in combination with C. albicans. In patients with duodenal ulcer activation of opportunistic microflora was observed in the periulcerous zone. More often H. pylori occurred in associations with fungi of the genus Candida, streptococci, staphylococci, enterobacteria, Pseudomonas and other microorganisms (of more than 30 genera). Quantitatively the dominating microorganisms (3.8-5.7 lg CFU/g) were H. pylori, fungi of the genus Candida, bacteria of the genera Streptococcus, Peptostreptococcus, Bacteroides, Gemella, Prevotella, Veillonella, Peptococcus, Bacillus, different species of opportunistic enterobacteria, as well as bacteria of the genera Staphylococcus, Micrococcus, Corynebacterium, Neisseria, Pseudomonas, etc. Opportunistic bacteria detected in the ulcerous zone, as a rule, expressed hemolytic, lecithinase, RNAase, caseinolytic, catalase and urease activity. Sonicated filtrates of such cultures produced a cytotoxic effect on cells HEp-2. Ulcer is an infected wound that needs sanitation.