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S Tola

Publications and source records attributed to S Tola.

At least 19 recordsLinked to original sources
PubMed2006

Two cases of human granulocytic ehrlichiosis in Sardinia, Italy confirmed by PCR.

In this work we report the first two cases of human granulocytic ehrlichiosis (HGE) in Sardinia. In early September 2004, a 69-year-old woman (patient 1) was admitted to the Infectious Diseases Institute of Sassari for rickettsiosis like-syndrome: high fever (39.5-40 degrees C), dyspnea, reduced consciousness, vomiting, and cutaneous rash. In late September 2004, a 30-year-old man (patient 2) with high fever was admitted for an evident palmar and oral erythema, edema of the labium, very intense arthralgia, myalgia, and dyspnea. In these two hospitalized patients, the diagnosis was made through indirect IgM and IgG immunofluorescent technique and confirmed by the presence of the specific DNA in the leukocytes. The two patients were A. phagocytophilum-PCR positive.

Adult↗
PubMed2005

Role of Chlamydophila abortus in ovine and caprine abortion in Sardinia, Italy.

Between 1999-2003, 14321 sera and 646 abortion samples (498 foetuses and 148 placentae) were analysed from 807 sheep and goat farms distributed all over the island of Sardinia. After notification of abortion in a flock, sera collected at random from adult animals were examined to detect antibodies specific to Chlamydophila (C.) abortus by ELISA, whereas foetuses and placenta were analysed by PCR assay. Specific IgG antibodies were detected in 611 (4.8%) sheep and 106 (5.8%) goats. From a total of 2050 ovine and 151 caprine fetal samples including muscle, liver, abomasum, spleen, brain and placenta, 29 (1.4%) ovine and 1 (0.6%) caprine samples were C. abortus PCR-positive. Placenta was the tissue with the highest detection rate. These results indicate that the seroprevalence of C. abortus infection in sheep and goats is very low in Sardinia, and PCR results demonstrate that C. abortus has no significant role in abortion, especially in goats.

Aborted Fetus↗
PubMed2005

Rapid differential diagnosis of Mycoplasma agalactiae and Mycoplasma bovis based on a multiplex-PCR and a PCR-RFLP.

The membrane-protein 81 gene (mb-mp81) of Mycoplasma bovis was cloned, sequenced and compared to membrane-protein 81 gene (ma-mp81) of Mycoplasma agalactiae. After alignment of both sequences, specific primers pairs were designed from variable or unchanging nucleotide segments. In this study, we describe the development and optimization of a multiplex-PCR (MPCR) for the rapid detection of M. agalactiae and M. bovis strains. In addition, a simple and rapid PCR-restriction fragment length polymorphism (RFLP) assay, using the restriction enzymes AluI, DraI, RsaI and XbaI, is described to distinguish between both species. The results suggest that MPCR and PCR-RFLP assays could be used as an alternative method in routine diagnosis for rapid and specific simultaneous detection of M. agalactiae and M. bovis.

Bacterial Proteins↗
PubMed2003

Survey of ovine and caprine toxoplasmosis by IFAT and PCR assays in Sardinia, Italy.

During the period 1999-2002, we have analyzed 9639 serum samples and 815 aborted samples (670 fetuses and 145 placenta) from 964 ovine and caprine farms distributed over all Sardinia island. After abortion notification, sera collected at random from adult animals were examined to detect simultaneously IgG and IgM antibodies specific to Toxoplasma gondii by indirect immunofluorescence assay, whereas fetuses and placenta were analyzed by a single tube nested PCR assay. Specific IgG antibodies were detected in 2048 (28.4%) sheep and 302 (12.3%) goats, specific IgM antibodies were found in 652 (9%) sheep and 139 (5.6%) goats. From a total of 2471 ovine and 362 caprine fetal samples including muscle, liver, abomasum, spleen, brain and placenta, 271 (11.1%) ovine and 23 (6.4%) caprine samples were T. gondii PCR-positive. Although T. gondii DNA was amplified from different types of tissues, placenta was the tissue with the highest detection rate. On the one hand, these results indicate that the seroprevalence of T. gondii infection in sheep and goats is relatively high, on the other PCR results demonstrate that T. gondii has a significant role in ovine and caprine abortion. Adequate management might be useful and essential to control the toxoplasmosis in the sheep and goats herds of Sardinia.

Aborted Fetus↗
PubMed2003

Ehrlichiosis and rickettsiosis in a canine population of Northern Sardinia.

Ehrlichiosis and rickettsiosis are very common, widespread diseases. These diseases are present in Sardinia year round because its temperate weather permits the survival of many kinds of tick vectors. A thousand dogs were subjected to physical, hematological, biochemical examinations and serological tests. All 1,000 sera were analyzed by indirect immunofluorescence antibody test to detect antibodies against E. canis and R. rickettsii. A high seroprevalence (about 50%) was detected for both etiological agents, without differences in relation to sex, breed, or usage. A high seroprevalence, corresponding to 62.5% for ehrlichiosis and 64% for rickettsiosis, was observed in the age group of 13-60 months. The mortality was greatest in the males in the age group, which manifested the disease in the chronic phase.

Animals↗
PubMed2001

Sequence, cloning, expression and characterisation of the 81-kDa surface membrane protein (P80) of Mycoplasma agalactiae.

Mycoplasma agalactiae, the causative agent of contagious agalactia in small ruminants, produces a protein, named P80, that is detectable in all wild-type isolates examined to date and that appears expressed during the early phase of infection. We describe here the identification, cloning and expression of the gene encoding P80 (ma-mp81). The deduced amino acid sequence is consistent with a hydrophobic and basic protein that possesses a lipoprotein signal peptide. Sequence analysis of gene ma-mp81 suggests that P80 is a membrane lipoprotein that shows significant homology with other putative lipoproteins of M. pneumoniae. An internal 1-kb fragment of ma-mp81 was expressed in Escherichia coli as a 6xHis-tagged protein. The purified recombinant protein greatly reacted with polyclonal anti-P80 sera raised in lamb.

Amino Acid Sequence↗
PubMed2001

A physical map of the Mycoplasma agalactiae strain PG2 genome.

We have constructed a physical map of the Mycoplasma agalactiae strain PG2 chromosome analyzing it by pulsed field gel electrophoresis in a contour-clamped homogeneous electric-field system. We mapped 33 cleavage sites generated with SmaI, XhoI, SalI, EclXI and BsiWI restriction endonucleases using double digestions, one- and two-dimensional pulsed electrophoresis, cross-hybridization and linking clones. We have also mapped the loci of some genes by Southern hybridization.

Blotting, Southern↗
PubMed1999

Experimental vaccination against Mycoplasma agalactiae using different inactivated vaccines.

Five sets of vaccines were prepared using Mycoplasma agalactiae washed cultures inactivated with phenol (1), formalin (2), heat-treatment (3), sodium hypochlorite (4) and saponin (5). All sets, except for saponin-vaccine, were adjuvated with aluminium hydroxide. Five groups of six sarda ewes were inoculated twice before pregnancy, once during pregnancy and challenged during the lactation period. Monthly blood samples were taken from the vaccinated sheep and from 12 controls: sera were assayed by immunoblotting, ELISA and growth inhibition tests. Four control sheep were infected by intracanalicular route with pooled mycoplasmas at concentrations of 10(4), 10(5), 10(6) and 10(7) CCU. The challenge involved using infected milk to contaminate the remaining sheep. All the controls and some ewes from groups 2, 3 and 4 developed contagious agalactia. Ewes vaccinated with phenol- and saponin-inactivated mycoplasmas resisted experimental challenge. These results suggest that these two vaccines are effective and that their use could limit the diffusion of M. agalactiae infection.

Animals↗
PubMed1999

Comparison of restriction pattern polymorphism of Mycoplasma agalactiae and Mycoplasma bovis by pulsed field gel electrophoresis.

Mycoplasma agalactiae and Mycoplasma bovis are closely related species in phylogenetic terms. Pulsed Field Gel Electrophoresis (PFGE) of SmaI, EclXI, Bsi WI, MluI, BssHII, SalI, XhoI, NruI and ApaI digested DNAs were used to analyse and to compare restriction fragment length polymorphism between M. agalactiae and M. bovis and to estimate their genome sizes. SmaI, EclXI and Bsi WI enzymes cleaved DNAs of both microrganisms. MluI, BssHII, SalI, XhoI and NruI digested only M. agalactiae DNA whereas ApaI cut only M. bovis DNA. The total DNA length was established to be 945 +/- 8.4 Kb for M. agalactiae and 961 +/- 18.9 Kb for M. bovis.

Animals↗
PubMed1997

Characterization of membrane surface proteins of Mycoplasma agalactiae during natural infection.

We have analyzed antigenic variation of seven M. agalactiae wild strains using different sera from naturally infected sheep. Only 30 day sera recognized all surface proteins and inhibited the growth of mycoplasmas. Furthermore, we have observed that two strongly immunogenic proteins: 55 and 35 kDa were digested using 500 micrograms/ml of trypsin. These two bands are immunoprecipitated together with four other proteins but only the 35 kDa protein is recognized by eluted antibodies.

Animals↗
PubMed1997

Detection of Mycoplasma agalactiae in sheep milk samples by polymerase chain reaction.

We developed a simple and rapid method for DNA extraction from sheep milk to use for polymerase chain reaction (PCR) diagnosis of Mycoplasma agalactiae. We tested 357 samples from 21 newly infected flocks (group 1) and 87 samples from 8 flocks infected in the past (group 2). PCR results were compared with those of conventional culture. By PCR we detected 175 positives in group 1, while by culture we detected only 153. Milk samples from group 2 were negative, both by PCR assay and by culture. Our PCR is much faster than culture and reduces the time required for diagnosis from several days to 5 h. The method could be used for the routine diagnosis of contagious agalactia caused by Mycoplasma agalactiae.

Animals↗
PubMed1996

Comparison of Mycoplasma agalactiae isolates by pulsed field gel electrophoresis, SDS-PAGE and immunoblotting.

We have analyzed 81 isolates of Mycoplasma agalactiae from four different regions of Italy between 1990 and 1995 in order to identify antigenic differences through SDS-PAGE and Western blotting and chromosomal DNA restriction endonuclease cleavage pattern differences. Antigenic variability in M. agalactiae isolates was investigated analyzing hydrophobic membrane protein fractions by immunoblotting using pooled sheep antiserum from naturally infected sheep. Large restriction fragments obtained cleaving genomic DNAs with SmaI, NruI, SalI, XhoI, BssHII and KpnI were analyzed by pulsed field gel electrophoresis. Genetic analysis indicates that isolates are all similar without intraspecific differences. This homogeneity was confirmed by immunoblotting: 80 and 50 kDa antigens are present in all strains analyzed.

Animals↗
PubMed1996

Rapid and specific detection of Mycoplasma agalactiae by polymerase chain reaction.

A polymerase chain reaction (PCR)-based test was developed for the detection of Mycoplasma agalactiae in sheep milk samples. Two oligonucleotide primers were designed to amplify a 375 bp fragment of M. agalactiae chromosomal DNA. Amplified products were analyzed by agarose gel electrophoresis and Southern blot hybridization using a fluorescein labeled 528 bp probe. The primers allowed the amplification of fragment of M. agalactiae DNA and did not amplify any specific fragment of other mycoplasmal DNAs (M. capricolum, M. mycoides subsp. mycoides, M. mycoides subsp. capri, M. putrefaciens, M. arginini and M. bovis) or other bacterial DNAs (S. aureus, S. epidermidis, P. haemalytica, E. coli, S. agalactiae, S. dysgalactiae, S. uberis, B. cereus, P. aeruginosa, S. durans, L. lactis, L. lactis var. diacetilactis, L. mesenteroides, S. thermophilus, L. bulgaricus and L. casei). The limit of detection of PCR assay was between 2.5 and 25 fg of purified DNA and 10(2) CCU ml-1 on mycoplasma cultures. These results indicate that the PCR technique can be used as a rapid and specific diagnostic method for detection of M. agalactiae.

Animals↗
PubMed1994

A species-specific DNA probe for the detection of Mycoplasma agalactiae.

A 4.5 kb M. agalactiae DNA fragment present in strains from different areas of Sardinia was cloned and used to detect M. agalactiae DNA in sheep milk samples by dot blot hybridization. The probe recognized only M. agalactiae strains and did not cross-hybridize with other mycoplasmas (M. capricolum, M. mycoides subsp. capri, M. mycoides subsp. mycoides, M. putrefaciens and M. arginini) or bacteria (E. coli, P. hemolytica, S. aureus, S. epidermis, L. casei, S. durans, S. lactis and S. thermophilus) which may also be found in sheep milk.

Animals↗
PubMed1994

Incidence of sciatic pain among men in machine operating, dynamic physical work, and sedentary work. A three-year follow-up.

The incidence of sciatic pain among 25-49 year-old men in three different types of work: machine operating, dynamic physical work (construction carpenters), and office work was assessed prospectively. Postal questionnaires both at the beginning and the end of the 3-year follow-up were answered by 1149 men who at baseline had no history of sciatic pain. The crude risk ratio of the incidence of sciatic pain was 1.6 (95% confidence interval 1.2-2.2) for the machine operators and 1.7 (1.3-2.4) for the carpenters when the office workers were referents. The adjusted risk ratios were 1.4 (1.0-1.9) and 1.5 (1.1-2.1), respectively. Previous history of severe lumbago or other low-back pain increased the risk fourfold. Frequent physical exercise and smoking were of borderline significance as predictors.

Adult↗