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.     

Effects of natural feeding stimulants and glutamic acid supplementation on the feed intake,growth performance and digestive enzyme activities of red sea bream (Pagrus major) fed fish meal‐free soy protein concentrate (SPC)‐based diet

Two experiments were conducted for red sea bream (Pagrus major). In experiment 1, the optimum level of glutamic acid and natural feeding stimulants to enhance feed intake were determined and found that glutamic acid level of 0.5% and fish meat hydrolysate (FMH) were effective. In experiment 2, fish were fed with soy protein concentrate (SPC)‐based diet with synthetic feeding stimulants (Basal diet), the Basal diet with FMH (FMH diet), the FMH diet with glutamic acid (FMHG diet) and with fish meal diet (FM diet) as a control until satiation for 8 weeks. Feed intake of FMHG‐fed fish was significantly higher than others (p < 0.05). Specific growth rate and the feed conversion ratio of FMHG were comparable to those of FM‐fed fish (p > 0.05). Relative visceral fat ratio and crude lipid content of any SPC‐based diet‐fed fish tended to be lower than those of FM diet‐fed fish. There were no significant differences in trypsin and lipase activities hepatopancreas among treatments. SPC can be utilized as a sole protein source in a diet for red sea bream. The lower growth performance in SPC‐based diet‐ fed fish was not due to poor digestive enzyme secretion but could be associated with lipid utilization disorder.     

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.     

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.     

Effects of feeding a fish meal‐free soy protein concentrate‐based diet on the growth performance and nutrient utilization of red sea bream (Pagrus major)

Two groups of red sea bream (average body weight 85 g) were fed to satiation fish meal (FM) or FM‐free soy protein concentrate (SPC)‐based diet (FM‐sat and SPC groups respectively). A third group (FM‐pair) was fed the same amount of FM as of SPC fed to SPC‐sat group for 6 weeks. Growth rate of SPC group was significantly slower than of FM‐sat and FM‐pair groups (p < 0.05). Feed conversion ratios of FM‐sat and FM‐pair groups were similar and were better than that of SPC group (p < 0.05). Visceral fat somatic index of SPC group was significantly lower than of the other groups (p < 0.05), while that of the latter two groups were similar. Plasma triglyceride of FM‐sat and FM‐pair groups was similar and lower than the level of SPC group (p < 0.05). Trypsin activity in hepatopancreas of SPC group was significantly lower than that of other groups (p < 0.05). We suggest a low growth of SPC group was not simply related to low‐feed intake. Feeding of a FM‐free SPC‐based diet may reduce lipid accumulation and consequently lead to low‐growth performance, in red sea bream.     

Antimicrobial susceptibilities and population structure of Staphylococcus epidermidis associated with ovine mastitis

Intramammary infections are a serious problem for dairy sheep farms, and Staphylococcus epidermidis is one of the main etiological agents of ovine mastitis. In this work, 131 S. epidermidis isolates, collected from 2201 dairy Sarda sheep belonging to 14 flocks with high somatic cell count scores, were studied. The flocks were located in diverse geographical areas of Sardinia, Italy. The aim of study was to assess the susceptibility of isolates to 13 antimicrobial agents, many of which are frequently used in mastitis therapy. Oxacillin was used for detecting methicillin-resistant S. epidermidis (MRSE) by disk diffusion test. Thirty-eight percent of the isolates (n = 50) were resistant to penicillin, 7.6% (n = 10) were resistant to tetracycline, and 2.3% (n = 3) were resistant to both penicillin and tetracycline (PTRSE). Two isolates were resistant to five antimicrobials including methicillin. Analysis of staphylococcal cassette chromosome mec (SCCmec) elements showed that both MRSE isolates harbored SCCmec type IVa. Based on pulsed-field gel electrophoresis (PFGE) typing by SmaI macrorestriction, S. epidermidis isolates were grouped into four clusters at 75% similarity level. The two multi-drug resistant MRSE isolates displayed distinct PFGE patters. This study indicates that S. epidermidis isolates from sheep milk samples may accumulate resistance markers for different antimicrobial agents. Furthermore, the occurrence of PTRSE and MRSE suggests to adopt adequate hygienic measures when handling animals with intramammary infections, in order to prevent spreading PTRSE and MRSE strains to humans through direct contact and/or consumption of contaminated food.     

Comparative diagnosis of parasitological,serological, and molecular tests in dourine-suspected horses

Study on comparative sensitivity of parasitological, serological, and molecular tests on 237 horses originating from two dourine-suspected districts of Arsi-Bale highlands of Ethiopia was conducted to determine the prevalence of the disease and degree of agreement of the diagnostic tests. Accordingly, the prevalence of the disease was found to be 4.6%, 36.7%, and 47.6% by parasitological Woo test, RoTat 1.2 and 18S PCR tests, respectively. The seroprevalence of the disease was 27.6% in CATT/Trypanosoma evansi test. In Ethiopia, it was for the first time that trypanosomes from dourine suspected horses were demonstrated in 4.6% of the animals using Woo test. The findings of the present study disclosed that dourine is highly prevalent and one of the major diseases of horses in the area. There was no statistically significant difference (P > 0.05) in prevalence of the disease between districts, sexes, and age groups of the animals. However, there was a statistically significant difference (P < 0.05) in the prevalence of the disease between emaciated and animals with good body condition. Assessment of the degree of agreement of the diagnostic tests employed revealed low to fair ( k = 0.1 - 0.4 ) \left( {k = 0.{1} - 0.{4}} \right) with significantly higher sensitivity by PCR than other tests.     

Detection of pathogens in ovine and caprine abortion samples from Sardinia, Italy, by PCR.

During 2003-2005, 399 abortion samples (315 fetuses and 84 placentae) were collected from 107 ovine and caprine farms in northern Sardinia. Tissues from aborted fetuses and placentae were examined by PCR assay to detect DNA from Coxiella burnetii, Chlamydophila abortus, Salmonella enterica Serovar abortusovis, Toxoplasma gondii, and Neospora caninum. The DNA from at least 1 of these 5 infectious agents was amplified in 41% of ovine fetuses, while only 17% of the caprine fetuses yielded a positive amplification result for at least 1 of the 5 agents. Out of a total of 366 ovine aborted samples, T. gondii DNA was detected most frequently (18.1% of fetuses and 13.1% of placentae), followed by S. abortusovis (13% of fetuses and 14.4% of placentae), C. burnetii (10.9% of fetuses, of 9.2% placentae), C. abortus (2.4% of fetuses, 6.5% of placentae), and N. caninum (2% of placentae). In 33 fetuses and 9 placentae, the simultaneous presence of pathogens with different associations was detected. Out of a total of 31 caprine aborted samples, T. gondii was detected most frequently (13% of fetuses and 25% of placentae), followed by C. abortus (12.5% of placentae), C. burnetii (12.5% of placentae), and N. caninum (8.6%).     

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

Between 1999–2003, 14 321 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.