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1.
Avian Pathol ; 53(4): 257-263, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38353105

RESUMEN

The aim of the present study was to monitor the dynamics and to measure the safety and efficacy of a live, attenuated, thermosensitive Mycoplasma anserisalpingitidis vaccine candidate, namely MA271, in geese breeder flocks under field conditions. Two rearing flocks were vaccinated with MA271 at 4 weeks of age and boosted at 24 weeks of age by cloaca inoculation (1 ml) and eye-dropping (60 µl). The geese then were transported to multi-aged breeding farms. Two breeding flocks served as controls. Colonization of the cloaca by MA271 showed 75% maximum prevalence between 4 and 6 weeks after the first vaccination. Then the prevalence decreased to 25% until the cooler, humid fall months which coincided with the booster vaccination. Boosting raised cloacal colonization to 100%. No clinical signs were observed in the vaccinated birds. After transportation to five multi-aged breeding farms, the wild-type strain appeared as well as MA271 in three flocks. In one flock, the wild-type strain completely displaced MA271, while in one flock only MA271 was detected. Only wild-type strains were detected in the control flocks; however, due to an HPAI outbreak, both flocks were exterminated before the end of the study. Based on the available data, the median percentage of infertile eggs was 3.7-5.1% in the MA271 vaccinated flocks, and 7.7% in the non-vaccinated flock. In conclusion, MA271 can colonize the cloaca of geese under field conditions. MA271 proved to be safe and presumably protects against M. anserisalpingitidis-induced reproduction losses.


Asunto(s)
Vacunas Bacterianas , Gansos , Infecciones por Mycoplasma , Enfermedades de las Aves de Corral , Vacunas Atenuadas , Animales , Enfermedades de las Aves de Corral/prevención & control , Enfermedades de las Aves de Corral/microbiología , Infecciones por Mycoplasma/veterinaria , Infecciones por Mycoplasma/prevención & control , Vacunas Atenuadas/inmunología , Vacunas Atenuadas/administración & dosificación , Vacunas Bacterianas/inmunología , Vacunación/veterinaria , Cloaca/microbiología , Mycoplasma/inmunología , Femenino , Granjas
2.
Acta Vet Hung ; 72(3): 155-160, 2024 Sep 12.
Artículo en Inglés | MEDLINE | ID: mdl-39213125

RESUMEN

Background: Mycoplasma hyopharyngis is a commensal bacterium in the upper respiratory tract of swine. As it is recognized to be apathogenic, examinations regarding this species are scarce, compared to other swine mycoplasmas. However, in a few cases, M. hyopharyngis was detected in lesions of different organs. This report presents a case study in which M. hyopharyngis (along with other bacteria) was isolated from the joint of a pig showing lameness. Case presentation: A Hungarian farm was repopulated with 250 gilts and 1,700 finishers after undergoing a complete depopulation and disinfection. Two days later, cases of diarrhoea and septicaemia caused by Salmonella enterica serovar typhimurium were seen in the finishers. At the same time, following the first farrowing, swollen joints were observed in 21-25 days old piglets. Joint samples were collected, and isolation of Mycoplasma sp. and other bacteria was attempted. Analysis of the joint samples revealed the presence of Staphylococcus haemolyticus, Staphylococcus hyicus, Aerococcus viridans, Trueperella pyogenes, Streptococcus agalactiae and M. hyopharyngis. Conclusions: This is the second isolation of M. hyopharyngis from joints, which highlights the necessity of a better understanding the biology of this often-overlooked species, and its role in the progress of arthritis or other lesions.


Asunto(s)
Infecciones por Mycoplasma , Mycoplasma , Enfermedades de los Porcinos , Animales , Infecciones por Mycoplasma/veterinaria , Infecciones por Mycoplasma/microbiología , Porcinos , Mycoplasma/aislamiento & purificación , Mycoplasma/clasificación , Enfermedades de los Porcinos/microbiología , Femenino , Articulaciones/microbiología
3.
Avian Pathol ; 51(4): 374-380, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35616517

RESUMEN

ABSTRACTMycoplasma iowae, a potential re-emerging avian pathogen mainly affecting turkeys, has been reported from many parts of the world. Poor hatchability, embryonic death, joint and skeletal abnormalities, poor ossification, runting-stunting, poor feathering and airsacculitis may be observed in infected flocks. The reduction of the severity of clinical signs and short-term control of M. iowae are performed by antibiotic treatment. However, M. iowae develops resistance more rapidly and is considered to be more resistant to antimicrobials than other avian pathogenic mycoplasmas. The aim of the present study was to determine the in vitro susceptibility of 101 M. iowae isolates and strains to ten clinically important antimicrobial agents, and to analyse and compare the susceptibility patterns of isolates of various origins and from a wide time-period. The examined reference strains showed high susceptibility to all antimicrobials except for spectinomycin. Low concentrations of tiamulin, florfenicol and oxytetracycline inhibited the growth of the clinical isolates. Nevertheless, slow tendency of increasing minimum inhibitory concentration (MIC) values was observed over time in the case of the above mentioned agents, while MIC values of enrofloxacin showed relatively rapid changes. Spiramycin, erythromycin, tilmicosin, tylosin, lincomycin and spectinomycin did not inhibit the bacterial growth in most of the cases. Isolates originating from captive game birds showed similar susceptibility profiles to isolates from industrial turkey hosts. The widely detected low susceptibility of M. iowae isolates to macrolides, lincomycin and spectinomycin, and the increase of MIC values of frequently used antimicrobials against this pathogen, emphasize the importance of targeted antibiotic therapy.RESEARCH HIGHLIGHTSAntimicrobial susceptibilities of 101 Mycoplasma iowae isolates were determined.Minimum inhibitory concentrations were determined by broth micro-dilution method.Tiamulin, oxytetracycline and florfenicol showed low MIC values.Isolates rapidly adapted to antimicrobial pressure.


Asunto(s)
Antiinfecciosos , Infecciones por Mycoplasma , Mycoplasma iowae , Oxitetraciclina , Enfermedades de las Aves de Corral , Animales , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Antiinfecciosos/farmacología , Lincomicina/farmacología , Lincomicina/uso terapéutico , Pruebas de Sensibilidad Microbiana/veterinaria , Infecciones por Mycoplasma/microbiología , Infecciones por Mycoplasma/veterinaria , Oxitetraciclina/farmacología , Oxitetraciclina/uso terapéutico , Enfermedades de las Aves de Corral/tratamiento farmacológico , Espectinomicina/farmacología , Espectinomicina/uso terapéutico
4.
Avian Pathol ; 51(6): 535-549, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-35866306

RESUMEN

Mycoplasma anserisalpingitidis is economically the most important pathogenic Mycoplasma species of waterfowl in Europe and Asia. The lack of commercially available vaccines against M. anserisalpingitidis had prompted this study with the aim to produce temperature-sensitive (ts+) clones as candidates for an attenuated live vaccine. The production of ts+ clones was performed by N-methyl-N'-nitro-N-nitrosoguanidine (NTG)-induced mutagenesis of Hungarian M. anserisalpingitidis field isolates. The clones were administered via eye-drop and intracloacally to 33-day-old geese. Colonization ability was examined by PCR and isolation from the trachea and cloaca, while the serological response of the birds was tested by ELISA. Pathological and histopathological examinations were performed in the eighth week after inoculation. Whole-genome sequence (WGS) analysis of the selected clone and its parent strain was also performed. NTG-treatment provided three ts+ mutants (MA177/1/11, MA177/1/12, MA271). MA271 was detected at the highest rate from cloacal (86.25%) and tracheal (30%) samples, while MA177/1/12 and MA271 elicited remarkable serological responses with 90% of the birds showing seroconversion. Re-isolates of MA271 remained ts+ throughout the experiment. Based on these properties, clone MA271 was found to be the most promising vaccine candidate. WGS analysis revealed 59 mutations in the genome of MA271 when compared to its parent strain, affecting both polypeptides involved in different cellular processes and proteins previously linked to bacterial fitness and virulence. Although further studies are needed to prove that MA271 is in all aspects a suitable vaccine strain, it is expected that this ts+ clone will contribute to the control of M. anserisalpingitidis infection.RESEARCH HIGHLIGHTS Three M. anserisalpingitidis ts+ vaccine candidates were produced by NTG-mutagenesis.Clone MA271 was able to colonize geese and induce a serological response.MA271 re-isolates remained ts+ during the 8-week-long experiment.WGS analysis revealed 59 mutations in the genome of MA271.


Asunto(s)
Infecciones por Mycoplasma , Mycoplasma , Enfermedades de las Aves de Corral , Animales , Infecciones por Mycoplasma/prevención & control , Infecciones por Mycoplasma/veterinaria , Enfermedades de las Aves de Corral/microbiología , Temperatura , Pollos/microbiología , Vacunas Bacterianas , Mycoplasma/genética , Metilnitronitrosoguanidina , Células Clonales
5.
Acta Vet Hung ; 2022 Sep 28.
Artículo en Inglés | MEDLINE | ID: mdl-36178765

RESUMEN

Several Mycoplasma species can form biofilm, facilitating their survival in the environment, and shielding them from therapeutic agents. The aim of this study was to examine the biofilm-forming ability and its potential effects on environmental survival and antibiotic resistance in Mycoplasma anserisalpingitidis, the clinically and economically most important waterfowl Mycoplasma species. The biofilm-forming ability of 32 M. anserisalpingitidis strains was examined by crystal violet assay. Biofilms and planktonic cultures of the selected strains were exposed to a temperature of 50 °C (20 and 30 min), to desiccation at room temperature (16 and 24 h), or to various concentrations of eight different antibiotics. Crystal violet staining revealed great diversity in the biofilm-forming ability of the 32 tested M. anserisalpingitidis strains, with positive staining in more than half of them. Biofilms were found to be more resistant to heat and desiccation than planktonic cultures, while no correlation was shown between biofilm formation and antibiotic susceptibility. Our results indicate that M. anserisalpingitidis biofilms may contribute to the persistence of the organisms in the environment, which should be taken into account for proper management. Antibiotic susceptibility was not affected by biofilm formation; however, it is important to note that correlations were examined only in vitro.

6.
Avian Pathol ; 50(6): 458-464, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34519598

RESUMEN

The objective of this study was to clarify whether the most common species of Mycoplasma can be detected in the reproductive organs and the cloaca, as well as in the semen of asymptomatic native Hungarian male geese. As it is necessary for the semen of that breed to be preserved pathogen-free in an in vitro gene-conservation programme, the presence of and sources of infection, as well as prevention of the survival of pathogens following semen cryopreservation, are key issues. Ten asymptomatic, 2-year-old ganders were tested. For the detection of mycoplasmas, samples were taken from both fresh and frozen/thawed semen, cloaca, phallus lymph, testes and vas deferens; that is five samples from each of the 10 ganders. The semen was statically frozen using dimethyl-formamide as a cryoprotectant and stored in liquid nitrogen at -196°C. Species-specific PCR systems targeting M. anserisalpingitidis, M. anseris and M. cloacale were used for screening and identification. Results of this study have shown, for the first time, that (1) among the three Mycoplasma species examined, all were detectable in the indigenous Hungarian ganders, with no clinical signs; (2) the pathogens could be detected in the cloaca, in both fresh and cryopreserved semen samples, but remained undetected within the inner reproductive organs; and (3) as pathogens were able to survive the freezing/storing/thawing procedures, the possibility of vertical transmission of the pathogens during artificial inseminations does exist, which causes problems in the in vitro gene-conservation programmes for this breed.


Asunto(s)
Mycoplasma , Preservación de Semen , Animales , Gansos , Genitales , Hungría , Masculino , Mycoplasma/genética , Preservación de Semen/veterinaria
7.
Avian Pathol ; 50(2): 161-173, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33291970

RESUMEN

Mycoplasma gallisepticum and Mycoplasma synoviae are bacterial pathogens that cause disease in poultry, adversely affecting their health and welfare, and are a financial burden on producers. This manuscript describes the results of the MycoPath project that is the first international antimicrobial susceptibility programme for mycoplasma pathogens isolated from poultry. Improved comparative analysis of minimal inhibitory concentration (MIC) results from participating countries was facilitated by using one laboratory determining all MICs. Chicken and turkey isolates were obtained from France, Germany, Great Britain, Hungary, Italy and Spain during 2014-2016. One isolate per farm was retained. The MIC of seven antimicrobial agents was determined using a broth microdilution method, with Friis Medium (M. gallisepticum) or Modified Chanock's Medium (M. synoviae). Of the 222 isolates recovered, 82 were M. gallisepticum and 130 were M. synoviae. M. gallisepticum MIC50/90 values were 0.12/0.5, 2/8, 0.5/4, 0.12/>64, 0.008/0.062, 0.008/32, 0.062/4 mg/l for doxycycline, enrofloxacin, oxytetracycline, spiramycin, tiamulin, tilmicosin and tylosin, respectively. For M. synoviae, the values were 0.5/1, 8/16, 0.5/1, 0.5/8, 0.25/0.5, 0.062/2 and 0.062/16 mg/l respectively. A bimodal MIC distribution for the fluoroquinolone (enrofloxacin) and the macrolides (spiramycin, tilmicosin and tylosin) indicate that both species have sub-populations that are less susceptible in vitro to those antimicrobials. Some differences in susceptibilities were observed according to host species, Mycoplasma species, and country of origin. This study provides a baseline of novel data for future monitoring of antimicrobial resistance in poultry Mycoplasma species. Additionally, this information will facilitate the selection of the antimicrobial agents most likely to be effective, thus ensuring their minimal use with targeted and correct therapeutic treatments.Highlights First large-scale pan-European collection of representative Mg and Ms isolates.MIC values assessed in central laboratory for Mg and Ms from chickens and turkeys.Range of MIC values for 82 Mg and 130 Ms isolates to seven licenced antibiotics shown.Data can be used to help determine Mg and Ms veterinary-specific breakpoints.


Asunto(s)
Antiinfecciosos/farmacología , Pollos/microbiología , Infecciones por Mycoplasma/veterinaria , Mycoplasma gallisepticum/efectos de los fármacos , Mycoplasma synoviae/efectos de los fármacos , Enfermedades de las Aves de Corral/microbiología , Pavos/microbiología , Animales , Farmacorresistencia Bacteriana , Europa (Continente) , Fluoroquinolonas/farmacología , Macrólidos/farmacología , Pruebas de Sensibilidad Microbiana/veterinaria , Infecciones por Mycoplasma/microbiología , Aves de Corral
8.
Acta Vet Hung ; 69(2): 105-109, 2021 06 08.
Artículo en Inglés | MEDLINE | ID: mdl-34106875

RESUMEN

Q fever is a disease of high zoonotic potential, but interest in its causative agent is rather low although it causes some public health problems in Hungary. The prevalence of Q fever is highly variable by country. The main reservoirs of the disease are the same domestic ruminant species everywhere, but the epidemiological profile depends on the features of the specific reservoir. The aim of this large-scale study was to demonstrate the importance of Q fever in different species as a possible source for human infection in most regions of Hungary. A total of 851 serum samples from 44 dairy farms, 16 sheep flocks, 4 goat farms and 3 zoos located in different parts of Hungary were tested. The presence of antibodies to Coxiella burnetii was surveyed in dairy cattle (n = 547), goats (n = 71), sheep (n = 200) and zoo animals (n = 33). The animal species tested in Hungary showed different seroprevalence values of C. burnetii infection. Seropositivity by the enzyme-linked immunosorbent assay was found in 258 out of 547 (47.2%) cows and in 69 out of 271 (25.5%) small ruminants, among them in 47 out of 200 (23.5%) sheep and in 22 out of 71 (31.0%) goats. Antibodies to C. burnetii were not detected in zoo animals. Seropositivity was demonstrated in 44 out of 44 (100%) dairy cattle farms, with at least one serum sample found to be positive on each farm. The seropositivity rate of small ruminant farms was 55.0% (11 positive out of 20 tested), with 9 out of 16 (56.3%) sheep flocks and 2 out of 4 (50.0%) goat herds showing seropositivity.


Asunto(s)
Enfermedades de los Bovinos , Enfermedades de las Cabras , Fiebre Q , Enfermedades de las Ovejas , Animales , Animales de Zoológico , Bovinos , Enfermedades de los Bovinos/epidemiología , Femenino , Enfermedades de las Cabras/epidemiología , Cabras , Hungría/epidemiología , Fiebre Q/epidemiología , Fiebre Q/veterinaria , Estudios Seroepidemiológicos , Ovinos , Enfermedades de las Ovejas/epidemiología
9.
Acta Vet Hung ; 69(2): 110-115, 2021 07 16.
Artículo en Inglés | MEDLINE | ID: mdl-34280127

RESUMEN

The control of Mycoplasma hyorhinis infection relies mainly on antimicrobial therapy. However, the antibiotic susceptibility testing of the bacteria is usually not performed before applying the treatment, and thus therapeutic failures are not uncommon. In the case of M. hyorhinis, several antibiotic-resistance-related single nucleotide polymorphisms (SNPs) are known but assays for their detection have not been described yet. The aims of the present study were to investigate macrolide- and lincomycin-resistance-related SNPs in Hungarian M. hyorhinis isolates and to develop mismatch amplification mutation assays (MAMA) to detect the identified resistance markers. Minimal inhibitory concentrations (MIC) of different drugs and whole genome sequences of 37 M. hyorhinis isolates were used to find the resistance-related mutations. One MAMA assay was designed to detect the mutation of the 23S rRNA gene at nucleotide position 2058 (Escherichia coli numbering). For further evaluation, the assay was challenged with 17 additional isolates with available MIC data and 15 DNA samples from clinical specimens. The genotypes of the samples were in line with the MIC test results. The developed assay supports the practice of targeted antibiotic usage; hence it may indirectly reduce some bacterial resistance-related public health concerns.


Asunto(s)
Infecciones por Mycoplasma , Mycoplasma hyorhinis , Animales , Antibacterianos/farmacología , Bioensayo/veterinaria , Farmacorresistencia Bacteriana/genética , Lincomicina/farmacología , Macrólidos/farmacología , Pruebas de Sensibilidad Microbiana/veterinaria , Infecciones por Mycoplasma/tratamiento farmacológico , Infecciones por Mycoplasma/veterinaria
10.
BMC Genomics ; 21(1): 403, 2020 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-32539834

RESUMEN

BACKGROUND: Mycoplasma anserisalpingitidis is a waterfowl pathogen that mainly infects geese, can cause significant economic losses and is present worldwide. With the advance of whole genome sequencing technologies, new methods are available for the researchers; one emerging methodology is the core genome Multi-Locus Sequence Typing (cgMLST). The core genome contains a high percentage of the coding DNA sequence (CDS) set of the studied strains. The cgMLST schemas are powerful genotyping tools allowing for the investigation of potential epidemics, and precise and reliable classification of the strains. Although whole genome sequences of M. anserisalpingitidis strains are available, to date, no cgMLST schema has been published for this species. RESULTS: In this study, Illumina short reads of 81 M. anserisalpingitidis strains were used, including samples from Hungary, Poland, Sweden, and China. Draft genomes were assembled with the SPAdes software and analysed with the online available chewBBACA program. User made modifications in the program enabled analysis of mycoplasmas and provided similar results as the conventional SeqSphere+ software. The threshold of the presence of CDS in the strains was set to 93% due to the quality of the draft genomes, resulting in the most accurate and robust schema. Three hundred thirty-one CDSs constituted our cgMLST schema (representing 42,77% of the whole CDS set of M. anserisalpingitidis ATCC BAA-2147), and a Neighbor joining tree was created using the allelic profiles. The correlation was observed between the strains' cgMLST profile and geographical origin; however, strains from the same integration but different locations also showed close relationship. Strains isolated from different tissue samples of the same animal revealed highly similar cgMLST profiles. CONCLUSIONS: The Neighbor joining tree from the cgMLST schema closely resembled the real-life spatial and temporal relationships of the strains. The incongruences between background data and the cgMLST profile in the strains from the same integration can be because of the higher probability of contacts between the flocks. This schema can help with the epidemiological investigation and can be used as a basis for further studies.


Asunto(s)
Genoma Bacteriano , Mycoplasma/clasificación , Mycoplasma/genética , Animales , Gansos/microbiología , Genotipo , Epidemiología Molecular , Tipificación de Secuencias Multilocus , Filogenia , Secuenciación Completa del Genoma
11.
Int J Syst Evol Microbiol ; 70(4): 2369-2381, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32068526

RESUMEN

In 1983, Mycoplasma sp. strain 1220 was isolated in Hungary from the phallus lymph of a gander with phallus inflammation. Between 1983 and 2017, Mycoplasma sp. 1220 was also identified and isolated from the respiratory tract, liver, ovary, testis, peritoneum and cloaca of diseased geese in several countries. Seventeen studied strains produced acid from glucose and fructose but did not hydrolyse arginine or urea, and all grew under aerobic, microaerophilic and anaerobic conditions at 35 to 37 ˚C in either SP4 or pleuropneumonia-like organism medium supplemented with glucose and serum. Colonies on agar showed a typical fried-egg appearance and transmission electron microscopy revealed a typical mycoplasma cellular morphology. Molecular characterization included analysis of the following genetic loci: 16S rRNA, 23S rRNA, 16S-23S rRNA ITS, rpoB, rpoC, rpoD, uvrA, parC, topA, dnaE, fusA and pyk. The genome was sequenced for type strain 1220T. The 16S rRNA gene sequences of studied strains of Mycoplasma sp. 1220 shared 99.02-99.19 % nucleotide similarity with M. anatis strains but demonstrated ≤95.00-96.70 % nucleotide similarity to the 16S rRNA genes of other species of the genus Mycoplasma. Phylogenetic, average nucleotide and amino acid identity analyses revealed that the novel species was most closely related to Mycoplasma anatis. Based on the genetic data, we propose a novel species of the genus Mycoplasma, for which the name Mycoplasma anserisalpingitidis sp. nov. is proposed with the type strain 1220T (=ATCC BAA-2147T=NCTC 13513T=DSM 23982T). The G+C content is 26.70 mol%, genome size is 959110 bp.


Asunto(s)
Gansos/microbiología , Mycoplasma/clasificación , Filogenia , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Femenino , Genes Bacterianos , Hungría , Mycoplasma/aislamiento & purificación , ARN Ribosómico 16S/genética , ARN Ribosómico 23S/genética , Análisis de Secuencia de ADN
12.
Avian Pathol ; 49(4): 317-324, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32181698

RESUMEN

Mycoplasma gallisepticum causes respiratory diseases and reproduction disorders in turkeys and chickens. The infection has considerable economic impact due to reduced meat and egg production. Because elimination programmes are not feasible in a large number of poultry farms, vaccination remains the only effective measure of disease control. Differentiating vaccine strains from field isolates is necessary in the control of vaccination programmes and diagnostics. The aim of this study was to develop a polymerase chain reaction based mismatch amplification mutation assay (MAMA) for the discrimination of K vaccine strain (K 5831, Vaxxinova Japan K.K.). After determining the whole genome sequence of the K strain, primers were designed to detect seven different vaccine-specific single nucleotide polymorphisms. After evaluating preliminary results, the MAMA-K-fruA test detecting a single guanine-adenine substitution within the fruA gene (G88A) was found to be the most applicable assay to distinguish the K vaccine strain from field isolates. The detected K strain-specific single nucleotide polymorphism showed genetic stability after serial passage in vitro, but this stability test should still be evaluated in vivo as well, investigating a large number of K strain re-isolates. The MAMA-K-fruA assay was tested on a total of 280 culture and field samples. The designed assay had 102 and 103 template copy number/µl sensitivity in melt-curve analysis based and agarose-gel based assays, respectively, and showed no cross reaction with other avian Mycoplasma species. The new MAMA provides a time- and cost-effective molecular tool for the control of vaccination programmes and for diagnostics.


Asunto(s)
Pollos/microbiología , Infecciones por Mycoplasma/veterinaria , Mycoplasma gallisepticum/genética , Polimorfismo de Nucleótido Simple/genética , Enfermedades de las Aves de Corral/microbiología , Pavos/microbiología , Animales , Vacunas Bacterianas/genética , Cartilla de ADN/genética , Mutación , Infecciones por Mycoplasma/diagnóstico , Infecciones por Mycoplasma/microbiología , Infecciones por Mycoplasma/prevención & control , Mycoplasma gallisepticum/inmunología , Mycoplasma gallisepticum/aislamiento & purificación , Reacción en Cadena de la Polimerasa/veterinaria , Enfermedades de las Aves de Corral/diagnóstico , Enfermedades de las Aves de Corral/prevención & control
13.
BMC Vet Res ; 16(1): 178, 2020 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-32503521

RESUMEN

BACKGROUND: Mycoplasma anserisalpingitidis causes significant economic losses in the domestic goose (Anser anser) industry in Europe. As 95% of the global goose production is in China where the primary species is the swan goose (Anser cygnoides), it is crucial to know whether the agent is present in this region of the world. RESULTS: Purulent cloaca and purulent or necrotic phallus inflammation were observed in affected animals which represented 1-2% of a swan goose breeding flock (75,000 animals) near Guanghzou, China, in September 2019. From twelve sampled animals the cloaca swabs of five birds (three male, two female) were demonstrated to be M. anserisalpingitidis positive by PCR and the agent was successfully isolated from the samples of three female geese. Based on whole genome sequence analysis, the examined isolate showed high genetic similarity (84.67%) with the European isolates. The antibiotic susceptibility profiles of two swan goose isolates, determined by microbroth dilution method against 12 antibiotics and an antibiotic combination were also similar to the European domestic goose ones with tylvalosin and tiamulin being the most effective drugs. CONCLUSIONS: To the best of our knowledge this is the first description of M. anserisalpingitidis infection in swan goose, thus the study highlights the importance of mycoplasmosis in the goose industry on a global scale.


Asunto(s)
Infecciones por Mycoplasma/veterinaria , Mycoplasma/aislamiento & purificación , Enfermedades de las Aves de Corral/microbiología , Animales , Antibacterianos/farmacología , China/epidemiología , Cloaca/microbiología , Femenino , Gansos , Masculino , Pruebas de Sensibilidad Microbiana/veterinaria , Mycoplasma/genética , Infecciones por Mycoplasma/epidemiología , Infecciones por Mycoplasma/microbiología , Pene/microbiología , Secuenciación Completa del Genoma
14.
Antonie Van Leeuwenhoek ; 113(7): 1067-1073, 2020 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-32318980

RESUMEN

Increasing amount of data attest that (in the context of vector-borne infections) birds are not only important as hosts of blood-sucking arthropod vectors, but also as reservoirs of vector-borne pathogens. From 2015 to 2019 cadavers of 100 birds (from 45 species, nine orders) were collected in Hungary, and their organs were screened for DNA from a broad range of vector-borne bacteria with PCR and sequencing. Molecular analyses revealed the presence of Anaplasmataceae, and sequencing identified bacteria closely related to Neorickettsia helminthoeca and Ehrlichia chaffeensis in a Eurasian teal (Anas crecca) and a song thrush (Turdus philomelos), respectively. All samples were PCR negative for rickettsiae, borreliae, Francisella and Coxiella spp., as well as for piroplasms. To our knowledge, this is the first report of a Neorickettsia and an Ehrlichia sp., which belong to the phylogenetic groups of N. helminthoeca and E. chaffeensis, respectively, from Europe. The potential presence of these two vector-borne bacteria needs to be taken into account during future studies on the eco-epidemiology of Anaplasmataceae in Europe.


Asunto(s)
Anaplasmataceae/clasificación , Aves/microbiología , Ehrlichia chaffeensis/clasificación , Neorickettsia/clasificación , Filogenia , Anaplasmataceae/genética , Anaplasmataceae/aislamiento & purificación , Animales , Técnicas de Tipificación Bacteriana , Enfermedades de las Aves/microbiología , Borrelia , ADN Bacteriano/genética , Ehrlichia chaffeensis/genética , Ehrlichia chaffeensis/aislamiento & purificación , Europa (Continente) , Hungría , Neorickettsia/genética , Neorickettsia/aislamiento & purificación , Reacción en Cadena de la Polimerasa , ARN Ribosómico 16S/genética , Rickettsia
15.
Acta Vet Hung ; 68(3): 305-309, 2020 09 30.
Artículo en Inglés | MEDLINE | ID: mdl-33156002

RESUMEN

Q fever is one of the commonest infectious diseases worldwide. A Coxiella burnetii prevalence of 97.6% has been found by ELISA and PCR tests of the bulk tank milk in dairy cattle farms of Hungary. The herd- and individual-level seroprevalence rates of C. burnetii in the examined dairy cows and farms have dramatically increased over the past ten years. Three high-producing industrial dairy farms were studied which had previously been found ELISA and PCR positive for C. burnetii by bulk tank milk testing. Coxiella burnetii was detected in 52% of the 321 cows tested by ELISA. Pregnancy loss was detected in 18% of the cows between days 29-35 and days 60-70 of gestation. The study found a higher seropositivity rate (80.5%) in the cows that had lost their pregnancy and a seropositivity of 94.4% in the first-bred cows that had lost their pregnancy at an early stage. The ELISA-positive pregnant and aborted cows were further investigated by the complement fixation test (CFT). In dairy herds an average of 66.6% individual seropositivity was detected by the CFT (Phase II) in previously ELISA-positive animals that had lost their pregnancy and 64.5% in the pregnant animals. A higher (Phase I) seropositivity rate (50.0%) was found in the cows with pregnancy loss than in the pregnant animals (38.5%). The high prevalence of C. burnetii in dairy farms is a major risk factor related to pregnancy loss.


Asunto(s)
Enfermedades de los Bovinos/epidemiología , Coxiella burnetii/aislamiento & purificación , Fiebre Q/veterinaria , Animales , Bovinos , Enfermedades de los Bovinos/microbiología , Pruebas de Fijación del Complemento/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Hungría/epidemiología , Embarazo , Prevalencia , Fiebre Q/epidemiología , Fiebre Q/microbiología , Estudios Seroepidemiológicos
16.
J Clin Microbiol ; 57(6)2019 06.
Artículo en Inglés | MEDLINE | ID: mdl-30971467

RESUMEN

Mycoplasma gallisepticum is among the most economically significant mycoplasmas causing production losses in poultry. Seven melt-curve and agarose gel-based mismatch amplification mutation assays (MAMAs) and one PCR are provided in the present study to distinguish the M. gallisepticum vaccine strains and field isolates based on mutations in the crmA, gapA, lpd, plpA, potC, glpK, and hlp2 genes. A total of 239 samples (M. gallisepticum vaccine and type strains, pure cultures, and clinical samples) originating from 16 countries and from at least eight avian species were submitted to the presented assays for validation or in blind tests. A comparison of the data from 126 samples (including sequences available at GenBank) examined by the developed assays and a recently developed multilocus sequence typing assay showed congruent typing results. The sensitivity of the melt-MAMA assays varied between 101 and 104M. gallisepticum template copies/reaction, while that of the agarose-MAMAs ranged from 103 to 105 template copies/reaction, and no cross-reactions occurred with other Mycoplasma species colonizing birds. The presented assays are also suitable for discriminating multiple strains in a single sample. The developed assays enable the differentiation of live vaccine strains by targeting two or three markers/vaccine strain; however, considering the high variability of the species, the combined use of all assays is recommended. The suggested combination provides a reliable tool for routine diagnostics due to the sensitivity and specificity of the assays, and they can be performed directly on clinical samples and in laboratories with basic PCR equipment.


Asunto(s)
Vacunas Bacterianas/inmunología , Tipificación Molecular , Infecciones por Mycoplasma/microbiología , Infecciones por Mycoplasma/prevención & control , Mycoplasma gallisepticum/genética , Mycoplasma gallisepticum/inmunología , Vacunas Bacterianas/genética , Tipificación de Secuencias Multilocus , Mycoplasma gallisepticum/aislamiento & purificación , Reacción en Cadena de la Polimerasa
17.
Antonie Van Leeuwenhoek ; 111(9): 1707-1717, 2018 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-29492770

RESUMEN

In Europe, several species of bats, owls and kestrels exemplify highly urbanised, flying vertebrates, which may get close to humans or domestic animals. Bat droppings and bird pellets may have epidemiological, as well as diagnostic significance from the point of view of pathogens. In this work 221 bat faecal and 118 bird pellet samples were screened for a broad range of vector-borne bacteria using PCR-based methods. Rickettsia DNA was detected in 13 bat faecal DNA extracts, including the sequence of a rickettsial insect endosymbiont, a novel Rickettsia genotype and Rickettsia helvetica. Faecal samples of the pond bat (Myotis dasycneme) were positive for a Neorickettsia sp. and for haemoplasmas of the haemofelis group. In addition, two bird pellets (collected from a Long-eared Owl, Asio otus, and from a Common Kestrel, Falco tinnunculus) contained the DNA of a Rickettsia sp. and Anaplasma phagocytophilum, respectively. In both of these bird pellets the bones of Microtus arvalis were identified. All samples were negative for Borrelia burgdorferi s.l., Francisella tularensis, Coxiella burnetii and Chlamydiales. In conclusion, bats were shown to pass rickettsia and haemoplasma DNA in their faeces. Molecular evidence is provided for the presence of Neorickettsia sp. in bat faeces in Europe. In the evaluated regions bat faeces and owl/kestrel pellets do not appear to pose epidemiological risk from the point of view of F. tularensis, C. burnetii and Chlamydiales. Testing of bird pellets may provide an alternative approach to trapping for assessing the local occurrence of vector-borne bacteria in small mammals.


Asunto(s)
Aves/microbiología , Quirópteros/microbiología , Heces/microbiología , Neorickettsia/genética , Anaplasma phagocytophilum/genética , Infecciones por Anaplasmataceae/microbiología , Animales , ADN Bacteriano/genética , Europa (Continente) , Neorickettsia/clasificación , Neorickettsia/aislamiento & purificación , Filogenia , ARN Ribosómico 16S/genética , Estrigiformes
18.
Cent Eur J Public Health ; 26 Suppl: S51-S55, 2018 12.
Artículo en Inglés | MEDLINE | ID: mdl-30817874

RESUMEN

OBJECTIVE: The aim of the study was to evaluate the seroprevalence of West Nile virus (WNV) among the variable population of Eastern Slovakia. METHODS: A serologic survey was conducted using 464 serum samples. The basic demographic, epidemiologic and clinical information was obtained for each serum sample at the time of specimen collection. The presence of antibodies against WNV was investigated using a commercial enzyme-linked immunosorbent assay (ELISA). All the ELISA positive samples were further analysed by a neutralization test with WNV and Usutu virus. RESULTS: Three serum samples (0.65%) from the participants (N = 464) were considered positive for antibodies to WNV. A 29-year-old female was repeatedly exposed to mosquito bites working as a shepherdess and participating in many outdoor activities. Two other females (61 and 76 years old) were treated at the Department of Neurology due to monoparesis of the upper extremity, vertigo; both had a significant epidemiological history with frequent tick and mosquito bites and stay in an endemic region. CONCLUSIONS: Although there was no evidence of WNV infection in the Slovak Republic, the epidemiological situation in the neighbouring countries warrants vigilance and appropriate measures, including the introduction of specific diagnostic tools into clinical practice. The constant monitoring of birds and mosquitoes also seems necessary.


Asunto(s)
Fiebre del Nilo Occidental/diagnóstico , Virus del Nilo Occidental/aislamiento & purificación , Animales , Anticuerpos Antivirales , Culicidae , Ensayo de Inmunoadsorción Enzimática , Femenino , Estudios Seroepidemiológicos , Eslovaquia/epidemiología , Fiebre del Nilo Occidental/epidemiología
19.
Acta Vet Hung ; 66(1): 107-115, 2018 03.
Artículo en Inglés | MEDLINE | ID: mdl-29580076

RESUMEN

Babesia vesperuginis is the only piroplasm known to infect bats. Unlike most members of the genus Babesia, it is probably transmitted by a soft tick species (i.e. Argas vespertilionis). Recently, two studies have been conducted to clarify the phylogenetic status of this species, and both agreed on placing it into a basal position among Babesia sensu stricto (s.s.). However, several important groups of piroplasms were not included in the already reported phylogenetic trees of B. vesperuginis isolates. Therefore, the aim of the present study was to amplify an approx. 950-bp fragment of the cytochrome c oxidase subunit 1 (cox1) gene of B. vesperuginis from A. vespertilionis specimens, and to compare its sequences with those from other piroplasmid groups in a broader phylogenetic context. Sequence comparisons focusing on either 18S rRNA or cox1 genes, as well as phylogenetic analyses involving separate and concatenated 18S rRNA and cox1 sequences indicate that B. vesperuginis is more closely related to the phylogenetic group of Theileriidae than to Babesia s.s. In particular, B. vesperuginis clustered closest to Cytauxzoon felis and the 'prototheilerid' B. conradae. The results of this study highlight that B. vesperuginis is a unique and taxonomically important species, which should be included in future studies aimed at resolving the comprehensive phylogeny of Piroplasmida.


Asunto(s)
Babesia/genética , Babesiosis/parasitología , Quirópteros/parasitología , Complejo IV de Transporte de Electrones/genética , Filogenia , ARN Ribosómico 18S/genética , Animales , Babesia/aislamiento & purificación , ARN Protozoario/genética
20.
Artículo en Inglés | MEDLINE | ID: mdl-27895010

RESUMEN

The molecular mechanisms of resistance to fluoroquinolones, tetracyclines, an aminocyclitol, macrolides, a lincosamide, a phenicol, and pleuromutilins were investigated in Mycoplasma bovis For the identification of mutations responsible for the high MICs of certain antibiotics, whole-genome sequencing of 35 M. bovis field isolates and 36 laboratory-derived antibiotic-resistant mutants was performed. In vitro resistant mutants were selected by serial passages of M. bovis in broth medium containing subinhibitory concentrations of the antibiotics. Mutations associated with high fluoroquinolones MICs were found at positions 244 to 260 and at positions 232 to 250 (according to Escherichia coli numbering) of the quinolone resistance-determining regions of the gyrA and parC genes, respectively. Alterations related to elevated tetracycline MICs were described at positions 962 to 967, 1058, 1195, 1196, and 1199 of genes encoding the 16S rRNA and forming the primary tetracycline binding site. Single transversion at position 1192 of the rrs1 gene resulted in a spectinomycin MIC of 256 µg/ml. Mutations responsible for high macrolide, lincomycin, florfenicol, and pleuromutilin antibiotic MICs were identified in genes encoding 23S rRNA. Understanding antibiotic resistance mechanisms is an important tool for future developments of genetic-based diagnostic assays for the rapid detection of resistant M. bovis strains.


Asunto(s)
Antiinfecciosos/farmacología , Mycoplasma bovis/citología , Mycoplasma bovis/genética , Animales , Antibacterianos/farmacología , Bovinos , Farmacorresistencia Microbiana/genética , Fluoroquinolonas/farmacología , Pruebas de Sensibilidad Microbiana , Mutación/genética , ARN Ribosómico 16S/genética
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