Mycoplasmas Isolated from the Respiratory Tract of Cattle and Goats in Tanzania

A microbiological study of the mycoplasma flora in the respiratory tracts of cattle and goats in selected regions of Tanzania is described. In the examination of cattle, mycoplasmas were isolated from 60 (17.8%) of the 338 examined lung samples, 8 (47.1%) of the 17 lymph nodes, 4 (13.3%) of the 30 pleural fluid samples and 4 (3.9%) of the 103 nasal swabs examined. All the isolates were identified as Mycoplasma mycoides subsp. mycoides, Small Colony type except for one isolate from pleural fluid which was identified as Mycoplasma arginini. M. mycoides subsp. mycoides, Small Colony type was isolated from samples originating from Dodoma, Iringa, Mbeya, Morogoro and Shinyanga regions where outbreaks of contagious bovine pleuropneumonia had been reported. In the examination of goats, mycoplasmas were isolated from 54 (34.0%) of the 159 examined lung samples, 41 (18.1%) of the 226 nasal swabs and 4 (40.0%) of the 10 pleural fluid samples. The species demonstrated were Mycoplasma capricolum subsp. capripneumoniae, M. mycoides subsp. mycoides, Small Colony type Mycoplasma ovipneumoniae and M. Capricolum subsp. arginini. The isolation of M. capripneumoniae in the Coast and Morogoro regions confirmed the presence of contagious caprine pleuropneumonia in the regions.     

The origin of the 'Mycoplasma mycoides cluster' coincides with domestication of ruminants

The 'Mycoplasma mycoides cluster' comprises the ruminant pathogens Mycoplasma mycoides subsp. mycoides the causative agent of contagious bovine pleuropneumonia (CBPP), Mycoplasma capricolum subsp. capripneumoniae the agent of contagious caprine pleuropneumonia (CCPP), Mycoplasma capricolum subsp. capricolum, Mycoplasma leachii and Mycoplasma mycoides subsp. capri. CBPP and CCPP are major livestock diseases and impact the agricultural sector especially in developing countries through reduced food-supply and international trade restrictions. In addition, these diseases are a threat to disease-free countries. We used a multilocus sequence typing (MLST) approach to gain insights into the demographic history of and phylogenetic relationships among the members of the 'M. mycoides cluster'. We collected partial sequences from seven housekeeping genes representing a total of 3,816 base pairs from 118 strains within this cluster, and five strains isolated from wild Caprinae. Strikingly, the origin of the 'M. mycoides cluster' dates to about 10,000 years ago, suggesting that the establishment and spread of the cluster coincided with livestock domestication. In addition, we show that hybridization and recombination may be important factors in the evolutionary history of the cluster.     

Relationships between members of the Mycoplasma mycoides cluster as shown by DNA probes and sequence analysis.

A gene probe, CAP-21, which demonstrated interrelationships between the members of the Mycoplasma mycoides cluster was developed. The probe easily differentiated mycoplasmas in this cluster by clear and predictable hybridization patterns in Southern blots and separated the cluster into four groups. Strains of M. mycoides subsp. mycoides which were capable of causing contagious bovine pleuropneumonia composed one group. Strains of M. mycoides subsp. mycoides which did not cause contagious bovine pleuropneumonia together with strains of M. mycoides subsp. capri composed the second group. Mycoplasma capricolum and the F38 mycoplasmas formed a third group, while the bovine group 7 mycoplasmas composed a separate, fourth group. Further support for the above grouping of the cluster was obtained when amplified DNA analogous to the probe from one representative strain of each of the cluster members was sequenced and these data were used to construct a phylogenic tree. Contagious caprine pleuropneumonia is recognized as an important disease, and the etiological agent of this disease is now known to be the F38 mycoplasma. The CAP-21 probe did not differentiate between M. capricolum and the closely related F38 mycoplasma. A second probe, F38-12, which was capable of distinguishing these two mycoplasmas was made.     

Genome sequence of Mycoplasma capricolum subsp. capripneumoniae strain M1601

Mycoplasma capricolum subsp. capripneumoniae is the causative agent of contagious caprine pleuropneumonia, a devastating disease of goats listed by the World Organization for Animal Health. Here we report the first complete genome sequence of this organism (strain M1601, a clinically isolated strain from China).     

Complete Genome Sequences of Mycoplasma leachii Strain PG50T and the Pathogenic Mycoplasma mycoides subsp. mycoides Small Colony Biotype Strain Gladysdale

Mycoplasma mycoides subsp. mycoides small colony biotype (SC) is the high-consequence animal pathogen causing contagious bovine pleuropneumonia. We report the complete genome sequences of the pathogenic strain M. mycoides subsp. mycoides SC Gladysdale and a close phylogenetic relative, Mycoplasma leachii PG50(T), another bovine pathogen of the M. mycoides phylogenetic clade.     

Electrophoretic Analysis of Isoenzymes of Mycoplasma Species

The purpose of this paper is to further characterize and determine relatedness among some Mycoplasma species or serogroups of significant practical veterinary interest. Twenty-four strains were examined for the presence of 35 enzymes by horizontal starch gel electrophoresis, revealing a total of 127 different electromorphs of 30 enzymes. Inter- as well as intraspecific differences were found demonstrating the application of isoenzyme studies in classification as well as epidemiology. It is concluded that the F38 group of MacOwan and group 7 of Leach constitute 2 new species, The elevation of the 2 subspecies of M. mycoides (mycoides and capri) to species level is favoured, but it is suggested that a decision be taken internationally, considering the practical consequences of a given nomenclature. Three alternative possibilities for classification are presented. Regarding identification the results suggest that the presence or absence of maltase and ornithine transcarbamylase indicate whether an isolate is related to the agent of contagious bovine pleuropneumonia or merely a representative of either the caprine LG type of M. mycoides subsp. mycoides or the classical caprine subspecies, M. mycoides subsp. capri.     

Isolation of Mycoplasma mycoides, mycoides (LC variant), from two naturally aborted caprine fetuses

Described in this study are 2 cases of caprine abortion associated with the LC type of Mycoplasma mycoides , subsp. mycoides . This mycoplasma and Mycoplasma mycoides , subsp. capri had been previously reported in adult goats in this herd. The abortion took place in the latter part of gestation. Samples from heart blood, lung and pleural exudate were collected for the isolation of mycoplasmas and other bacterials in both fetuses. Two strains of Mycoplasma mycoides , subsp. mycoides (LC variant) were isolated. The only gross lesion of the internal organs in the aborted fetuses was congestion of the lungs. Microscopic lesions were encountered in the lungs, and these were characterized by patchy to diffuse pneumonia. Exfoliated cells, many alveolar macrophages, scattered neutrophils and lymphocytes were seen in the lumen of the terminal airways and alveolar spaces. This report appears to be the first isolation of Mycoplasma mycoides , subsp. mycoides (LC variant) from aborted caprine fetuses.     

Comparative genomics analysis of Mycoplasma capricolum subsp. capripneumoniae 87001

Mycoplasma capricolum subsp. capripneumoniae (Mccp), belongs to Mycoplasma mycoides cluster and is a causal pathogen of contagious caprine pleuropneumonia (CCPP). This paper presents the complete annotated genome sequence of Mccp Strain 87001—a strain that was isolated from pneumonia affected goats on a farm in China, and comparative genomics analysis of five Mccp genomes in addition to comparative genomics within Mycoplasma mycoides cluster. The Mccp strain 87001 genome consists of a single circular chromosome 1017333 bp in length and encodes 898 open reading frames (orfs) averaging 944 bp in length. Fifty eight potential virulence genes were identified, including variable surface lipoproteins, hemolysin A, and P60 surface lipoprotein. Comparative genomic analysis revealed eight virulence genes and four extracellular genes which remained unchanged in five Mccp genomes for forty years, which can be used as potential target for drug development and vaccine design. We revealed 183 Mccp unique genes as markers to distinguish Mccp with other mycoplasma strains from goats, and different virulence factors contributing to host specificity and different syndrome of bovine pathogens and caprine pathogens.     

Antibodies to Brucella spp. among blue wildebeest and African buffalo in Kenya

A serologic survey of blue wildebeest (Connochaetes taurinus Burchell) and African buffalo (Syncerus caffer Sparrman) in the Masai Mara area was conducted. Antibodies to Brucella spp. were found in 18% of the blue wildebeest and 30% of the African buffalo examined. There were titers in all age groups and in both sexes. Hygromata were seen in both species. The increase in numbers of blue wildebeest and African buffalo which share grazing and watering areas with cattle of the Masai people, makes the presence of infections by Brucella spp. in wildlife an important consideration in any program for control of brucellosis.     

Genomic maps of some strains within the Mycoplasma mycoides cluster

Genomic restriction maps for the small colony (SC) strains (PG1, KH3J, Gladysdale, and V5) of Mycoplasma mycoides subsp. mycoides (the agent of contagious bovine pleuropneumonia) and for Mycoplasma strain PG50 (classified as bovine serogroup 7), with respective sizes of 1,280, 1,280, 1,260, 1,230, and 1,040 kbp, were compared with the map (1,200 kbp) for a large colony strain (Y goat) of M. mycoides subsp. mycoides. The number and order of all mapped restriction sites were fully conserved in the SC genomes, as were the approximate positions of mapped loci. A number of these restriction sites in the Y genome and some, but fewer, in the PG50 genome appeared to be conserved. The SC and large colony strains shared conservation in the relative positions of the mapped loci, except for rpoC.     

Severity of arthritis is predicted by antibody response to gp135 in chronic infection with caprine arthritis-encephalitis virus.

Antibody titers to caprine arthritis-encephalitis virus surface glycoprotein gp135 and core protein p28 in synovial fluid and serum from 35 goats infected for 3 years were compared with the histologic severity of arthritis in these animals. Anti-gp135 antibody titers in synovial fluid and serum directly reflect the severity of carpal arthritis in chronically infected goats.     

Electrophoretic analysis of proteins from Mycoplasma capricolum and related serotypes using extracts from intact cells and from minicells containing cloned mycoplasma DNA

The acidic proteins of six different mycoplasma serotypes causing bovine or caprine pleuropneumonia were compared by two-dimensional gel electrophoresis of extracts of 35S-labelled cells. The organisms investigated were Mycoplasma mycoides subsp. mycoides (PG1), M. mycoides subsp. mycoides (Y-goat), M. mycoides subsp. capri (PG3), M. capricolum (California kid), the unclassified bovine serogroup 7 of Leach (PG50) and the F38-like group (F38). The results suggested a close relationship between M. capricolum and F38 and a similarly close relationship between the different M. mycoides subspecies, whereas the two M. mycoides subspecies appeared to be quite distant from M. capricolum and F38. The representative strain of the bovine serogroup 7 of Leach was equally distant from F38, M. capricolum and the three strains of M. mycoides. Strikingly, all six mycoplasma strains apparently shared six proteins in the two-dimensional gels. In Escherichia coli minicells, DNA from strain PG50 cloned in the vector pBR325 gave rise to incorporation of radioactive label into proteins which were identified as mycoplasma proteins by two-dimensional electrophoresis and immunoprecipitation.     

丝状支原体丝状亚种SC型脂蛋白Q N末端基因的表达、纯化与抗原性分析

为了表达丝状支原体丝状亚种SC型(MmmSC)中国分离株HVRIⅩ脂蛋白Q(LppQ)N末端基因,将该基因经PCR扩增后克隆至原核表达载体pET32a中,经酶切、PCR、测序证实获得了重组表达质粒,转化Escherichia coliBL21(DE3)菌,经IPTG诱导后获得可溶性融合蛋白,表达量占菌体总蛋白的53.7%,用Ni-NTAHis.Bind纯化试剂盒纯化后,蛋白纯度达95%以上。表达蛋白经Western blot检测其抗原活性,结果表明纯化蛋白可与CBPP标准阳性血清发生强烈的反应,而与阴性血清不发生反应。     

Prevalence of contagious caprine pleuropneumonia in the Borana pastoral areas of Ethiopia

The study was conducted in selected districts of Borana pastoral areas of Ethiopia, namely Yabello, Dire, Moyale and Liban, during June–September 2004 to determine the status of contagious caprine pleuropneumonia (CCPP). The study includes a retrospective study, fieldwork, serology, an abattoir investigation and isolation of the causative Mycoplasma. Outbreaks of CCPP were reported from almost all regions of the country, especially from lowland areas, which are known goat-rearing regions, with the highest in 2002. The predominant clinical findings observed in affected flocks were coughing, nasal discharge, weakness, reluctance to move, abduction of elbow and laboured breathing. A total of 35 sheep and goats, of 183 examined, were sero-positive for MCCP F₃₈ antibodies using CFT. From the total of 217 goats examined in the abattoir for the presence of CCPP lesions, 21 (9.7%) showed pathological lesions, inflamed lungs with marbled appearance, fibrous pleuropneumonia, yellowish-coloured pleural fluid and swollen bronchial and mediastinal lymph nodes. The causative agent (Mycoplasma capricolum sp. capripneumoniae) was isolated and identified from tissue samples and thoracic fluid treated bacteriologically. In conclusion, the study indicates that CCPP is becoming a very important goat disease in Borana pastoral areas.     

Classification and Identification of Ovine and Caprine Mycoplasmas

The purpose of this investigation was to give a survey of the classification of ovine/caprine mycoplasmas as a basis for the identification of strains isolated from sheep and goats. A total of 13 strains representing 13 species and/or serogroups were biochemically examined, and serological cross-titrations were performed using metabolism inhibition, growth inhibition and immunofluorescence. Serogroup 6 (Al-Aubaidi) was found to be identical with Mycoplasma capricolum. The results of identification of 57 isolates, sent to the reference centre from different countries, are given. On the basis of the above investigations and a comparison of some of the classification systems described in the literature, it is concluded that the following species have been isolated from goats and/or sheep: M. agalactiae, M. arginini, M. bovis, M. capricolum, M. conjunctivae, M. mycoides subsp. capri, M. mycoides subsp. mycoides, M. ovipneumoniae, M. putrefaciens, Acholeplasma granularum, A. laidlawii and A. oculi. In addition, both Ureaplasmas and strains representing 6 serogroups (groups 5, 7, 10 and 11 of Al-Aubaidi and groups 17 and 18 of Cottew) have been isolated. These serogroups ought to be finally species-classified as soon as possible. kw|Keywords|k]ovine/caprine mycoplasmas; k]classification; k]identification     

The F38-Like Group,a New Group of Caprine Mycoplasmas?

This paper concerns the taxonomic status of the F38-like group (MacOwan), a prime determinant of contagious caprine pleuropneumonia (CCPP). Extensive biochemical and serological investigations on strain F38 are reported. Some complex serological relationships with other mycoplasma species are revealed. The results, taken in conjunction with earlier published work on geno-typic characters, lead to the conclusion that final classification of these organisms should await further comparative studies of a number of field strains with a related group of strains classified as M. capri-colum. The characterization of F38 confirms its partial relationship to the “M. mycoides group” of ovine/caprine/bovine mycoplasmas, and has also revealed a very close phenotypic relationship to the bovine mycoplasma serogroup 7, a finding of potential diagnostic and epidemiological importance.     

Influence of cohabitation between domestic goat (Capra aegagrus hircus) and Iberian ibex (Capra pyrenaica hispanica) on seroprevalence of infectious diseases

We have investigated the prevalence of infection with different pathogens in domestic goat (Capra hircus) and Iberian ibex (Capra pyrenaica hispanica) populations from two neighbouring geographical zones (with or without cohabitation between wild and domestic species) in Spain. No ibexes were found to be seropositive for Brucella melitensis, Mycoplasma mycoides supspecies mycoides (Mycoplasma mycoides), bovine leukaemia virus, and Chlamydophila abortus or bovine herpes virus type 1 (BHV-1). Domestic goats from both zones were detected with antibodies to Chlamydophila abortus and BHV-1. The percentage seroprevalence against Coxiella burnetii, Mycoplasma agalactiae, Mycobacterium avium subspecies paratuberculosis (MAP), caprine arthritis encephalitis virus (CAEV) and bluetongue virus (BTV) was clearly higher in domestic goats compared to wild ibexes in both zones, although CAEV showed negative results in a zone where species were isolated from each other. Moreover, Borrelia burgdorferi and Pestivirus infection showed moderate seroprevalence differences between domestic and wild goats in cohabitation zones, with the highest prevalence found in wild ibexes from a non-cohabitation zone. Our results showed that cohabitation between species was a risk factor (P?<?0.05) in Q fever infection.     

Observations on the suitability and importance of the domestic intermediate hosts of Echinococcus granulosus in Uttah Pradesh, India

The present study investigated the suitability and importance of buffaloes, camels, sheep, goats and pigs in maintaining the life-cycle of Echinococcus granulosus in Aligarh, India. A total of 565 (36%) of 1556 buffaloes, 20 (2%) of 1208 goats, 5 (1%) of 559 pigs, 6 (6%) of 109 sheep and two of three camels were found to harbour hydatid cysts. The frequency distribution of the hydatid cysts in each intermediate host species was over-dispersed and in buffaloes cyst fertility increased with increasing cyst size. Of 2171, 95 and four buffalo, goat, and camel cysts examined 327 (15%), two (2%) and three cysts respectively were fertile. No pig or sheep cysts were found to contain protoscoleces. The unfenced buffalo abattoir and the large number of dogs allowed access to the abattoir coupled to the number of buffalo slaughtered in comparison to the other potential hosts, indicates that the buffalo is the most significant host for maintaining the life-cycle of the parasite in this area of India. Applicable control measures for the region are suggested.