Broth cultures

Vet Microbiol, 1994 Nov, 42(2-3), 239 - 44
Detection of verotoxigenic Escherichia coli in bull semen using the polymerase chain reaction; Gradil C et al.; Oligonucleotide primers used in a polymerase chain reaction (PCR) protocol detected the verotoxin 2 (VT2) gene in E . coli present in experimentally contaminated bull semen . The VT2 (Shiga-like toxin II {SLT-II}) primers targeted a 346-bp fragment of the gene coding for the A subunit of the toxin . PCR products, corresponding to the VT2 gene sequence, were amplified from template E . coli nucleic acid extracted from 18-h broth culture and from E . coli in contaminated semen in the undiluted state, diluted in egg yolk-Tris and diluted in milk . The sensitivity of the assay to detect E . coli was determined to be 1 pg of nucleic acid, and as few as 10-20 E . coli organisms/ml could be detected in raw and diluted semen . Preliminary confirmation of the PCR product was accomplished by slot blot hybridization to a radiolabeled specific oligoprobe . Sequencing of the PCR products identifying VT2 gene sequence revealed 99.7% homology with published gene sequences for VT2 . This study demonstrates the feasibility of applying PCR technology for the detection of E . coli in bovine semen . This technique may find wide application for the detection of other pathogens that may be present in semen.

J Bacteriol, 1994 Oct, 176(19), 5929 - 37
An amplifiable DNA region from the Mycoplasma hyorhinis genome; Deng G et al.; A novel amplifiable genomic region that displays variability in the number of tandem copies of a 1,368-bp DNA sequence (designated RS-2) was discovered among individual clonal derivatives within Mycoplasma hyorhinis broth-grown cell populations . Clonal isolates representing variant subpopulations from the original broth culture were of a single size variant, and although continued culture under a variety of growth conditions did not result in further amplification of RS-2, evidence for deletion events which reduced RS-2 copy number, presumably by homologous recombination, was obtained . RS-2 homologous sequences were identified in all M . hyorhinis strains tested, but only the tissue culture-derived strains GDL-1 and GDL-2 showed variability in genomic dosage . The RS-2 nucleotide sequence established that each tandem copy is flanked by direct repeats of a 20-bp sequence and suggested a possible mechanism for its original duplication as the initial phase of a genetic amplification process . The coding strand was defined by PCR amplification of a reverse transcriptase-generated cDNA, and its sequence revealed that RS-2 encodes a 456-residue internal, highly cysteine-rich domain of a larger M . hyorhinis protein whose coding sequence initiates and terminates in unique genomic sequences several hundred base pairs from RS-2 on either side of it . Changes in RS-2 copy number maintain the reading frame, and therefore the coding capacity, for this predicted size-variant protein.

Immunol Cell Biol, 1994 Oct, 72(5), 398 - 405
Cloning, sequencing, expression and inflammatory activity in skin of ovine interleukin-8; Seow HF et al.; Ovine IL-8 (oIL-8) cDNA was obtained by probing a spleen cell cDNA library with human IL-8 (hIL-8) cDNA . The oIL-8 cDNA was 1434 base pairs long with a single open reading frame encoding a 101 amino acid precursor protein of relative molecular mass 11,268 . The inferred amino acid sequence has 78, 82, 84 and 67% similarity with human, rabbit, porcine and guinea-pig IL-8, respectively . By analogy with the most prevalent form of hIL-8, a 72 amino acid form of oIL-8 was expressed as a fusion protein containing glutathione-S-transferase and purified by affinity chromatography on a glutathione-Sepharose column yielding 8 mg IL-8/L broth culture . The fusion protein lacked chemotactic activity for ovine neutrophils, whereas the 72 amino acid form of oIL-8 was equipotent with rhIL-8 . At 6 and 24 h after intradermal injection of 10(-9) mol oIL-8, there was intense accumulation of neutrophils, and very mild accumulation of eosinophils, CD5, CD4 and T19 (a gamma delta TCR subset) cells but not CD8 cells . The availability of roIL-8 and its cDNA probes will permit the role of this important member of the IL-8 family of chemotactic cytokines to be determined in inflammatory diseases of sheep.

Gene, 1994 Sep 15, 147(1), 47 - 54
Hypertransposing derivatives of the streptomycete insertion sequence IS493; Solenberg PJ et al.; Transposons derived from the Streptomyces lividans insertion sequence IS493 are useful for the genetic analysis and manipulation of a number of Streptomyces spp . Tn5099-10, an IS493 derivative that contains a spontaneous deletion terminating in the left inverted repeat (IR-L), transposed at a 1000-fold higher frequency in Streptomyces griseofuscus, and at a tenfold higher frequency in Streptomyces fradiae, than the IS493 derivatives, Tn5096 and Tn5099 . The IR-L from Tn5099-10 was used to construct a cassette which hypertransposes from plasmids containing the transposon genes, ORFA and ORFB, outside of the inverted repeats . The target sequences of two Tn5099-10 insertions conformed to the consensus target sequence of the other IS493 derivatives, gNCaNTgNNy (where lower-case letters indicate that other nt have been observed at this position and N is any nt) . Transposition mutant libraries of S . griseofuscus and S . fradiae can be easily prepared in broth culture by using the hypertransposing elements and a temperature-sensitive delivery plasmid.

Klin Lab Diagn, 1994 Sep-Oct, (5), 45 - 6
{Method of storage and transportation of bacterial culture using paper disks}; Fel'dman IuM et al.; A method for bacterial culture storage and transportation is suggested consisting in impregnation of filter paper disks with broth culture and drying . Bacterial cultures can be stored in this mode for at least a year at 4 degrees C.

Mol Cell Probes, 1994 Apr, 8(2), 125 - 30
Evaluation of the detection limits of PCR for identification of Mycoplasma pneumoniae in clinical samples; Leng Z et al.; The detection limits of the polymerase chain reaction (PCR) for Mycoplasma pneumoniae were determined using specimens from persons known to have had M . pneumoniae pneumonia . Four primers were selected from the known sequence of the P1 gene . The primer pair (P1-178 and P1-809) which generates a 631 fragment gave the lowest detection limit . Nineteen of 21 throat swabs, which contained between 0.06 and 2 colony-forming units (CFU) per microlitre, from culture positive patients, were positive by PCR . The fact that M . pneumoniae grows in broth culture in spherules causes problems for determining the number of CFU detected in PCR . Filtering broth cultures through a 0.6 micron polycarbonate filter increased the number of CFUs two-to-ten-fold compared to unfiltered cultures . The lysis method needed to assay throat swabs differed from that necessary for broth cultures in that proteinase K treatment for 18 h increased the detection limit 10- to 100-fold when compared to NaOH digestion.

Am J Vet Res, 1993 Nov, 54(11), 1874 - 80
Aerosol vaccination of pigs against Mycoplasma hyopneumoniae infection; Murphy D et al.; Aerosol vaccination is used effectively to immunize poultry against Newcastle disease, but to the authors' knowledge, this vaccination procedure is not well studied in other species . The efficacy of IM and aerosol vaccination of pigs against Mycoplasma hyopneumoniae infection was evaluated . Twenty-one pigs from a Mycoplasma-free herd were randomly allotted by litter and body weight into 3 groups . One group was given aerosolized phosphate-buffered saline solution (PBSS) by inhalation . The second group (AERO) was given aerosolized M hyopneumoniae vaccine by inhalation . The third group (IM) was given the same vaccine by IM injection . Vaccination by IM administration was repeated once, and aerosol vaccination was repeated twice at 2-week intervals . Two weeks after the last vaccination, all pigs were intratracheally challenge-exposed with 3 ml of broth culture containing 10(7) color-changing units (CCU) of a low-passage strain of virulent M hyopneumoniae . Pigs were observed daily for coughing . Four weeks after challenge exposure, all pigs were necropsied . Percentage of lung affected by gross pneumonia was measured, bronchioalveolar lavage fluid (BALF) cells were counted, and quantitative culture for mycoplasmas was performed on lung sections . Additionally, M hyopneumoniae-specific antibodies were measured in prevaccination, postvaccination, and postchallenge-exposure serum and BALF by use of indirect ELISA . Mean prevalence of persistent coughing in pigs of the AERO group (4.6 d/pig) was not different from that in pigs of the PBSS group (3.7 d/pig) . Prevalence of coughing in IM vaccinated pigs (1.0 d/pig) was lower (P < 0.05) than that in pigs of the PBSS group.(ABSTRACT TRUNCATED AT 250 WORDS)

J Clin Microbiol, 1992 Dec, 30(12), 3043 - 9
Western immunoblot analysis and serologic characterization of Blastomyces dermatitidis yeast form extracellular antigens; Hurst SF et al.; A major 98-kDa extracellular protein antigen of Blastomyces dermatitidis was shown in Western blot (immunoblot) analysis to be highly reactive with serum antibodies from patients with blastomycosis . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of yeast form B . dermatitidis culture filtrates and cell extracts demonstrated over 50 proteins, only 24 of which were immunoreactive . Of these, a 98-kDa protein was found to be the most specific and was isolated . This protein was found in both broth culture filtrates and extracts of yeast forms . Western blot tests with this antigen detected serum antibodies in 91% of 32 patients with clinically proven blastomycosis, in contrast to an enzyme-linked immunosorbent assay (ELISA) with DEAE-purified antigen, which detected 88% of the cases . The Western blot test also exhibited lower reactivity with a panel of sera from patients with heterologous fungal infections that were cross-reactive in the ELISA with DEAE-purified B . dermatitidis antigen . The 98-kDa protein electroeluted from polyacrylamide gels was identical to the diagnostic A antigen used in the blastomycosis immunodiffusion test . Comparison of the 98-kDa antigen with a previously described 120-kDa yeast form cell wall protein antigen of B . dermatitidis showed that these two antigens are almost identical.

J Med Microbiol, 1992 Nov, 37(5), 332 - 4
A biphasic system for primary isolation of mycobacteria compared to solid medium and broth culture; Hoffner SE et al.; A biphasic culture system, the MB Check, was compared with conventional culture on Lowenstein-Jensen egg (LJ) solid medium and with Bactec broth culture for primary isolation of mycobacteria from clinical samples . A total of 104 mycobacterial isolates was detected from 985 samples examined by the three methods . The most sensitive primary isolation was with LJ culture and MB Check; these methods detected 93% and 87% of all positive cultures, respectively . MB Check allowed a somewhat more rapid detection than LJ culture . The presence of atypical mycobacteria was demonstrated most rapidly with the Bactec system.

Avian Dis, 1992 Oct-Dec, 36(4), 956 - 63
Mycoplasma gallisepticum vaccination-challenge: an egg-production model; Evans RD et al.; Specific-pathogen-free layer hens in maximum lay were exposed by aerosol to a broth culture of Mycoplasma gallisepticum R' strain . Egg-production loss of greater than 50% was evident 7-14 days following challenge of unvaccinated chickens, with a gradual recovery during the next several days . Various vaccine preparations were tested to determine the effect in the model . All vaccinated chickens exhibited significantly (P < or = 0.05) lower egg-production loss than the unvaccinated controls . The model provides a method for testing treatment effects on egg-production losses resulting from controlled exposure to M . gallisepticum and may be useful in simulating field exposure.

Trans R Soc Trop Med Hyg, 1992 Sep-Oct, 86(5), 554 - 6
Blood cultures from Bangladeshi children with septicaemia: an evaluation of conventional, lysis-direct plating and lysis-centrifugation methods; Saha SK et al.; The use of a laboratory-made lysis-direct plating and lysis-centrifugation (LDP/LC) device for blood cultivation has been compared with the conventional broth culture method in respect of speed and sensitivity in detecting organism(s) and cost effectiveness . 400 blood cultures yielded 95 clinically significant isolates . Both methods recovered 73 organisms (76.8%); 20 (21%) were detected by LDP/LC methods only, and 2 (2.1%) were isolated by the conventional method only . All the 93 isolates (97.8%) recovered by LDP/LC were isolated within 48 h, whereas the broth culture method took 7 d to isolate a total of 75 organisms (78.9%) . The LDP/LC method, with our laboratory-made device, costs one-fourth of the cost of the conventional broth culture system.

Hum Pathol, 1992 Sep, 23(9), 1004 - 10
Characterization of HeLa cell vacuoles induced by Helicobacter pylori broth culture supernatant; Cover TL et al.; Helicobacter pylori broth culture supernatants induce eukaryotic cell vacuolation in vitro, a phenomenon that has been attributed to cytotoxic activity . We sought to characterize further the vacuolation of HeLa cells that occurs in response to H pylori culture supernatant . Nascent vacuoles were detectable by electron microscopy after 90 minutes of incubation with H pylori supernatant and were not associated with any identifiable organelle . After 6 days of incubation with H pylori supernatant, vacuoles were membrane-bound structures filled with electron-dense debris, which resembled secondary lysosomes . Acid phosphatase activity was detected within the vacuoles . The vacuoles induced by H pylori supernatant were then compared with vacuoles induced by trimethylamine, a weak base known to induce lysosomal swelling . Neutral red dye rapidly entered the vacuoles induced by either H pylori supernatant or trimethylamine, and both types of vacuoles were reversible . Compared with trimethylamine-induced vacuoles, the vacuoles induced by H pylori supernatant were larger and typically lacked a limiting membrane . In the early stages of formation, vacuoles induced by trimethylamine were labeled by lucifer yellow, a pinocytotic marker, whereas H pylori cytotoxin-induced vacuoles were not . These data suggest that trimethylamine-induced vacuoles arise directly from endocytic compartments, whereas H pylori cytotoxin induces vacuole formation via an autophagic mechanism.

J Med Microbiol, 1992 Aug, 37(2), 118 - 22
Cytopathic effects of Helicobacter pylori on cultured mammalian cells; Konishi H et al.; Cytopathic effects of broth-culture filtrates from eight clinical isolates and one reference strain of Helicobacter pylori on three cultured mammalian cell lines were investigated . All the strains, including NCTC 11637, produced cytotoxic factors that caused intracellular vacuolation on these cell lines . AGS and SflEp cells were more sensitive than HEp-2 cells . To examine the role of urease in the cytotoxic effect, a urease-negative mutant was produced . Filtrates from both wild-type and mutant strains produced similar vacuolation on SflEp cells in the absence of urea, suggesting that H . pylori produces a cytotoxic substance other than urease . In contrast, ammonia alone, or jack bean urease with urea, also induced rounding and detachment of SflEp cells, whereas ammonium salts induced the production of small vacuoles . The combination of the broth filtrate of the wild-type strain and urea induced vacuolation followed by rounding and detachment of SflEp cells . Evidence is presented that the latter changes are due to ammonia produced during incubation . Nevertheless, the amounts produced were less than that needed to induce cytopathic effects by itself . These results suggest that the cytotoxic substance induces intracellular vacuolation, and that the vacuolated cells are more susceptible to killing by ammonia . Thus both the cytotoxic substance and urease may contribute to the lethal cytotoxicity of H . pylori in vitro.

Zentralbl Veterinarmed B, 1992 Jul, 39(5), 353 - 61
Preparation and characterization of monoclonal antibodies against Mycoplasma bovis; Berthold E et al.; Monoclonal antibodies (mabs) against Mycoplasma (M.) bovis were prepared for use in diagnosis of bovine mastitis . From the original 32 hybridomas actively secreting mabs against M . bovis, 6 stable lines were cloned . Two of them, Mb 5D8 and Mb 4F6, recognized M . bovis antigens of estimated molecular weights of 33 and 26 kDa, respectively . They showed no cross-reaction to other bovine mycoplasmas, thus rendering them useful for specific detection of this pathogen . All mabs investigated cross-reacted with M . agalactiae which is known to be closely related to M . bovis, but does not occur in cattle . Two other mabs, Mb 5D4 and Mb 1F6, exhibited further cross-reactions to a number of bovine mycoplasma species . Finally, mabs Mb 5D5 and Mb 2G5 reacted with all mycoplasmas tested . The possibility that they recognized constituents of the broth culture medium is discussed.

Vet Microbiol, 1992 Jun 1, 31(2-3), 251 - 61
Detection of entero- and verocyto-toxin genes in Escherichia coli from diarrhoeal disease in animals using the polymerase chain reaction; Woodward MJ et al.; Oligonucleotide primers were designed for the specific polymerase chain reaction (PCR) amplification of the enterotoxins STIa and LTI and of the verocytotoxins VT1 and VT2 . All of 184 E . coli isolates from cases of diarrhoea from pigs, cattle and sheep gave identical toxin profiles by PCR and gene probe . Differentiation between VT2 and VT2v was achieved using two oligonucleotide primers pairs in PCR and showed that all of 34 VT2+ porcine isolates, of which 23 were 0138:K1, harboured VT2v whereas 20 VT2+ bovine and ovine isolates harboured VT2 . No isolate harboured both VT2 polymorphs . Simplified methods for sample preparation for PCR were examined and showed that PCR was not inhibited by direct addition of broth culture to the reaction mixture.

Am J Gastroenterol, 1991 Nov, 86(11), 1596 - 603
New small animal model for human gastric Helicobacter pylori infection: success in both nude and euthymic mice; Karita M et al.; In order to develop an experimental rodent model, we administered 2 ml (10(8) organisms/ml) broth culture of any of four human isolated strains of Helicobacter pylori by the oral route on a one-time basis to both BALB/c nude and BALB/c euthymic mice . After 20-wk examination, we have successfully demonstrated that the gastric mucosa of nude mice was continuously, and that of euthymic mice, temporarily (for 2 wk), colonized by orally administered, freshly isolated strains obtained from humans with gastritis, gastric ulcer, and duodenal ulcer, but never by the established strain . After colonization, gastritis and duodenitis were produced, and the presence of bacteria in gastric mucus was pathologically proved in all infected animals . We have confirmed that such colonization is always established when a large dose of a freshly isolated strain of H . pylori is administered at one time . We believe that this is the first reported rodent model of H . pylori-associated gastritis.

Microb Pathog, 1991 Jun, 10(6), 481 - 6
Production of type 1 fimbriae by Escherichia coli HB101; Elliott SJ et al.; Escherichia coli HB101 is frequently used as a host in the cloning of bacterial virulence genes because of its reported lack of virulence determinants such as fimbriae, adhesins and haemagglutinins . However, passage of HB101 in standing broth culture rapidly induced the production of fimbriae which mediated adhesion to HEp-2 cells and mannose-sensitive haemagglutination of human and guinea-pig erythrocytes . Fimbrial serology, morphology and pilin molecular mass of 18 kDa were consistent with those of type 1 fimbriae.

Infect Immun, 1991 Apr, 59(4), 1264 - 70
Effect of urease on HeLa cell vacuolation induced by Helicobacter pylori cytotoxin; Cover TL et al.; Concentrated broth culture supernatants from 50 to 60% of Helicobacter pylori strains induce eukaryotic cell vacuolation in vitro . A quantitative assay for cell vacuolation was developed on the basis of the rapid uptake of visibly vacuolated HeLa cells was significantly greater than that of nonvacuolated cells . By using the rapid NRU assay, we sought to determine the roles of H . pylori cytotoxin, urease, and ammonia in the vacuolation of HeLa cells . The NRU of HeLa cells incubated in medium containing ammonium chloride or ammonium sulfate was significantly greater than that of cells incubated in medium alone . In addition, ammonium salts augmented the NRU induced by H . pylori supernatants . The NRU induced by jack bean urease was augmented by the addition of urea to cell culture medium; this suggests that urease-mediated NRU occurs via the generation of ammonia . Acetohydroxamic acid blocked the NRU induced by jack bean urease and urea but failed to block the uptake induced by H . pylori supernatants . Supernatant from a non-urease-producing H . pylori mutant strain induced NRU identical to that of the urease-positive parental strain . These observations indicate that the vacuolating activity in H . pylori supernatants is not mediated solely by urease activity but that it may be potentiated by urease-mediated ammonia production.

Infect Immun, 1991 Jan, 59(1), 337 - 45
Effects of Bordetella pertussis infection on human respiratory epithelium in vivo and in vitro; Wilson R et al.; Bordetella pertussis infection probably involves attachment to and destruction of ciliated epithelial cells, but most previous studies have used animal tissue . During an epidemic, nasal epithelial biopsy specimens of 15 children (aged 1 month to 3 1/2 years) with whooping cough were examined for ciliary beat frequency, percent ciliation of the epithelium, and ciliary and epithelial cell ultrastructure . In addition, the in vitro effects of filtrates from a 24-h broth culture and of tracheal cytotoxin derived from B . pertussis on human nasal tissue organ culture were measured . B . pertussis was cultured from nasal swabs from 12 children . The mean ciliary beat frequency of their nasal biopsy specimens, 11.3 Hz (range, 10.4 to 13.0 Hz) was similar to that found in biopsy specimens from 10 normal children (mean, 12.5 Hz; range, 11.8 to 13.5 Hz) . The abnormalities of the epithelium observed in 14 of 15 patients were a reduction in the number of ciliated cells, an increase in the number of cells with sparse ciliation, an increase in the number of dead cells, and extrusion of cells from the epithelial surface . In vitro, neither culture filtrate nor tracheal cytotoxin had any acute effect on ciliary function, but culture filtrate and tracheal cytotoxin (1 and 5 microM, respectively) caused extrusion of cells from the epithelial surface of turbinate tissue, loss of ciliated cells, an increased frequency of sparsely ciliated cells, and toxic changes in some cells . These changes were dose dependent and progressive, and between 36 and 90 h ciliary beating ceased . The observations made with patient tissue confirm that B . pertussis infection damages ciliated epithelium, and the in vitro experiments suggest that tracheal cytotoxin may be responsible for the abnormalities observed in vivo.

J Infect Dis, 1990 Nov, 162(5), 1189 - 92
Pulmonary clearance and phagocytic cell response in a murine model of Branhamella catarrhalis infection; Verghese A et al.; C5-sufficient Swiss-Webster mice (C5+) and C5-deficient DBA/2J mice (C5-) when challenged endotracheally with 2 x 10(7) cfu of Branhamella catarrhalis rapidly clear the lungs of viable bacteria in 48 h . This rapid clearance correlates with a striking influx of polymorphonuclear neutrophils that is of equal magnitude in both C5+ and C5- animals . Supernatant from Todd-Hewitt broth culture of B . catarrhalis exhibits in vivo chemotactic activity in the murine lung and in vitro chemotactic activity in Boyden chambers . The ability of B . catarrhalis to recruit granulocytes, independent of complement, may suggest a role for this organism in promoting protease-mediated lung destruction.

Infect Immun, 1990 Sep, 58(9), 3056 - 60
In vitro surface properties of the newly recognized gastric pathogen Helicobacter pylori; Smith JI et al.; There appears to be a particular association between Helicobacter pylori and the gastric antrum, but the mechanisms by which the organism adheres to and colonizes the gastric mucosa are unclear . Surface hydrophobicity and surface charge mediate the adherence of other bacterial pathogens to mucosal epithelial cell surfaces . Therefore, in this study we characterized both the surface hydrophobicity and the surface charge of 10 H . pylori strains grown in broth culture . Four complementary methods were used to determine hydrophobicity: hydrophobic interaction chromatography, the salt aggregation test, comparison of bacterial adherence to polystyrene with adherence to sulfonated polystyrene, and measurement of contact angle with droplets of water . Three of the methods (salt aggregation test, adherence to polystyrene, and contact angles) indicated that each of the 10 strains expressed a relatively hydrophilic cell surface . In contrast, hydrophobic interaction chromatography determinations with both phenyl- and octyl-Sepharose suggested that the H . pylori strains were relatively hydrophobic . However, tetramethyl urea (0.4 M) did not reduce the binding of H . pylori to phenyl-Sepharose columns . DEAE-cellulose ion-exchange chromatography showed that each of the 10 strains of H . pylori had a surface which, overall, was highly negatively charged . We conclude that H . pylori expresses an overall relatively hydrophilic and negatively charged surface in vitro.

Infect Immun, 1990 Mar, 58(3), 603 - 10
Characterization of and human serologic response to proteins in Helicobacter pylori broth culture supernatants with vacuolizing cytotoxin activity; Cover TL et al.; Helicobacter pylori infection is strongly associated with histologic gastritis and peptic ulcer disease . Broth culture supernatants from a subset of H . pylori strains induce vacuolization in cultured cells, a phenomenon that has been attributed to cytotoxin activity . Concentrated culture supernatants from 15 of 28 (53.6%) H . pylori strains tested induced vacuolization in HeLa cells in titers ranging from 1:10 to 1:180 . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining of supernatants from these 28 strains and 2 control strains demonstrated an 82-kilodalton (kDa) protein band in 3 of 16 supernatants with vacuolizing activity, but in none of 14 supernatants without vacuolizing activity . By immunoblotting with human sera, a 128-kDa band was recognized in all 16 supernatants with vacuolizing activity, compared with 9 of 14 (64%) supernatants without vacuolizing activity (P = 0.014) . Serologic recognition of the 128-kDa band in H . pylori culture supernatants was more prevalent among persons infected with vacuolizing H . pylori strains than among persons infected with nonvacuolizing strains, but the difference was not statistically significant (80 versus 45%; P = 0.079); human serologic recognition of the 82-kDa band was less common . The 128-kDa band was recognized by 100% of 31 serum samples from H . pylori-infected patients with duodenal ulcer disease, compared with 60.8% of 74 serum samples from H . pylori-infected persons without peptic ulcer disease (P = 0.0001) . These data indicate that antigenic 128- and 82-kDa proteins are present in H . pylori broth culture supernatants with vacuolizing activity and that serologic responses to the 128-kDa protein are more prevalent among H . pylori-infected persons with duodenal ulceration than among infected persons without peptic ulceration.

Infect Immun, 1990 Mar, 58(3), 740 - 5
Surface antigens from Escherichia coli O2 and O78 strains of avian origin; Dho-Moulin M et al.; Fimbriae from O2 and O78 virulent strains of avian Escherichia coli were compared with type 1A fimbriae with regard to the apparent molecular weights of their subunits and their antigenic relationships . Under static broth culture conditions, most O78 strains expressed fimbriae closely related to those of type 1A . Under the same culture conditions, another type of fimbriae, sharing some common properties with type 1A fimbriae, was observed only on O2 strains; however, these fimbriae differed from type 1A fimbriae in the apparent molecular weights of their subunits and in the expression of specific epitopes . They were called type 1-like fimbriae . Homologies in lipopolysaccharide and outer membrane protein profiles were also demonstrated among the strains expressing type 1-like fimbriae, which suggests the existence of a clonal relationship among O2:K1 avian E . coli strains . The O78 strains studied did not appear to be clonally related.

Am J Vet Res, 1990 Feb, 51(2), 191 - 6
Comparative characterization of the leukocidic and hemolytic activity of Moraxella bovis; Hoien-Dalen PS et al.; The cytotoxic effect of Moraxella bovis 118F on bovine neutrophils was evaluated and characterized by use of a 51Cr release assay . Neutrophils harvested from healthy adult cattle were labeled with 51Cr . The leukocidic activity produced by M bovis 118F, a hemolytic strain of M bovis, was heat-labile . A live culture of strain 118F, at a ratio of 100 bacteria/neutrophil, released 97.7% of the 51Cr from labeled neutrophils . Neither a heat-killed preparation of M bovis 118F nor a live or heat-killed preparation of M bovis IBH63 (a nonhemolytic and nonpathogenic strain) induced significant (P greater than 0.05) release of 51Cr . Moraxella bovis 118F broth culture filtrates prepared for evaluation of leukocidic activity also were evaluated for hemolytic activity . These 2 toxic activities had several characteristics in common . Both were filterable, heat-labile, produced by a hemolytic strain, and were released during early logarithmic phase growth from broth cultures . Leukocidic and hemolytic activities were protected from degradation by phenylmethyl sulfonyl fluoride, a serine protease inhibitor . Leukocidic and hemolytic activities were dependent on calcium ions . Filtrate resulted in 54.1% 51Cr release from labeled neutrophils and contained 646.7 hemolytic U/ml, respectively, when saline (0.85% NaCl) + 10 mM CaCl2 solution was used as diluent . Neither saline solution nor saline + 10 mM MgCl2 solution supported leukocidic or hemolytic activity . Serum, obtained from several calves 10 to 38 days after M bovis inoculation, substantially neutralized leukocidic and hemolytic activities, compared with paired preinoculation serum samples.(ABSTRACT TRUNCATED AT 250 WORDS)

Rocz Panstw Zakl Hig, 1990, 41(1-2), 58 - 62
{Modification of the test for determining bacterial capacity for nitrate reduction}; Pogorzelska E et al.; For facilitation of the detection of bacteria able to reduce nitrates the presently used method of 24-hour broth culture with KNO3 and the method of growing on slant agar cultures with KNO3 (standard method) were compared, with certain modifications introduced . It was found that shortening of the incubation period from 24 to 6 hours and reduction of the volume of broth with KNO3 from 10 to 0.5 ml made possible obtaining of a greater number of correct results as compared to the standard method . In the 24-hour broth cultures with KNO3 done as yet in case of negative results it is indispensable to confirm them in the test with zinc dust . The placement of Griess reagents on points onto colonies growing on KNO3 agar may lead to false positive results.

Differentiation, 1989 Mar, 40(1), 1 - 9
Differential regulation of glycoprotein sulfation and fucosylation during growth of Dictyostelium discoideum; Tschursin E et al.; During early starvation-induced development, amoebae of Dictyostelium discoideum have been previously shown to increase sulfation and fucosylation of glycoprotein-linked oligosaccharides to levels above those observed in axenically growing cells . We report here that the axenic broth culture itself induces generation of high levels of fucosylated glycoprotein-linked oligosaccharides at all stages in the growth curve . However, when grown on bacteria, amoebae of both the axenic strain and the wild type show dramatic depression in fucose incorporation during early exponential growth . In mid- and late-exponential stages of growth, fucosylation rises to the levels found at all stages of axenic culture . Sulfation also increases during early development, but, in contrast to fucosylation, oligosaccharide sulfation is not altered by growth in axenic medium and does not increase during growth on bacteria . Starvation of bacterially grown cells results in increased sulfation and a further rise in fucosylation, as is also characteristic of broth-grown cells . The ability of axenic culture to uncouple control of these two classes of glycan-modification steps suggests that the synchronous increases during early development actually reflect responses to different regulatory signals, even though they participate in the same metabolic process . The increase in in vivo fucosyltransferase activity, which can act on many substrate glycoproteins, may alter many characteristics of the cells.

J Antibiot (Tokyo), 1987 Nov, 40(11), 1627 - 35
Hygromycin A, an antitreponemal substance . II . Therapeutic effect for swine dysentery; Nakagawa A et al.; This study was conducted to evaluate hygromycin A fed to growing swine at 1, 5, 10 or 20 g/ton feed for the control of Treponema hyodysenteriae-caused dysentery . Pigs provided carbadox at 50 g/ton feed served as an infected treatment control group . All pigs were orally, via stomach intubation, administered 100 ml of a T . hyodysenteriae broth culture . During the in vivo test, rectal swabs were taken for T . hyodysenteriae isolation, body weights of all pigs and the feed consumption was determined . All pigs were euthanized and necropsied at study end; the large intestine was cultured for T . hyodysenteriae and gross intestinal lesions were noted . T . hyodysenteriae-caused swine dysentery was successfully controlled by feeding hygromycin A at 5 g/ton . Hygromycin A medicated pigs performed as well as or better than carbadox-medicated pigs.

Vet Microbiol, 1987 Jun, 14(2), 137 - 44
Pathogenicity of Mycoplasma capricolum in sheep after experimental infection; Taoudi A et al.; Mycoplasma (M.) capricolum has been frequently isolated from diseased as well as from healthy sheep in Morocco . In order to determine its pathogenicity for sheep, experiments were performed in three trials with the Moroccan isolate 012 . The following results were obtained: Ewes inoculated intramammarily developed acute mastitis; the organism was transmitted to lambs suckling these ewes . Only a mild mastitis appeared after a second inoculum, performed 5 weeks after the first . Young lambs, 1 month of age, fed four times with M . capricolum broth culture, died during the septicemic phase, showing a generalized septicemia, polyarthritis and a diffuse interstitial pneumonia . Lambs, 2 and 3 months of age inoculated intrabronchially developed a disease, which appeared to be age and weight dependent . The more susceptible animals died within 1-2 weeks after infection . The older lambs recovered gradually from the disease, which was characterized by pneumonia, conjunctivitis and arthritis . A significant increase of antibodies against M . capricolum developed in the older animals in the complement fixation test.

J Med Vet Mycol, 1987 Apr, 25(2), 107 - 14
Determination of viability of Histoplasma capsulatum yeast cells grown in vitro: comparison between dye and colony count methods; Kwon-Chung KJ et al.; The viability of Histoplasma capsulatum yeast cells grown under different conditions was determined by dye tests with Eosin-Y and Janus Green B and by colony counts of cells plated on brain-heart infusion agar supplemented with histoplasma growth factor and bovine serum albumin (BHI-SAG) . The test samples included cells grown on brain-heart infusion agar at 37 degrees C for 2-7 days, cells grown in glucose-cysteine broth medium for 1-31 days, and cells grown on brain-heart infusion agar for 3 days at 37 degrees C and then irradiated by ultraviolet light . The colony count indicated that the viability of the yeast cells grown on brain-heart infusion agar for 2 or 3 days varied between 68 and 100% depending on the isolates . The viability, however, dropped from 16 to 29% by day 7 . The results of dye tests showed 78 to 99% dye-negative cells among the 2- and 3-day-old cultures while the number of dye-negative cells dropped to 32-36% on day 7 . The colony count with the cells grown in the broth culture showed 100% viability until day 7 and dropped significantly by day 9 . The results of dye tests showed no correlation with the colony count findings . The survival curve of ultraviolet-irradiated cells determined by colony count showed that irradiation at 180 erg mm-2 killed more than 50% of cells; fewer than 10% of cells survived 360 erg mm-2 . The results of the dye test showed no difference between the irradiated and control populations.(ABSTRACT TRUNCATED AT 250 WORDS)

J Infect Dis, 1987 Feb, 155(2), 304 - 8
An agar culture technique to quantitate Trichomonas vaginalis from women; Philip A et al.; In subjects with trichomoniasis the number of Trichomonas vaginalis in vaginal discharges or secretions is unknown . The presence of T . vaginalis was evaluated in 177 consecutive female patients attending an inner city sexually transmitted disease clinic by patient history, wet mount, and broth culture . T . vaginalis was quantitated by a novel agar culture technique . Of the 177 women, 86 (49%) were positive for T . vaginalis by either wet mount or culture . Clinical symptoms were not reliable for making an accurate diagnosis of trichomoniasis . Culture on modified Diamond's medium was more sensitive (98%) than the wet mount method (38%) in detecting T . vaginalis . Of the 86 patients who were positive for trichomoniasis, quantitation was obtained for 81 patients, with 70% yielding greater than 10(4) colony-forming units (cfu)/ml . The number of T . vaginalis ranged from 40 to greater than 10(6) cfu/ml . The wet mount method was very insensitive for detecting T . vaginalis and was positive only in patients yielding greater than 10(5) cfu/ml.

Arch Gerontol Geriatr, 1986 Dec, 5(4), 343 - 9
Influence of environmental stress on lipofuscin production; Aloj Totaro E et al.; The Corollospora maritima, a marine ascomycete, has been used as an experimental model to investigate the possibility that age pigments can be considered indicators also of environmental stress . Synthetic sea water enriched with iron or copper has been inoculated in a broth culture of the fungus . After 5 days of incubation the mycetes were assayed for lipofuscin fluorescent pigment and malondialdehyde content . The presence in the culture medium of the heavy metal ions results in an increase of the lipofuscin and malondialdehyde production . The same evidence has been obtained with sea water samples collected at seven sites along the coast of the Gulf of Naples (Italy): the lipofuscin and malondialdehyde production increases proportionally with the copper and iron pollution in the sea water.

Biken J, 1986 Dec, 29(3-4), 73 - 5
Application of Biken test (modified Elek test) for sampling of heat-stable enterotoxin of Escherichia coli isolated in Bangladesh; Akhtar SQ; The usefulness of Biken Agar 2 as a source of heat-stable toxin for the suckling mouse assay was examined . Escherichia coli (E . coli) strains isolated in Bangladesh from patients with gastroenteritis found to produce heat-stable toxin (n = 152), both heat-stable and heat-labile toxin (n = 60) and not to produce heat-stable or heat-labile toxin (n = 25) by standard suckling mice assay using broth culture were tested . Sampling from Biken Agar 2 gave comparable results to those obtained using standard broth cultures . This is the first field survey of evaluation of the Biken test for sampling heat-stable toxin of E . coli . The result further clarifies the applicability of the Biken test for sampling heat-stable toxin, and the usefulness of the Biken test for detections of heat-labile and heat-stable toxin produced by E . coli.

Diagn Microbiol Infect Dis, 1986 Apr, 4(4), 299 - 306
Semiquantitative cultures of intravascular catheters from cancer patients; Jones PG et al.; Three-hundred seventy-nine catheter tips were prospectively cultured by both a semiquantitative method and by broth culture, over a 2-month period . One hundred eleven of the catheters were culture-positive in broth, and 47 of these were also culture-positive by the semiquantitative method . Clinical signs of infection were reviewed for the 111 culture-positive catheters and for 50 of the 268 culture-negative catheters . Both culture-positive and culture-negative catheters were infrequently associated with local signs of infection (10% and 12%, respectively) . Culture-positive catheters, however, were more likely to be associated with systemic signs of infection than were culture-negative catheters (15% and 2%, respectively) . Among the culture-positive catheters, those that yielded greater than or equal to 15 colonies on semiquantitative culture were more likely to be associated with septicemias than were those with less than 15 colonies (22% and 6%, respectively) . Nevertheless, there were five catheter-related bacteremias that were associated with catheters which were culture-negative on semiquantitative culture but culture-positive in broth . The proportion of patients with culture-positive and culture-negative catheters who were febrile was similar (30% and 42%, respectively) . Semiquantitative cultures of catheters from cancer patients are useful, but the result should be interpreted with some caution.

Appl Environ Microbiol, 1986 Jan, 51(1), 74 - 9
Interactions between clay minerals and siderophores affect the respiration of Histoplasma capsulatum; Lavie S et al.; The reduction in the respiration of Histoplasma capsulatum in broth culture caused by montmorillonite appeared to be the result, in part, of the interference by the clay with the iron nutrition of the fungus . This interference was apparently the result of the adsorption by the clay of the iron-transporting siderophore (deferricoprogen B) produced by the fungus, as the reduction in respiration was partially alleviated by the addition of foreign siderophores . Neither kaolinite nor attapulgite (palygorskite) appeared to adsorb significant amounts of the siderophores, probably because of the low cation exchange capacity and specific surface area of kaolinite and the inaccessibility of adsorption sites in the fibrous attapulgite . These observations, in addition to the adhesion of montmorillonite to the hyphae, suggest mechanisms that may explain the discrete geographic distribution of this fungus, which is pathogenic to humans and which has been isolated essentially only from soils that do not contain montmorillonite.

J Hyg (Lond), 1985 Aug, 95(1), 59 - 68
The weak immunogenicity of Fusobacterium necrophorum; Smith GR et al.; Three of four extreme methods of immunization completely failed to protect mice against challenge with the homologous strain of Fusobacterium necrophorum . Unsuccessful vaccines included (1) broth culture killed by mild heat and emulsified with Freund's complete adjuvant, and (2) a homogenate of heavily infected mouse brains, inactivated by mild heat and given in two doses . Also unsuccessful as a method of immunization was the production of a severe subcutaneous infection with F . necrophorum, followed by curative treatment with metronidazole . Slight but significant protection against subcutaneous challenge resulted, however, from two such infections given in rapid succession . It would appear that the main virulence factors of F . necrophorum are only weakly immunogenic, and the experiments give little encouragement to the prospect of an effective necrobacillosis vaccine.

Am J Vet Res, 1985 Mar, 46(3), 637 - 42
Effect of virus-induced destruction of villous epithelium on intestinal secretion induced by heat-stable Escherichia coli enterotoxins and prostaglandin E1 in swine; Whipp SC et al.; Villous atrophy and crypt hyperplasia were induced in the jejunal epithelium of thirteen 3-week-old pigs by inoculation with transmissible gastroenteritis virus . The responses (changes in net fluid movement) induced in ligated intestinal loops of these pigs by intraloop injections of prostaglandin E1 (PGE1) or Escherichia coli broth culture filtrates containing either or both E coli heat-stable enterotoxins (STa and STb) were compared with the responses induced by these preparations in littermates not inoculated with virus . Villous atrophy was associated with a marked decrease in response to preparations containing STa, STb, or STa + STb, but the response to PGE1 was undiminished . These results were consistent with the reports of others that the response to cyclic adenosine monophosphate-mediated secretogogues (PGE1) is a function of crypt epithelium; however, the present results also suggest that the secretory response to STa and to STb is dependent on the integrity of the villous epithelium . In the present study, loss of villous epithelium was associated with loss of response to STa and STb, but not to PGE1.

Vet Med Nauki, 1985, 22(6), 46 - 55
{Morphological and biochemical changes in the blood of lambs with experimental Erysipelothrix polyarthritis}; Koichev K et al.; Polyarthritis was induced in lambs via the i/v infection with 2 cm3 of 24-hour Erysipelothrix rhusiopathiae broth culture, which led to distinctive morphologic and biochemical changes in the peripheral blood . The hemoglobin content, the erythrocyte count, and the hematocrit value dropped, while ESR rose with the development of the infection process . The white blood picture presented transient and slight drop of the leukocyte count followed by leukocytosis with shifting to the left, aneosinophilia, lympho- and monopenia in the acute stage, and well manifested eosinophilia in the chronic stage of the infection . The changes in the total protein and the protein fractions consisted in hypoproteinemia in the first days following infection, hypoalbuminemia during the entire period, and hyperproteinemia and hypergammaglobulinemia in the chronic stage . The changes in the blood electrolites consisted in the rise of Ca and K, the drop of Na, and transient changes in the level of P, tending toward a rise in the chronic stage . It was also established that the values of sialic acid were raised in the entire period of polyarthritis development, while those of aspartate aminotransferase and alanine aminotransferase were higher in the first seven-day period only.

Dev Biol Stand, 1985, 61, 103 - 11
Bordetella pertussis tracheal cytotoxin; Goldman WE et al.; The most consistent pathological feature of pertussis is the selective colonization and subsequent destruction of ciliated cells in the respiratory epithelium . Phase I B . pertussis can reproduce the human respiratory tract cytopathology during in vitro infection of hamster tracheal organ cultures . However, most isolated biologically active components produced by B . pertussis (lymphocytosis-promoting factor, adenylate cyclase, dermonecrotic toxin, etc.) have no apparent cytopathic effect on the respiratory epithelium . We have purified a glycopeptide from the culture supernatant of virulent B . pertussis that mimics completely the ciliated cell pathology characteristic of pertussis (5) . Tracheal cytotoxin (TCT) is released during log phase broth culture and consists of 15 amino acid residues as well as two amino sugars . The selective biological activity of TCT has been studied in tracheal organ cultures by light and electron microscopy . A series of pathological changes precedes the eventual extrusion of ciliated cells, while all other epithelial cell types appear ultrastructurally normal . TCT also causes a dose-dependent inhibition of DNA synthesis in cultured hamster trachea epithelial cells, providing a quantitative bioassay to monitor TCT activity during purification steps . Previously, TCT could be completely purified from oxidized glutathione (a major contaminant from the culture medium) only by high-voltage paper electrophoresis, a procedure not well suited for large scale work . We have now substituted a final column chromatography step that separates TCT from oxidized glutathione and other contaminating peptides . This change and other preparative scale adaptations now allow us to purify 150-250 nmol of biologically active TCT from one liter of culture supernatant (3-4 X 10(13) bacteria), a ten-fold increase over our previous batch size with no increase in processing time.(ABSTRACT TRUNCATED AT 250 WORDS)

J Bacteriol, 1984 Nov, 160(2), 691 - 5
Immunofluorescence study of fimbrial phase variation in Escherichia coli KS71; Nowicki B et al.; An immunofluorescence assay was developed to study fimbrial phase variation in a pyelonephritogenic Escherichia coli strain, KS71 . By using fluorochrome-labeled antibodies specific for either P, type-1C, or type-1 fimbriae of strain KS71, it was shown that in a broth culture of strain KS71 the fimbrial types mostly occurred on different cells . Only 9% of the cells carried more than one fimbrial type . The KS71 cell population was fractionated into subpopulations expressing only one of the fimbrial types or lacking fimbriae . Immunofluorescence assay of the subpopulations revealed a rapid phase variation in fimbrial synthesis . Kinetic analyses of a nonfimbriated cell population suggested that a change from one fimbrial phase to another was not totally random.

Diagn Microbiol Infect Dis, 1984 Apr, 2(2), 139 - 43
Growth of toxigenic Escherichia coli in oral rehydration solutions; Keusch LS et al.; The ability of three strains of toxigenic Escherichia coli to grow in glucose-electrolyte oral rehydration solution (ORS) used for the treatment of dehydration due to diarrhea was studied . The purpose was to determine the potential risk that such home-based therapy might pose for transmission of infection if the ORS were contaminated with potential pathogens . Inocula of 10(5) unwashed organisms from broth culture were rapidly killed in ORS made with chlorinated tap water and increased by 1.5-2 log10 in ORS in triple-distilled laboratory water . When media constituents were first removed by washing, one strain failed to grow in ORS-distilled water, but multiplied to a level of 10(6) organisms/ml in ORS in water obtained from a rural village in the highlands of Guatemala . Multiplication of organisms in ORS depends on the presence of a nitrogen source . These limited data support current guidelines for preparation and use of ORS in the field employing available drinking water, as bacterial growth is restricted by the nitrogen content of the water.

Avian Dis, 1984 Jan-Mar, 28(1), 224 - 34
Evaluation of inactivated Mycoplasma gallisepticum oil-emulsion bacterins for protection against airsacculitis in broilers; Yoder HW Jr et al.; Broiler chicks were vaccinated subcutaneously in the neck at various ages with a single 0.5-ml dose of beta-propiolactone-inactivated Mycoplasma gallisepticum (MG) oil-emulsion bacterin . Four weeks later, vaccinated and control chicks were placed in cold environmental cabinets, infected with infectious bronchitis virus intratracheally, and 2 days later challenged by aerosol exposure to live MG broth culture . All chicks were killed 21 days later and scored postmortem for the rate and severity of airsacculitis produced in each group . Broiler chicks vaccinated at 1 day of age had only slight protection against the development of airsacculitis . Results were variable when chicks were vaccinated at 7 days of age, with little evidence of resistance to airsacculitis . However, when broiler chicks were vaccinated with MG bacterins at 11 to 15 days of age, they acquired significant protection against airsacculitis compared with controls . Viable MG organisms were readily isolated from most of the sampled tracheas and air-sac lesions cultured 21 days post-challenge, indicating a lack of protection against infection of the respiratory tract . MG-vaccinated chicks generally produced antibodies readily detectable by the rapid serum-plate test, tube-agglutination, and hemagglutination-inhibition (HI) tests . Some of the vaccinated chicks, but none of the unvaccinated control chicks, developed positive reactions to agar-gel-precipitin tests following challenge . Low HI titers at challenge were not necessarily indicative of lack of protection against the development of airsacculitis, since good protection was often observed in chickens with low to moderate HI titers.

JAMA, 1983 Oct 28, 250(16), 2185 - 8
Clinical evaluation of a lysis-centrifugation technique for the detection of septicemia; Kelly MT et al.; A commercially available lysis-centrifugation blood culture system was compared with a two-bottle broth-culture system employing 100 mL of broth and 10 mL of blood per bottle to analyze 1,913 blood specimens . Of 154 clinically significant isolates, 89% were detected by the lysis-centrifugation technique, and 73% were detected by the broth-culture method . Twenty-seven percent of the organisms were detected only by the lysis-centrifugation technique, and 11% were detected only by the broth system . Fifteen polymicrobial cultures were encountered; the lysis-centrifugation technique detected 93% of the organisms in these cultures, while the broth-culture method detected only 20% . Isolated colonies of clinically important organisms were available 30 hours earlier with the lysis-centrifugation technique . These results suggest that the lysis-centrifugation technique may provide a substantial improvement over conventional methods for blood cultures.

Can J Comp Med, 1983 Oct, 47(4), 464 - 70
Comparison of the enzyme-linked immunosorbent assay and the indirect hemagglutination and complement fixation tests for detecting antibodies to Mycoplasma hyopneumoniae; Armstrong CH et al.; Caesarean-derived, colostrum-deprived swine were exposed to a broth culture of a low passage field isolate of Mycoplasma hyopneumoniae by intranasal inoculation . The intranasal-inoculated swine subsequently were commingled with their litter-mates to effect transmission via contact-exposure . Sera were collected from the swine at two to four week intervals for approximately one year postexposure and evaluated by the enzyme-linked immunosorbent assay (ELISA), indirect hemagglutination and complement fixation tests . The intranasal-exposed swine seroconverted earlier, developed higher titers and remained indirect hemagglutination and complement fixation positive longer than the contact-exposed swine . It was concluded that the antibody response of intranasal-exposed swine was artificially high and that sera from such swine were not suitable for evaluating the sensitivity of mycoplasmal pneumonia of swine serodiagnostic tests . The indirect hemagglutination test was relatively insensitive and technically cumbersome and the least promising as a practical field test . The complement fixation test appeared to be slightly more sensitive in detecting early antibody production (especially in contact-exposed swine) but it was the least sensitive in detecting late antibodies . The ELISA was generally the most sensitive procedure . Individual high ELISA titers were from ten to 32 times greater than maximum complement fixation and indirect hemagglutination titers . The most striking difference among the three tests was the persistence of high ELISA titers late in the study . All swine were ELISA positive at necropsy approximately one year postexposure despite the fact that lungs were devoid of lesions and culturally and immunofluorescent negative for M . hyopneumoniae.

Yale J Biol Med, 1983 Sep-Dec, 56(5-6), 623 - 9
Monoclonal antibodies to Mycoplasma hyorhinis surface antigens: tools for analyzing mycoplasma-lymphoid cell interactions; Wise KS et al.; A library has been constructed of approximately 50 monoclonal antibodies that recognize antigens of Mycoplasma hyorhinis . Characteristics of six antibodies are discussed . Each reacts with a discrete determinant borne on a protein-containing molecule of distinct molecular size . Three of these respective antigens are expressed at the surface of mycoplasmas colonizing lymphoblastoid cells in culture . Of these three surface antigens, two are selectively expressed on strain GDL but not on strain BTS-7 of M . hyorhinis, thus defining strain-restricted immunological specificities within these species . One monoclonal antibody of the IgM class (mu, kappa) mediated marked complement-dependent growth inhibition of M . hyorhinis in broth culture . These monoclonal reagents should facilitate analysis of mycoplasma surface architecture, and the molecular interactions of these organisms with the host cell surface.

Trop Anim Health Prod, 1983 Aug, 15(3), 144 - 8
Effect of some cultural factors on T1 broth vaccine for contagious bovine pleuropneumonia; Miles RJ; The effect of some aspects of buffering capacity, glucose concentration, length of incubation and extent of exposure to air during the preparation of live Mycoplasma mycoides var . mycoides strain T1 broth culture vaccine was investigated . A consistently good vaccine was obtained using 0.07 M phosphate buffered medium containing 0.1% glucose and incubating in stoppered bottles . The shelf-life of this vaccine stored at 4 degrees C was up to 12 weeks . The effect of other storage temperatures on the viability of the vaccine is given.

J Gen Microbiol, 1983 Jun, 129 (Pt 6), 1921 - 8
Obligatory coupling of colicin release and lysis in mitomycin-treated Col+ Escherichia coli; Pugsley AP; Previous work has shown that Escherichia coli K12 strains carrying the small, high copy number ColE2-P9 plasmid produce large amounts of colicin and then lyse and release colicin when grown in broth culture containing mitomycin C . Strains carrying the larger, low copy number ColIa-CA53 plasmid produced much less colicin and did not lyse or discharge more than 15% of their colicin when grown under the same conditions . Naturally-occurring Col+ strains and E . coli K12 derivatives carrying different Col plasmids could be classified either as ColE2+-like or ColIa+-like according to whether or not they produced large amounts of colicin and lysed and discharged colicin when grown in the presence of mitomycin, and also by the size and presumed copy number of the Col plasmid they carried . Strains carrying multiple copies of the cloned colicin Ia structural gene produced large amounts of colicin but did not lyse or release colicin when grown in the presence of mitomycin . This result rules out the possibility that high level accumulation of colicin is sufficient to cause lysis . Conditions were sought under which colicin Ia could be released from the producing cells . It was found that mitomycin-treated cultures of strains carrying both ColE2 and ColIa plasmids released both colicins when they lysed, although colicin Ia release occurred later than colicin E2 release . It was also noted that colicin Ia-laden cells released their colicin when diluted into fresh culture medium.

Am J Clin Pathol, 1983 Mar, 79(3), 379 - 81
Evaluation of a commercial exoantigen test system for the rapid identification of systemic fungal pathogens; Denys GA et al.; Seventy-nine mycelial-form stock cultures of Blastomyces dermatitidis, Coccidioides immitis, Histoplasma capsulatum, and morphologically similar fungi were extracted and tested by using commercial macroimmunodiffusion exoantigen test kits and the Centers for Disease Control (CDC) reference system for identifying fungal isolates . Results showed 100% correlation between the two systems . Specific exoantigens of C . immitis and H . capsulatum extracted from agar slant cultures (slant extraction method) readily were identified . In eight of 26 cultures of B . dermatitidis, broth culture filtrates (broth-shake-flask method) were required to demonstrate the specific bands of identity . No false-negative reactions or cross-reactivity among the pathogens and other fungi were observed . The commercial test kits provided a rapid and specific method for identifying or confirming suspected fungal pathogens.

Acta Pathol Microbiol Immunol Scand {B}, 1983 Feb, 91(1), 89 - 91
Legionella extracellular protease activity on chromogenic peptides: a simplified procedure for biochemical enzyme identification; Berdal BP et al.; Legionella strains produce extracellular proteases . One method to demonstrate these is through the activity of broth culture filtrates on para-nitroanilide (pNA)-derivatized peptides . Previously, this method has comprised a 100-times concentration of the filtrates, in order to bring the protease activity, as measured by the liberation of free pNA from hydrolysed peptides, up to easily recordable values . By introducing a diazotation step, the sensitivity of the test has been increased sufficiently to omit the time-consuming concentration procedure . Accordingly, the analysis of extracellular Legionella proteases by pNA-derivatized peptides has become rapid and straight-forward.

Med Biol, 1982 Apr, 60(2), 116 - 20
Isolation and preliminary characterisation of mycoplasmavirus 20-P; Jansson E et al.; A mycoplasmavirus, MV 20-P, was isolated from the fastidious, slow-growing human mycoplasma 20-P, previously identified by the growth inhibition test as Mycoplasma hominis type 2 . Its broth culture was ultracentrifuged into 35% saccharose, the pellet was resuspensed in buffer, and the suspension was dropped onto laws of the virus indicator Acholeplasma laidlawii BCL-13 . The virus was washed off the plaque areas by buffer . MV 20-P appeared in electron microscopy as spherical, enveloped particles about 80 nm in diameter . It was sensitive to lipid solvents . These preliminary results indicate that MV 20-P is a MVL2 type mycoplasmavirus and possibly the first of human origin.

Microbiol Immunol, 1982, 26(1), 1 - 14
Comparison of ciliostasis by mycoplasmas in mouse and chicken tracheal organ cultures; Araake M; The cilia-stopping effect of mycoplasmas of human and various animal origin in mouse and chicken tracheal organ cultures was studied . From the results in mouse tracheal organ cultures, the mycoplasma strains tested were divided into three groups: Mycoplasma pulmonis m53, M pulmonis JB, M . pulmonis OK, M . mycoides subsp . Mycoides PGl and M . Gallisepticum S6 showed a strong cilia-stopping effect; M . pulmonis PG22, M . mycoides subsp . capri PG3, M . meleagridis 19729, M . neurolyticum Type A and M . arthritidis PG6 showed a mild effect; and M . pneumoniae FH, M . salivarium Hup, M . hominis type 1-C and M . orale N-C human origin and Acholeplasma laidlawii PG8 showed a weak effect . On the other hand, in chicken tracheal organ cultures, only M . gallisepticum S6 showed a strong effect, M . meleagridis 19729 was affected to a lesser degree, and the other mycoplasma strains showed a weak or no effect . The results indicate that some murine and poultry mycoplasmas showed a cilia-stopping tendency in mouse and chicken tracheal organ cultures, respectively, while human mycoplasmas showed weak or little effect in both organ cultures . In mouse tracheal organ cultures, M . pulmonis m53 treated with heat, trypsin or formaldehyde, and the sterile filtrate of an m53 broth culture showed no cilia-stopping effect . The relationship of the pathogenicity of mycoplasmas for their natural hosts to that for cultured respiratory cells is discussed.

J Clin Microbiol, 1982 Jan, 15(1), 28 - 34
Use of arginine aminopeptidase activity in characterization of arginine-utilizing mycoplasmas; Ball HJ et al.; The aminopeptidase activity of arginine-utilizing mycoplasmas was investigated with 20 aminoacyl beta-naphthylamide substrates . High levels of arginyl-beta-naphthylamide hydrolysis were demonstrated in 6 of 11 species when extracts of concentrated washed organisms were used . Relatively low arginine aminopeptidase activity was demonstrated with similar extracts from 22 species not utilizing arginine . The high level of arginine aminopeptidase activity could also be demonstrated with unwashed, unconcentrated samples of the same 6 species and also with Mycoplasma arthritidis . The procedure for preparing the extract of M . arthritidis appeared to remove the arginine aminopeptidase activity which was demonstrated to be present in the untreated culture . Fluorogenic and chromogenic tests were developed whereby this distinctive arginine aminopeptidase activity could be demonstrated within 4 h with the use of small volumes of broth culture (10 microliter) or single colonies, thus providing a rapid test for early characterization of some Mycoplasma species.

Avian Dis, 1981 Oct-Dec, 25(4), 821 - 6
Immunogenic potency of oil-emulsified Mycoplasma gallisepticum bacterins; Panigraphy B et al.; Immunogenicity of an aqueous Mycoplasma gallisepticum (MG) bacterin and two oil-emulsified bacterins with aqueous-phase-to-oil-phase ratios of 1:2 and 1:4 were evaluated in 3 groups of 5-week-old MG-free White Leghorn chickens . Group 4 chickens were nonimmunized controls . Group 1, 2, and 3 chickens received primary immunizations with 0.5 ml of bacterin subcutaneously (SC) . Six weeks later, half of the vaccinated chickens received a second immunization (0.5 ml SC) . Six weeks after the last dose of vaccine, all 4 groups of chickens were challenged intraocularly with a 24-hr broth culture of pathogenic MG ("R' strain) . The geometric mean hemagglutination-inhibition antibody titers (GMT) of non-immunized, once-immunized, and twice-immunized chickens were compared at 2-week intervals following primary immunization, secondary immunization, and challenge . Both oil-emulsified bacterins stimulated significantly higher (P less than or equal to 0.05) levels of antibody than the aqueous bacterin after the primary and secondary immunizations . Following challenge with pathogenic MG, the GMT in the immunized and non-immunized chickens approached comparable values . The highest GMT in groups 1, 2, 3, and 4 respectively were: a) primary response, 19, 99, 121, and 11; b) secondary response, 105, 423, 793, and 21; and c) challenge response, 171, 278, 300, and 160 . After challenge there was a further rise of antibody level in non-immunized chickens, those immunized with the aqueous bacterin, and those immunized only once with the oil-emulsified bacterins . This suggested a lack of protection against challenge . However, there was no increase of GMT in chickens immunized twice with the oil-emulsified bacterins.

Am J Vet Res, 1980 Sep, 41(9), 1396 - 401
Intramammary coliform infection after heavy external contamination of teats; Schultze WD et al.; Teats of lactating dairy cows were exposed to massive, repeated external contamination by application of a freshly prepared broth culture of Escherichia coli that was maintained in wet contact with the teat ends during 14 or 21 successive entire intermilking periods . When contamination was restricted to the intermilking periods by dipping teats in an iodophor germicide before each milking, 18 new intramammary infections occurred among the 93 mammary quarters at risk . The rate of infection achieved was 1/89 milkings . Transient residence of E coli in streak canals and frequent occurrence of sterile inflmmation of mammary quarters made diagnosis of new infection difficult . Teats also were exposed to E coli without sanitization before milking . The milking machine was modified to maximize the likelihood of contaminated milk droplets impacting on teat orifices through back-jetting . Diagonally opposed milking maching inflations were fitted with rifled bore short milk tubes designed to be protective against back-jetting . The rate of new E coli infections was 1/291 milkings among control and protected quaters . When contamination was extended into the milking operation, the new infection rate was not greater than that achieved when exposure was limited to intermilking periods; therefore, the protective value of rifled-bore short milk tubes was not adequately tested and a role of the machine as a vector was not demonstrated.

Poult Sci, 1980 Jul, 59(7), 1548 - 9
Effect of sex and Mycoplasma synoviae infection on chicken red blood cells used for hemagglutination-inhibition test; Drott JH et al.; A total of 10 male and 10 female 9-week-old commercial Mycoplasma-free broiler chickens was used in this test . Five males and 5 females were artificially infected by aerosol with a broth culture of Mycoplasma synoviae (MS) F10-2AS . The other 5 males and 5 females were used as noninfected controls . At 3, 9, and 12 weeks postinfection all birds were bled, and the blood was pooled into 4 groups: infected male, noninfected male, infected female, and noninfected female . Hemagglutination-inhibition (HI) tests for MS and Mycoplasma gallisepticum (Mg) were run on a selected group of known positive serums using the 4 pools as red blood cell (RBC) sources . The geometric mean HI titers of the 4 groups showed no significant differences between male and female or MS-infected and noninfected chickens as a source of RCB's for the MS or Mg HI test . These results indicate that if procedures are correct, there is no danger in using RBC's from MS-infected birds.

Biull Eksp Biol Med, 1980 Apr, 89(4), 426 - 8
{Changes in the serum acid phosphatase activity of guinea pigs poisoned with C1 perfringens type A toxin and a mixture of toxin and a broth culture filtrate}; Tselukh AV et al.; It was shown in experiments on guinea pigs that the injection of C1 . perfringens type A toxoid induced an increase in acid phosphatase activity of animal blood serum . The action of the toxoid increased under the effect of C1 . butyricum cultural filtrate, which gave rise to an earlier enhancement of the specific activity of the enzyme as compared to the injection of the toxoid alone . Increased activity of acid phosphatase may play a pathogenetic role in cases of anaerobic infection caused by association of C1 . perfringens and C1 . butyricum.

Am J Clin Pathol, 1979 Dec, 72(6), 980 - 4
A semiquantitative culture technic for identifying infection due to steel needles used for intravenous therapy; Band JD et al.; Using steel intravneous needles obtained from patients with hematologic malignancies, the authors evaluated the efficacy of a semiquantitative method for culturing vascular cannulas on solid medium, comparing it with conventional broth culture . Of 148 needles studied, 140 (95%) were negative on semiquantitative culture (less than 15 colonies on the plate) although 38 produced growth in broth or on the plate; none of these needles caused septicemia . Eight needles were positive on semiquantitative culture (greater than or equal to 15 colonies), and two of these caused septicemia (P = .002) . Positive semiquantitative cultures were associated with local inflammation (P = .02) . Semiquantitative culturing was equal to the broth method in sensitivity (100%) in the diagnosis of needle-related septicemia; specificity (96% vs . 92%) and the predictive value of a positive needle culture (25% vs . 14%) were both enhanced with the semiquantitative method . The semiquantitative technic differentiates infection from contamination and offers other major advantages compared with the broth method, and is recommended for culturing steel needles as well as plastic catheters.

Am Rev Respir Dis, 1979 Mar, 119(3), 337 - 43
A fiberoptic bronchoscopy technique to obtain uncontaminated lower airway secretions for bacterial culture; Wimberley N et al.; Seven types of brush catheter were studied in vitro to determine the optimal catheter design for obtaining specimens for bacterial culture using fiberoptic bronchoscopy . The various catheters were inserted through the inner channel, which was contaminated with fresh saliva . The specimen was then obtained by inserting the brush into a broth culture of a marker organism at the distal end of the bronchoscope . Relative merits were based on quantitative bacterial counts of salivary "contaminants" and the marker organism . The catheter that proved superior had telescoping cannulas with a distal occlusion . Twenty-six samplings using catheters with this design uniformly showed no contaminants and high counts of the marker organism . The catheter with telescoping cannulas and a distal occlusion composed of polyethlene glycol was then tested with bronchoscopy performed on 8 healthy volunteers and 6 patients with clinical evidence of a lower airway infection . The specimens yielded likely pulmonary pathogens in high concentrations from patients with pneumonia or lung abscess . Possible contaminants were recovered in low concentrations from 2 of 14 subjects . These in vitro studies and the preliminary clinical trial support the use of this type of catheter for obtaining bronchoscopic specimens for bacterial culture.

Folia Microbiol (Praha), 1979, 24(3), 247 - 52
The decreased in vitro chemotactic activity of polymorphonuclear leukocytes of newborns and infants; Miler I et al.; Using Boyden's technique, a statistically significant decrease in the chemotactic activity of polymorphonuclear (PMN) leukocytes was found during the early postnatal period, i.e . in the cord blood and in blood of newborns within the first 10-15 d of life after stimulation of cells with both zymosan-activated adult serum (ZAS) and with an abacterial filtrate of Escherichia coli broth culture (ECF) . After this period, the responsiveness of leukocytes to both chemotactic agents increased and remained at the same level during the whole observation period, i.e . up to the age of 6 months . Nevertheless even then it did not reach fully the responsiveness of the leukocytes of mothers and pregnant women . Zymosan-activated serum was shown to be a more potent chemotactic stimulus to leukocytes of infants as compared to the E . coli filtrate.

Int Arch Allergy Appl Immunol, 1979, 60(2), 140 - 7
Antigens of Micropolyspora faeni strains; Kurup VP et al.; Exocellular antigens from Micropolyspora faeni and Micropolyspora rectivirgula were prepared by growing them in a synthetic broth culture at 50 degrees C for 1 week under continuous shaking . The broth was separated from the organisms by filtration and the filtrate was dialyzed and freeze-dried . The cross-reactivity of these antigens were studied by antigen-antibody crossed-immunoelectrophoresis and the presence of similar antigens in different strains by tandem crossed-immunoelectrophoresis using rabbit anti-M . faeni and anti-M . rectivirgula sera . The results indicate that a number of antigenic components are common to the different strains of M . faeni and M . rectivirgula, while each strain demonstrates several specific antigenic determinants . Of the 26 clearly demonstrable precipitin arcs formed by crossed-immunoelectrophoresis, only 1 was detected in all the strains while 2 more were detected frequently . This study indicates that M . rectivirgula and M . faeni strains are not distinguishable from each other by morphological, biochemical or immunological criteria.

Dev Biol Stand, 1978, 41, 361 - 5
The T1 vaccine used in the control of contagious bovine pleuropneumonia in Nigeria; Onoviran O et al.; A lyophilised T1 vaccine was produced by direct lyophilisation of a 72-hour broth culture to which 10% sterile sucrose solution was added . Each vial contained 2 ml to make a total of 10 doses when reconstituted . Each ml of the product contained an average of 109 colony forming units of the organism . When injected at the tip of the tail or subcutaneously behind the shoulder of 2,000 zebu and 1,0000 Ndama cattle, no outward reactions were observed . 12 zebu cattle were vaccinated and challenged 6 months later by direct contact with artificially infected cattle for 4 months . All the animals were completely protected against the disease.

Vet Med Nauki, 1978, 15(2), 65 - 71
{Fluctuations in susceptibility of pigs to erysipelas depending on age}; Arsov R et al.; The results of experiments have shown that up to their second month of age pigs are not susceptible to Erysipelothrix rhusiopathiae infection . The invasion of the infective agent into the body of sich pigs neither causes an outbreak of the disease nor brings about changes in the immunologic indices under investigation, which shows that no infection process develops in these animals . At a later age infecting the pigs both with a broth culture of the pathogen and with material from organs, using the same doses as in the case of younger pigs has led to the development of a clinical process followed by enhancement of phagocytosis and rise of agglutination titers.

Vet Med Nauki, 1978, 15(4), 11 - 3
{Production of live attenuated vaccine against colibacillosis in pigs}; Stoianov V et al.; Two strains of beta-hemolytic Escherichia coli, p-0141 and i-0149 were used to obtain mutant E . coli organisms with unchanged cultural, morphological, biochemical, and serological properties . The mutants proved apathogenic for albino mice at intravenous injection of a 6-hour broth culture at the rate of 0.3 cm3 or an endotoxin in the same dose . Neither were they enteropathogenic for pigs when inoculated into intestinal segments . They could be used as vaccinal strains.

J Virol, 1978 Jan, 25(1), 253 - 62
Biochemical and morphological characterization of mycobacteriophage R1; Soloff BL et al.; Large-scale propagation of mycobacteriophage R1 in broth culture has allowed the isolation of quantities of virus sufficient for characterization of its nucleic acid and lipid components as well as investigation of its ultrastructural attributes . Analysis of R1 DNA indicates that it is double stranded and possesses a molecular weight of 2.5 X 10(7) and a guanine-plus-cytosine content of 65.7 +/- 0.5% . The lipid fraction of R1 accounts for 14% of the total dry weight of the virus, 20% of which was identified as free or esterified sterols . A rapid loss of viral titer occurred after seconds of exposure to organic solvents . This result suggests that the lipid fractions of R1 is essential for its infectivity . Electron microscopic investigation of solvent-extracted R1 showed extensive deterioration of its normal morphology, including nucleocapsid disintegration and base plate separation . Routine phosphotungstate preparations demonstrated a particle with an oval head and a noncontractile tail . Altering the pH of the phosphotungstate negative stain from neutrality damage the viral particles . Uranyl formate-contrasted specimens displayed an elongated hexagonal nucleocapsid with a neck region; the cross-striated tail possessed a starlike base plate.

Science, 1977 Dec 23, 198(4323), 1262 - 3
Spiroplasmavirus citri 3: propagation, purification, proteins, and nucleic acid; Cole RM et al.; SVC3 is a short-tailed polyhedral virus particle morphologically detectable in many spiroplasmas . It was isolated from two different spiroplasmas (Spiroplasma citri and the suckling mouse cataract agent) by infecting lawns and broth culture of another strain of Spiroplasmavirus citri . Virions from either donor strain had a buoyant density of 1.26 grams per cubic centimeter (metrizamide) or 1.45 grams per cubic centimeter (cesium chloride), and contained five proteins and linear double-stranded DNA with a molecular weight of 14 X 10(6) . Other spiroplasmaviruses have not been propagated, and the molecular weights of double-stranded DNA from other mycoplasma (Acholeplasma) viruses are unknown.

Biull Eksp Biol Med, 1977 Sep, 84(9), 372 - 5
{Mycoplasma hominis as an agent of respiratory tract diseases}; Gusman BS et al.; Respiratory organs of newborn rats inoculated intranasally with broth culture of Myc . hominis were studied by the histological, histochemical and immunofluorescence methods . Tracheitis and development of purulent and interstitial pneumonia with a hemorrhagic component were revealed 24 hours after the infection . These changes were observed up to the 7th day of the experiment . At the same period a specific fluorescence of the Myc . hominis antigen was found by the antibody fluorescent test . The present study pointed to the pathogenicity of the Myc . hominis for the respiratory tract of the newborn rats.

J Clin Microbiol, 1977 Sep, 6(3), 314 - 6
Rapid screening method for enterotoxigenic Escherichia coli; Gurwith M; A rapid screening method for detection of Escherichia coli producing heat-labile enterotoxin is described . Single colonies are transferred directly from primary culture plates into 96-well microculture plates containing 0.3 ml of brain heart infusion broth in each well . After 24 h at 37 degrees C, each brain heart infusion broth culture is assayed by the miniculture method in the corresponding well of a microculture plate in which Y-1 mouse adrenal tumor cells have been grown . All enterotoxigenic isolates detected by this method were confirmed in the assay but with culture supernatants.

N Engl J Med, 1977 Jun 9, 296(23), 1305 - 9
A semiquantitative culture method for identifying intravenous-catheter-related infection; Maki DG et al.; We evaluated a semiquantitative culture technic for identifying infection due to intravenous catheters: rolling the catheter segment across blood agar . This method was compared to broth culture . Of 250 catheters studied, 225 (90%) had low-density colonization on semiquantitative culture (less than 15 colonies on the plate) although 49 (19.6%) of these grew some organisms in broth or on the plate . None of these catheters led to septicemia . Twenty-five catheters (10%) grew greater than or equal to 15 colonies by the semiquantitative technic; most gave confluent growth . Septicemia originated from four of these catheters (P = 0.008) . Of 37 catheters exposed to bacteremias from distant foci of infection, four yielded matching growth in broth, whereas none were concordant with the blood isolate on semiquantitative culture . Local inflammation was associated with high-density colonization semiquantitative culture (P less than 0.001) . The semiquantitative technic distinguishes infection (greater than or equal to 15 colonies) from contamination and is more specific in diagnosis of catheter-related septicemia than culture of the catheter in broth.

J Dairy Sci, 1976 Dec, 59(12), 2152 - 4
Coliform contaminated bedding and new infections; Natzke RP et al.; Ten Holstein cows were bedded on fresh, uncured sawdust seeded with coliform broth culture . Escherichia coli concentrations were maintained at 10(6) colony-forming units/g for the 4-wk experimental period . Ten control cows were bedded with dry shavings . The elevated bacterial contamination caused an increase in contamination of teat ends; however, no new coliform infections occurred.

Am J Vet Res, 1976 Sep, 37(9), 1025 - 9
Etiologic diagnosis of diarrheal disease of calves: frequency and methods for detecting enterotoxin and K99 antigen production by Escherichia cola; Moon HW et al.; Escherichia coli isolated from calves in Minnesota and Montana were tested for enterotoxigenicity via bio-assay of cell-free broth culture fluid and for K99 antigen via a serum agglutination test . Infant mice were used to assay for heat-stable enterotoxin (ST), and adrenal cells in culture were used to assay for heat-labile enterotoxin (LT) . Forty-six of the 345 E coli isolates produced ST enterotoxin, but none produced LT enterotoxin . Thirty-five of the 46 enterotoxigenic isolates had K99 antigen, and only 9 of 66 nonenterotoxigenic isolates so tested had this antigen . The enterotoxigenicity of 28 additional E coli isolates known or suspected to be calf enteropathogens and provided by investigators from 3 different laboratories was also tested . All isolates from 2 laboratories produced ST but not LT . All isolates from the 3rd laboratory produced LT but not ST . Escherichia coli organisms that were positive in the infant mouse assay also caused positive ligated, jejunal-loop responses in calves and in 9-day-old (but not in 5-week-old) pigs . It was concluded that the infant mouse and adrenal cell tests for ST and LT, combined with the agglutination test for K99, would be useful in the diagnosis of enteric enterotoxic colibacillosis of calves.

Vet Rec, 1976 Aug 21, 99(8), 137 - 41
Mycoplasmas and ovine keratoconjunctivitis; Jones GE et al.; The clinical course of an outbreak of keratoconjunctivitis in housed lambs and their dams was followed . Signs were transient generally and became severe in only a small proportion of lambs . The outbreak became most obvious when the lambs were 46 to 55 days old, when 46.9 per cent were affected . Mycoplasma conjunctivae isolations, confirmed by comparison with the type strain by biochemical and serological reactions, increased to 62.1 per cent of all eyes swabbed, but no correlation could be demonstrated between presence of the organism and clinical status . The reasons for this are discussed . Mycoplasma ovipneumoniae was also recovered from the eyes of a small number of lambs . Instillation of a broth culture of M conjunctivae into the conjunctival sacs of four hoggs produced a transient keratoconjunctivitis similar to that observed in the field, but no effect was observed in animals inoculated intravenously . M conjunctivae may therefore be the aetiological agent of non-follicular infectious ovine keratoconjunctivitis, although further work in gnotobiotic or specific pathogen free lambs is required to establish the fact beyond doubt.

Can J Comp Med, 1976 Jul, 40(3), 241 - 6
The prevalence of enterotoxigenic Escherichia coli in the feces of calves with diarrhea; Sivaswamy G et al.; A study was undertaken to determine the prevalence of enterotoxigenicity among Escherichia coli isolated from calves with diarrhea and from a control group of normal calves . The test organisms consisted of 200 E . coli recovered from scouring calves less than two weeks of age and 100 E . coli from normal calves . The enterotoxigenicity of the cultures was evaluated by three methods, namely, injection of ligated segments of piglet intestine, injection of ligated segments of calf intestine and oral inoculation of suckling mice . Live cutures of all the test organisms were used for the ligated intestine studies whereas sterile broth culture supernatants were used in the suckling mouse tests . Of the isolates from scouring calves, 36% were enterotoxigenic in the piglet intestine and 28% in the calf intestine . Amongst the isolates from normal calves, none was enterotoxigenic in the piglet intestine and one was enterotoxigenic in the calf test system . The ligated piglet intestine was considered unsuitable for determining the enterotoxigenicity of bovine E . coli, whereas the ligated calf intestine test was satisfactory and correlated completely with the suckling mouse test . The enterotoxigenic E . coli of bovine origin produced an enterotoxin that resembled the heat stable enterotoxin of typical porcine enteropathogenic E . coli.

Res Vet Sci, 1976 Jul, 21(1), 28 - 35
The experimental infection of specific pathogen free lambs with Mycoplasma ovipneumoniae; Foggie A et al.; Six colostrum-deprived SPF lambs inoculated endobronchially with a second passage broth culture of a Scottish strain of Mycoplasma ovipneumoniae, were killed in batches of two at seven, 14 and 28 days post-inoculation . One lamb from each batch showed macroscopic and microscopic lung lesions similar to but milder than those described for respiratory mycoplasmoses in other species of animals and exhibited minor clinical symptoms . Mycoplasma were recovered from all infected but from no control animals: five infected lambs yielded mycoplasma from lung tissue . Two lambs infected with M ovipneumoniae by endobronchial intubation were placed in contact with six other SPF lambs . M ovipneumoniae was recovered from the upper respiratory tract only of all six contact lambs, but no pathological changes were noted in their lungs . Both donor lambs yielded mycoplasma from lung tissue, but microscopic lesions were detected in only one of them, and these were minimal . No seroconversion due to the infection could be demonstrated in any of the lambs by either the indirect haemagglutination or metabolic inhibition tests.

J Dairy Sci, 1976 Jun, 59(6), 1124 - 30
Effect of coliform challenge at milking time on new udder infections; DeHart DA et al.; This project was designed to study rates of infection in udders of cows exposed to an Escherichia coli broth culture at milking time . Forty Holstein cows of varied stages of lactation were divided randomly into three treatment and one control group of ten cows each . The treatment groups were exposed for 3 wk to an Escherichia coli broth of 10(9) colony forming units per ml at milking time by either 1) dipping teat ends in broth before milking, 2) spraying the udder and leaving it dripping wet during milking, or 3) dipping teat ends in broth after milking . Eleven of 30 treated cows became infected in one or more quarters; all control cows remained uninfected . The infection rate of the three Escherichia coli-treated groups was higher than the controls . However, there was no difference among treated groups . Exposure to the broth culture of Escherichia coli increased the infection rate, but the time at which the udder was exposed to the organisms was unimportant . All infections were of the same type with the same O and H group antigens as the Escherichia coli broth.

Ann Sclavo, 1976 Mar-Apr, 18(2), 286 - 90
{An antigenic extract analyzed by bidimensional immunoelectrophoresis (author's transl)}; Piacentini I et al.; An antigenic extract from broth culture of Aspergillus fumigatus has been analyzed by bidimensional immunoelectrophoresis of Clark and Freeman against rabbit antiserum . The analysis has show the extract to be constituted by seven different antigens.

Vet Med Nauki, 1976, 13(3), 74 - 8
{Production and testing of a live lyophilized bivalent vaccine against Aujeszky's disease and erysipelas in swine}; Vasilev V et al.; A live freeze-dried bivaccine against swine erysipelas and Aujeszky's disease in pigs was produced, its antigen titers reaching 10(8) and 10(4.67) for the two components, respectively . The immunity conferred to the vaccinated pigs (so far as Aujeszky's disease is concerned) had an antibody titer of 1:4, and in sheep the titer was up to 1:16, these animals showing resistance at a control infection with 5000 LD50 per head . The vaccinated pigs likewise withstood a control infection with a mixed 24-48-hour broth culture of virulent strains causing swine erysipelas.

Poult Sci, 1976 Jan, 55(1), 268 - 73
The resistance and carrier status of meat-type hens exposed to Mycoplasma synoviae; Verdaman TH; Forty-eight 32-week-old meat-type females, free from Mycoplasma synoviae (Ms.) and Mycoplasma gallisepticum (Mg.) were aerosol exposed with a 24-hour broth culture of Ms . 1331 and placed in 3 pens . 16 birds per pen . Two males were placed in each of the 3 pens and used as contact birds . All birds were bled at 2, 4, 7, 10, 14, 24, 26, and 30 weeks post Ms . exposure . After the 24-week bleeding, the females were equally divided into 4 pens . All females in 2 of the pens were given a foot pad injection of 0.3 ml . of Ms . 1331 broth culture . Ms . isolation attempts were made from the trachea of all birds at 2, 14, and 24 weeks post Ms . exposure . Ms . isolation attempts were made from all eggs produced, either from the allantoic fluid and egg yolk of 17- or 18-day-old embryos, dead embryos, and infertile eggs, or from the tracheas of day-old progenies . Each day-old progeny was bled . Ms . and Mg . serum plate tests were conducted on the serums from the progeny and adult birds from each bleeding . Ms . hemagglutination-inhibition (HI) tests were conducted on the serums of the adult birds . In the Ms.-exposed birds, the geometric mean HI titer of the serums from each bleeding rose significantly over the previous bleeding . The percentages of Ms . serum plate reactions increased with length of time after Ms . exposure . There were false Mg . serum plate reactions in the early stages of Ms . infection . A foot pad challenge with a broth culture of Ms . 1331 24 weeks after Ms . exposure did not significantly increase the geometric mean HI titer over the Ms . exposed birds that did not have a Ms . foot pad challenge . Ms . was isolated from the trachea of 91.6%, 100%, and 100% of the Ms.-exposed birds at 2, 14, and 24 weeks after Ms . exposure, respectively . There were 35 Ms . isolates from 575 attempts from 17- or 18-day embryos, dead embryos, or infertile eggs, and 7 Ms . isolates from 67 attempts from the trachea of day-old progenies . All isolates were made from eggs collected from 6 through 31 days afer Ms . exposure . No isolations were made from 1760 attempts made from eggs collected from 32 through 210 days after Ms . exposure, including eggs collected from Ms . foot pad-challenged birds at 168 days after Ms . exposure . There were 312 broilers reared from eggs collected from the 22 through 24 weeks after Ms . exposure . The broilers were marketed at a federally inspected poultry processing plant . Their records showed 3 birds condemned for septicemia-toxemia, but none were condemned for air-sacculitis or synovitis . Twenty-four serums from the broilers that were positive to Ms . or Mg . serum plate tests were all negative to the Ms . and Mg . HI tests . Ms . serum plate reactors began to show up in the day-old progeny from eggs collected beginning on the 76th day after Ms . exposure . There were 16.0% (80/500) positive to the Ms . serum plate test from eggs collected from the 76 through 126 days after Ms . exposure and 14.0% (52/372) from eggs collected from the 169 through 210 days after Ms . exposure.

Biull Eksp Biol Med, 1976, 82(11), 1305 - 10
{Changes in the electrical activity of the cerebral cortex under the combined influence of Cl . perfringens type A toxin and products of Cl . butyricum activity}; Tselukh AV et al.; A study of the changes of the electrocorticogram (ECoG) and the electrocardiogram (ECG) in combined action of Cl . perfringens, type A, and of the Cl . butyricum broth culture filtrate showed that desynchronization of the cortical electrical activity and its subsequent depression occurred at earlier periods than in the case of isolated administration of Cl . perfringens toxin . The general character of the changes in the cortical rhythmic activity remained the same as in intoxication caused by Cl . butyricum toxin alone . The ECoG and ECG changes occurred at shorter intervals . Cl . butyricum filtrates induced no ECoG and ECG changes . It is supposed that the effect of the products of the Cl . butyricum vital activity consisted in increase in the tissue barrier permeability and, in this connection, in a greater penetration of Cl . perfringens toxin into the tissues, including the central nervous system.

Avian Dis, 1976 Jan-Mar, 20(1), 118 - 25
Efficacy of Linco-Spectin water medication on Mycoplasma synoviae Airsacculitis in broilers; Hamdy AH et al.; Three trials were conducted to evaluate the efficacy of Linco-Spectin (LS) water medication on Mycoplasma synoviae (MS) air-sacculitis in broilers under controlled experimental conditions . Day-old chicks were vaccinated against infectious bronchitis and Newcastle disease virus and exposed to a broth culture of MS by the respiratory route . In each trial, one-half of the flock was treated with 2 g of LS per gallon of drinking water for the first five days of life, and the other half was kept as a control . At two and eight weeks postinoculation (PI) birds were weighted individually and examined serologically, culturally, and grossly for MS airsacculitis . Linco-Spectin water medication was effective in controlling MS airsacculitis in broilers.

Biull Eksp Biol Med, 1975 Nov, 80(11), 57 - 60
{Determination of choleragen activity in tadpoles}; Ermol'eva EV et al.; A method of titration of the V . cholerae toxin obtained from the broth culture filtrates is presented . In difference from the earlier methods, the larvae of Rana temporaria were used as a test object . The tadpoles were placed in the vessels with water containing graded concentrations of the toxin . The titer of the toxin activity was evaluated by the percentage of dead tadpoles . The concentration causing the death of 50 per cent of the tadpoles in the course of the 24 hours was taken as a unit of toxin activity . The method is simple, easily reproducible and economical.

Zentralbl Bakteriol {Orig A}, 1975 Aug, 232(4), 477 - 81
{Protective oral immunization against coli enterotoxemia in swine (author's tranls)}; Illes J; The issue of immunity in the case of enteric infections caused by bacteria is still widely unresolved . To clarify this, weaned piglets 6-7 weeks of age were used as a model, because of the particular susceptibility to colienterotoxemia shown by them at this particular age . The experimental design was as follows . The animals were immunized orally and parenterally (i.v . and i.p . routes) by means of a vaccine consisting of bacteria incapable of reproduction . 14 days later, the animals were challenged orally with virulent EC-O 141 B 85 and EC-O 139 B 82 . The results have shown that a) parenteral immunization did not induce protection against oral infection with virulent E . coli; b) repeated oral immunization produced measurable immunity against oral infection; and c) 1 7-hour broth culture proved to be particularly suitable for oral infection . Results also point to the fact local immunity in the intestine as present after enteric infection by bacteria may be artificially induced only by protective oral immunization.

Aust Vet J, 1975 Jan, 51(1), 28 - 31
Isolation of Mycoplasma hyopneumoniae and its association with pneumonia of pigs in Australia; Furlong SL et al.; Nine strains of mycoplasmas were isolated from the lungs of 5 pigs with clinical signs of naturally acquired enzootic pneumonia . Mycoplasma hyopneumoniae was isolated from the lungs of 1 pig and M . hyorhinis from the lungs of 4 . An unidentified mycoplasma, which utilized arginine, grew rapidly in broth culture and produced centred colonies on solid medium, was isolated from the lungs of 4 pigs . The pathogenicity of the isolated strain of M . hyopneumoniae was determined by inoculation of pigs from an enzootic pneumonia-free herd . Enzootic pneumonia was produced in the lungs of all 5 pigs inoculated intranasally and intratracheally with broth cultures of the organism isolatied by limit dilution techniques . Enzootic pneumonia was produced in 3 of 6 pigs inoculated intranasally and intratracheally with M . hyopneumoniae purified by the passage of colonies on agar blocks . M . hyopneumoniae was isolated in pure culture from the lungs of all pigs with induced pneumonic lesions.

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