Large Colony Type Research Articles

Bovine and ovine mycoplasmal arthritis and synovitis

Extract The pathology of mycoplasmal gonitis in a Friesian cow aged 15 months, and polyarthritis in a 3-week-old East Friesian sheep was described. Both stifle joints of the cow, which was 1/63 calves that developed polyarthritis after being fed milk contaminated with Mycoplasma mycoides mycoides large colony type (MmmLC), were distended with yellow flocculent fluid. The capsular and synovial tissues and their associated tendon sheaths were greatly thickened due to chronic active fibrinopurulent inflammation with ulceration of synovial membranes and progressive organisation of the exudate by granulation tissue and fibrosis. Nodular aggregates of lymphocytes and plasma cells studded the membrane. Articular surfaces were unremarkable. Mycoides cluster organisms were demonstrated on polymerase chain reaction (PCR) analysis of the joint tissues and these were confirmed on culture as MmmLC.

Characterization of a chromosomal region of Mycoplasma sp. bovine group 7 strain PG50 encoding a glycerol transport locus (gtsABC).

Mycoplasma species bovine group 7, represented by the type strain PG50, is one of six members of the Mycoplasma mycoides cluster and has been implicated in sporadic and outbreak cases of polyarthritis and mastitis in Australian dairy cattle. This study describes cloning and sequencing a 7.9 kb region of the PG50 chromosome and identification of genes involved in glycerol transport (gtsA, gtsB and gtsC) that are followed by a putative lipoprotein gene lppB and a genomic locus containing two ORFs encoding putative membrane proteins. Long range PCR using primers spanning gtsABC and downstream flanking genes, and Southern hybridization analyses using a suite of probes derived from M. mycoides subsp. mycoides small colony type (SC) strain Afadé for gtsA, gtsB and gtsC, lppB and the two downstream genes confirmed that these genes were conserved among Mycoplasma sp. bovine group 7 isolates and mycoplasmas belonging to the M. mycoides subcluster [M. mycoides subsp. mycoides SC, M. mycoides subsp. mycoides large colony type (LC) and M. mycoides subsp. capri] but were absent in mycoplasmas belonging to the Mycoplasma capricolum subcluster (M. capricolum subsp. capricolum and M. capricolum subsp. capripneumoniae). M. capricolum subsp. capricolum type strain California kid did not hybridize with the probe for gtsA and gave only weak or no hybridization signals with probes derived from the loci downstream of gtsABC, suggesting that this region has diverged in mycoplasmas belonging to subspecies of M. capricolum. It is shown that PG50, after the addition of a physiological concentration of glycerol to the growth medium, generates H(2)O(2) at levels comparable with strain Afadé, implying that the glycerol transport system is functional in Mycoplasma sp. bovine group 7. This suggests that in PG50, as in M. mycoides subsp. mycoides SC, glycerol uptake is followed by phosphorylation to glycerol 3-phosphate and then conversion to dihydroxyacetone phosphate, catalysed by L-alpha-glycerophosphate oxidase, resulting in the production of H(2)O(2). The ability of Mycoplasma sp. bovine group 7 to generate significant amounts of hydrogen peroxide may be important in pathogenesis.

Changes in lymphocyte subsets in the bronchus-associated lymphoid tissue of goats naturally infected with different Mycoplasma species.

The distribution of cells containing lysozyme, S-100 protein, CD3, CD4, CD8, major histocompatibility complex class II antigen and immunoglobulin G (IgG) was analysed in the bronchus-associated lymphoid tissue (BALT) of goats naturally infected with three Mycoplasma species. This study included the immunohistochemical characterization of the pneumonic lesions of 18 goats (3-5 months old) infected with one of the following Mycoplasma species: M. mycoides ssp. mycoides, Large Colony type (goats no. 1-6), M. mycoides ssp. capri (goats no. 7-12) and M. capricolum ssp. capricolum (goats no. 13-18). Microscopically, infected animals showed a moderate broncho-interstitial pneumonia, characterized by lymphoid hyperplasia of the BALT and infiltration of mononuclear cells in the alveolar walls and airways. The main cellular type in the BALT was represented by CD3+ T lymphocytes, and the ratio of CD4+:CD8+ cells was > 2. The BALT showed large germinal centres mainly composed of IgG+ B lymphocytes, with numerous S-100+ follicular dendritic cells. The presence of follicular dendritic cells confirmed the high degree of organization of this lymphoid tissue. The immunohistochemical results showed that activated T lymphocytes, particularly in the CD4 subset, and IgG+ B cells, play a major role in the immune response of the caprine lung infected with these species of mycoplasmas.

Mastitis Caused by Mycoplasma mycoides subspecies mycoides (Large Colony Type) in Goat Flocks in Spain

We describe three different outbreaks of mastitis caused by M. mycoides subspecies mycoides LC type (Mmm LC) in three goat flocks from the Extremadura Region of south-west Spain. Thirty-two fast-growing isolates were obtained on Hayflick's and Friis's media with inhibitors from different specimens. All were identified as Mmm LC in spite of their cultural, biochemical and serological features.

A pathological and immunohistochemical study of goat kids undergoing septicaemic disease caused by Mycoplasma capricolum subsp. capricolum, Mycoplasma mycoides subsp. capri and Mycoplasma mycoides subsp. mycoides (large colony type).

This paper studies the pathological and immunohistochemical findings in 12 kids experimentally infected with Mycoplasma capricolum subsp. capricolum (Mcc), M. mycoides subsp. capri (Mmc) and M. mycoides subsp. mycoides (large colony type), (MmmLC). For the demonstration of Mcc, Mmc and MmmLC antigens an immunoperoxidase technique based on the labelled streptavidin biotin method was used in the 12 kids inoculated with mycoplasmas and the three control kids inoculated with mycoplasma medium. All 12 kids, inoculated by different routes, developed subcutaneous swelling at the point of inoculation and terminated in fatal septicaemia from 1 to 5 days post inoculation. The histopathological findings consisted of cellulitis at the point of inoculation, acute diffuse interstitial pneumonia, arthritis and multifocal necrotic purulent splenitis in some kids. The Mcc, Mmc and MmmLC antigens were detected immunohistochemically in all kids with specific punctiform labelling inside the cytoplasm of the leucocytes or extracellularly at the inoculation point, respiratory airways, spleen, liver, joints, tonsils and lymph nodes. The results obtained in this study showed that the inoculation of these mycoplasmas by parenteral routes caused mycoplasmaemia. Moreover, the immunohistochemical results appear fully to confirm that the mycoplasmas were the cause of the death of the kids because of a septicaemic state.

Contagious agalactia of small ruminants: current knowledge concerning epidemiology, diagnosis and control.

Contagious agalactia of small ruminants is a syndrome which principally affects the mammary glands, joints and eyes. The main causal agents are Mycoplasma agalactiae in sheep, and M. agalactiae, M. mycoides subsp. mycoides large colony type and M. capricolum subsp. capricolum in goats. In addition, M. putrefaciens can produce a similar clinical picture, particularly in goats. Contagious agalactia occurs on all five continents and is often enzootic. The evolution of the infection tends to be chronic in affected animals and herds. Symptomless shedding of mycoplasmas, mainly in the milk, may persist for a long time. These insidious infections, associated with carriage in the ears of healthy animals, are difficult to diagnose and to control. The main mode of transmission between flocks is related to the sale of carrier animals and contact during transhumance, whereas transmission within a flock occurs through contact, suckling and milking. This review discusses the clinical features, epidemiology, treatment, prevention and control of the disease.

Agalactie contagieuse des petits ruminants: épidémiologie, diagnostic et contrôle

Contagious agalactia of small ruminants is a syndrome which affects mainly the mammary glands, joints and eyes. The principal causal agents are Mycoplasma agalactiae in sheep and M. agalactiae, M. mycoides subsp. mycoides large colony type and M. capricolum subsp. capricolum in goats. In addition, M. putrefaciens can produce a similar clinical picture, particularly in goats. Contagious agalactia occurs on all five continents and is often enzootic. These infections are chronic in animals and in flocks. Symptomless shedding of mycoplasmas, mainly in the milk, may persist for a long time. Associated with carriage in the ears of healthy animals, these insidious infections are difficult to diagnose and control. The sale of carrier animals and contact during transhumance are the main modes of transmission between flocks, while transmission within a flock occurs through contact, suckling and milking. This review discusses clinical features, epidemiology, treatment, prevention and control.

Caprine mycoplasmal arthritis in the Sultanate of Oman

An outbreak of caprine arthritis in a small ruminant research facility in northern Oman is reported. Synovial fluid samples were analyzed, and a non-randomized clinical trial was undertaken to determine response to several antibiotics. Results suggest varying susceptibility to arthritis between breeds of goats and indicate a greater effectiveness of lincospectin/spectinomycin and tylosine over other antibiotics in such cases. The presence of both Mycoplasma mycoides subsp. mycoides large colony type and M. mycoides subsp. capri in Oman is confirmed for the first time and it is suggested that they both may play a role in the polyarthritis syndrome seen there.

Cytokines induced in vitro by Mycoplasma mycoides ssp. mycoides, large colony type

Septicemic disease in goats and sheep caused by the large colony type of Mycoplasma mycoides ssp. mycoides has clinical and pathological features in common with septic endotoxemia. We studied the ability of the mycoplasma to induce mediators of biological responses to endotoxin, such as TNFα, IL-1α, IL-6 and nitric oxide. Heat-killed suspensions of mycoplasmas in a concentration of 25 μg protein ml −1 induced all mediators in macrophages from peritoneal cavity and bone marrow of both C3H HeN (LPS responsive) and C3H HeJ (LPS low-responsive) mice. Lipopolysaccharide (LPS) in a concentration of 100 ng ml −1 induced the mediators in C3H HeN derived macrophages, only. Simultaneous stimulation of macrophages with interferon-γ enhanced the secretion of nitric oxide (measured as nitrite) but not the cytokines. We conclude that heat-killed Mycoplasma mycoides ssp. mycoides, large colony type, has cytokine inductive activity similar to bacterial endotoxin, but with an induction mechanism different from LPS.

High mortality in goats associated with the isolation of a strain of Mycoplasma mycoides subsp. mycoides (Large Colony type).

A goat pleuropneumonia outbreak occurring in a herd of 800 animals in the province of Ciudad Real, Spain, is described in this paper. Severe respiratory signs and high mortality were the most significant clinical observations. The adult goats presented mainly respiratory symptoms and/or mastitis, whereas the young animals died showing arthritis and/or keratoconjunctivitis. The most significant lesions were found in the thoracic cavity. A focal extensive fibrinonecrotic pleuropneumonia was macroscopically seen, and the histopathological analysis confirmed a fibrinopurulent and necrotic pleuropneumonia with areas of acute pyogenic bronchopneumonia and fibrinous pericarditis associated with a multifocal purulent mastitis and/or a fibrinopurulent arthritis in some goats. Microbiologically, the mycoplasmas isolated grew rapidly (18-24 h), and, after 48 h, there were colonies of 1-1.5 mm diameter. These isolations were biochemically characterized as Mycoplasma mycoides spp. and showed serological characteristics corresponding to Mycoplasma mycoides subsp. mycoides Large Colony. One of the isolations, the strain 2/93, experimentally showed its pathogenicity causing acute interstitial pneumonia and arthritis when it was inoculated in 1-week-old kids.

Agglutination of "Streptococcus milleri" by lectins.

The agglutination of 218 clinical isolates and three ATCC type strains of "Streptococcus milleri" was tested with 25 different lectins from plants and fungi. An agglutination reaction with one or more lectins was observed with 42 isolates when the cells were untreated. After trypsinisation of the bacteria, 109 strains yielded a positive reaction and after boiling the bacterial cells at pH2, 218 isolates were agglutinated. As an overall result of our experiments with untreated, trypsinised and boiled cells, 17, 37 and 45 different agglutination patterns, respectively, were obtained. The lectins from Datura stramonium, Robinia pseudoacacia and Dolichos biflorus agglutinated isolates belonging only to Lancefield group C, being non-reactive with other isolates. These lectins were also found to be specific for "large colony type" streptococci of group C. The use of lectin agglutination in epidemiological and ecological studies of "S. milleri" is discussed.

Effects of fatty acids on the growth and composition of Malassezia pachydermatis and their relevance to canine otitis externa

Malassezia pachydermatis is one of the many infective agents involved in canine otitis externa. Two major phenotypes (large and small colony types), which had different fatty acid compositions, were found among strains of M pachydermatis isolated from canine ears or skin. The large-colony type contained palmitic, stearic, oleic and linoleic acids, whereas the small-colony type contained myristic, palmitic, stearic, linoleic acids and five other unidentified fatty acids. Exogenous saturated fatty acids in the medium had no effect on the growth of a large colony strain but oleic and linoleic acids were mycostatic. Adding fatty acids to the medium changed the fatty acid composition of the strain. Saturated fatty acids were incorporated as such into the cell, but the unsaturated fatty acids caused complex changes in composition, including the appearance of a new fatty acid. It is suggested that these changes in the fatty acid composition of M pachydermatis could explain the mycostatic effect of unsaturated fatty acids like oleic and linoleic acids at least in vitro.

Application of dot immunobinding on membrane filtration (MF dot) to the study of relationships within “ M. mycoides cluster” and within “glucose and arginine-negative cluster” of ruminant mycoplasmas

A total of 189 isolates from cattle, sheep and goats, allocated to two groups on biochemical grounds, have been examined by a dot immunobinding membrane-filtration (MF dot) method. Seventy glucose fermenting isolates, showing relationships with the “ Mycoplasma mycoides cluster”, have been compared by MF dot against polyclonal hyperimmunesera prepared against the following reference strains: M. mycoides subsp. mycoides, small colony type (SC), strain PG1 and large colony type (LC) strain Y Goat; M. capricolum strain California Kid (CK); M. species bovine serogroup 7 strain PG50, and, ovine/caprine serogroup 11 strain 2-D. The isolates fell into 5 main groups: (a) 14 serologically homogeneous isolates similar to subsp. mycoides SC PG1 (b) 4 homogeneous isolates similar to PG50 (c) 14 isolates all serologically similar to Y Goat, but heterogeneously reactive with subsp. capri PG3 and M. capricolum CK antisera (d) 7 isolates serologically similar to subsp. capri PG3, but heterogeneously reactive with subsp. mycoides SC PG1, M. capricolum CK and 2-D antisera (e) 28 isolates strongly reactive with both M. capricolum CK and serogroup 7 PG50 antisera. 119 isolates that were all glucose and arginine negative were also compared by the MF dot method with the reference strains. Most of these could be classified definitely as M. bovis (78 isolates), M. agalactiae (21 isolates) and serogroup 11 (5 isolates). 13 isolates gave a strong reaction with both M. bovigenitalium and serogroup p11 antisera. 2 isolates showed an unclassifiable pattern. The results confirm that the glucose and arginine-negative cluster strains that reacted with 2-D antiserum, also share serological relationships with the “ M. mycoides cluster”, albeit with a very heterogeneous pattern.

Pathogenicity of Mycoplasma mycoides subsp. mycoides (large colony type) for goat kids

Pathogenicity of Mycoplasma mycoides subsp. mycoides (large colony type) was studied in goat kids by intraperitoneal, intravenous, intramuscular, intratracheal and oral inoculations of 48 hr log phase culture of organisms isolated from spontaneous cases. All experimental animals developed fever, anorexia, acute pain, swollen joints with lameness, recumbency and dyspnea 20 to 72 h post inoculation. All kids died of septicemia within 8 days. Leucopenia due to decrease of neutrophils, increase of blood sugar, alkaline phosphatase and transaminase activity, and decrease of total serum protein with simultaneous reduction of albumin to globulin ratio were recorded. Mycoplasmas were recovered in relatively high numbers from heart blood, synovial fluid and infected lung, liver, kidney and joint tissues. Animals had cellulitis at the sites of inoculation, interstitial pneumonia, pleural haemorrhages, fibrinous hepatitis and pericarditis, myocarditis, necrotic enteritis, lymphadenitis, splenic necrosis, acute nephritis, adreno-cortical necrosis, orchitis, mild meningitis and fibrino-purulent polyarthritis. Vasculitis and widespread intravascular thrombi were also characteristic, indicative vascular damage, possibly produced by toxic effect of mycoplasmas. The results suggested that large colony type of Mycoplasma mycoides subsp. mycoides produces severe pathogenic effects in goat kids.

The external ear canal of goats and other animals as a mycoplasma habitat

In an experimental study Mycoplasma capricolum or M. mycoides subsp. mycoides (large colony type) was cultured from the external ear canal of ear mite-free goats, sheep and a pig and isolated from 19 of 20 animals at necropsy whenever a state of mycoplasmal septicemia existed. In M. putrefaciens infected goats, the mycoplasma could not be recovered at necropsy from the ear canal 7–10 days following a state of septicemia. In a field study, M. mycoides subsp. mycoides was culturable from the external ear canal of a clinically normal dairy goat herd with a prior history of infection, but nares and milk did not yield the organism. Cultures of the external ear canal of a small herd of immature and mature Holstein cows without history of mycoplasmal infection, however, resulted in isolation of M. bovoculi from the external ear canal, but no mycoplasma isolations were made from the nares or milk. The significance of ear mites in the external ear canal and their role in mycoplasmal transfer between animals is currently unknown. Mycoplasmemia explains the seeding of the ear canal by bloodborne mycoplasma, but is an unlikely cause for the finding of up to five Mycoplasma sp. from an ear swab culture. For the moment, the precise explanation for the seeding of mycoplasma in the ear canal must await documentation, although a likely reason appears to be the seeding of the canal by mites as they transfer from goat to goat, which apparently occurs with ease.

PHENOTYPIC PLASTICITY IN SCENEDESMUS COMMUNIS HEGEW. (CHLOROPHYCEAE). II. EXAMPLES OF ALGAL CYCLO‐ AND NONCYCLOMORPHOSIS

A new dimension to the well‐known phenotypic plasticity in Scenedesmus is presented. Large colony type Scenedesmus communis Hegew. reproduced in a typical fashion in batch culture in standard media but produced small colonies (SCT) resembling S. komarekii Hegew. and S. subspicatus Chod. at low cell densities in dilute media. Highest frequency of production occurred after cells had been pretreated in a concentrated (inorganic nutrients) medium. Examples of both cyclic and noncyclic behavior in the life history of Scenedesmus are presented. The noncyclic behavior resembles previous reports on induced heritable changes in flax and tobacco due to different treatment applications of fertilizers. An analysis of gross morphological features using scanning electron microscopy showed that isolated strain SCT1 was similar to S. komarekii (strain UTEX 1236) and isolated strain SCT2 was similar to S. subspicatus (strain UTEX 1358). Growth and morphological expression of the induced SCT strains were highly similar to those of the respective UTEX species. The current state of taxonomy and the implications of phenotypic plasticity on taxonomy in Scenedesmus are discussed.

Phenotypic Plasticity in Scenedesmus communis Hegew. (Chlorophyceae). II. Examples of Algal Cyclo- and Noncyclomorphosis

A new dimension to the well-known phenotypic plasticity in Scenedesmus is presented. Large colony type Scenedesmus communis Hegew. reproduced in a typical fashion in batch culture in standard media but produced small colonies (SCT) resembling S. komarekii Hegew. and S. subspicatus Chod. at low cell densities in dilute media. Highest frequency of production occurred after cells had been pretreated in a concentrated (inorganic nutrients) medium. Examples of both cyclic and noncyclic behavior in the life history of Scenedesmus are presented. The noncyclic behavior resembles previous reports on induced heritable changes in flax and tobacco due to different treatment applications of fertilizers. An analysis of gross morphological features using scanning electron microscopy showed that isolated strain SCT1 was similar to S. komarekii (strain UTEX 1236) and isolated strain SCT2 was similar to S. subspicatus (strain UTEX 1358). Growth and morphological expression of the induced SCT strains were highly similar to those of the respective UTEX species. The current state of taxonomy and the implications of phenotypic plasticity on taxonomy in Scenedesmus are discussed.

Goat flea (order Siphonaptera) as a possible vector for the transmission of caprine mycoplasmal polyarthritis with septicaemia

Goat fleas of the order Siphonaptera were collected from the body surfaces of naturally infected polyarthritic goat kids with septicaemia. These fleas and the infected kids were positive for Mycoplasma mycoides subsp. mycoides (large colony type) identified by cultural, morphological, biochemical and growth inhibition tests. Blood from the infected fleas contained 1 × 10 2−1 × 10 5 viable subsp. mycoides ml −1. Experimental transmission of the disease to other kids was established by placing 120 infected fleas on each kid's body surfaces. All experimentally infected kids developed characteristic clinical signs and showed leucopenia with neutropenia, an increased amount of fibrinogen and mortality with lesions of suppurative polyarthritis associated with septicaemia. The organisms were also recovered in high numbers from heart blood, body fluids, and infected organs and joints. The results suggested that fleas of the order Siphonaptera acted as vectors for the transmission of the spontaneous disease in kids.

Biochemical Alterations in the Blood, Milk and Tissues of Sheep Mammary Gland After Experimental Mycoplasmal Mastitis

home Acta Veterinaria Brno Journal of the University of Veterinary Sciences Brno, Czech Republic Search:

Isolation and restriction endonuclease analysis of a mycoplasma plasmid

A 1.7-kb plasmid was isolated from a large colony type strain of Mycoplasma mycoides subsp. mycoides. It was cloned into the Escherichia coli vector M13mp19, and DNA from the resultant recombinant was used to construct a restriction map of the plasmid. Because mycoplasmas show deviation from normal coding patterns, the availability of a mycoplasma plasmid could be useful in the development of a cloning vector for them.