Mycoplasma Diseases Research Articles

Research Progress in the Development of Vaccines against Mycoplasma gallisepticum and Mycoplasma synoviae.

Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) are the primary agents responsible for mycoplasma disease in poultry. MG has been identified as a significant cause of chronic respiratory disease in chickens, while MS has been linked to the development of tenosynovitis, joint swelling and other symptoms in chickens, leading to considerable economic losses for the poultry industry. Unfortunately, there is no specific drug for treatment and vaccination is the most important way to control the disease. There are some different types of vaccines, including live vaccines, inactivated vaccines, sub-unit vaccines and vector vaccines. This paper provides a comprehensive review of the development of vaccines for MG and MS.

LYME AND DENGUE FEVER: FROM BANGLADESH TO ITALY

Abstract Background Dengue fever (Flavivirus) and Lyme disease (Borrelia burgdorferi) are rare tropical infectious diseases transmitted by hematophagous vectors (Aedes mosquitoes and ticks, respectively), and they are extremely uncommon in Italy. In European countries, both represent a public health concern as they manifest as imported diseases. Case Report A 54–year–old man from Bangladesh was admitted with symptoms of fever, dyspnea, and pericarditic chest pain. Laboratory and imaging examinations revealed severe acute respiratory failure secondary to left lower lobe pneumonia and pleuropericardial effusion. Broad–spectrum antibiotic therapy and anti–inflammatory treatment were initiated, suspecting para–pneumonic pleuropericarditis. During hospitalization, there was a gradual improvement with a reduction in inflammatory indices and resolution of lung consolidation. The patient was discharged after 14 days with anti–inflammatory therapy (ibuprofen and colchicine). Approximately 7 days later, the patient returned to the emergency department complaining of a recurrence of symptoms and fatigue. The EKG showed phases of 2:1 AV block, EAS and EPS, complete de novo left bundle branch block (LBBB), alternating with an escape atrial rhythm, not evident in previous EKG. During hospitalization, complete AV block emerged with prolonged asystolic phases, requiring urgent placement of a temporary pacemaker. Concurrently, blood tests revealed anemia and increased levels of inflammatory markers, troponin, creatinine, transaminases, cholestatic indices, LDH, and lipase, indicative of multiple organ failure (MOF). Acute pathologies or active bleeding were ruled out by total body CT with contrast. Blood cultures and serological tests were performed to exclude infections by Trichinella, Mycoplasma, Borrelia, Dengue, and other rare infectious diseases. Due to progressive deterioration, the patient was transferred to a tertiary care hospital. In the following days, laboratory results showed positive IgG and IgM for Dengue (without serological evidence of viral RNA) and positive IgG and IgM antibodies for Borrelia burgdorferi. Conclusion In Italy, the incidence of these diseases is low, and their coexistence in the same host is even rarer (if not anecdotal). However, coinfection cannot be excluded in patients from areas where these diseases are endemic, such as Bangladesh.

The relationship between Mycoplasma and Kawasaki disease in pediatric patients: An updated systematic review and meta-analysis.

This study aimed to clarify the relationship between Mycoplasma pneumoniae (M. pneumoniae) and Kawasaki disease by conducting an updated systemic review and meta-analysis of published studies. Studies mentioning M. pneumoniae and Kawasaki disease before October 2022 were included in this meta-analysis. The pooled prevalence was calculated, and the log odds ratio in the random effects model was applied to estimate the pooled prevalence of M. pneumoniae infection in pediatric patients with Kawasaki disease. In addition, the clinical parameters, such as hemoglobin and erythrocyte sedimentation rate, were analyzed. Six studies with a total of 1,859 pediatric patients with Kawasaki disease were enrolled. The focused outcome was the pooled prevalence and clinical parameters. The pooled prevalence of M. pneumoniae infection was statistically significant in pediatric patients with Kawasaki disease. In addition, the values of hemoglobin and erythrocyte sedimentation rate were significantly different between M. pneumoniae-infected and non-M. pneumoniae-infected patients with Kawasaki disease. Other clinical parameters were not significantly different between M. pneumoniae-infected and non-M. pneumoniae-infected patients with Kawasaki disease. The results suggest that M. pneumoniae infection is significantly prevalent in pediatric patients with Kawasaki disease. The lower values of hemoglobin and erythrocyte sedimentation rate in M. pneumoniae-infected patients with Kawasaki disease might be needed to investigate further.

БИОЛОГИЧЕСКИЕ И ХОЗЯЙСТВЕННЫЕ ОСОБЕННОСТИ ОБРАЗЦОВ ТОМАТА С РАЗЛИЧНОЙ СТЕПЕНЬЮ ДЕТЕРМИНАНТНОСТИ

The research was carried out with the aim of studying new tomato varieties with a determinate bush type selected by Federal Scientific Center for Vegetable Growing (VNIISSOK) and identifying samples with economically valuable traits for subsequent cultivation in plain and foothill conditions. Experimental work was carried out in Terek-Sulak subprovince of the Republic of Dagestan in 2021-2022. The material for the study was 8 semi-standard tomato varieties - Blagodatny, Viking, Voskhod VNIISSOK, Magnat, Severyanka, Sodruzhestvo, Perst, Patris, control - variety Fakel. The repetition is 3-fold, the area of the registration plot is 15 m2, the placement of options is systematic. We used the seedling method of growing tomatoes. Planting pattern 150×25 cm. The experiment was carried out on alluvial-meadow medium-salt soil. According to the timing of ripening, the samples were included in three groups: very early (97...100 days) – Severyanka, Blagodatny, Magnat, Voskhod VNIISSOK; early (102...105 days) – Patris, Viking, Perst; mid-early (111...114 days) – Commonwealth, Torch. All samples, except for the Viking variety, exceeded the control in total yield by 8.8...44.1 t/ha. The first group with a yield of 68.2...86.3 t/ha included Patris, Sodruzhestvo, Voskhod VNIISSOK, Blagodatny; in the second (51.0...54.5 t/ha) – Severyanka, Magnat, Perst; in the third (41.4...42.2 t/ha) – Viking, Torch. High-yielding varieties Patris, Blagodatny, Voskhod VNIISSOK were distinguished by the highest yield of marketable products, 91...92% of the total harvest. The weight of the fruit varied from 60 g (Perst) to 130 g (Sodruzhestvo). The varieties Patrice and Perst had very good keeping quality. The fruits were characterized by a high dry matter content of 5.3...6.4%. A weak spread of viral and mycoplasma diseases at a level of 0.7...1.0 points was observed on plants of the Blagodatny, Voskhod VNIISSOK, Perst, and Patris varieties. According to the type of bush, 5 ordinary and semi-standard samples with a height of no more than 72 cm were identified. - Patrice, Perst, Voskhod VNIISSOK, Blagodatny, Sodruzhestvo.

КОМПЛЕКСНАЯ СИСТЕМА ЗАЩИТЫ КАРТОФЕЛЯ ОТ СОРНЯКОВ, БОЛЕЗНЕЙ И ВРЕДИТЕЛЕЙ

Potatoes are among the crops that are heavily affected by diseases. To a large extent, this is due to the peculiarities of the biology of the potato plant. In the conditions of the North Caucasus region, the probability of the spread of late blight and various viral diseases is high. In addition, potatoes, like other crops, high-yielding varieties are most sensitive to climatic factors, their potential yield cannot be realized if they are not resistant to extreme conditions. In the southern regions of Russia, including in the Republic of Alania, such limiting factors should include, first of all, high summer temperatures and rather long dry or long rainy periods during the growing season of potatoes. Under the influence of high temperatures during the formation and growth of tubers, prolonged hot weather causes metabolic disorders, a decrease in yield; weakened plants are more affected by viral, viroid and mycoplasma diseases. In this regard, the most urgent issue is the identification of drought-resistant varieties and hybrids. In recent years, along with the immunological characteristics of varieties, more and more attention has been paid to their qualitative consumer properties, therefore, resistance to diseases, pests and weeds should be combined with the main economically useful qualities - yield, starchiness and others. The weed control system in landscape farming should be considered as an integral part of the management of the entire agricultural system. To build a weed control system, it is necessary, first of all, to know the actual and potential contamination of the object under examination (plot, field, crop rotation). The analysis of factors shows that one of the main reasons is a strong contamination of potato plantings with the floral composition of perennial, annual cereal and broad-leaved weeds, infestation with pests and diseases. Weeds, pests and diseases, having complex biological features, are characterized by strong adaptability to potato cultivation conditions, high competitive ability, harmfulness and on this basis reduce the yield of tubers, their quality in the absence of a scientifically-based comprehensive system for protecting plantings from them. In this regard, potato plants are characterized by low competitive ability compared to weeds, pests and diseases, and vulnerability to them, which necessitates the integrated use of protection methods at all stages of crop cultivation technology. At the same time, it is advisable to direct the agricultural technology of culture cultivation to create favorable conditions for the growth and development of potato plants, the dominant component of agrophytocenosis, increasing its competitive ability against harmful objects and reducing their harmfulness. Agroecological assessment of the conditions affecting the clogging of potato plantings, their infestation with pests and diseases allows us to note that this is facilitated by a decrease in the material and technical base and financial capabilities of agriculture, violation of farming systems, crop rotations, established technologies, lack of monitoring of the species composition of weeds, weak implementation of the results of scientific developments

Infectious human diseases: Regions, habitats, threats, and mitigation strategies: The issues—Part II

Abstract Globally, vaccines against 47 infectious etiologic agents are approved by at least one regulatory authority as of December 2022. The numbers of viral diseases, infections caused by mycoplasma, bacterial and protozoal diseases, mycoses, diseases caused by parasitic worms, and other infectious diseases of unknown etiology run into thousands. Therefore, more vaccines are required to be developed to keep more populations disease free. Climate change and global increase in temperature may promote sea level rises and an increase in the intensity of rains, causing an upsurge in certain infectious diseases in regions of the human population and even causing movement of habitats. In societies where the expenditure on health as a percentage of gross domestic product is higher, people are more capacitated to tackle the treatment and spread of infectious diseases. As several of such conditions are zoonotic, well-planned strategies for controlling the spread would go a long way in the proper direction. Poverty and contagious diseases are infallibly linked. Major infectious diseases of poverty include tuberculosis, human immunodeficiency virus (HIV)/ acquired immuno deficiency syndrome (AIDS), malaria, measles, pneumonia, diarrheal diseases, and several neglected tropical diseases. The world is not yet unified in action to jointly work to contain the global problem of infectious diseases in every human habitat. There was a visible divide between and among the rich countries versus the developing countries regarding accessibility and deployment of vaccines against COVID-19 flu. To contain the spread of infectious diseases in the future, the whole world would have to work together, raising funds, strengthening epidemiological surveys, inventing effective vaccines, and vaccinating the eligible population, as also treating the infected with therapy in time, besides resorting to other preventive measures for overall human progress. The developmental efforts are to be pursued jointly and together to benefit all people, respecting the world as one.

Programmed Host Cell Death and Infectious Process of Mycoplasma

A common characteristic of Mycoplasma diseases is their chronicity, Mycoplasma spp. infection have been frequently associated with upper respiratory infections, chronic lung disease , asthma, meningeal encephalitis, mastitis, arthritis, heart problems, sterility, bone problems , oviduct dysfunctions and death in human and different animal species. In former years, changeable expression of membrane antigens has been detected in a number of Mycoplasma spp., resulting in the assumption that immune prevarication may be an important part of the their infection and pathogenesis. It has been determined how the attachment organelle, which mediates the complicated interactions between various adhesins and auxiliary adhesion proteins to mediate the critical first stage of cytoadherence to respiratory tract epithelium. Additionally, it has been demonstrated that inflammatory cytokines cause tissue damage by intracellular localization, direct cytotoxicity, and activation of the inflammatory cascade via Toll-like receptors (TLRs), and inflammosome activation, which causes air passage inflammation. All of these play crucial roles in the infectious process. This paper seeks to provide a thorough assessment of recent developments in our understanding of Mycoplasma pathogenesis with the understanding of its virulence mechanisms.

Immune and Oxidative Response against Sonicated Antigen of Mycoplasma capricolum subspecies capripneumonia-A Causative Agent of Contagious Caprine Pleuropneumonia.

Vaccines are vital for prevention and control of mycoplasma diseases. The exploration of a vaccine candidate for the development of a vaccine is imperative. The present study envisages the evaluation of immune and oxidative response against an adjuvanted, sonicated antigen of Mycoplasma capricolum subsp. capripneumonia in male Angora rabbits (1 year old, 2 kg) divided in four groups, each having six animals. Group 1 was the healthy control and received 1 mL PBS via subcutaneous route. Group 2 was administered 1 mL of saponin-adjuvanted and -sonicated antigen, Group 3 was given 1 mL of montanide ISA 50-adjuvanted and-sonicated antigen, and Group 4 was given 1 mL of standard vaccine via subcutaneous route. Animals were evaluated for cellular and humoral immune response and oxidative parameters at 0, 7, 14, 21, and 28 days of the study. Total leukocytic, neutrophilic, and basophilic counts showed a significant (p < 0.05) increase in vaccinated groups compared to the healthy group on most of the intervals. TNF-α levels were significantly (p < 0.05) higher in the Group 2 than the Group 1 at all the time intervals and more comparable to Group 4 than Group 3. IL-10 levels were significantly (p < 0.05) higher in vaccinated groups compared to the healthy group on days 14, 21, and 28, but were lower in Group 3 than in Group 2 and Group 4. More hypersensitivity as inflammation and histopathological cellular infiltration in the ear was produced in Group 2 and Group 4 than in Group 3. IgG levels were significantly (p < 0.05) higher in Group 2 and Group 4 than in Group 3 on days 14 and 21. Antibody titers were comparatively higher in Group 4, followed by Group 2 and 3, than Group 1. Significantly (p < 0.05) higher oxidant and lower antioxidant values were noted in Group 2 and 4 compared to Group 3 and Group 1 on most of the intervals. The TLC and antibody titer showed increasing trend throughout the trial, whereas TNF-α, IgG, L, M and E started decreasing from day 14, and IL-10, N and B started decreasing from day 21. This study concludes that the saponin-adjuvanted and-sonicated antigen induces comparatively higher immune response than montanide but is associated with oxidative and inflammatory reactions.

Genome Engineering of the Fast-Growing Mycoplasma feriruminatoris toward a Live Vaccine Chassis.

Development of a new generation of vaccines is a key challenge for the control of infectious diseases affecting both humans and animals. Synthetic biology methods offer new ways to engineer bacterial chassis that can be used as vectors to present heterologous antigens and train the immune system against pathogens. Here, we describe the construction of a bacterial chassis based on the fast-growing Mycoplasma feriruminatoris, and the first steps toward its application as a live vaccine against contagious caprine pleuropneumonia (CCPP). To do so, the M. feriruminatoris genome was cloned in yeast, modified by iterative cycles of Cas9-mediated deletion of loci encoding virulence factors, and transplanted back in Mycoplasma capricolum subsp. capricolum recipient cells to produce the designed M. feriruminatoris chassis. Deleted genes encoded the glycerol transport and metabolism systems GtsABCD and GlpOKF and the Mycoplasma Ig binding protein-Mycoplasma Ig protease (MIB-MIP) immunoglobulin cleavage system. Phenotypic assays of the M. feriruminatoris chassis confirmed the corresponding loss of H2O2 production and IgG cleavage activities, while growth remained unaltered. The resulting mycoplasma chassis was further evaluated as a platform for the expression of heterologous surface proteins. A genome locus encoding an inactivated MIB-MIP system from the CCPP-causative agent Mycoplasma capricolum subsp. capripneumoniae was grafted in replacement of its homolog at the original locus in the chassis genome. Both heterologous proteins were detected in the resulting strain using proteomics, confirming their expression. This study demonstrates that advanced genome engineering methods are henceforth available for the fast-growing M. feriruminatoris, facilitating the development of novel vaccines, in particular against major mycoplasma diseases.

EFFECT OF VIRUS AND MYCOPLASMA DISEASES ON PRODUCTIVITY OF ALFALFA HAY CULTIVARS

Purpose: To evaluate the productivity of different alfalfa hay cultivars under the effect of local virus and mycoplasma diseases. Methodology: The study was conducted under center pivot irrigation system similar to TADCO mechanized alfalfa hay crop production. Results: Imported alfalfa hay cultivars were found infected with three virus and mycoplasma diseases: alfalfa enation virus, alfalfa mosaic virus and alfalfa witches' broom. Infections with these diseases lead to a severe reduction on the productivity of alfalfa hay cultivars in the third cropping season. Alfalfa enation virus was found to cause the most economic damage to the alfalfa hay crop, and the percentage yield reduction from this disease reached 17.6, 36.8% in the second and third seasons of cultivation respectively. Unique Contribution to Theory, Practice and Policy: The authors recommended to grow alfalfa hay cultivars with less susceptibility to virus diseases infections as we found variability in the productivity of the tested cultivars. The study also recommends to grow alfalfa hay cultivars under mechanized system for two seasons as we observed significant yield reduction in the third season due to high infestation with weeds and grasses which affected the hay quality. In the first field trial (1999 - 2002), productivity of alfalfa hay cultivars CUF 101 and Siriver reached above 30 M.T. /Ha under aphid-controlled conditions. In the second field trial (2002 - 2004), alfalfa hay productivity of the cultivars Fast Grow, SW 9500, Magic (SW 9301) and Super Supreme reached above 30 M.T./Ha in the first season 2003; and in the second season 2004, productivity of Magic, Fast Grow and Super Supreme reached above 26 M.T./Ha. Productivity over the two seasons 2003 and 2004 shown Fast Grow and Magic were the highest with 60.38, 59.85 M.T./Ha. respectively.

Vaccines for Mycoplasma Diseases of Small Ruminants: A Neglected Area of Research.

Mycoplasmas cause some of the most economically important diseases of sheep and goats, including diseases listed by the World Organisation for Animal Health (OIE) such as contagious caprine pleuropneumonia (CCPP) and contagious agalactia (CA). Other important mycoplasma diseases include chronic respiratory and arthritic syndrome (CRAS) and atypical pneumonia, both present on all continents where small ruminants are farmed. Unfortunately, owing to a lack of investment, most commercial vaccines for these diseases are of poor quality, being mostly composed of killed bacteriocins of dubious or unknown efficacy. Several Mediterranean laboratories produce autogenous vaccines, but these can only be used on farms where outbreaks have been officially declared, and consequently have limited impact on disease nationally. Effective live vaccines are available, but their use is often restricted because of safety concerns. With the necessary safeguards in place, we argue for their greater use. This review examines reported vaccines for mycoplasma diseases of small ruminants and attempts to identify new candidate antigens that may enable the development of improved products. Vaccines for CCPP are covered elsewhere.

Correlation between Vitamin A Deficiency and the Severity of Mycoplasma Pneumoniae Pneumonia in Children

Mycoplasma pneumoniae pneumonia is a problem that has attracted the attention of children’s respiratory department in recent years. The relationship between vitamin A deficiency (VA) and the severity of Mycoplasma disease (MPP) in children was studied. In this paper, the hospitalized children with Mycoplasma pneumoniae pneumonia (MPP) in our hospital were selected as the research objects, and they were divided into common type and refractory type (RMPP) for comparative experiment. The VA and immunoglobulin levels of the two groups were compared. Attention should be paid to the normal ratio of VA deficiency (CVAD) and sub deficiency (svad) in the two groups. In this paper, the relationship between infection, immunoglobulin level and VA level was analyzed. The results showed that CVAD was 27.27% in MPP group and 63.75% in RMPP group. Comparison of immunoglobulin levels between the two groups: the levels of IgM, IgA and IgG in the normal MPP group were significantly lower than those in the RMPP group. On the other hand, the detection rate of CVAD was 81.48% in RMPP with infection and 54.72% in RMPP without infection. The detection rate of CVAD in patients with infection was significantly higher than that in patients without infection. There was a correlation between VA Deficiency and MPP classification.

Contemporary Direction of Eggplant Selection for the South of Russia

Eggplant selection is crucial for the south of Russia due to the concentration of production and processing capacities in this region. When selecting eggplants, regional characteristics, in particular specific stressors, should be taken into account. The main stressors are low temperatures in the initial period of plant growth and development, sharp changes in soil and air temperature and humidity during the growing season, high temperatures during crop formation, and harmful fungal and mycoplasma diseases. It is possible to reduce the dependence on imports by developing production in spring greenhouses, which will extend the consumption period by 3-4 months. This requires varieties and hybrids with specific properties: high productivity, high adaptability, compact habit of plants, lack of thorns, low pubescence, etc. F1 hybrids combining high productivity with adaptability and product quality can grow in greenhouses. When collecting linear materials for creating hybrids, it is advisable to use hard provocative backgrounds with a subsequent assessment of the combinational ability. This research aimed to develop new varieties and hybrids of eggplant for the south of Russia. The study was conducted in the Rostov Selection and Seed Center in 2010-2019 on fields and in spring greenhouses. Materials from different regions of the world were collected. Varieties and hybrids were evaluated by their valuable traits and properties. Linear materials were created for sources and donors of characters that are in demand. The collection of the best varieties was involved in selection programs for creating an assortment for the south of Russia. More than 10 eggplant varieties and hybrids with high productivity and adaptive qualities were obtained.&#x0D; Keywords: eggplant, source material, variety, hybrid, selection, signs, applications, stability, heterosis

Emergence of new insect pests on vegetables during the last decade: A case study

With the changes in cropping system, climate and introduction of highly input-intensive high-yielding varieties/hybrids are the root cause for a shift in insect pest status in time and space, resulting in enhanced damage caused by them in the world. Many of them also act as vectors for several viral and mycoplasma diseases, aggravating the problem further. In India, yield loss due to major insect pests is varying from 30–40%. In addition to the regular pests, recently, many exotic and invasive insect pests have invaded in many parts of the countries. South American pin worm (Tuta absoluta Meyrick), solenopsis mealy bug (Phenacoccus solenopsis Tinsley) are few such insects. Similarly, mirid bugs (Nesidiocoris cruentatus (Ballard) and Metacanthus pulchellus Dallas), melon weevil [Acythopius curvovistris citrulli (Marshall)], white plume moth [Sphenarches caffer (Zeller)], cucumber moth (Diaphania indica) and moringa fruit borer (Noorda blitealis Walker), tortoise beetle (Cassida circumdata Herbst) are the insects which have come up in bigger way in current decade either by expanding their host horizon or increase their severity. Therefore, these emerging insect pests in vegetable ecosystem in current decade, their suitable control measures and some issues/challenges in their ecofriendly management are discussed.

Effect of Virus and Mycoplasma Diseases on The Productivity of Alfalfa Hay Cultivars

Purpose: To evaluate the productivity of different alfalfa hay cultivars under the effect of local virus and mycoplasma diseases.Methodology: The study was conducted under center pivot irrigation system similar to TADCO mechanized alfalfa hay crop production. Results: Imported alfalfa hay cultivars were found infected with three virus and mycoplasma diseases: alfalfa enation virus, alfalfa mosaic virus and alfalfa witches’ broom. Infections with these diseases lead to a severe reduction on the productivity of alfalfa hay cultivars in the third cropping seasons. Alfalfa enation virus was found to cause the most economic damage to the alfalfa hay crop, and the percentage yield reduction from these disease reached 17.6, 36.8% in the second and third seasons of cultivation respectively.Unique contribution to theory, practice and policy: The authors recommended to grow alfalfa hay cultivars with less susceptibility to virus diseases infections as we found variability in the productivity of the tested cultivars. The study also recommends to grow alfalfa hay cultivars under mechanized system for two seasons as we observed significant yield reduction in the third season due to high infestation with weeds and grasses which affected the hay quality. In the first field trial (1999 – 2002), productivity of alfalfa hay cultivars CUF 101 and Siriver reached above 30 M.T. /Ha under aphid-controlled conditions. In the second field trial (2002 – 2004), alfalfa hay productivity of the cultivars Fast Grow, SW 9500, Magic (SW 9301) and Super Supreme reached above 30 M.T./Ha in the first season 2003; and in the second season 2004, productivity of Magic, Fast Grow and Super Supreme reached above 26 M.T./Ha. Productivity over the two seasons 2003 and 2004 shown Fast Grow and Magic were the highest with 60.38, 59.85 M.T./Ha. respectively

Transcriptome sequencing analysis of porcine MDM response to FSL-1 stimulation

Mycoplasma infection can cause many diseases in pigs, resulting in great economic losses in pork production. Innate immune responses are thought to play critical roles in the pathogenesis of mycoplasma disease. However, the molecular events involved in immune responses remain to be determined. Hence, the object of this study was to use RNA-Seq to investigate the gene expression profiles of the innate immune response mediated by FSL-1 in pig monocyte-derived macrophages (MDMs). The results revealed that 1442 genes were differentially expressed in the FSL-1 group compared with the control groups, of which 777 genes were upregulated and 665 genes were downregulated. KEGG pathway analysis showed that the upregulated genes were mainly involved in innate immune-related pathways including the TNF signaling pathway, cytokine-cytokine receptor interaction, Toll-like receptor signaling pathway, Jak-STAT signaling pathway, chemokine signaling pathway, NOD-like receptor signaling pathway and NF-kappa B signaling pathway. The downregulated genes were only involved in the cGMP-PKG signaling pathway and glycerophospholipid metabolism. Our results showed that FSL-1 stimulation activated the TLR2 signaling pathway and resulted in diverse inflammatory responses. FSL-1 induced the transcription of numerous protein-coding genes involved in a complex network of innate immune-related pathways. We speculate that TNF, IL1B, IL6, NFKB1, NFKBIA, CXCL2, CXCL8, CXCL10, CCL2, CCL4 and CCL5 were the most likely hub genes that play important roles in the above pathways. This study identified the differentially expressed genes and their related signaling pathways, contributing to the comprehensive understanding of the mechanisms underlying host-pathogen interactions during mycoplasma infection and providing a reference model for further studies.

Phenotypic diversity in an emerging mycoplasmal disease

The avian pathogen Mycoplasma gallisepticum (MG) is a known pathogen of poultry, and newly emerged pathogen of house finches wherein it is associated with lethal conjunctivitis. Factors present in MG that are known to mediate virulence include cytadherence, sialidase activity, peroxide production, and biofilm formation. We have quantitatively assessed these factors for MG isolates from house finches from a temporal and geographic distribution across the continental United States that show differing capacity for virulence in vivo. Statistically significant (P < 0.05) differences were observed across strains for sialidase activity, cytadherence, and hydrogen peroxide production. Sialidase activity increased over time in geographically static populations, but did not correlate with time overall. All strains were able to bind α-2,6-linked sialic acid. No strains were found to bind α-2,3-linked sialic acid. All strains produced biofilms in vitro; however, no significant differences were observed in the density of biofilms across strains. Quantitative variance in virulence-associated traits is consistent with within-host evolutionary adaptation in response to a change in ecological niche by a parasitic pathogen.

Evidence for the Cytoplasmic Localization of the L-α-Glycerophosphate Oxidase in Members of the "Mycoplasma mycoides Cluster".

Members of the “Mycoplasma mycoides cluster” are important animal pathogens causing diseases including contagious bovine pleuropneumonia and contagious caprine pleuropneumonia, which are of utmost importance in Africa or Asia. Even if all existing vaccines have shortcomings, vaccination of herds is still considered the best way to fight mycoplasma diseases, especially with the recent and dramatic increase of antimicrobial resistance observed in many mycoplasma species. A new generation of vaccines will benefit from a better understanding of the pathogenesis of mycoplasmas, which is very patchy up to now. In particular, surface-exposed virulence traits are likely to induce a protective immune response when formulated in a vaccine. The candidate virulence factor L-α-glycerophosphate oxidase (GlpO), shared by many mycoplasmas including Mycoplasma pneumoniae, was suggested to be a surface-exposed enzyme in Mycoplasma mycoides subsp. mycoides responsible for the production of hydrogen peroxide directly into the host cells. We produced a glpO isogenic mutant GM12::YCpMmyc1.1-ΔglpO using in-yeast synthetic genomics tools including the tandem-repeat endonuclease cleavage (TREC) technique followed by the back-transplantation of the engineered genome into a mycoplasma recipient cell. GlpO localization in the mutant and its parental strain was assessed using scanning electron microscopy (SEM). We obtained conflicting results and this led us to re-evaluate the localization of GlpO using a combination of in silico and in vitro techniques, such as Triton X-114 fractionation or tryptic shaving followed by immunoblotting. Our in vitro results unambiguously support the finding that GlpO is a cytoplasmic protein throughout the “Mycoplasma mycoides cluster.” Thus, the use of GlpO as a candidate vaccine antigen is unlikely to induce a protective immune response.

Relationship between Antimicrobial Susceptibility and Multilocus Sequence Type of Mycoplasma bovis Isolates and Development of a Method for Rapid Detection of Point Mutations Involved in Decreased Susceptibility to Macrolides, Lincosamides, Tetracyclines, and Spectinomycin.

Mycoplasma bovis isolates belonging to the sequence type 5 (ST5) group, the dominant group in Japan since 1999, were low susceptible to 16-membered macrolides and tetracyclines and were confirmed to have a guanine-to-adenine transition mutation at position 748 in the 23S rRNA gene (rrl) and adenine-to-thymine transversion mutations at positions 965 and 967 in the 16S rRNA gene (rrs) (Escherichia coli numbering). Moreover, isolates of ST93 and ST155, members of the ST5 group, were low susceptible to lincosamides and azithromycin and showed an adenine-to-guanine transition mutation at position 2059 of rrl Isolates of ST93 were additionally low susceptible to spectinomycin and showed a cytosine-to-adenine transversion mutation at position 1192 of rrs Strains of the ST5 group seem to spread to Japan and Europe from North America with imported cows, while strains of ST93 and ST155 originated in Japan. Melting curve analysis using hybridization probes revealed the existence of point mutations involved in decreased susceptibility to macrolides, lincosamides, and spectinomycin, as demonstrated by changes in the melting curve shape and/or decreases in the melting peak temperature, so the susceptibility to these antimicrobials can be assessed on the same day. For decreased susceptibility to fluoroquinolones to exist, nonsynonymous mutations in the DNA gyrase gene (gyrA) and topoisomerase IV gene (parC) had to coexist. The combination of amino acid substitutions of serine at position 83 in gyrA and serine at position 80 in parC resulted in particularly low susceptibility to fluoroquinolones.IMPORTANCEMycoplasma bovis is the main causal species of bovine mycoplasmal disease and leads to significant economic losses because of its severe symptoms, strong infectivity, and refractoriness. As for mastitis, culling cows with intramammary infections is a general countermeasure to prevent spreading. The conventional antimicrobial susceptibility test for mycoplasma is time-consuming and troublesome, but no quick and easy method for grasping the antimicrobial susceptibility of the causal strain exists at present. Treatment without antimicrobial susceptibility information may be one reason why M. bovis infection is refractory. Detecting a mutation involved in decreased susceptibility to antimicrobial agents of the causal strain makes it possible to easily select suitable antimicrobials for treatment, and this technique will help improve the cure rate and prevent the overuse of ineffective antimicrobial agents. In this study, we developed a technique to quickly and easily assess antimicrobial susceptibility based on the genetic characteristics of M. bovis strains in Japan.

Diagnosis of mycoplasmal infections on common mushroom in artificial agroecosystems

Purpose. To screen and diagnose common mushroom (Agaricus bisporus (J. Lge) Imbach) for infection with mycoplasmas to obtain mushroom spawn free of infection. Methods. Visual, microbiological (obtaining pure fungus culture, studying cultural properties of colonies, determining sensitivity to antibiotics), electron microscopy, statistical. The studies were conducted during 2015–2019 at the Department of Ecobiotechnology and Biodiversity of the National University of Life and Environmental Sciences of Ukraine. There were studied spawn (strain Sylvan A-15, included in the State Register of Ukraine), casing material, substrate, and fruit bodies of common mushroom (40 samples) selected in mushroom farms in Vasylkiv and Kyiv-Sviatoshyn districts of Kiev region. Results. In 17 samples out of 40, symptoms of mycoplasma diseases were found. The symptoms inherent to them were diagnosed: acquisition of gray coloring by young fruit bodies, premature cap opening. Occurrence of exudates in the samples which was observed in 36% of the studied samples also indicates mycoplasmal nature. In the case of infection of casing material, the symptoms coincided with the data on the isolation and identification of the disease. The electron diffraction pattern confirms the presence of mycoplasma conglomerate in the deep spawn of common mushroom. Mycogone and mummification samples appeared to be highly sensitive to antibiotics (the sterile area around indicator disc with the antibiotic was 29 mm), Сoprinus and bacterial rust pathogen were the sensitive ones (19 mm), and bacterial lesion of substrate was the sensitive one (12 mm). The method for diagnosing mycoplasmas on fungi of the species A. bisporus has been improved for specific symptoms: necrosis and exudate. Conclusions. Primary screening of common mushroom samples in greenhouse conditions was performed using visual method and electron microscopy to diagnose mycoplasma infection of fruit bodies and spawn. Microbiological methods for the detection of mycoplasma infections in mushroom culture have been optimized.