Draft Genome Sequences of Mycoplasma mycoides subsp. mycoides Strains APF9 and AP108, Isolated in Nigeria in 2014 to 2016.

Mycoplasma mycoides subsp. mycoides is the etiological agent of contagious bovine pleuropneumonia (CBPP). While several findings on CBPP prevalence in Nigeria were documented, no data were reported about the genomic characterization of Nigerian M. mycoides subsp. mycoides strains. Here, we present the draft genome sequences of two novel M. mycoides subsp. mycoides strains isolated in Nigeria.

Canine Leishmania infantum infection: an imported case in UK after staying in the Canary Islands.

Leishmaniosis is reported in the Iberian Peninsula and the Balearic Islands, but the Canary Islands are deemed free. In the present communication, we report a clinical leishmaniosis due to Leishmania infantum in a dog that was presumptively infected during its stay on Tenerife. The result of Leishmania serology (whole-cell based ELISA with L. infantum antigen) was high positive (test score of 82.2 at a cut-off value of 12.0). This result was further confirmed with quantitative real-time polymerase chain reaction (qPCR) for Leishmania spp. on a blood sample. A medium load of parasites was detected (48 parasites/ml blood). L. infantum was identified by RFLP analysis of the ITS-1 PCR product. Confirmation that leishmaniosis is endemic to the Canary Islands would further require study on local dogs with no travel history as well as reassessment on frequency and distribution of Phlebotomus spp. as well as Leishmania spp. detection in the sand fly vector. However, this case strongly suggests that L. infantum is present on the Canary Islands. Although transmission seems to be still exceptional, preventive measures in dogs travelling to the Canaries should be considered.

Whole-Genome Sequencing of Mycoplasma mycoides subsp. mycoides Italian Strain 57/13, the Causative Agent of Contagious Bovine Pleuropneumonia.

Mycoplasma mycoides subsp. mycoides is generally considered one of most pathogenic Mycoplasma species, and it is the etiological agent of contagious bovine pleuropneumonia (CBPP). Here, we present the annotated genome sequence of M. mycoides subsp. mycoides Italian strain 57/13, isolated in 1992 during CBPP outbreaks in Italy.

IgG antibodies from dourine infected horses identify a distinctive Trypanosoma equiperdum antigenic pattern of low molecular weight molecules.

Diagnosis and control of dourine is strongly based on serological evidence, but knowledge of the humoral response of horses during infection is limited. In this study we developed a chemiluminescent immunoblotting (cIB) assay to characterise the Trypanosoma equiperdum antigen pattern recognised by IgGs from naturally or experimentally dourine-infected horses and analyse the kinetics of IgG humoral response following the infection. One compounding factor is that sera from uninfected animals often cross-react with T. equiperdum antigens. Development of the cIB assay was based on the hypothesis that serum IgGs from healthy and infected animals recognise different T. equiperdum antigen patterns. We used sera from 8 naturally infected horses which had recovered from Italian outbreaks and 2 experimentally infected mares. In addition, sera from 10 healthy control animals, eight of which were CFT positive but IFA negative for dourine, were collected from disease free regions. Sera were compared by the complement fixation test (CFT), indirect immune fluorescence (IFA) and the cIB assay. cIB analysis revealed that IgGs from infected horses, in contrast to IgGs from healthy horses, specifically recognise a T. equiperdum antigenic profile with low molecular weight bands ranging between 16 and 35 kDa. A time course experiment indicated that IgGs specific for the 16-35 kDa parasite protein fraction appear 17 days post-infection. The cIB assay confirmed all ten infected animals as positive and all controls as negative. This study demonstrated that analysis of IgGs by cIB can provide clear confirmation of trypanosome infection in horses, suggesting that this technique can be applied as a confirmatory serological test for dourine infection.

One test microbial diagnostic microarray for identification of Mycoplasma mycoides subsp. mycoides and other Mycoplasma species.

The present study describes the use of microarray technology for rapid identification and differentiation of Mycoplasma mycoides subsp. mycoides from other mycoplasmas that may be pathogenic to ruminants, including those of the Mycoplasma mycoides cluster, genetically and antigenically strictly correlated with Mycoplasma mycoides subsp. mycoides. A microarray containing genetic sequences of 55 different bacterial species from Acholeplasma, Mycoplasma, Spiroplasma and Ureaplasma genera was constructed. Sequences to genes of interest were collected in FASTA format from NCBI. The collected sequences were processed with OligoPicker software. Oligonucleotides were then checked for their selectivity with BLAST searches in GenBank. The microarray was tested with ATCC/NCTC strains of Mycoplasma spp. of veterinary importance in ruminants including Mycoplasma belonging to the mycoides cluster as well as Mycoplasma mycoides subsp. mycoides and Mycoplasma mycoides subsp. capri field strains. The results showed that but one ATCC/NCTC reference strains hybridized with their species-specific sequences showed a profile/signature different and distinct from each other. The heat-map of the hybridization results for the nine genes interrogated for Mycoplasma mycoides subsp. mycoides demonstrated that the reference strain Mycoplasma mycoides subsp mycoides PG1 was positive for all of the gene sequences spotted on the microarray. CBPP field, vaccine and reference strains were all typed to be M. mycoides subsp. mycoides, and seven of the nine strains gave positive hybridization results for all of the nine genes. Two Italian strains were negative for some of the genes. Comparison with non-Mycoplasma mycoides subsp. mycoides reference strains showed some positive signals or considerable homology to Mycoplasma mycoides subsp. mycoides genes. As expected, some correlations were observed between the strictly genetically and antigenically correlated Mycoplasma mycoides subsp. mycoides and Mycoplasma mycoides subsp. capri strains. Specifically, we observed that some Italian Mycoplasma mycoides subsp. mycoides strains were positive for two out of the three Mycoplasma mycoides subsp. capri genes, differently from what has been observed for other European or African Mycoplasma mycoides subsp. mycoides strains. This study highlighted the use of microarray technology as a simple and effective method for a single-step identification and differentiation of Mycoplasma mycoides subsp. mycoides from other mycoplasmas that may be pathogenic to ruminants, including those of the Mycoplasma mycoides cluster, genetically and antigenically strictly correlated with Mycoplasma mycoides subsp. mycoides. The opportunity to discriminate several mycoplasmas in a single analysis enhances diagnostic rapidity and may represent a useful tool to screen occasionally mycoplasmas affecting animal farming in territories where diagnostic laboratory support is limited. The heat-map of the hybridization results of the comparative genomic hybridizations DNA-designed chip clearly indicates that the microarray performs well for the identification of the tested Mycoplasma mycoides subsp. mycoides reference and field strains, discriminating them from other mycoplasmas.

Determination of haematological reference intervals in healthy adult greyhounds.

OBJECTIVE: The purpose of this study was to establish breed-specific reference intervals for haematological measurands in non-racing greyhounds. Suitability of the data for partitioning according to sex was also examined. METHODS: Haematological data were collected from 304 healthy non-racing greyhounds and analysed using non-parametric methods. Results were compared with non-breed-specific canine reference intervals and also with greyhound reference intervals obtained by other investigators. RESULTS: Compared with non-breed-specific reference intervals, the results showed comparable mean and upper limit and higher lower limit for erythrocyte count; higher values for haemoglobin, haematocrit and mean corpuscular volume; and lower values for total leucocyte count and absolute concentration of neutrophils, lymphocytes, monocytes, eosinophils and platelets. Partitioning according to sex was recommended by the statistical analysis for all analytes except haematocrit and total leucocyte count. CLINICAL SIGNIFICANCE: In this study the reference intervals were derived from a large sample size. The results are in general agreement with previous reports, although higher values for low reference limits have been noted for the erythroid parameters, and lower values for upper reference limits have been observed for the total and different leucocyte counts. Breed-specific reference intervals provide a useful clinical tool for haematological evaluations.

Determination of serum biochemistry reference intervals in a large sample of adult greyhounds.

BACKGROUND: As large breed, highly muscled dogs, greyhounds are regarded as physiologically different to other breeds. Biochemistry reference intervals have previously been determined using small numbers of greyhounds or based on the observations from racing dogs. OBJECTIVES: The purpose of this study was to develop statistically defined reference intervals for biochemical analytes in healthy non-racing greyhounds. Partitioning according to gender was also examined. METHODS: Biochemical analytes from a population of non-racing healthy greyhounds, including 269 males, 202 females and 28 dogs where gender had not been recorded, were examined using a non-parametric statistical approach. The dogs were aged between one and nine years old. RESULTS: Total protein, albumin, globulin and creatinine reference intervals differed from generic reference intervals used for dogs. The reference intervals for greyhounds in this study are similar to those obtained by other studies of greyhounds, but often had a narrower range of values, likely reflecting more accurate estimation associated with larger numbers of reference individuals. Recommended methods for assessment of partitioning do not indicate a need to partition according to gender.

Clinical, humoral and IFNgamma responses of cattle to infection with Mycoplasma mycoides var. mycoides small colony and attempts to condition the pathogenesis of the infection.

Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides var. mycoides small colony (MmmSC), is one of the most important diseases of cattle in Africa. The role of innate or acquired cell mediated and humoral immunity in conferring protection against MmmSC infection has not yet been elucidated. On the other hand, the pathological lesions caused by the aetiological agent have been considered indicative of an immunopathological process. In this study ten naïve cattle were exposed to in-contact infection with animals infected by intubation with a strain of MmmSC. Clinical signs, antibody response, IFNgamma release and pathological changes at necropsy were analysed and compared with the events following in-contact infection of an equal number of animals kept under daily treatment with cyclosporine for the entire observation period of 84 days. Cyclosporine is a suppressor of the immune response related to the T-cell system. Under the conditions of the experiment, cyclosporine appeared to condition the pathogenesis of CBPP by delaying the events that follow infection, bringing further support to the possibility that the immune response may have an impact on the disease outcome.